the m&m police: a study in paper chromatographythe m&m police: a study in paper...
TRANSCRIPT
The M&M Police: A Study in Paper Chromatography
Benjamin Morgan
Salk Middle School
Spokane, WA
and
Randall Stephens
Mabton High School
Mabton, Wa
Washington State University Advisors
Neil Ivory
Dept. of Chemical Engineering
&
Jeff Burke
Graduate Research Assistant
July 2009
The Project herein was supported by the National Science Foundation Grant No. EEC-
0808716. Dr. Richard L. Zollars, Principle Investigator and Dr. Donald C. Orlich, co-PI.
The module was developed by the authors and does not represent an official endorsement
by the National Science Foundation.
Table of Contents
Project summary…………………………………………………………………….....
Overview of project……………………………………………………………………
Intended audience………………………………………………………………….......
Estimated duration………………………………………………………………..........
Introduction……………………………………………………………………………
Rationale for module……………………………………………………………….....
Science…………………………………………………………………………………
Engineering…………………………………………………………………………….
Goals…………………………………………………………………………………...
Prerequisite student skills/knowledge…………………………………………………
Procedures …………………………………………………………………………….
M&M Middle School Chromatography Project:
Safety
precautions……………………………………………………………………..
Lab Equipment………………………………………………………………
Lesson #1………………………………………………………………………
Lesson #2………………………………………………………………………
Lesson #3………………………………………………………………………
Lesson #4………………………………………………………………………
Lesson #5………………………………………………………………………
Lesson #6………………………………………………………………………
Lesson #7………………………………………………………………………
Lesson #8………………………………………………………………………
Lesson #9………………………………………………………………………
Lesson #10………………………………………………………………………
High School Leaf Chromatography Project
Safety precautions…………………………………………………………………..
Lab Equipment………………………………………………………………
Lesson #1………………………………………………………………………
Appendix A…………………………………………………………………
Appendix B…………………………………………………………………
Appendix C……………………………………………………………
Appendix D…………………………………………………………………
Appendix E…………………………………………………………………
Appendix F…………………………………………………………………
References……………………………………………………………………………..
3
3
3
4
4
16
17
18
18
25
13
26
26
26
28
35
42
44
47
57
63
68
73
79
98
98
98
98
99
113
123
125
128
131
147
162
PROJECT SUMMARY:
Overview of project
This project has been designed to enhance the interest in engineering amongst middle
school and high school students through the use of paper chromatography to solve an
M&M Cartoon Character Crime, and an exploration of pigments in leaves. The notes and
lab activities provide opportunities to better understand mixtures, solutions, separation
techniques, and unknown identifications; thus, giving students a better understanding of
the principles of engineering. Furthermore, the unit activities, labs, notes, and concepts
incorporate many of the Essential Academic Learning Requirements promoted by the
Office of the Superintendent of Public Instruction in Washington State.
Intended audience
The first projects’ intended audience is middle school 7th
-8th
grade. It was written with
the Grades 6-8 Chemistry Science Standards in mind. However, teachers of students in
high school could also utilize this module. Students with minimal prior understanding of
chemistry, mixtures, solutions, and/or chromatography can perform these activities. The
activities can be modified to fit the state standards for the high school student.
Much effort was put into the notes, worksheets, and activities/labs so that the instructor
could easily see the goal of the activity and our actual lab results. It is our hope that this
would reduce teacher preparation time as much as possible. Most of the materials are
found at the local school supply store, grocery store, or are things most middle school
and/or high school teachers have in their classroom or supply rooms. Chromatography
4
paper will be the one item that will need to be preordered. Laboratory filter paper is
cheaper and also works, but the results are not as tight or as neat. All results in this paper
are given using chromatography paper (about $20/100 sheets).
The second projects’ intended audience is high school level Biology students. The leaf
chromatography activity in designed to enhance their understanding of the different leaf
pigments and the concept of energy transfers. This is part of a several week long unit on
plants, including a separate greenhouse growth study.
Estimated Duration:
The middle school module is designed to build on each preceding activity beginning with
an understanding of what density is and how to calculate it; then moving into the
concepts of mixtures, solutions, and chromatography; finally culminating in solving a
crime the students must use utilizing the concepts of chromatography (mixture
separation) and density utilizing some algebra. Two weeks is the estimated time
necessary to complete the entire module.
The high school module will take approximately 2 class periods, with the extensions
adding several days if the teacher desires to utilize all of them.
Introduction:
Chromatography can be a challenging subject to comprehend for a middle school student,
most of whom are getting their first introduction to the topic. Hands on activities, a
5
visually appealing medium, and an interesting project problem can help alleviate those
difficulties and stimulate interest and learning. Students learn the basics of density,
mixtures, and solutions through basic chromatography techniques in a CSI slanted
interactive learning module.
Substances are different from other substances due to their own inherent physical
properties. We use these physical properties to classify, organize, and identify
substances. Some of these physical properties are seen on the surface like size and color.
Others, we have to measure like density. Students will learn how physical properties help
us to categorize and identify things and how they are inherent to a particular substance.
In the culminating project these physical properties will help them narrow down their
suspect list and solve a CSI M&M Cartoon Character case. The color of the ―blood‖ left
behind by the M&M suspect will help to identify which color of M&M did it. By
examining a footprint in the mud left behind by the suspect students will notice where the
water level rose to as a result of the mass of the M&M character. Thus, they will know
the density of the M&M who committed the crime.
One physical property not yet discussed is solubility. This is where chromatography is
utilized: to separate a mixture into its components by how soluble it is in a solvent. A
mixture is a material composed of two or more elements or parts. Many mixtures are
solutions which contain a solvent (which does the dissolving) and a solute (which is the
substance that is dissolved). How readily that solute dissolves into the solvent is a
6
measure of its solubility. In any mixture the substances mixing do not combine. They
only mix. Each constituent retains its own physical and chemical properties.
All mixtures can be separated back out into their distinctive parts. Sand and iron filings
can be separated by using a magnetic. Students can easily understand why this is a
mixture: they can visually see two distinct components. They can also clearly see that the
two sections are not combined in any way (reacted together) so they will maintain their
separate properties. Thus, the physical property of magnetic attraction can be utilized to
separate them.
Solutions provide students with a dilemma. The constituents are much smaller so they
cannot actually see the separate entities. As a result, many students find it much more
difficult to not only believe that there is more than one part in the solution, but that these
parts are actually separate and have their own unique (separate) characteristics.
Chromatography allows the teacher to visually show all of these things in a similar
fashion to the sand and iron filing situation and, I believe, in an interesting approach to
the eighth grade mind.
Paper chromatography is a separation technique for mixtures. It is especially geared
toward separating dyes. In paper chromatography a small sample dye from a marker,
food coloring, etc. is placed on the bottom of chromatography paper (about 1 cm from the
bottom edge). The chromatography paper is called the stationary phase. Usually, you
will be putting multiple sample dots on your chromatography paper. As a result, use a
7
pencil to create a line across the entire bottom surface about 1 cm up so they are lined up.
I will describe how I like to create larger chromatograms later.
In a beaker, or other container, a small amount of solvent is placed. In this module
distilled water and 50% rubbing alcohol were used as solvents. A good solvent should be
able to dissolve the samples’ components in different amounts. As a result, the
components of the mixture will be separated for the student to see. Distilled water was
used for all water based samples: water based markers (crayola – non washable kind, Vis-
à-vis, Mr. Sketch), food coloring, and Kool Aid. The 50% rubbing alcohol was used to
separate the permanent markers: Sharpies and EXPO. However, as it will be noted later
the EXPO’s did not dissolve in this either.
The amount of solvent used is very important. You do not want the solvent to touch the
sample dots you created when you place them in the beaker. I prefer to put together
larger chromatograms (the graphic record produced by the chromatography) because it is
both quicker and they do not fold over so easily. Place this next to the beaker and
add/remove water as needed prior to putting in your chromatogram.
8
Once you place your chromatogram in the beaker the solvent will begin to rise up the
stationary phase (chromatography paper) due to the affinity between the two substances.
Capillary action, the movement of liquid within spaces of porous material due to forces
of adhesion, cohesion, and surface tension also plays a major role in this. This is why the
solvent is called the mobile medium.
Once the solvent front (top edge of the solvent) reaches the sample dots you prepared the
dots/mixtures the individual components will begin to dissolve and rise with the mobile
Container
Water Level is below dots
Black
C M V
Brown
C M V
Red
C M V
Orange
C M V
Your Name: Markers dipped in Water
9
medium. Those parts of the mixture with greater solubility will rise higher that those
with less solubility; this, we have our separation technique via a physical property
difference of it’s mixture components (solubility). Let the mobile medium continue to
rise until all parts are separated or until the solvent front is nearing the top. You will get
a better feel for when to stop after you have done a few. When it is done pull the
chromatogram out by the dry top and let dry on a paper towel.
Here is an example chromatogram I made:
Black
C M V
Brown
C M V
Red
C M V
Orange
C M V
Folded paper towel to make it thicker
Your Name: Markers dipped in Water
10
Other clues left behind besides density (foot print in mud) and blood color as discussed
earlier is; a ransom note written in marker; blood splatter; and urine sample. Students
will create a master chromatogram of the population of M&M suspects like the one
pictured above for urine (made from Kool Aid), blood (made from food coloring), and
markers. They will then test their samples to determine which exact M&M committed
the crime.
For the high school project, a greater depth of knowledge is demanded of the students,
including an assessment on the history and different kinds of chromatography, the
practical uses of it, real world examples, how it works, and a more technical analysis of
the results. Background knowledge includes the following.
Briefly, chromatography is a technique for separating mixtures into their components in
order to analyze, identify, purify, and/or quantify the mixture or components.
Chromatography is used by scientists to:
Analyze – examine a mixture, its components, and their relations to one another
Identify – determine the identity of a mixture or components based on known
components
Purify – separate components in order to isolate one of interest for further study
Quantify – determine the amount of the a mixture and/or the components present
in the sample
11
Toxicologists use the process to identify the toxins found in the tissues of plants
or animals exposed to polluted environments
Real-life examples of uses for chromatography:
Pharmaceutical Company – determine amount of each chemical found in new
product
Hospital – detect blood or alcohol levels in a patient‘s blood stream
Law Enforcement – to compare a sample found at a crime scene to samples from
suspects
Environmental Agency – determine the level of pollutants in the water supply
Manufacturing Plant – to purify a chemical needed to make a product
In detail, chromatography is a method used by scientists for separating organic and
inorganic compounds so that they can be analyzed and studied. By analyzing a
compound, a scientist can figure out what makes up that compound. Chromatography is a
great physical method for observing mixtures and solvents. A good example of an
industrial separation technique is the fractional distillation of raw petroleum in an oil
refinery to produce diesel fuel, gasoline, lubricants, and so on.
The word chromatography means "color writing" which is a way that a chemist can test
liquid mixtures. While studying the coloring materials in plant life, a Russian botanist
invented chromatography in 1903. His name was M.S. Tswett.
12
Chromatography is used in many different ways. Some people use chromatography to
find out what is in a solid or a liquid. It is also used to determine what unknown
substances are. The Police, F.B.I., and other detectives use chromatography when trying
to solve a crime. It is also used to determine the presence of cocaine in urine, alcohol in
blood, PCB's in fish, and lead in water.
How it works:
Chromatography is based on differential migration. There is a mobile phase and a
stationary phase, and the solutes in the mobile phase go through the stationary phase
picking up the compounds to be tested. At different points in the stationary phase the
different components of the compound are going to be absorbed and are going to stop
moving with the mobile phase. This is how the results of any chromatography are
achieved, and is called chromatographic development.
In paper and thin-layer chromatography the mobile phase is the solvent. The stationary
phase in paper chromatography is the strip or piece of paper that is placed in the solvent.
In thin-layer chromatography the stationary phase is the thin-layer cell. Both these kinds
of chromatography use capillary action to move the solvent through the stationary phase.
What is the Retention Factor, Rf ?
The retention factor, Rf, is a quantitative indication of how far a particular compound
travels in a particular solvent. The Rf value is a good indicator of whether an unknown
compound and a known compound are similar, if not identical. If the Rf value for the
13
unknown compound is close or the same as the Rf value for the known compound then
the two compounds are most likely similar or identical. Solubility of a particular
chemical in a given solvent is similar to periodic trends in the table of the elements.
The retention factor, Rf, is defined as
Rf = distance the solute (D1) moves divided by the distance traveled by the solvent front
(D2)
Rf = D1 / D2 where
D1 = distance that color traveled, measured from center of the band of color to the point
where the food color was applied
D2 = total distance that solvent traveled
14
The Different Types of Chromatography
Liquid Chromatography is used in the world to test water samples to look for pollution in
lakes and rivers. It is used to analyze metal ions and organic compounds in solutions.
Liquid chromatography uses liquids which may incorporate hydrophilic, insoluble
molecules.
Gas Chromatography is used in airports to detect bombs and is used is forensics in many
different ways. It is used to analyze fibers on a persons body and also analyze blood
15
found at a crime scene. In gas chromatography helium is used to move a gaseous mixture
through a column of absorbent material.
Thin-layer Chromatography uses an absorbent material on flat glass or plastic plates. This
is a simple and rapid method to check the purity of an organic compound. It is used to
detect pesticide or insecticide residues in food. Thin-layer chromatography is also used in
forensics to analyze the dye composition of fibers.
Paper Chromatography is one of the most common types of chromatography. It uses a
strip of paper as the stationary phase. Capillary action is used to pull the solvents up
through the paper and separate the solutes.
16
The table below summarizes the information from above.
Type of Chromatography Applications in the
Real World Why and What is it
Liquid Chromatography
test water samples
to look for
pollution,
Used to analyze metal ions
and organic compounds in
solutions. It uses liquids
which may incorporate
hydrophilic, insoluble
molecules.
Gas Chromatography
detect bombs in
airports, identify
and quantify such
drugs as alcohol,
used in forensics to
compare fibers
found on a victim
Used to analyze volatile
gases. Helium is used to
move the gaseous mixture
through a column of
absorbent material.
Thin-Layer
Chromatography
detecting pesticide
or insecticide
residues in food,
also used in
forensics to analyze
the dye composition
of fibers
Uses an absorbent material
on flat glass plates. This is
a simple and rapid method
to check the purity of the
organic compound.
Paper Chromatography
separating amino
acids and anions,
RNA fingerprinting,
separating and
testing histamines,
antibiotics
The most common type of
chromatography. The
paper is the stationary
phase. This uses capillary
action to pull the solutes up
through the paper and
separate the solutes.
RATIONALE FOR MODULE:
The goal of the originators of the grant funding this project is to increase middle and high
school students’ exposure to and understanding of the various fields of engineering. A
―best fit‖ for this introduction to engineering would seem to be in the science curriculum
17
of middle and high schools. With this in mind the developers of this module strived to
combine existing curriculum with new activities and a new angle so as to fulfill the goals
of the project while at the same time avoiding the necessity of removing existing material
from the science curriculum.
The module introduces students to (or builds upon existing knowledge, depending at
what level this module is used) key scientific principles such as mixtures, solutions,
separation techniques, physical properties, and chemical properties. Depending on the
science background of the instructor and students, this module could easily be used at the
introductory level in a middle school physical science course or with slight modification,
in a high school physical science, biology, or chemistry course.
SCIENCE:
The scientific basis of this module is the concept of mixtures, solutions, solubility, and
physical properties. Chromatography, allows the teacher to more concretely show
students that solutions are separate components that do not combine. These individual
components retain their physical and chemical properties. Without chromatography this
idea remains very abstract. In addition, chromatography offers the teacher an excellent
opportunity to show students how mixtures can be separated using solubility (an essential
academic learning requirement).
To the student many new terms relating to the science of chromatography, mixtures,
solutions, and physical properties are introduced. Terms such as; solvent, solute,
18
chromatogram, solubility, mixture, solution, stationary medium, mobile medium, along
with several other will, by design, become part of the students’ vocabulary during this
module. For the high school project, an understanding of photosynthesis, pigments,
plant structure, and energy flows is required.
ENGINEERING:
Arguably the key to this module is the melding of engineering and science in such a way
so as to not have to remove existing curriculum from the science course but rather to
teach the same concepts from an engineering view. The culminating projects involve the
students analyzing, categorizing, and problem solving crime scene clues to solve a case.
They will have to take what they learn and then apply it to a new situation. They will
need to take the clues and measure density, test the urine sample, blood sample, ransom
note sample of an M&M Cartoon Character to rule out some possible suspects and
narrow their suspect list down…until they have the culprit. In essence, during this
project they will learn to become a chemical engineer; albeit, on a small part time basis.
This chromatography application of chemical engineering is also shown to apply to the
biological sciences through the separation of pigments in leaves. This will stimulate an
interest in the student in cross-curricular applications of engineering technology.
GOALS:
The goals of this project match the GLE’s. The GLE’s here are from the most recent
2008 revised EALR’s. They are the Standards for Grades 6-8 since this is the focus of
19
my classroom and the project. Bear in mind that you may focus on which GLE/EALR
fits your needs.
EALR 4: Physical Science
Big Idea: Matter: Properties and Change (PS2)
Core Content: Atoms and Molecules In prior grades students learned the scientific
meaning of the word matter, and about changes of state. In grades 6-8 students learn the
basic concepts behind the atomic nature of matter. This includes the idea that elements
are composed of a single kind of atom. Atoms chemically combine with each other or
with atoms of other elements to form compounds. When substances are combined in
physical mixtures, their chemical properties do not change; but when they combine
chemically, the new product has different physical and chemical properties from any of
the reacting substances. When substances interact in a closed system, the amount of mass
does not change. Atomic theory also explains the ways that solids, liquids, and gases
behave. These concepts about the nature of matter are fundamental to all sciences and
technologies.
Content Standards Performance Expectations
Students know that: Students are expected to:
6-8
PS2A
Substances have characteristic
intrinsic properties such as density,
solubility, boiling point, and melting
point, all of which are independent
of the amount of the sample.
Use characteristic intrinsic
properties such as density, boiling
point, and melting point to identify
an unknown substance.
6-8
PS2B
Mixtures are combinations of
substances whose chemical
properties are preserved.
Separate a mixture using differences
in properties (e.g., solubility, size,
magnetic attraction) of the
substances used to make the
mixture.
EALR 2: Inquiry
20
Big Idea: Inquiry (INQ)
Core Content: Questioning and Investigating In prior grades students learned to plan
investigations to match a given research question. In grades 6-8 students learn to revise
questions so they can be answered scientifically and then to design an appropriate
investigation to answer the question and carry out the study. Students learn to think
critically and logically to make connections between prior science knowledge and
evidence produced from their investigations. Students can work well in collaborative
teams and communicate the procedures and results of their investigations, and are
expected to critique their own findings as well as the findings of others.
Content Standards Performance Expectations
Students know that: Students are expected to:
6-8
INQA
Question
Scientific inquiry involves
asking and answering questions
and comparing the answer with
what scientists already know
about the world.
Generate a question that can be
answered through scientific
investigation. This may involve refining
or refocusing a broad and ill-defined
question.
6-8
INQB
Investig
ate
Different kinds of questions
suggest different kinds of
scientific investigations.
Plan and conduct a scientific
investigation (e.g., field study,
systematic observation, controlled
experiment, model, or simulation) that
is appropriate for the question being
asked.
Propose a hypothesis, give a reason for
the hypothesis, and explain how the
planned investigation will test the
hypothesis.
Work collaboratively with other
students to carry out the investigations.
21
6-8
INQC
Investig
ate
Collecting, analyzing, and
displaying data are essential
aspects of all investigations.
Communicate results using pictures,
tables, charts, diagrams, graphic
displays, and text that are clear,
accurate, and informative. *a
Recognize and interpret patterns – as
well as variations from previously
learned or observed patterns – in data,
diagrams, symbols, and words.*a
Use statistical procedures (e.g., median,
mean, or mode) to analyze data and
make inferences about relationships.*b
6-8
INQD
Investig
ate
For an experiment to be valid, all
(controlled) variables must be
kept the same whenever
possible, except for the
manipulated (independent)
variable being tested and the
responding (dependent) variable
being measured and recorded. If
a variable cannot be controlled,
it must be reported and
accounted for.
Plan and conduct a controlled
experiment to test a hypothesis about a
relationship between two variables. *c
Determine which variables should be
kept the same (controlled), which
(independent) variable should be
systematically manipulated, and which
responding (dependent) variable is to be
measured and recorded. Report any
variables not controlled and explain
how they might affect results.
6-8
INQE
Model
Models are used to represent
objects, events, systems, and
processes. Models can be used to
test hypotheses and better
understand phenomena, but they
have limitations.
Create a model or simulation to
represent the behavior of objects,
events, systems, or processes. Use the
model to explore the relationship
between two variables and point out
how the model or simulation is similar
to or different from the actual
phenomenon.
6-8
INQF
Explain
It is important to distinguish
between the results of a
particular investigation and
general conclusions drawn from
these results.
Generate a scientific conclusion from an
investigation using inferential logic, and
clearly distinguish between results (e.g.,
evidence) and conclusions (e.g.,
explanation).
Describe the differences between an
objective summary of the findings and
an inference made from the findings.*c
22
6-8
INQG
Commu
nicate
Clearly
Scientific reports should enable
another investigator to repeat the
study to check the results.
Prepare a written report of an
investigation by clearly describing the
question being investigated, what was
done, and an objective summary of
results. The report should provide
evidence to accept or reject the
hypothesis, explain the relationship
between two or more variables, and
identify limitations of the
investigation.*c
6-8
INQH
Intellect
ual
Honestl
y
Science advances through
openness to new ideas, honesty,
and legitimate skepticism.
Asking thoughtful questions,
querying other scientists'
explanations, and evaluating
one’s own thinking in response
to the ideas of others are abilities
of scientific inquiry.
Recognize flaws in scientific claims,
such as uncontrolled variables,
overgeneralizations from limited data,
and experimenter bias.*c
Listen actively and respectfully to
research reports by other students.
Critique their presentations respectfully,
using logical argument and evidence. *c
Engage in reflection and self-evaluation.
6-8
INQI
Conside
r Ethics
Scientists and engineers have
ethical codes governing animal
experiments, research in natural
ecosystems, and studies that
involve human subjects.
Demonstrate ethical concerns and
precautions in response to scenarios of
scientific investigations involving
animal experiments, research in natural
ecosystems
EALR 3: Application
Big Idea: Application (APP)
Core Content: Science, Technology, and Problem Solving In prior grades students
learned to work individually and collaboratively to produce a product of their own
design. In grades 6-8 students work with other members of a team to apply the full
process of technological design, combined with relevant science concepts, to solve
problems. In doing so they learn to define a problem, conduct research on how others
23
have solved similar problems, generate possible solutions, test the design, and
communicate the results. Students also investigate professions in which science and
technology are required so they can learn how the abilities they are developing in school
are valued in the world of work.
Content Standards Performance Expectations
Students know that: Students are expected to:
6-8
APPA
People have always used technology
to solve problems. Advances in
human civilization are linked to
advances in technology.
Describe how a technology has
changed over time in response to
societal challenges.
6-8
APPB
Scientists and technological
designers (including engineers) have
different goals. Scientists answer
questions about the natural world;
technological designers solve
problems that help people reach
their goals.
Investigate several professions in
which an understanding of science
and technology is required. Explain
why that understanding is necessary
for success in each profession.
6-8
APPC
Science and technology are
interdependent. Science drives
technology by demanding better
instruments and suggesting ideas for
new designs. Technology drives
science by providing instruments
and research methods.
Give examples to illustrate how
scientists have helped solve
technological problems (e.g., how
the science of biology has helped
sustain fisheries) and how engineers
have aided science (e.g., designing
telescopes to discover distant
planets).
6-8
APPD
The process of technological design
begins by defining a problem and
identifying criteria for a successful
solution, followed by research to
better understand the problem and
brainstorming to arrive at potential
solutions.
Define a problem that can be solved
by technological design and identify
criteria for success.
Research how others solved similar
problems.
Brainstorm different solutions.
6-8
APPE
Scientists and engineers often work
together to generate creative
solutions to problems and decide
which ones are most promising.
Collaborate with other students to
generate creative solutions to a
problem, and apply methods for
making trade-offs to choose the best
24
solution.*a
6-8
APPF
Solutions must be tested to
determine whether or not they will
solve the problem. Results are used
to modify the design, and the best
solution must be communicated
persuasively.
Test the best solution by building a
model or other representation and
using it with the intended audience.
Redesign as necessary.
Present the recommended design
using models or drawings and an
engaging presentation.*b
6-8
APPG
The benefits of science and
technology are not available to all
the people in the world.
Contrast the benefits of science and
technology enjoyed by people in
industrialized and developing
nations.
6-8
APPH
People in all cultures have made and
continue to make contributions to
society through science and
technology.
Describe scientific or technological
contributions to society by people in
various cultures.
The following are EALRs specific to the leaf chromatography activities and are taken
from the 10th
grade standards.
EALR 1 — SYSTEMS: The student knows and applies scientific concepts and principles
to understand the properties, structures, and changes in physical, earth/space, and living
systems.
Forms of Energy
1.1.4 Analyze the forms of energy in a system, subsystems, or parts of a system. W
Explain the forms of energy present in a system (i.e., thermal energy, sound energy,
light energy, electrical energy, kinetic energy, potential energy, chemical energy, and
nuclear energy).
25
Component 1.2 Structures: Understand how components, structures, organizations,
and interconnections describe systems.
Energy Transfer and Transformation
1.2.2 Analyze energy transfers and transformations within a system, including
energy conservation.
Distinguish conditions likely to result in transfers or transformations of energy from
one part of a system to another (e.g., a temperature difference may result in the flow of
thermal energy from a hot area to a cold area).
Prerequisite student skills and knowledge:
Students should know what physical properties are. They should also have a basic idea
of dissolving, what atoms are, and the difference between a solid a liquid. Students
should understand that we use physical properties to categorize substances. These
physical properties are both size, shape, etc. as well as things we can measure boiling
point, melting point, and density. For project 2, the students should already have a basic
understanding of the structure of leaves, especially the light reactions, and the processes
of photosynthesis.
26
M&M Middle School Chromatography Project
Safety precautions:
Though none of the activities carried out in this module pose any undo risk of injury,
common laboratory safety practices should be observed. Protective eyewear should be
worn at all times during the lab activities. Though unlikely, the possibility of eye damage
due to contact with rubbing alcohol should be noted to the students. We are only using
water, food dye, Kool Aid, and markers. The only real danger is posed to student
clothing.
Lab equipment:
Each activity will have a detailed list of the necessary equipment and supplies. The
following is a general list of the equipment and supplies needed to conduct all of the
activities included in this module:
1. Chromatography paper: I would suggest ordering this in large sheets. It’s cheaper
and you can cut it into the size you need.
2. Candy for Density Experiments: Small snickers and 3 musketeers candy bars.
Almond M&M’s, Regular Chocolate M&M’s, Peanut M&M’s, and Peanut Butter
M&M’s.
3. Electronic Scales
4. Plastic Pipets
5. Food Coloring:
a. Black Food Coloring (sold in big bottles…one is enough)
b. Boxed Food Coloring (comes with red, blue, green, and yellow)
27
c. Boxed Neon Food Coloring (comes in Neon Green, Pink, Neon Purple,
and…)
6. Toothpicks
7. Sepup Trays: for mixing things. Test tubes work, but are longer and more difficult to
use. Toothpicks should fit into it for mixing and getting samples out with. You
probably have something in your store room
8. Deionized Water: We only used deionized water in our labs due to rules of our lab;
however, I don’t think using regular water will hurt.
9. 50% Rubbing Alcohol: We used 40% isopropyl alcohol for our tests. I believe that
rubbing alcohol is 75-80% isopropyl alcohol.
10. 500 mL beakers or maybe some other large glassware to do the chromatography in
11. Markers: Each should come with eight colors. Black, brown, red, orange, yellow,
green, blue, and violet are the ones needed.
a. Mr. Sketch
b. Crayola (nonwashable kind)
c. Vis-à-vis
d. Sharpies
e. Expos
28
ACTIVITIES AND LESSONS:
Lesson #1
Teacher Notes:
This is the background on the M&M Planet. They need to do this worksheet first in order
to understand some basic biology about the M&M’s so they will be able to do the
labs/project. There is a worksheet that goes with the reading at the end.
Student Handout on Next Page:
29
M&M Cartoon Characters:
In this project a crime will be committed. You will learn how to solve that crime (like a
Crime Scene Investigator). Before starting the project you need to have some basic
background about this ―imaginary planet‖ where the crime will be committed. We will
then learn some techniques that CSI investigators use to solve the cases.
It is 4000 AD on Earth. You are a 13-14 year old on Earth and are studying a distant
culture living on another planet: Planet Mars Candy Bar. This planet is not as advanced
as Earth and they do not realize they are being studied.
On planet Mars Candy Bar resides living and breathing M&M characters. These are
characters (not candies). You, being a fan of M&M candy watch this culture carefully
and identify different physical properties of this M&M culture. Here are some things you
learned.
There are four types of M&M species: Almond M&M’s, Peanut M&M’s, Chocolate
M&M’s, and Peanut Butter M&M’s: just like the candy you buy at the store…absolutely
amazing. Each type of M&M has the exact same physical properties in appearance
(except one). All Almond M&M’s have a height of 6 feet 5 inches and are shaped just
like an almond…how odd!! That’s probably why they are the best M&M basketball
players. All Peanut M&M species have a height of 5 foot 8 inches. They, of course, are
perfectly round. Peanut Butter M&M’s are all 5 foot 6 inches and are also round. And,
finally, the chocolate M&M’s are the smallest at 4 foot 6 inches. They are like little mini
M&M’s. They seem to have a short M&M complex at times, but they are nice M&M’s.
Almond M&M below Peanut M&M pictured
Below
Peanut Butter M&M on the left and
smaller regular Chocolate M&M on
the right.
And, of course there are many female M&M’s.
30
For example, all Almond M&M’s have the same height, width, mass, foot size, hand size,
etc. as the other Almond M&M’s. The only thing in physical appearance that
distinguishes one Almond M&M from another is the color of that M&M. There are six
M&M colors: Red, Orange, Yellow, Green, Blue, and Brown. But, ALL Almond
M&M’s look exactly the same except for their hard shell color. This is also true for
Peanut M&M’s, Chocolate M&M’s, and Peanut Butter M&M’s.
The shell of the M&M is like a human’s blood. And, just as humans have different blood
types so do M&M’s. Red M&M’s have red colored ―blood‖. But, you learn that there
are two types of Red Blood: Type 1 and Type 2. Orange M&M’s have orange colored
blood. There are two types of Orange Blood: Type 1 and Type 2. This pattern is true for
the other colored M&M’s: Yellow, Green, and Blue.
As you watch the M&M’s you notice some other oddities of this society. For example,
they eat only M&M candy which is the same size in their hands as M&M’s are in our
hands on Earth. M&M’s only eat candy M&M’s (not living M&M characters…they
aren’t M&M cannibals). As a result, candy M&M’s is like money to an M&M species.
A rich M&M would have lots of candy M&M’s. This allows him/her to have a luxury
car with spinners, an M&M mansion, etc. An M&M character with lots of M&M candy
is living the high life.
They also only drink Kool Aid. In fact, they are so particular about their Kool Aid that
they only drink certain kinds of punch. If they drink a Kool Aid of a different type they
get sick and throw up. This is why when they urinate they actually urinate the Kool Aid
they drink. Yes, they pee Kool Aid.
There are 6 different colors of Kool Aid: Red, Orange, Yellow, Green, Blue, and Violet.
However, there are two types of each colored Kool-Aid: Type 1 and Type 2. As a result,
at the party there are twelve Kool Aid’s for the M&M’s to drink. There is Type 1 Blue
Kool Aid, Type 2 Blue Kool Aid, Type 1 Red Kool Aid, Type 2 Red Kool Aid, etc.
31
Each M&M must drink the Kool-Aid Type that matches their shell (blood) color. In
addition, since there are two types of each colored Kool Aid they not only must drink the
correct color, but also the correct type. For example, a Blue M&M must drink a Blue
Kool-Aid, but he/she must drink the Kool Aid that matches his/her urine type (Type 1
Kool Aid or Type 2 Kool Aid). If the M&M drinks the wrong Kool-Aid type he/she gets
sick and throws up the Kool Aid.
Finally, there are only 5 types of writing utensils on Planet Mars Candy Bar. There are
Crayola Markers, Sharpies, Expo Markers, Vis a Vis Markers, and Mr. Sketch Markers.
There are eight different colored markers of each type: Red, Orange, Yellow, Green,
Blue, Violet, Brown, and Black. Each M&M is very particular and will ONLY own and
write with a certain type of marker. They can write with any color they want, but it must
be the brand they own. Asking an M&M to write with another brand of marker would be
like asking you to cut off your finger and write with your blood.
32
Name: _______________________________ Worksheet #
Planet Mars Candy Bar and the M&M Characters
Question Answer
1. What will you be doing in this
project (at the end of the project)?
2. What year is it on Earth?
3. What is the other planet called
that you are studying?
4. Which planet is the more
advanced civilization?
5. What lives on this other planet?
6. How many different type of
species live on Planet Mars
Candy Bar?
7. What are the four different
species called?
8. Do all almond M&M species
have the same height, width,
mass, foot size, and hand size?
9. Do all peanut butter M&M
species have the same height,
width, mass, foot size, and hand
size?
10. Do all peanut M&M species have
the same height, width, mass, foot
size, and hand size?
11. Do all chocolate M&M species
have the same height, width,
mass, foot size, and hand size?
12. What makes one chocolate M&M
species LOOK different than
another chocolate M&M species
(besides how they are dressed)?
13. Do chocolate M&M species and
Peanut M&M species have the
same height, width, mass, foot
size, and hand size?
14. Are there any female M&M’s?
33
15. How tall are Chocolate M&M
species?
16. How tall are Peanut Butter
M&M’s?
17. How tall are almond M&M’s?
18. How tall are Peanut M&M’s?
19. What one M&M visual physical
property makes one Peanut Butter
M&M different than another
Peanut Butter M&M?
20. How many different colors does
each different M&M species
come in?
21. List what these different colors
are called
22. Do M&M’s have something
similar to blood?
23. If yes, what is the ―blood‖ of an
M&M character?
24. How many colors of M&M blood
are there?
25. What are the different colors of
M&M blood?
26. How many ―Types‖ of blood are
there for each color?
27. So…how many TOTAL blood
types are there in the M&M
world?
28. What do M&M’s eat?
29. What is money in M&M land?
30. What do M&M’s drink?
31. What happens when they drink
the wrong Kool Aid?
32. What do M&M’s urinate?
33. How many different colors of
Kool Aid are there?
34. What are the colors?
35. How many types of each colored
Kool Aid are there?
34
36. So… how many total different
Kool Aid juices are there?
37. What are the four (4) M&M’s that
must drink the Kool Aid that
matches there shell (blood) color?
38. What are the two (2) M&M’s that
must brink the Kool Aid that does
NOT match their shell color?
39. What colored Kool Aid must
these M&M’s drink?
40. How many types of writing
markers are there?
41. What is each marker brand
called?
42. How many different colors does
each brand of markers come in?
43. What are these colors?
44. Each M&M character owns how
many different brands of
markers?
45. Can an M&M character write
with a different brand of marker
than the one he/she own?
46. What are two physical properties of each M&M listed below:
2 physical properties of Almond M&M Characters:
2 physical properties of Chocolate M&M Characters:
2 physical properties of Peanut Butter M&M Characters:
2 physical properties of Peanut M&M Characters:
Explain why these are considered to be physical properties:
47. What are two physical properties of each M&M listed below:
35
Lesson #2
Notes to Teacher:
An introduction to physical properties and density should be given. These notes are
followed by a lab on density calculation. It is suggested that you do some basic algebra
as well where students are given the density and volume and they must determine the
mass of the substance. This is something they will be doing in the final culminating
project.
Notes:
Slide 1
Notes
Physical Property
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 2 • All substances have their own
characteristics.
– Example:
• Apples:
– Red
– White on the inside
– Odorless until cut
– Juicy.
• Grapes:
– Purple
– Round
– Etc.
– These characteristics are called physical
properties
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 3 • Physical Property:
– Characteristic of a substance
– All substances have their own physical properties
• Examples of things that are physical properties:
• Easy ones:– Odor (Smell)
– Taste
– Color (sight)
– Shape (sight)
– Texture (Feel)
• More difficult ones:– Density
– Boiling point
– Melting point
– Solubility (Chromatography)
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
36
Slide 4 • Physical Properties are used to:
– Describe a substance
– Categorize a substance
– Identify an unknown substance
• We can use one of two things when we
categorize substances:
– Data Table
• Visually organizes properties of substance into
columns and rows.
– Dichotomous Key
• Visually organizes properties of substances into a
web
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 5
LargeOrangeOrange
LargeRedApple
SmallRedCherry
SizeColor
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 6 All Fruits
Large Small
Red Color Orange Color Red Color
Cherry FruitOrange FruitApple Fruit
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 7 1) Density:
1) How much space there is between the
molecules in a substance
2) We can measure density by:mass of the substance divided by Volume of the substance
3) Every substance has it’s own density
number.
4) So … density is a physical property.
5) Density can be used to identify
substances
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
37
Candy Bar Density Determination
Materials: (for each pair of students)
• 3 Musketeers bar (small size)
• Snickers bar (small size)
• Electronic scale
• Metric Ruler
• Beaker (250 mL) … big enough to float
the candy in
• Water
• 2 Plastic Baggies (to seal the candy in)
Teacher Preparation:
a) Prior to starting this lab go through the notes on density. This should be a review for
the students.
b) Put the candy bars in the plastic baggies. Seal them and make sure there is no air in
them.
Teacher Procedure:
a) Make sure each pair of students has a set of materials.
b) Have students follow the student procedure, fill in all data tables and answer all
questions (student page attached) Students should work as independently as possible
even though they share some materials.
Answers:
1. The Snickers bar is denser than the 3 Musketeers bar. We found that Snickers has a
density of 1.07 g/cm3 and the 3 Musketeers bar has a density of 0.96 g/cm
3. This
means that there must be heavier stuff packed inside a Snickers bar. Visual
38
observation and eating the bars revealed that Snickers bars have peanuts and caramel
inside which seem thicker and denser than the nougat stuff which seems to have
bubbles in it.
2. If the bars are wet when massing them, the water would cause you to measure a
higher mass which would lead to a higher density than the true density.
3. For one thing, we noticed that the Snickers bar sinks and the 3 Musketeers bar floats
when put in water. But even without doing that we would be able to predict which
would float by comparing the density of the candy bar to the density of water which is
exactly 1.0 g/ml. Snicker's density is greater than 1.0 and 3 Musketeer's density is less
than 1.0.
Notes to Teacher:
1. This should give students a better understanding of density. Students in 8th
grade
have not measured density yet (at least in my district), but they do have a general
understanding of the difference between mass and density.
2. The other goal is to get students to understand that each object has it‘s own density.
Therefore, making this a physical property of that substance. This can be used to
identify a substance. They will use this skill with the M&M candies so this shows
students how this can be accomplished.
3. Students should work individually on this. They will have there table partners to
compare results at the end, but each should measure everything themselves for both
candies.
4. Have one candy bar cut open on a plate for students to see (of each typ
39
Candy Bar Density Assessment
(Student Handout)
Materials: (for each pair of students)
• 3 Musketeers bar (small size)
• Snickers bar (small size)
• Electronic scale
• Metric Ruler
• Beaker (250 mL) … big enough to float the candy in
• Water
• 2 Plastic Baggies (to seal the candy in)
Procedure:
1) Find the mass of the candy bar you were given. (Do not remove from baggie.) Make
sure the baggie is dry.
Mass of Snickers = _________________ Mass of 3 Musketeers =
_________________
2) Without removing the candy bar from the baggie, find the volume of the candy bar by
measuring the length, width, and height of the candy bar.: Remember: Volume =
Length x Width x Height
Snickers Candy Bar Results
Length of Candy Bar
Width of Candy Bar
Height of Candy Bar
Volume of Candy Bar
3 Musketeers Candy Bar Results
Length of Candy Bar
Width of Candy Bar
Height of Candy Bar
Volume of Candy Bar
40
3) Calculate the density of each candy bar.
Candy Bar Mass Volume Density
(g/mL)
Show how you
determined density
here:
3 Musketeers
Snickers
4) Put each candy bar in a different plastic bag. Seal it and make sure you get all of the
air out of the baggie. Now, drop the baggies with the candy bars in them in a beaker
of water. Describe what you observe:
3 Musketeers:
Snickers:
Post Lab Questions:
1. Which candy bar is more dense? Explain why. What do you think makes it more
dense? (Support your explanation with data you measured and observations you
made.)
2. Explain why each type of candy bar sinks or floats in water. How would you be able
to predict whether each candy bar would float without even putting it in water?
41
3. If a candy bar is denser than water then it will sink or float? Which candy bar is
denser?
4. Look at the results of everybody else. Did everyone else in class have a similar
density number for the 3 musketeers and for the snickers? If so, is density a physical
property? Explain why.
42
Lesson #3
Notes to Teacher:
For students to have a clear understanding of chromatography they need to know what
mixtures and solutions are. The following notes are intended for that purpose.
Slide 1
Notes #
Mixtures vs. Pure Substance
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 2 1. Pure Substance:
– Definition: A substance made of only one
type of molecule
– Example: Salt, Water, etc.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 3 2. Mixture:
– Definition: Substance made of two different types
of molecules. Mixtures are solids, liquids, or
gases
– Examples: Salt and Pepper, Sand and Water
– Important: The parts are mixed together, but they
did NOT combine into one substance.
– Example:
• Red Marbles and Blue M&M’s are mixed together.
They are still separate parts. They did NOT combine or
change. So…red marbles are still not chocolate
3. Components:
– The individual parts of a mixture.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 4 4. Solution:
– A mixture where a solute is dissolved in a solvent.
– Solutions are liquids.
– **The parts are mixed together, but they did NOT
combine into one substance.
– Example:
• Salt and Water are mixed together. They are still separate parts.
They did NOT combine. So…the salt part tastest like salt and the
water part tastes like water.
5. Solvent:
– Definition: A substance that dissolves another substance
to form a solution.
6. Solute:
– Definition: The substance in a solution that is dissolving
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
43
Slide 5 7. Solubility:
– The more easily a substance dissolves is it’s
solubility.
– Example: Salt is more soluble in water than sand
8. A solution is a mixture, but a mixture
doesn’t have to be a solution.
9. Density:
– The more solute you add to a solvent the higher
the density.
– Why: More solute means there is less space
between the molecules.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 6 Adding sugar or another solid that dissolves to water:
Water molecules
bonded together
Sugar
Sugar molecules fill in the spaces
between the water molecules.
The density is now increased.
NOTICE how the salt and water don’t
combine…they are separate.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 7
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 8
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
44
Lesson #4
Teacher Notes:
Here’s some basic information students need to understand about Paper Chromatography.
Without it, what they do during the labs will not make sense.
Notes:
Slide 1
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 2 What is Chromatography?
- Chromatography is a technique for separating mixtures into their different parts by how soluble the substance is in a solvent.
- Once separated the different parts can be analyzed, identified, and purified
Separate
• Analyze
• Identify
• Purify
Solutes
Separated out
Mixture
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 3 Uses for Chromatography
Chromatography is used by scientists to:
• Analyze – examine a mixture and what it is made out of
• Identify – determine the identity of a mixture or components based on what the mixture is made of
• Purify – separate components in order to isolate one of interest for further study
• Quantify – determine the amount of the a mixture and/or the components present in the sample
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 4 Separation of Mixtures Occur
If the different substances in the mixture have a different:
– Magnetic Attraction (magnets can be used)
– Particle Size (filter paper can be used)
– Boiling Point (Then boiling one off and catching the vapor
will separate them)
– Density (using water and adding salt to it can separate them)
– Solubility – how easily a solute dissolves in the solvent. The
more it dissolves the higher up the paper it goes. This is paper
chromatography.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
45
Slide 5 Uses for Chromatography
Real-life examples of uses for chromatography:
• Pharmaceutical Company – determine amount of each chemical found in new product
• Hospital – detect blood or alcohol levels in a patient’s blood stream
• Law Enforcement – to compare a sample found at a crime scene to samples from suspects
• Environmental Agency – determine the level of pollutants in the water supply
• Manufacturing Plant – to purify a chemical needed to make a product
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 6 Definitions
• Stationary Phase: The nonmoving section (for us…the chromatography paper)
• Solvent: What we will be dipping the stationary phase in. This will do the dissolving.
• Solvent Front: As the solvent rises up the stationary phase you will see a line signifying the front end…the solvent front.
• Mobile Phase: This is the moving solvent as it goes up the paper (stationary phase)
• Sample: – The dot/mark you put on the stationary phase
to examine. • Components: The individual parts of a mixture
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 7 Definitions
• Mixture: Substance made out of two different types of components
• Solubility: – How easily a component or particle dissolves in
the solvent. – The more soluble the component the higher up
the particle will move.• Chromatogram:
– The graphic record produced by chromatography
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 8
• Small sample is placed on stationary phase/paper (small dot)
• Solvent passes through the paper (could be water, rubbing alcohol, hydrogen peroxide, etc.)
• Solvent rises up the paper
• Solvent dissolves the small sample (small dot you made)
• Solvent carries the individual components in the small sample a certain distance through the stationary phase, depending on their attraction to the solvent.
• Basically, the more soluble the component the higher it will go.
• If the sample is a mixture with two parts. The more soluble part is carried further up. The less soluble part is near the bottom. You’ll see both.
Basics of Chromatography
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
46
Slide 9 Illustration of Chromatography
More soluble (dissolvable) in solventRed
Most soluble (dissolvable) in solventYellow
Slightly soluble (dissolvable) in solventBlack
Insoluble in SolventBlue
ATTRACTION TO SOLVENTMIXTURE OF
Mixture Components
Separation
Stationary
Phase
(paper)
Mobile Phase
Solvent Front
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 10
Solvent… this
is the mobile
phase that will
travel up
3 samples
being
examined for
their
components
Stationary Phase
None were
mixtures…red
was made of
only red dye.
Blue was made
of only blue dy.
Green was made
of only green dye
Red was least
soluble
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 11 Red Left: Made of only one
type of component. Not a
mixture.
Red Middle: Made of two
components (red and blue). Red
was least soluble so rose the least.
This is a mixture.
Red Right: Made of two
components (red and green).
This is a mixture.
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 12
• Liquid Chromatography – separates liquid samples with a liquid solvent (mobile phase) and a column composed of solid beads (stationary phase)
• Gas Chromatography – separates vaporized samples with a carrier gas (mobile phase) and a column composed of a liquid or of solid beads (stationary phase)
• Paper Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a paper strip (stationary phase)
• Thin-Layer Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a glass plate covered with a thin layer of alumina or silica gel
(stationary phase)
Types of ChromatographyTypes of Chromatography
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
Slide 13 Example of What it looks like
___________________________________
___________________________________
___________________________________
___________________________________
___________________________________
47
Lesson #5
Notes to Teacher:
During the next few labs students will be making sheets that they will compare the
final CSI M&M Cartoon Character crime to. These will be kept by the students and
used again during that lab.
During this lab take the pre-cut square chromatography paper and cut them in half.
Each student will receive two of these.
Also, some common mistakes are to make the dots too big, not putting them on the
line, putting them too close together, or putting too much water into the beaker.
Another mistake is to not put a gap between the paper causing the marks to blead
together. See step 9.
Make sure students do not have the paper touching the sides of the glass container. I
try to stand them up on end with the folds somehow.
The last common mistake is just plopping it in the water. They need to check
beforehand to make sure the water level is below their dots.
You will pass out one more smaller piece of chromatography paper to students to do
permanent markers. They are just doing a test to see what will happen. Of course,
since they aren’t water soluble they won’t go anywhere. Just make sure they get
enough paper to try it.
Students will be keeping and using these for the ending project. If there’s bled badly,
is unreadable, etc. have them start again. They will be unable to identify unknowns
with their chromatograms if that is the case.
Each student will be working individually.
48
Somewhat fuzzy picture, but here are the results of the lab: I did not put in the
permanent ones…nothing happens…not soluble
Materials:
Markers: Mr. Sketch, Crayola (not the washable kind, the kind teachers prefer), Vis
a Vis, Sharpies, and Expo
49
Each brand should have eight different colors in it (Black, Brown, Red, Orange,
Yellow, Green, Blue, and Violet). Some brands are sold in packages of 10 only.
Others in eight. Either way just have them test the above colors.
2 Mason jars per student or beakers big enough to fit the chromatography paper.
1 Chromatography Paper per student pre cut into two equal sized rectangles (thus 2
papers per student). Lab filter paper works and is cheaper, but doesn’t separate out as
neatly or clearly. I prefer the chromatography paper.
As students get the hang of it pass out one more really small piece of chromatography
paper. This one will be used for the permanent markers.
Paper towels (to dry the wet and completed chromatograms).
50
Directions to Student:
Step #1:
Take the rectangular chromatography paper and fold
it in half so it looks like a square.
Step #2:
Open the chromatography paper so you can see the
crease in the center. Now fold each side to the crease
so you have four equally sized sections.
Step #3:
Now open the paper. You should have four sections
of equal size. Repeat steps 1-3 with your other
chromatography paper. So you have two papers like
the one pictured below.
Step #4:
We will be labeling now. Be sure you label small, but
neatly. Use a pen or pencil (no markers).
The “C” stands for Crayola
The “M” stands for Mr.
Sketch
The “V” stands for Vis-a-Vis
Black
C M V
Brown C M V
Red C M V
Orange C M V
Your Name: Markers dipped in Water
Opened Chromatography
Paper with crease from
fold in step #1
51
Step #5:
With a ruler and a pencil only measure ½ inch up
on the bottom of each side. Put a little x there.
Now connect them with a line.
Step #6:
Now, grab the crayola markers. Put them in the
same order as the paper marked below (Black, brown,
red, orange.). Put one small dot on the line for each
crayola marker color under the letter ―C‖. Be careful
here. Do not put it in the wrong spot.
Step #7:
Now, grab the Mr. Sketch markers. Put them in the
same order as the paper marked below (Black, brown,
red, orange.). Put one small dot on the line for each
color under the letter ―M‖.
Step #8:
Now, grab the Mr. Sketch markers. Put them in the
same order as the paper marked below (Black, brown,
red, etc.). Put one small dot on the line for each color
under the letter ―M‖.
Black
C M V
Brown
C M V
Red
C M V
Orange
C M V
Your Name: Markers dipped in Water Black
C M V
Brown
C M V
Red
C M V
Orange
C M V
Your Name: Markers dipped in Water
Black
C M V
Brown
C M V
Red
C M V
Orange C M V
Your Name: Markers dipped in Water
Black
C M V
Brown
C M V
Red
C M V
Orange C M V
Your Name: Markers dipped in Water
52
Step #9:
Fold the paper into a rectangle that can stand up
on it’s own. Do not fold two dots together so they
mix.
The dots will be on the inside. Be careful NOT to
touch the marker spots with your fingers.
Leave a gap between the ends. You do NOT want
them touching.
Put some tape at the top only to hold the gap in
place.
Step #10:
1. Take your beaker and add a small amount of water
to it.
2. Have your folded chromatography paper next to it.
Make sure that the water level will be BELOW the
where the dots are. If the water level is above then
pour some water out.
Step #11:
1. Carefully, place the chromatography paper inside
the beaker.
2. It will take about 10-15 minutes to separate out all
of the dyes. Answer the questions on the next
page while you wait.
Step #12: (in about 10-15 minutes)
1. Pull out the paper (by only touching the dry top
part of the paper) when the water has travelled up
80% of the paper. Ask your teacher if you’re
unsure when.
2. Put the paper on a paper towel folded a few times
to make it thicker. Cut the tape off and let it air
dry. Do not pat it, touch the water section, etc.
Black
C M V
Brown
C M V
Red
C M V
Orange C M V
Folded paper towel to make it thicker
Your Name: Markers dipped in Water
Container
Water Level is below dots
53
Step #13: (while waiting to do step #12)
1. While you are waiting to do Step #12 set up the
other chromatography paper like this one with the
Yellow, Green, Blue, and Violet
2. It should be just like the one you did previously.
Step #14: (while waiting to do step #12)
3. While you are waiting to do Step #12 set up the
other chromatography paper like this one with the
Yellow, Green, Blue, and Violet
4. It should be just like the one you did previously.
Step #15:
1. Your teacher will be coming around with one
small strip of chromatography paper. Take that
strip and fold it in ½ then fill it in like pictured
below
Step #16:
1. Fold the paper in ½ so the dots are on the inside.
Be careful to not touch the dots together so they
mix.
Yellow
C M V
Green
C M V
Blue
C M V
Violet C M V
Fold and tape it so it looks just like the one on the left.
(remember dots on the inside). Then put it in the other
container with water below the dots.
Your Name: Markers dipped in Water Yellow
C M V
Green
C M V
Blue
C M V
Violet C M V
Fold and tape it so it looks just like the one on the left.
(remember dots on the inside). Then put it in the other
container with water below the dots.
Your Name: Markers dipped in Water
Sharpie Expo
Black Red Black Red
Name: Markers dipped in Water
54
Name: _______________________________
Post-Lab Questions:
1. Answer the questions below:
What is a solution:
What is solubility:
Identify the solvent in this experiment:
Identify the stationary phase in this experiment:
Identify the mobile phase in this experiment:
Over time what happens to the water level on the paper:
The leading edge of the rising water level on the paper is called the:
The more soluble dyes should be where on the stationary phase: (higher up or lower
down)
What are the samples that we are testing:
What is a chromatogram?
2. Examine the chromatogram pictured below. It is of a silver color. The samples were
placed into a water solvent.
Mr. Sketch
Crayola
Explain how the different components of the
Crayola Marker were able to rise up and separate
out: (Be complete here…look how much space
you have)
Silver
Orange
Blue
Rose
Yellow
55
3. Examine the black chromatogram for the Mr. Sketch, Crayola, and Vis a Vis Brand
Markers.
Are all the black marker brands made with the same dyes?
Explain how you know:
How many components is the Mr. Sketch black marker made of: Identify
what these are:
Identify the most soluble component/dye in Mr. Sketch:
Identify the 2nd
most soluble component/dye in Mr. Sketch:
Identify the 3rd
most soluble component/dye in Mr. Sketch:
Identify the least soluble component/dye in Mr. Sketch:
4. Examine the Orange chromatogram.
Identify which marker brand (Mr. Sketch, Crayola, or Vis a Vis) is made with one dye
only:
Identify which marker brand is not made with any orange dye:
Explain how they made their marker orange if they didn’t use orange dye:
5. Examine the yellow chromatogram.
Does each yellow dye/component start and end in the same place:
Do the three brands use the same yellow dye or different yellow dyes:
Why do you think so:
6. Examine the Sharpie and Expo Marker Chromatograms.
Describe what occurred when water was used as a solvent:
Explain why:
56
7. You mark your sleeve with a marker. You dab it with water to try to remove it.
If the marker was a crayola would the mark begin to lessen:
Explain why:
If the marker was a Sharpie would the mark begin to lessen:
Explain why:
8. Answer the questions below about physical properties:
What is a physical property:
What physical property was used to separate out the vis-à-vis marker dyes:
9. Answer the questions below about solubility:
What is solubility?
What components make up the green crayola:
Which dye is most soluble, 2nd
most soluble, etc. in the green crayola:
Is solubility a physical property: Explain why:
Does each different dye color component have the same solubility or different solubility:
57
Lesson #6
Note to Teacher:
During this lab take the pre-cut square chromatography paper and cut them in half.
Each student will receive one of these.
Keep in mind the mistakes/tips from the last lab.
Students will be keeping and using these chromatograms for the ending project. If
there’s bled badly, is unreadable, etc. have them start again. They will be unable to
identify unknowns with their chromatograms if that is the case.
Here is what the results should look like with 40% isopropyl alcohol (rubbing alc is
around 80%)…you’ll use 50% rubbing alc.
58
Materials:
Markers: Sharpies and Expos
Each brand should have eight different colors in it (Black, Brown, Red, Orange,
Yellow, Green, Blue, and Violet)
2 Mason jars per student
1 Chromatography Paper per student pre cut into two equal sized rectangles (thus 2
papers per student)
Paper towels (to dry the chromatograms)
Rubbing Alcohol (50%)
59
Directions to Student:
Step #1:
Create your four equal fold sections like we did
before so it looks like the diagram below.
Step #2:
1. Now, we are going to make 8 equal sections.
Fold side A to the ―fold‖ at B so you can put in a
crease like the dashed line. (don’t draw a dashed
line, just create a fold/crease.
2. Do the same thing on the other side by folding
side E to the fold at letter D.
3. Find the ½ way point for the other two sections so
you have 8 equal sections with creases separating
them.
Step #3:
Now, fill in the chromatogram like the one below. (Remember to use
pencil for the line)
Then, put small marker dots in the correct location.
A B C D E
Black
E S
Brown
E S
Red
E S
Orange
E S
Yellow
E S
Green
E S
Blue
E S
Violet
E S
Your Name: Markers dipped in 50% Rubbing Alc.
E = Expo Marker
S = Sharpie Marker
60
Step #4:
Fold the paper into a rectangle (probably look
more like an octagon) that can stand up on it’s
own. Do not fold two dots together so they mix.
The dots will be on the inside. Be careful NOT to
touch the marker spots with your fingers.
Leave a gap between the ends. You do NOT want
them touching.
Put some tape at the top only to hold the gap in
place.
Step #5:
3. Take your beaker and add a small 50% rubbing
alcohol to it. Do NOT put water in it.
4. Have your folded chromatography paper next to it.
Make sure that the water level will be BELOW the
where the dots are. If the water level is above
then pour some water out.
5. Put you paper into the beaker.
6. This one will take a while to rise. 25-30 min.
Then dry on a paper towel as before
Container
Water Level is below dots
61
Name: _______________________________ Pen Chromatography Lab 2
Post-Lab Questions:
1. Answer the questions below:
Identify the solvent in this experiment:
Identify the stationary phase in this experiment:
Identify the mobile phase in this experiment:
Over time what happens to the rubbing alcohol level on the paper:
The leading edge of the rising water level on the paper is called the:
The more soluble dyes should be where on the stationary phase: (higher up or lower
down)
What are the samples that we are testing:
What is a chromatogram?
2. Examine the EXPO and Sharpie Chromatograms.
Which marker is more soluble (EXPO or Sharpie):
Explain how you know:
What liquid is this brand more soluble in (water or rubbing alcohol):
3. Examine all the sharpie chromatograms.
Are most sharpie’s made of multiple dyes like crayola markers:
Which two Sharpie colors are made of two components (dyes) AND what are those two
components for each:
(1)
(2)
What physical property was used to separate these markers into their components:
Did each different dye color component have it’s own specific solubility or does each
different dye component have the exact same solubility:
62
4. Examine the EXPO chromatograms.
Are EXPO markers soluble in water:
Are EXPO markers soluble in 50% rubbing alcohol:
Explain how you know:
If you were to write on your sleeve with an expo would you be able to get it out easily:
5. Examine the red and black sharpie chromatograms.
Which one rose higher on the mobile phase:
Explain why:
6. Joe is in you Social Studies class and likes to mess around with his sharpies. He
―accidentally‖ puts a mark on the sleeve of you favorite white sweatshirt.
Knowing what you learned from this lab can you remove the sharpie mark?
Explain how you would remove it & why this would remove the sharpie mark on your
sweatshirt:
63
Lesson #7
Note to Teacher:
1. The blood types need to be prepared. Using the actual candy coating will get results,
but they only yield 1 color per candy coating color. This is why I use food dye. You
will need three types of food dye: regular food dye colors or red, blue, yellow, and
green, black food dye (they sell at the store in big bottles), and neon food dye package
of four (pink, neon purple, neon green, and something else comes with it)
2. For each sample I add as many drops of water as I add drops of food dye. Students
will be dipping into it with toothpicks and marking there chromatography paper so it
should last. I have 2 students per station, but they each gather their own
chromatography data.
3. Follow the data table for making the M&M ―blood‖ which is the shell color. Also,
some colors won‘t look like the actual color until a small spot is marked on the paper
due to the concentration of the food coloring. Remember, to add drops of water at the
end (1:1 ratio) to make it last longer
Red M&M Orange
M&M
Yellow
M&M
Green M&M Blue M&M Brown
M&M
Type
1
Red food dye 5 drops
yellow
1 drop red
Yellow food
dye and
yellow vis-à-
vis
3 drops green
1 drop red
Blue Food
dye
4 drops red, 2
yellow, and 2
blue
Type
2
4 drops
yellow
6 drops red
8 drops
yellow
2 drops Neon
Purple
Yellow food
dye
Green food
dye
Neon Blue
Food dye
4 drops Neon
Green
1 drop Pink
64
For the Brown: the type 2 below is not used. Type 3 is Type 2 (mislabeled)
4. To create the yellow put the yellow food dye drop in a container. Then draw with the
yellow vis-à-vis on a paper. Cut out the part and then put in test tube. Add 1 drop
water and then combine with the food dye. This was the only way I could get two
yellow colors that look different on the chromatogram.
65
5. During this lab students must each have 2 pre-cut chromatography papers they will
fold into four sections like before. They will keep this for their culminating project so
they must be done carefully or they will not have a good reference to look at.
6. At each station there should be:
6.1.Two students
6.2.One Sepup tray with 10 mixing holes to put ―blood/shell‖ samples (food dye)
6.3.Fill each of the mixing holes with the different blood types and label them. I put
paper underneath and tape it there.
6.4.Have 12 clean toothpicks for each station. Be sure to emphasize they do NOT
mix them up or throw away.
6.5.Four cut chromatography paper per station (2 per student)
6.6.Four mason jars
7. Troubleshooting:
7.1.Be sure students don‘t mix toothpicks up.
7.2.Too small of dots. They should be the size of the marker dots from a sharpened
(new) crayola.
7.3.These are quite concentrated so they do not need to put several dots on after
drying here.
Red Type 1 Orange Type 1 Yellow Type 1 Green Type 1 Blue Type 1 Brown Type 1
Red Type 2 Orange Type 2 Yellow Type 2 Green Type 2 Blue Type 2 Brown Type 2
66
8. Pictures of Chromatography Answers:
Name: _________________________ Determining Blood/Shell Type in M&M’s
Background to Student:
You will be solving an M&M character crime case (i.e. CSI investigation). Just as humans have blood so do
M&M Characters. There ―blood‖ is there shell. There are six shell colors: red, orange, yellow, green, blue, and
brown. Each shell/blood color has two types called Type 1 and Type 2. So, there are actually 12 blood/shell
types: Red Type 1, Red Type 2, Orange Type 1, and so on. You need to determine the exact components of
each blood type. This will be a reference point for you later. You will need two chromatography papers. One
will be created with four sections like we‘ve done in the past. The other you just need to fold in ½ (see the one
on the bottom right). Once you have them folded correctly make them look like the ones below after you have
set them up.
1. Problem: Correctly, create 6 chromatograms (12 samples) of the different M&M blood types. Your grade will be based on how
many you got correct out of six.
2. Gather Information: 2.1. How many different types of M&M blood are there? _______________
2.2. What is M&M blood? ___________________________________
3. Steps to do the Plan: Write the steps to doing your plan. Include what materials you will be using.
Step #1: _ _______________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
4. Diagram of Solution: Draw a diagram of your solution. Label the materials you used.
5. Test Solution: Describe how you will test your solution to see if it was correct or not.
6.
7. Data: Share how your solution worked. Tape your chromatograms on the back page.
Red M&M
Type1 Type2
Orange M&M
Type1 Type2
Yellow M&M
Type1 Type2
Green M&M
Type1 Type2
Blue M&M
Type1 Type2
Brown M&M Type1 Type2
Name: M&M Shell/Blood Name: M&M Shell/Blood
Lesson #8
Name: _____________________________URINALYSIS OF M&M CHARACTERS
Note to Teacher:
1. The urine for the M&M‘s need to be prepared. It is only Kool Aid. Some Kool Aid
has food dye added to it to make the different types. I was only able to make four
different Kool Aid types. Each color needed to have two possible types and I was
unsuccessful with producing two (Orange Kool Aid Drink and Chocolate Kool Aid
Drink). Thus, these two drink Blue Kool Aid. In the future I hope to have time to
correct this or you can find the solution. Not as easy as it seems though.
2. I add as much water will completely dissolve a few pinches of Kool Aid. So…not
very much. At the end I had about 5-8 mL of Kool Aid (before adding any dye).
Remember, kids will be spotting it on the Chromatography paper with toothpicks so
you don‘t need a huge amount.
3. When I add the food dye I would also add a drop of water on top of it.
4. Here‘s what I prepared:
Red M&M Orange
M&M
Yellow
M&M
Green M&M Blue M&M Brown
M&M
Type
1
Tropical
Punch
Orange Lemonade Lemon Lime Ice Blue
Raspberry
Lemonade
+ 2 Blue
Grape + 6
Red + 4 Blue
Type
2
Pink
Lemonade
+ 4 Neon
Pink
+ 4 Red
Orange + 4
Neon Green +
1 Red
Lemonade
+ 2 Yellow
Lemon Lime
+
2 Ne Violet
Ice Blue
Raspberry
Lemonade
Grape + 4
black
69
Mixtures in order (R1, R2, O1, O2, Y1, Y2, G1, G2, B1, B2, Br1, Br2)
5. During this lab students must each have 2 pre-cut chromatography papers they will
fold into four sections like before. They will keep this for their culminating project so
they must be done carefully or they will not have a good reference to look at.
6. At each station there should be:
6.1.Two students
6.2.One Sepup tray with 10 mixing holes to put ―blood/shell‖ samples (food dye)
6.3.Fill each of the mixing holes with the different blood types and label them. I put
paper underneath and tape it there.
70
6.4.Have 12 clean toothpicks for each station. Be sure to emphasize they do NOT
mix them up or throw away.
6.5.Four cut chromatography paper per station (2 per student)
6.6.Four mason jars
7. Troubleshooting:
7.1.Be sure students don‘t mix toothpicks up.
7.2.Too small of dots. They should be the size of the marker dots from a sharpened
(new) crayola.
7.3.These are quite concentrated so they do not need to put several dots on after
drying here.
Red Type 1 Orange Type 1 Yellow Type 1 Green Type 1 Blue Type 1 Brown Type 1
Red Type 2 Orange Type 2 Yellow Type 2 Green Type 2 Blue Type 2 Brown Type 2
71
8. Pictures of Chromatography Answers:
Name: ___________________________ Determining Urinalysis Type in M&M’s
Background to Student:
You will be solving an M&M character crime case (i.e. CSI investigation). Just as humans have urine so do
M&M Characters. This will be just like the blood type. There are two types of urine for each colored M&M.
The urine is actually Kool Aid.
1. Problem: Correctly, create 6 chromatograms of the different M&M urine types. Your grade will be based on how many you got
correct out of six.
2. Gather Information: 2.1. How many different types of M&M urine are there? _______________
2.2. What is M&M urine? ___________________________________
3. Steps to do the Plan: Write the steps to doing your plan. Include what materials you will be using.
Step #1: ________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
4. Diagram of Solution: Draw a diagram of your solution. Label the materials you used.
5. Test Solution: Describe how you will test your solution to see if it was correct or not.
6. Data: Share how your solution worked. Tape your chromatograms on the back page.
Red M&M
Type1 Type2
Orange M&M
Type1 Type2
Yellow M&M
Type1 Type2
Green M&M
Type1 Type2
Blue M&M
Type1 Type2
Brown M&M
Type1 Type2
Lesson #9
Note to Teacher:
In this lab students will learn how to take a sample from a crime scene (mark from one of
the markers) and determine what pen it belongs to by doing some chromatography of the
mark and comparing it to the chromatograms that they did earlier. I run this lab as a
thinking/problem solving lab and help students as needed. Here are some things to keep
in mind:
1. When writing the letters write thick letters
2. Students work in small groups
3. Make five different notes from the five different pens. Different groups have
different notes.
4. Give student groups the following materials:
4.1.Note with letters on it. You can just use thick marks on paper too. This would
be easier for students. Give several so they can try different ideas.
4.2.Sepup Tray
4.3.Water Dropper/Plasic Pippet.
4.4.50% rubbing alcohol (in large sepup tray container)
4.5.Water (in large sepup tray container)
4.6.Toothpicks
4.7.Chromatography paper cut into small strips so they can try different ideas.
5. Picture of Chromatograms of Pens here:
74
75
6. Solution:
6.1.Students need to cut out some letters. Then dissolve them in the sepup tray holes
(smallest ones). Test tubes work…just harder to work with due to depth.
6.2.They should add no more than two drops of water to the letters. One preferable.
It’s amazing how far that can go and it keeps the concentration much higher
which equals better results.
6.3.After trying water then use pure rubbing alcohol (1-2 drops) on new dry letters if
the water does not dissolve the letters.
76
6.4.Use a toothpick to transfer marker liquid to prepared chromatography paper like
previous labs. Let the mark dry and then add another drop. Do this at least five
times to increase concentration and effectiveness. The biggest issue is have
chromatograms that are so light to look at from the concentration of the dot
being too small.
7. Common mistakes…
7.1.Dropping drops all over the paper so they can’t get a clear picture. Students just
need to go slow and be careful.
7.2.I have students do three separate dots that way if one gets messed up they have
two others. If students are not getting results be sure to have them increase the
concentration by using more dots.
Name: ________________________________ Who Wrote that Note Activity
Background:
You go to lunch when a friend gives you a note he/she received from someone who has a ―crush‖ on you. Your
heart goes pitterpat… pitterpat … pitterpat … as you read the note. You secretly hope it’s Candidate A
(pitterpat … pitterpat … pitterpat), the love of your short life so far. But, you think it could also be Candidate
B, Candidate C, Candidate D, or Candidate E based off of some keen observations. You tell your Mom and she
wonders why so many people. Unfortunately, you get to the bottom of the page and there is no name given.
Bummer. You pester your friend, but he/she won’t tell you. Bummer again.
You notice that whoever wrote the note wrote in a black marker. And, you know that each of the five
―suspects‖ you believe could have wrote the note only write in markers. Suspect A writes with nothing but Vis-
à-vis, Suspect B writes with a Sharpie marker, and Suspect C writes with a crayola marker, Suspect D writes
with a Mr. Sketch marker, and Suspect E writes with an EXPO marker.
Luckily, you remember Mr. Morgan stating that you can determine what marker wrote what note by doing
paper chromatography on it and comparing it to the chromatograms of the markers we did in class. You have
all of the marker chromatograms from Mr. Morgan’s class so you decide to scientifically determine who is in
love with you. The ethics of this don’t seem to bother you. You share your idea with Mr. Morgan and he won’t
stop laughing, but he gives you what you need anyway: paper towels, chromatography paper, , plastic
pippets/droppers, scissors, sepup tray, water and rubbing alcohol in the sepup tray big dishes, toothpicks, and a
mason jar. You, of course, provide the love letter. Remember, your immediate dating life depends on you!
Don’t let yourself down.
1. Problem: Determine who wrote the note to you.
2. Gather Information:
A. What brand marker does each suspect write with?
Suspect #1: ________________________________________
Suspect #2: ________________________________________
Suspect #3: ________________________________________
Suspect #4: ________________________________________
Suspect #5: ________________________________________
B. What does soluble mean? __________________________________________________________________________________________
_______________________________________________________________________________________________________________
C. What three marker brands are soluble in water? (1) _______________________ (2)______________________ (3)_______________
D. What marker brand is soluble in rubbing alcohol, but not in water? ________________________________
E. What marker brand is not soluble in rubbing alcohol or water? ___________________________________
3. Explore Ideas: Describe two ideas to solve the problem. Each idea should be explained in a paragraph (3-4 sentences).
Idea #1: _________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
Idea #2: _________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
78
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
4. Plan Summary: Which idea will you choose to use? Include reasons for choosing this selection.
________________________________________________________________________________________________________________________
________________________________________________________________________________________________________________________
5. Steps to do the Plan: Write the steps to doing your plan. Include what materials you will be using.
Step #1: ________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
_______________________________________________________________________________________________________
6. Diagram of Solution: Draw a diagram of your solution. Label the materials you used
7. Test Solution: Describe how you will test your solution to see if it works or not.
8. Data:
Paste your chromatogram
here.
Who wrote the note and how do you know:
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
_____________________________________________________________________________________________
Lesson #10
Note to Teacher:
1. The data sheets will be needed for this lab. See the data tables at the end of this
lesson. I‘ve set it up so there are four different scenarios in the classroom. This way
different teams of students have to solve/find different kidnappers. Students will run
chromatograms to find the kidnapper and cross reference with the suspects.
2. There is one density problem where they have to determine which type of M&M was
involved: Almond or Chocolate. I have not used all four types of M&M‘s because
the densities are too close together. However, you can use all four types of M&M‘s if
you want. The data table below shows the densities of several different types of
M&M‘s. I calculated the volume by doing a length x width x height (they are not
cubes, but it is the most accurate one to use with 8th
graders). Be sure to premeasured
10-15 M&M‘s. Some densities might throw the students off.
Mass
(grams) L (cm) H (cm) W (cm) Volume
(cm3) Density
Chocolate – 1 0.86 1.3 1.3 0.7 1.183 0.726965
Chocolate – 2 0.84 1.4 1.4 0.6 1.176 0.714286
Chocolate – 3 0.78 1.3 1.3 0.6 1.014 0.769231
Chocolate – 4 0.81 1.3 1.3 0.6 1.014 0.798817
Chocolate – 5 0.84 1.4 1.3 0.6 1.092 0.769231
Chocolate – 6 0.84 1.3 1.3 0.6 1.014 0.828402
Chocolate – 7 0.87 1.3 1.3 0.6 1.014 0.857988
Chocolate – 8 0.86 1.3 1.3 0.6 1.014 0.848126
Chocolate – 9 0.86 1.3 1.3 0.6 1.014 0.848126
Chocolate – 10 0.83 1.3 1.3 0.6 1.014 0.81854
PB-1 1.48 1.6 1.5 0.9 2.16 0.685185
PB-2 1.8 1.6 1.5 1.1 2.64 0.681818
PB-3 1.97 1.6 1.5 1.2 2.88 0.684028
PB-4 1.62 1.6 1.5 1 2.4 0.675
PB-5 1.75 1.5 1.6 1 2.4 0.729167
PB-6 1.6 1.6 1.5 1 2.4 0.666667
PB-7 1.75 1.6 1.5 1.1 2.64 0.662879
PB-8 1.73 1.5 1.6 1 2.4 0.720833
PB-9 1.42 1.5 1.5 0.9 2.025 0.701235
PB-10 1.76 1.6 1.5 1 2.4 0.733333
80
Almond – 1 4.15 2.4 1.7 1.5 6.12 0.678105
Almond – 2 3.1 2.5 1.5 1.2 4.5 0.688889
Almond – 3 2.67 2 1.5 1.3 3.9 0.684615
Almond – 4 2.69 2 1.5 1.4 4.2 0.640476
Almond – 5 2.94 1.5 2.3 1.2 4.14 0.710145
Almond – 6 3.27 2.1 1.6 1.5 5.04 0.64881
Almond – 7 2.74 2.1 1.6 1.3 4.368 0.627289
Almond – 8 2.48 2 1.4 1.3 3.64 0.681319
Almond – 9 3.56 2.6 1.5 1.2 4.68 0.760684
Almond – 10 2.86 1.5 1.9 1.3 3.705 0.77193
Peanut – 1 2.21 1.7 1.4 1.3 3.094 0.714286
Peanut – 2 2.58 1.8 1.5 1.3 3.51 0.735043
Peanut – 3 2.25 1.6 1.5 1.3 3.12 0.721154
3. Each student or group of students will receive a piece of paper and some paper towels
with the following on them:
3.1.The density of the culprit.
3.2.Blotch of blood (food coloring) on a paper towel. Paper towels seem to work
better for the food coloring than a piece of paper.
3.3.Blotch of urine (Kool Aid) on a paper towel. Paper towels seem to work better
for the food coloring than a piece of paper.
3.4.Writing in marker. Use large letters or just give thick marks on a piece of paper.
4. Here is an example of what the scenarios might look like: (of course, don‘t give the
kidnapper away…you may use as many different ones as you wish.
Kidnapper Scenario #1
Denstiy of Shoe Imprint = 0.8 g/cm3
Blood Splotch = Green Type 1
Urine Splotch = Green Type 2
Marker = Mr. Sketch
Kidnapper = Jennifer Chocolate Green
Kidnapper Scenario #2
Volume of Shoe Imprint = 0.65 g/cm3
Blood Splotch = Blue Type 1
Urine Splotch = Blue Type 1
Marker = Vis-à-vis
Kidnapper = Abby Almond Blue
81
5. Here is the different mixtures ―splots‖ you will be working from:
Blood/Shell Types Red M&M Orange
M&M
Yellow
M&M
Green M&M Blue M&M Brown
M&M
Type
1
Red food dye 5 drops
yellow
1 drop red
Yellow food
dye and
yellow vis-à-
vis
3 drops green
1 drop red
Blue Food
dye
4 drops red, 2
yellow, and 2
blue
Type
2
4 drops
yellow
6 drops red
8 drops
yellow
2 drops Neon
Purple
Yellow food
dye
Green food
dye
Neon Blue
Food dye
4 drops Neon
Green
1 drop Pink
Urine/Kool Aid Types Red M&M Orange
M&M
Yellow
M&M
Green M&M Blue M&M Brown
M&M
Type
1
Tropical
Punch
Orange Lemonade Lemon Lime Ice Blue
Raspberry
Lemonade
+ 2 Blue
Grape + 6
Red + 4 Blue
Type
2
Pink
Lemonade
+ 4 Neon
Pink
+ 4 Red
Orange + 4
Neon Green +
1 Red
Lemonade
+ 2 Yellow
Lemon Lime
+
2 Ne Violet
Ice Blue
Raspberry
Lemonade
Grape + 4
black
6. Materials students will need:
6.1.Sepup tray with water and 50% rubbing alcohol in the larger containers.
6.2.Two different plastic pipets.
6.3.Toothpicks
6.4.Small strips of chromatography paper
6.5.Paper and Paper towels with the information/clues from the crime
6.6.Handout – shown later in this section/lesson
82
Student Handout:
To Catch an M&M Investigation
Last night was the 500th
annual Independence Day Bash. Each M&M had to sign in at
the door when they entered. This door was monitored.
Three hours into the party somebody noticed that one of the M&M character‘s was
missing. It was Aaron Blue Almond oh no. They checked the last place he was and
noticed a note stating the following:
―If I don‘t get 5 million M&M candies delivered to Bank M&M in three days then Aaron
Blue Almond will be fed to the wild Kit Kats‖
Wild kit kats are very dangerous cookie nuggets (they‘re not just candy on this planet)
that live in the far reaches of Planet Mars Candy Bar.
The following clues were left behind by the M&Mknapper:
1. There was a footprint left behind in the mud. Based off of how much the water
rose in the footprint the density of the culprit was calculated.
2. The note was written with a marker
3. The M&Mknapper left behind some blood (shell color) in his/her haste
4. The toilet was flushed, but some kool aid urine was left behind
You have been doing lots of research in Future Mr. Morgan‘s class (remember this is
4000 AD) so you believe you can help solve this M&Mknapping case. However, going
as a human won‘t work so you gather all of your chromatography data from Future Mr.
Morgan‘s class and drink the M&M elixir so you can become an M&M character and
solve the M&Mknapping.
83
Name: ____________________________ M&M Character
To work on the case you become a living breathing M&M cartoon character. Draw what
you look like below in color. Don‘t forget to give yourself a name that matches what
their names are like. Don‘t get graphic here…these are M&M cartoon characters… be
school appropriate for credit….
Name of M&M: _________________________________________________
Type of M&M: ____________________________________
Place of Birth: Across the Mars Bar Ocean in the city of __________________________
Name: _____________________________ Project Paper: M&M CSI
1. Problem: Determine who kidnapped the M&M‘s.
2. Pre-Lab Questions: The questions below should be answered in 2-3 paragraphs. You should have an
introductory sentence to the paragraph and restate the question for each question below:
There is substance A and Substance B. Substance A is a liquid with a boiling point of 75 degrees.
Substance B is a solid with a boiling point of 300 degrees. The two substances are mixed together so
that a solution forms.
A. What is a solution?
B. What is the solute? Explain why
C. What is the solvent? Explain why
D. Is this a mixture or pure substance? Explain why
E. What is one physical property of Substance A and Substance B? Explain why
F. When the two substances are mixed together into a solution do the components combine into one
substance or do they stay as separate substances?
G. When Substance A is mixed with substance B so a solution forms does the boiling point of substance A
or Substance B change? Explain why.
H. Do all chocolate M&M‘s have the same density? Do all almond M&M‘s have the same density? Is this
density the same or different than the chocolate M&M‘s?
I. Is density a physical property of different types of M&M‘s? Explain why.
3. Steps to do the Plan: Write the steps to doing your plan. Include what materials you will be using.
Step #1:
Step #2:
Step #3:
Etc.
4. Diagram of Solution: Draw a diagram of your solution. Label the materials you used
5. Test Solution: Describe how you will test your solution to see if it works or not.
6. Data: Share what happened in your project. All of your chromatograms and/or calculations should be
organized here so your teacher can clearly see how you came up with who stole the M&M candies. I should be
able to tell the following:
Share the color (before doing chromatography) of the urine
Share the color (before doing chromatography) of blood
Share the color (before doing chromatography) of the marker
Chromatogram of suspect’s blood/shell color and the known chromatograms of ALL of the
different blood/shell colors.
Chromatogram of suspect’s urine/kool aid sample and the known chromatograms of ALL of
the different urine/kool aid samples..
Chromatogram of the suspest’s marker and the known chromatograms of ALL of the different
marker colors and types.
Calculations of density. (Mass, length, width, height, and show your work for how you
determined the density.
85
9. Conclusion: In several paragraphs answer the following:
A. What M&M type did the crime (Peanut Butter, Peanut, Chocolate, or Almond)? Explain how you know
B. What color of M&M did the crime (Red, Orange, Yellow, Green, Blue, or Brown)? Explain how you
know
C. What M&M blood type did the crime? Explain how you know
D. What M&M urine type did the crime? Explain how you know
E. What marker type/brand wrote the letter? Explain how you know
F. Based off of this information who committed the crime? Explain how you know
10. Post Lab Questions:
A. What color was the ―blood/shell‖ blot from your crime scene?
a. Is the ―blood/shell‖ blot a mixture or pure substance? Explain how you know
b. What would be the solvent?
c. What would be the solute(s)?
d. You were able to separate this ―blood/shell‖ blot because it is made out of different substances with
what different physical property?
B. What color was the ―kool aid urine‖ blot?
a. What different components make up this ―kool aid urine‖ blot color?
b. Explain how these different components were able to separate out.
c. The components were able to separate out because of what physical property being different between
each component?
C. What color was the marker that was used?
a. What different components make up this marker color?
b. Do these different components combine in the marker or is it a mixture of separate dyes?
c. Identify which dye was most soluble, 2nd
most soluble, 3rd
most soluble, etc. Explain how you
know.
Red M&M's Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Alm
Red
Density
of M&M Needs to be calculated Blood Type
Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Choc Red
Density of M&M Needs to be calculated
Blood Type
Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
87
Red M&M's Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Peanut
Red
Density
of M&M Needs to be calculated Blood Type
Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
PB Red PB Red PB Red
PB Red
PB Red
PB Red PB Red
PB Red
PB Red
PB Red
PB Red PB Red PB Red
PB Red PB Red
PB Red
PB Red
PB Red
PB Red
PB Red
Density of M&M Needs to be calculated
Blood Type
Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 1 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2 Red 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
Red Kool
Aid 1
Red Kool
Aid 2
88
Orange M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Alm
Orange
Density
of M&M Needs to be calculated Blood
Type
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Choc
Orange
Density
of M&M Needs to be calculated
Blood Type
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
89
Orange M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Peanut
Orange
Density
of M&M Needs to be calculated Blood
Type
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
1
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Orange
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
PB
Orange
Density
of M&M Needs to be calculated
Blood Type
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 1
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Orange 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
Orange
Kool Aid 1
Orange
Kool Aid 2
90
Yellow M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Alm
Yellow
Density
of M&M Needs to be calculated Blood
Type
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Choc
Yellow
Density
of M&M Needs to be calculated
Blood Type
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
91
Yellow M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Peanut
Yellow
Density
of M&M Needs to be calculated Blood
Type
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
1
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Yellow
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Yellow
Kool Aide 1
Yellow
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
PB
Yellow
Density
of M&M Needs to be calculated
Blood Type
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 1
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Yellow 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
Yellow
Kool Aid 1
Yellow
Kool Aid 2
92
Green M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Alm
Green
Density
of M&M Needs to be calculated Blood
Type
Green
1
Green
1
Green
1
Green
1
Green
1
Green 1 Green
1
Green
1
Green
1
Green
1
Green
2
Green 2 Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Green
Kool Aide 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Choc
Green
Density of M&M Needs to be calculated Blood
Type
Green
1
Green
1
Green
1
Green
1
Green
1
Green 1 Green
1
Green
1
Green
1
Green
1
Green
2
Green 2 Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
93
Green M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Peanut
Green
Density
of M&M Needs to be calculated Blood
Type
Green
1
Green
1
Green
1
Green
1
Green
1
Green 1 Green
1
Green
1
Green
1
Green
1
Green
2
Green 2 Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Green
Kool Aide 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Green
Kool Aid 1
Green
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
PB
Green
Density of M&M Needs to be calculated Blood
Type
Green
1
Green
1
Green
1
Green
1
Green
1
Green 1 Green
1
Green
1
Green
1
Green
1
Green
2
Green 2 Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Green
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
Green
Kool
Aid 1
Green
Kool
Aid 2
94
Blue M&M's Name Aaron
Blue
Almond
Allen
Blue
Almond
Abby
Blue
Almond
Allison
Blue
Almond
Ben
Blue
Almond
Brian
Blue
Almond
Beth
Blue
Almond
Becky
Blue
Almond
Betsy
Blue
Almond
Carrie
Blue
Almond
Dan
Blue
Almond
Dave
Blue
Almond
Deanna
Blue
Almond
Darlene
Blue
Almond
Erica
Blue
Almond
Ellen
Blue
Almond
Anne
Blue
Almond
Frank
Blue
Almond
Gary
Blue
Almond
Helen
Blue
Almond
Density
of M&M Needs to be calculated Blood
Type
Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Blue
Kool Aid 1
Blue
Kool Aid 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aid 1
Blue
Kool Aid 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Name Irene
Choc
Blue
Janet Choc
Blue
Jill Choc
Blue
Karen Choc
Blue
Ken Choc
Blue
Larry Choc
Blue
Louise Choc
Blue
Mary Choc
Blue
Megan Choc
Blue
Mark Choc
Blue
Nicole Choc
Blue
Nancy Choc
Blue
Oleta Choc
Blue
Oscar Choc
Blue
Ryan Choc
Blue
Rennae Choc
Blue
Rachel Choc
Blue
Summer Choc
Blue
Sandy Choc
Blue
Sara Choc
Blue
Density
of M&M Needs to be calculated
Blood
Type
Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2
Pen type
owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Blue
Kool Aid 1
Blue
Kool Aid 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aid 1
Blue
Kool Aid 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
Blue
Kool Aide 1
Blue
Kool Aide 2
95
Name Aaron
Peanut
Blue
Allen
Peanut
Blue
Abby
Peanut
Blue
Allison
Peanut
Blue
Ben
Peanut
Blue
Brian
Peanut
Blue
Beth
Peanut
Blue
Becky
Peanut
Blue
Betsy
Peanut
Blue
Carrie
Peanut
Blue
Dan
Peanut
Blue
Dave
Peanut
Blue
Deanna
Peanut
Blue
Darlene
Peanut
Blue
Erica
Peanut
Blue
Ellen
Peanut
Blue
Anne
Peanut
Blue
Frank
Peanut
Blue
Gary
Peanut
Blue
Helen
Peanut
Blue
Density
of M&M Needs to be calculated Blood Type
Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2
Pen Type Owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Blue Kool
Aid 1
Blue Kool
Aid 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aid 1
Blue Kool
Aid 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Name Irene
PB Blue
Janet
PB Blue
Jill PB
Blue
Karen
PB Blue
Ken
PB Blue
Larry
PB Blue
Louise
PB Blue
Mary
PB Blue
Megan
PB Blue
Mark
PB Blue
Nicole
PB Blue
Nancy
PB Blue
Oleta
PB Blue
Oscar
PB Blue
Ryan
PB Blue
Rennae
PB Blue
Rachel
PB Blue
Summer
PB Blue
Sandy
PB Blue
Sara
PB Blue
Density
of M&M Needs to be calculated
Blood Type
Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 1 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2 Blue 2
Pen type owned
Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a Vis
Vis a Vis
Mr. Sketch
Mr. Sketch
Expo Expo Sharpie Sharpie
Urinalysis Blue Kool
Aid 1
Blue Kool
Aid 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aid 1
Blue Kool
Aid 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
Blue Kool
Aide 1
Blue Kool
Aide 2
96
Brown M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Alm
Brown
Density
of M&M Needs to be calculated
Blood
Type
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Choc
Brown
Density
of M&M Needs to be calculated Blood Type
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
97
Brown M&M Name Aaron Allen Abby Allison Ben Brian Beth Becky Betsy Carrie Dan Dave Deanna Darlene Erica Ellen Anne Frank Gary Helen
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Peanut
Brown
Density
of M&M Needs to be calculated
Blood
Type
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
1
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Brown
2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Name Irene Janet Jill Jake Jerry Jennifer Kelly Larry Ken Louise Megan Mellissa Nicole Nelly Nancy Oleta Ryan Sam Sara Taylor
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
PB
Brown
Density
of M&M Needs to be calculated Blood Type
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 1
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Brown 2
Pen Type
Owned
Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie Crayola Crayola Vis a
Vis
Vis a
Vis
Mr.
Sketch
Mr.
Sketch
Expo Expo Sharpie Sharpie
Urinalysis Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
Brown
Kool Aid 1
Brown
Kool Aid 2
High School Leaf Chromatography Project
Separating Leaf Pigments Using Paper Chromatography
Safety precautions:
Though none of the activities carried out in this module pose any undo risk of injury,
common laboratory safety practices should be observed. Protective eyewear should be
worn at all times during the lab activities. Though unlikely, the possibility of eye damage
due to contact with methanol and/or acetone should be noted to the students, and caution
should be exercised when working with the hot water baths and hot plate.
Lab Equipment:
test tubes
beakers
chromatography paper
methanol
acetone
scissors
leaves of spinach, Oregon grape, purple flowers, and any dark green leaf
tooth picks or capillary tubes
hot plate
hot water bath
sand
99
ruler
tape
plastic pipettes
Lesson #1
Objective:
In this project, you will uncover the changing colors of fall leaves by separating plant
pigments with paper chromatography.
Introduction:
We all enjoy the colors of autumn leaves. Did you ever wonder how and
why a fall leaf changes color? Why a maple leaf turns bright red? Where do the yellows
and oranges come from? To answer those questions, we first have to understand what
leaves are and what they do.
Leaves are nature's food factories. Plants take water from the ground through their roots.
They take a gas called carbon dioxide from the air. Plants use sunlight to turn water and
carbon dioxide into glucose. Glucose is a kind of sugar. Plants use glucose as food for
energy and as a building block for growing. The way plants turn water and carbon
dioxide into sugar is called photosynthesis. That means "putting together with light."
Tree leaves have pigments or colorful molecules, inside them. These pigments make food
for the tree when the energy of the sun is absorbed by the plant pigments. A chemical
100
called chlorophyll helps make photosynthesis happen. Chlorophyll is what gives plants
their green color.
Chlorophyll molecule
As summer ends and autumn comes, the days get shorter and shorter. This is how the
trees "know" to begin getting ready for winter.
During winter, there is not enough light or water for photosynthesis. The trees will rest,
and live off the food they stored during the summer. They begin to shut down their food-
making factories. The green chlorophyll disappears from the leaves. As the bright green
fades away, we begin to see yellow and orange colors that we call accessory pigments.
101
Small amounts of these colors have been in the leaves all along. We just can't see them in
the summer, because they are covered up by the green chlorophyll.
The bright reds and purples we see
in leaves are made mostly in the fall. In some trees, like maples, glucose is trapped in the
leaves after photosynthesis stops. Sunlight and the cool nights of autumn cause the leaves
turn this glucose into a red color. The brown color of trees like oaks is made from wastes
left in the leaves.
It is the combination of all these things that make the beautiful colors we enjoy in the fall.
Chlorophyll (green) - Chlorophyll is necessary for photosynthesis, which is the
chemical reaction that enables plants to use sunlight to manufacture sugars for
their food. Trees in the temperate zones store these sugars for their winter
dormant period and so do not need to actively make food during the winter. Tree
leaves must constantly make chlorophyll when they need it for photosynthesis, so
once they stop, the green goes away.
102
Xanthophylls (yellow) and Carotenoids (orange) - These pigments are also used
for photosynthesis, and are there all summer long, you just don't see them because
they are usually masked by the strong color of the green chlorophyll. These
pigments do not break down as fast as chlorophyll, so they hang around longer
during the fall. These pigments are also commonly found in such things as corn,
carrots, and daffodils, as well as rutabagas, buttercups, and bananas.
Anthocyanins (red) -The anthocyanins are different, because they begin to appear
after the plant stops making chlorophyll. So these pigments are not there during
the summer, and they appear latest in the fall succession of color. Anthocyanins
are very intense color molecules, and they also give color to such familiar things
as cranberries, red apples, concord grapes, blueberries, cherries, strawberries, and
plums. They are water soluble and appear in the watery liquid of leaf cells.
When you look at a leaf, you see the result of all of these colors mixing together. But if
you separate the pigment molecules, you can see each individual pigment color on its
own. One method for separating molecules is called paper chromatography. In this
method, a solvent is used to dissolve the molecules of interest that could be water or an
organic solvent such as methanol or acetone. Then the solution containing the dissolved
molecules is passed through a strip of strong paper. The fibers of the paper trap the
molecules as the solvent carries them through the paper. Larger molecules get trapped by
the paper fibers first, and smaller molecules can travel farther along through the paper
fibers. With this method, a colorful mixture of pigment molecules can be separated by
size.
103
In this project you will use paper chromatography to separate the colored pigment
molecules from fall leaves. By collecting leaves at different stages of turning, you will be
able to capture all of the colors of fall. Will you be able to uncover the hidden colors, and
tell the full fall story?
Terms, Concepts and Questions to Start Background Research:
To do this type of experiment you should know what the following terms mean. Have an
adult help you search the Internet or take you to your local library to find out more.
Pigment
Photosynthesis
Chlorophyll
Xanthophyll
Carotenoid
Anthocyanin
Paper chromatography
Solvent
PRE-LAB QUESTIONS
What makes a leaf look so colorful?
Can I extract the pigment from a leaf?
104
Why do leaves turn fall colors?
Are plant pigments involved in leaves turning fall colors?
Why is energy required for life?
How does energy enter the living world?
Visible white light is composed of what
Procedure:
Part I: Leaf Pigment Preparation
1. Obtain a spinach leaf, tear it into small pieces, and grind it using a mortar and pestle.
Add a few grains of sand to facilitate grinding. Add 8-10 drops of acetone to extract the
pigments from the leaf.
2. Continue grinding until you have a few drops of dark green liquid. It might be
necessary to add a little more acetone.
3. Pipette the liquid only into a test tube, screw on the cap, and place into an
approximately 150 degree F hot water bath for several minutes.
4. Obtain a strip of chromatography paper which is about 1 inch wide and 10 inches long.
Fold the paper so that it will stand upright in a beaker. Do not handle the flat part of the
paper with your fingers. Oils from your skin will alter how the solvent reacts with the
105
paper and affect your results. Just handle the edges of the paper and/or put on a pair of
latex gloves. Label the top of the paper ―Spinach‖.
5. Using a PENCIL, draw a line across the paper approximately 1 inch from the bottom.
This is your baseline.
6. Using pipette tip or a toothpick, apply a small dot of the pigment extract at the center
of the line. Let dry and repeat four or five times. You should have a small, very dark spot
of pigment. If the spot is not dark, apply more extract.
7. Pour the required amount of methanol into a beaker or mason jar so that the top of the
solvent is below your baseline (about ½ inch deep), then place the chromatography paper
in the methanol.
8. Allow the chromatogram to run until the solvent almost reaches the top of the paper
strip (5-15 minutes).
9. Remove the paper and use a pencil to mark a line at the highest point the solvent
reached. This is your solvent front.
10. Examine the chromatogram for the presence of different bands of color. Each color
band is a different pigment. Chlorophyll a appears blue-green, chlorophyll b appears
yellow-green, carotene appears bright yellow/orange, and xanthophyll appears pale
yellow-green. You may not see all of these bands.
11. Once dry, delineate each of the colored bands with a pencil, and record the color and
the distance each band moved from the starting line or origin (the pencil line you drew at
106
the beginning). Describe the colors as specifically as you can (olive green, grass green,
yellow orange, purple, etc). Record the distance of the solvent front from the origin.
12. Label your chromatogram and save it to attach to your lab report
13. Calculate the Rf values for each of your pigments
Rf = distance traveled by pigment / distance traveled by solvent
Part II: Data Analysis
For each of your pigments you have an Rf value and a description of the color of each
band. Pigments may be identified by their color and the Rf values.
1. Use your chromatogram to fill in the following table (report data in your note book)
Pigment / solvent band (Color) Distance traveled (cm) Rf
Chlorophyll a
Chlorophyll b
Anthocyanins
Carotenoids
Xanthophylls
107
2. What do the Rf values indicate about the relative attraction of the pigments to the
more polar paper or the more non-polar (hydrophobic) solvent? Review the introduction
to this lab if necessary…
3. From your chromatogram, what is the order in which the pigments separated?
4. Is each band one pure pigment? Explain.
Extensions: Repeat the above procedure using Oregon grape leaves and a four-way mix
of leaves and flowers.
108
Extension 1. Oregon grape. Use your chromatogram to fill in the following table.
Pigment / solvent band (Color) Distance traveled (cm) Rf
Chlorophyll a
Chlorophyll b
Anthocyanins
Carotenoids
Xanthophylls
Extension 2. 4-way mix. Use your chromatogram to fill in the following table.
Pigment / solvent band (Color) Distance traveled (cm) Rf
Chlorophyll a
Chlorophyll b
Anthocyanins
Carotenoids
Xanthophylls
109
Discussion Questions:
1. What is the value of chromatography?
2. Which pigments are present in the smallest amounts in the leaf?
3. Which pigments are present in the greatest amount?
4. a) What is the role of chlorophyll a? b) What are the roles of carotene and xanthophyll?
5. Which pigment(s), chlorophyll a, chlorophyll b, and/or carotenoids, will travel the
farthest on
the chromatography paper?
110
6. Which pigment(s) is least soluble in the solvent?
7. Besides leaves, name 2 other sources of natural pigments.
111
Additional Questions:
1. Describe what happened to the original line of spinach juice that you painted onto the
chromatography paper.
________________________________________________________________________
________________________________________________________________________
______________________________________________________________
2. Based on the colors of the bands, which plant pigments were you able to separate and
visualize?
________________________________________________________________________
________________________________________________________________________
3. Besides leaves, name 2 other sources of natural pigments.
________________________________________________________________________
_______________________________
4. How might an environmental scientist use chromatography? How could he/she use the
results?
________________________________________________________________________
________________________________________________________________________
112
5. What is the name for the pattern that is produced as a result of the different plant
pigments
separating into different locations on the paper?
____________________________________
6. Which 3 of your lab materials are the most important and the ones that you should use
first?
________________________________________________________________________
________________________________________________________________________
8. Many leaves change color in the fall. Explain. (Hint: chlorophyll a and b are easily
broken down in the cooler autumn temperatures.)
Bibliography
www.science buddies.org/science-fair-projects
www.sciencemade simple.com/leaves
113
APPENDIX A: Additional leaf chromatography activities.
Activity 1: (adapted from www.science buddies.org/science-fair-projects)
Materials and Equipment
Leaves at different stages of turning colors (30-40, 10 per color group)
Scissors
Good, strong glasses (3-4)
Rubbing alcohol (isopropyl alcohol)
Wooden spoon
Fork
Very small bowls for evaporating and concentrating extract
Strong, white, heavy-weight, ultra-absorbent paper towels
Ruler
Pencil
Toothpicks
Plate
Tall mason jars (3-4)
Clothes pins (3-4)
Experimental Procedure
114
1. Go on a nice walk with an adult and collect some leaves from different stages of
color change during fall. It is best for all of your leaves to come from the same
tree, so look for a tree with a variety of leaves at different stages. Here are some
leaves I collected from a tree in my neighborhood:
2. Separate and group the leaves into color groups, with ten good leaves in each
group (unless you are using a tree with small leaves, like aspen or birch, then you
should use a higher number of leaves). Try to form groups from colors that are as
different as possible. For example, I made a green group, a yellow group, and a
red group. In each group, I chose leaves in the deepest colors possible:
115
3. Cut the leaves into small pieces with your scissors and put each group into the
bottom of a good, strong glass:
4. Add 1 Tbsp. of rubbing alcohol to each glass.
5. Using the blunt end of a wooden spoon, macerate (soften) the chopped leaves by
squashing them into the rubbing alcohol at the bottom of the cup.
6. As you squish the leaves, you will notice that the rubbing alcohol will start to turn
the color of the leaves. This is called extraction, and the rubbing alcohol is called
the solvent.
7. Continue until the liquid turns a deep shade of the color of the leaves, about 5
minutes per glass.
8. Let the macerated suspensions sit for 30 minutes in a dark, room-temperature
place to allow the color molecules to fully extract.
9. Using a fork, lift out the bits and pieces of leaf material and set them aside. Take
care to remove any liquid by gently pressing the leafy bits against the glass before
116
you remove them. You should be left with a dark suspension of leafy color in
rubbing alcohol at the bottom of your cup.
10. Pour each extract into a very small bowl (I used tealight candle holders), and
leave in a dark room-temperature place to evaporate off some of the rubbing
alcohol. This will concentrate your extract, and make the color even more intense.
In the meantime, you should prepare your paper towel strips.
11. Cut up a good, thick piece of paper towel into long, 1-inch thick strips. Make sure
they are long enough to reach the bottom of the mason jars and still drape over the
top. You will need a few strips (2-3) for each color group.
12. Measure up from the bottom of each strip and using a pencil, gently draw a line
that is 1 inch from the bottom of the strip. This will be where you apply the color
extract to the paper towel strip for your separation.
13. When your color extracts have concentrated, they will be gooey when stirred by a
toothpick. Stir each color thoroughly with a toothpick to blend and loosen any bits
117
of dried up pigment from the side of the bowl. Be sure to use a different toothpick
for each separate color so you don't mix them!
14. Using the toothpick, "paint" some of the colored extract onto the pencil line on
your paper towel strip. Some plant pigments can stain, so you should do this on a
plate so that the color won't seep through and stain your work surface. Try to
apply the extract as smoothly and evenly as you can along the line. Repeat with 2-
3 more strips, using the same color extract, so that you have duplicates for each
color pigment.
15. Repeat with the other colored extracts. Be sure to use a new toothpick for each
different color! Allow the strips to dry.
16. While the strips are drying, pour a small amount of rubbing alcohol into each
mason jar, just enough to cover the bottom. About 2 Tbsp. will usually do the
trick. You will need one jar for each color extract you have.
17. When the strips are dry, carefully lower the pigmented end of the strip down into
the jar until the bottom edge of the strip just touches the alcohol. Drape the top of
the strip over the mouth of the jar and secure it with a clothes pin. Hang the other
strips similarly. You can put strips of the same color extract together in the same
jar, just be sure to keep them from touching each other and drape the tops
separately over the mouth of the jar, each strip secured with a different clothes
pin. However, different colored extracts should be in different jars.
18. Make sure that the strips do not come into contact with the sides of the jar, except
for at the top where they are secured, so that they are all hanging freely. Leave the
jars undisturbed.
118
19. Set the glasses aside for about 30 minutes, and watch as the colors separate along
the length of the strip. Stop when one of the colors reaches the top. As soon as
one of the colors reaches the top of a strip, remove ALL of the strips and allow
them to dry.
20. Compare the colors found in the different strips. What happened to the colors?
Did the different groups of leaves have unique colors, or shared colors, or both? Is
each color found in the same place along each strip, or in different places? Are the
colors in the same order, or in a different order of separation along the strip?
Variations
If you find a really good tree, you can include all of the intermediate stages of leaf
turning in your experiment. An especially good source of a wide variety of colors
are aspen trees!
119
There are many other natural sources of color, and you can use the same rubbing
alcohol extraction technique to see them. How do the color molecules of different
plant sources compare? Some ideas to try are: red cabbage, blueberries,
cranberries, carrots, beets, spinach, flowers, and practically any other intensely
colored plant you can get a hold of.
This experiment uses paper towels to separate the colors, but if you want a more
precise and advanced way of separating the colors you can use laboratory filter
paper or thin layer chromatography.
120
Activity 2:
Autumn Leaves Science Projects
NOTE: ADULT SUPERVISION IS REQUIRED. Please read all instructions completely
before starting. Observe all safety precautions.
PROJECT 1 - Separate Colors in a Green Leaf using Chromatography
What you need:
leaves, small jars (baby food jars work well)
covers for jars or aluminum foil or plastic wrap
rubbing alcohol, paper coffee filters
shallow pan, hot tap water, tape, pen
plastic knife or spoon, clock or timer.
What you do:
1. Collect 2-3 large leaves from several different trees. Tear or chop the leaves into
very small pieces and put them into small jars labeled with the name or location
of the tree.
2. Add enough rubbing alcohol to each jar to cover the leaves. Using a plastic knife
or spoon, carefully chop and grind the leaves in the alcohol.
SAFETY NOTE: Isopropyl rubbing alcohol can be harmful if mishandled or
misused. Read and carefully follow all warnings on the alcohol bottle.
121
3. Cover the jars very loosely with lids or plastic wrap or aluminum foil. Place the
jars carefully into a shallow tray containing 1 inch of hot tap water.
SAFETY NOTE: Hot water above 150 F can quickly cause severe burns. Experts
recommend setting your water heater thermostat no higher than 125 F.
4. Keep the jars in the water for at least a half-hour, longer if needed, until the
alcohol has become colored (the darker the better). Twirl each jar gently about
every five minutes. Replace the hot water if it cools off.
5. Cut a long thin strip of coffee filter paper for each of the jars and label it.
6. Remove jars from water and uncover. Place a strip of filter paper into each jar so
that one end is in the alcohol. Bend the other end over the top of the jar and secure
it with tape.
7. The alcohol will travel up the paper, bringing the colors with it. After 30-90
minutes (or longer), the colors will travel different distances up the paper as the
alcohol evaporates. You should be able to see different shades of green, and
possibly some yellow, orange or red, depending on the type of leaf.
8. Remove the strips of paper, let them dry and then tape them to a piece of plain
paper. Save them for the next project.
PROJECT 2 - Separate Colors in a Fall Leaf using Chromatography
What you need: same as Project 1.
What you do:
122
1. Repeat step (1)-(8) from Project 1, this time using leaves that have changed color.
You may have to wait much longer in steps (4) and (7). There is normally much
less of the other colors in the leaves compared to the green chlorophyll.
Bibliography
This project was adapted from "How To Do Paper Chromatography With Leaves" at
About.com: Chemistry:
Helmenstine, A.M. (2007). How To Do Paper Chromatography With Leaves. About.com:
Chemistry. The New York Times Company. Retrieved December 6, 2007 from
http://chemistry.about.com/cs/howtos/ht/paperchroma.htm
123
APPENDIX B
History
Main article: History of chromatography
(http://en.wikipedia.org/wiki/History_of_chromatography)
The history of chromatography begins during the mid-19th century. Chromatography,
literally "color writing", was used—and named— in the first decade of the 20th century,
primarily for the separation of plant pigments such as chlorophyll. New types of
chromatography developed during the 1930s and 1940s made the technique useful for
many types of separation process.
Some related techniques were developed during the 19th century (and even before), but
the first true chromatography is usually attributed to Russian botanist Mikhail
Semyonovich Tsvet, who used columns of calcium carbonate for separating plant
pigments during the first decade of the 20th century during his research of chlorophyll.
Chromatography became developed substantially as a result of the work of Archer John
Porter Martin and Richard Laurence Millington Synge during the 1940s and 1950s. They
established the principles and basic techniques of partition chromatography, and their
work encouraged the rapid development of several types of chromatography method:
paper chromatography, gas chromatography, and what would become known as high
performance liquid chromatography. Since then, the technology has advanced rapidly.
Researchers found that the main principles of Tsvet's chromatography could be applied in
many different ways, resulting in the different varieties of chromatography described
124
below. Simultaneously, advances continually improved the technical performance of
chromatography, allowing the separation of increasingly similar molecules.
125
APPENDIX C:
Chromatography terms:
The analyte is the substance that is to be separated during chromatography.
Analytical chromatography is used to determine the existence and possibly also
the concentration of analyte(s) in a sample.
A bonded phase is a stationary phase that is covalently bonded to the support
particles or to the inside wall of the column tubing.
A chromatogram is the visual output of the chromatograph. In the case of an
optimal separation, different peaks or patterns on the chromatogram correspond to
different components of the separated mixture.
Plotted on the x-axis is the retention time and plotted on the y-axis a signal (for
example obtained by a spectrophotometer, mass spectrometer or a variety of other
detectors) corresponding to the response created by the analytes exiting the
system. In the case of an optimal system the signal is proportional to the
concentration of the specific analyte separated.
A chromatograph is equipment that enables a sophisticated separation e.g. gas
chromatographic or liquid chromatographic separation.
Chromatography is a physical method of separation in which the components to
be separated are distributed between two phases, one of which is stationary
126
(stationary phase) while the other (the mobile phase) moves in a definite
direction.
The effluent is the mobile phase leaving the column.
An immobilized phase is a stationary phase which is immobilized on the support
particles, or on the inner wall of the column tubing.
The mobile phase is the phase which moves in a definite direction. It may be a
liquid (LC and CEC), a gas (GC), or a supercritical fluid (supercritical-fluid
chromatography, SFC). A better definition: The mobile phase consists of the
sample being separated/analyzed and the solvent that moves the sample through
the column. In one case of HPLC the solvent consists of a carbonate/bicarbonate
solution and the sample is the anions being separated. The mobile phase moves
through the chromatography column (the stationary phase) where the sample
interacts with the stationary phase and is separated.
Preparative chromatography is used to purify sufficient quantities of a
substance for further use, rather than analysis.
The retention time is the characteristic time it takes for a particular analyte to
pass through the system (from the column inlet to the detector) under set
conditions. See also: Kovat's retention index
The sample is the matter analysed in chromatography. It may consist of a single
component or it may be a mixture of components. When the sample is treated in
the course of an analysis, the phase or the phases containing the analytes of
interest is/are referred to as the sample whereas everything out of interest
127
separated from the sample before or in the course of the analysis is referred to as
waste.
The solute refers to the sample components in partition chromatography.
The solvent refers to any substance capable of solubilizing other substance, and
especially the liquid mobile phase in LC.
The stationary phase is the substance which is fixed in place for the
chromatography procedure. Examples include the silica layer in
Chromatography#Thin layer chromatography
Bibliography: http://en.wikipedia.org/wiki/Chromatography
128
APPENDIX D:
LEARN MORE ABOUT:
HOW PLANTS PREPARE FOR WINTER
All summer, with the long hours of sunlight and a good supply of liquid water, plants are
busy making and storing food, and growing. But what about wintertime? The days are
much shorter, and water is hard to get. Plants have found many different ways to get
through the harsh days of winter.
Some plants, including many garden flowers, are called "annuals," which means they
complete their life cycle in one growing season. They die when winter comes, but their
seeds remain, ready to sprout again in the spring. "Perennials" live for more than two
years. This category includes trees and shrubs, as well as herbaceous plants with soft,
fleshy stems. When winter comes, the woody parts of trees and shrubs can survive the
cold. The above ground parts of herbaceous plants (leaves, stalks) will die off, but
underground parts (roots, bulbs) will remain alive. In the winter, plants rest and live off
stored food until spring.
As plants grow, they shed older leaves and grow new ones. This is important because the
leaves become damaged over time by insects, disease and weather. The shedding and
replacement continues all the time. In addition, deciduous trees, like maples, oaks and
elms, shed all their leaves in the fall in preparation for winter.
129
"Evergreens" keep most of their leaves during the
winter. They have special leaves, resistant to cold and moisture loss. Some, like pine and
fir trees, have long thin needles. Others, like holly, have broad leaves with tough, waxy
surfaces. On very cold, dry days, these leaves sometimes curl up to reduce their exposed
surface. Evergreens may continue to photosynthesize during the winter as long as they
get enough water, but the reactions occur more slowly at colder temperatures.
During summer days, leaves make more glucose than the plant needs for energy and
growth. The excess is turned into starch and stored until needed. As the daylight gets
shorter in the autumn, plants begin to shut down their food production.
Many changes occur in the leaves of deciduous trees before they finally fall from the
branch. The leaf has actually been preparing for autumn since it started to grow in the
spring. At the base of each leaf is a special layer of cells called the "abscission" or
separation layer. All summer, small tubes which pass through this layer carry water into
the leaf, and food back to the tree. In the fall, the cells of the abscission layer begin to
swell and form a cork-like material, reducing and finally cutting off flow between leaf
and tree. Glucose and waste products are trapped in the leaf. Without fresh water to
renew it, chlorophyll begins to disappear.
The bright red and purple colors come from anthocyanin (an-thuh-'si-uh-nuhn) pigments.
These are potent antioxidents common in many plants; for example, beets, red apples,
130
purple grapes (and red wine), and flowers like violets and hyacinths. In some leaves, like
maple leaves, these pigments are formed in the autumn from trapped glucose. Why would
a plant use energy to make these red pigments, when the leaves will soon fall off? Some
scientists think that the anthocyanins help the trees keep their leaves a bit longer. The
pigments protect the leaves from the sun, and lower their freezing point, giving some
frost protection. The leaves remain on the tree longer, and more of the sugars, nitrogen
and other valuable substances can be removed before the leaves fall. Another possible
reason has been proposed: when the leaves decay, the anthocyanins seep into the ground
and prevent other plant species from growing in the spring.
Brown colors come from tannin, a bitter waste product. Other colors, which have been
there all along, become visible when the chlorophyll disappears. The orange colors come
from carotene ('kar-uh-teen) and the yellows from xanthophyll ('zan-thuh-fil). They are
common pigments, also found in flowers, and foods like carrots, bananas and egg yolks.
We do not know their exact role in leaves, but scientists think they may be involved
somehow in photosynthesis. Different combinations of these pigments give us a wide
range of colors each fall.
As the bottom cells in the separation layer form a seal between leaf and tree, the cells in
the top of the separation layer begin to disintegrate. They form a tear-line, and eventually
the leaf is blown away or simply falls from the tree.
One more important question remains. What causes the most spectacular display? The
best place in the world for viewing fall colors is probably the Eastern United States. This
is because of the climate there, and the wide variety of deciduous trees. The brightest
131
colors are seen when late summer is dry, and autumn has bright sunny days and cool (low
40's Fahrenheit) nights. Then trees make a lot of anthocyanin pigments. A fall with
cloudy days and warm nights brings drab colors. And an early frost quickly ends the
colorful display.
www.sciencemade simple.com/leaves
132
APPENDIX E:
Adapted from:
http://www.google.com/imgres?imgurl=http://employees.csbsju.edu/hjakubowski/classes
/ch331/oxphos/photosynth.gif&imgrefurl=http://employees.csbsju.edu/hjakubowski/class
es/ch331/oxphos/olphotsynthesis.html&h=850&w=705&sz=39&tbnid=YzmOhpbia04RE
M:&tbnh=145&tbnw=120&prev=/images%3Fq%3Dphotosynthesis%2Blight%2Breactio
ns&hl=en&usg=__HXPAbIHGPaYnB8nYGaR-
uTz3dEw=&ei=kFpvSr32F4jIsQOfvJj7Ag&sa=X&oi=image_result&resnum=2&ct=ima
ge
The Photosynthetic Light Reactions:
We have just seen how we can transduce the chemical potential energy stored in
carbohydrates into chemical potential energy of ATP - namely through coupling the
energy released during the thermodynamically favored oxidation of carbon molecules
through intermediaries (high energy mixed anhydride in glycolysis or a proton gradient in
aerobic metabolism) to the thermodynamically uphill synthesis of ATP. There is a
situation that occurs when we wish to actually reverse the entire process and take CO2 +
H2O to carbohydrate + O2. This process is of course photosynthesis which occurs in
plants and certain photosynthetic bacteria and algae. Given that this process must by
nature be an uphill thermodynamic battle, let us consider the major requirements that
must be in place for this process to occur:
An strong oxidizing agent must be formed which can take water and oxidize it to
dioxygen. We know that redox reactions occur in the direction of stronger to
weaker oxidizing agent (just as acid base reactions are thermodynamically
favored in the direction of strong to weak acid). Somehow we must generate a
133
stronger oxidizing agent than dioxygen, which often has the most positive
standard reduction potential in tables.
Plants must have high concentrations of a reducing agent for the reductive
biosynthesis of glucose from CO2. The reducing agent used for most biosynthetic
reactions in nature is NADPH, which differs from NADH only by the addition of
a phosphate to the ribose ring. This phosphate differentiates the pool of
nucleotides in the cells used for reductive biosynthesis (NADPH/NADP+) from
those used for oxidative catabolism (NADH/NAD+)
Finally, plants need an abundant source of ATP which will be required for
reductive biosynthesis.
We will discuss only the light reaction of photosynthesis which produces these three
types of molecules. The dark reaction , which as the name implies can occur in the dark,
involves that actual fixation of carbon dioxide into carbohydrate using the ATP and
NADPH produced in the light reaction.
THE LIGHT REACTION
Obviously, the energy to power the light reactions comes directly from sunlight. Clue two
is that plants have an organelle that animal cells don't - the chloroplast. Its structure is in
many ways similar to a mitochondria in that it has internal membranes (thylacoid
membranes) surrounding enclosed compartments.
Plants have many pigments (chlorophyll, phycoerthryins, carotenoids, etc.) whose
absorption spectra overlap that of the solar spectra. The main pigment, chlorophyll, has a
134
protophorphryin IX ring (same as in heme groups) with Mg instead of Fe. When the
chlorophyll absorbs light, the excited electrons must relax eventually to their ground
state. It can do this by either radiative or nonradiative decay. In radiative decay, a photon
of lower energy is emitted (after some energy has already been lost by vibrational
transitions) in a process of either fluorescence or phosphorescence. In nonradiative
decay, the energy of an excited electron can be transferred to another similar molecule (in
this case other chlorophyll molecules) in a process which excites the energy of an
electron in the second molecule to the same excited state. (It is as if a photon is released
by the first excited molecule which then is absorbed by an electron in a second molecule
to excite it to the same exited state, although the excitation occurs without photon
production). In this fashion, energy is transferred from one chlorophyll to another. This
type of energy transfer is called resonance energy transfer or exciton transfer.
Figure: resonance energy transfer
One type of chlorophyll has slightly different characteristics, however. Because of its
unique environment, the energy level of the excited state electron is lower than in the rest
135
of the chlorophyll molecules, in much the same way that pKa's of amino acid side chains
differ with environment, and the standard reduction potential of FADs that are tightly
bound to enzymes differ due to the different environment of FAD/FADH2 These unique
chlorophyll centers are called reaction centers.
Figure: reaction centers
The rest of the chlorophyll molecules act as antennas which transfer energy to the
reaction centers. An electron in an adjacent excited state chlorophyll (which is at a higher
level than the excited state energy of the reaction center) can then be transferred to this
lower energy state level in the reaction center, in a process which forms a positively
charged ion from the first excited state molecule and an anion from the recipient. This
process of energy transfer is called electron transfer.
136
Figure: electron transfer
Photoexcitation of the non-reaction center chlorophyll turns that molecule into a good
reducing agent, which transfers its electron to the excited state level of the reaction center
chlorophyll. If you count both step together, the non-reaction center chlorophyll gets
"photooxidized", in the process producing the "strong" oxidizing agent which is the
positively charged chlorophyll derivative. The extra electron passed onto the second
molecule will eventually be passed on to NADP+ to produce NADPH. The light reaction
of photosynthesis in green plants is shown below. In this process, in a scheme that is
reminiscent of electron transport in mitochondria, water is oxidized by photosystem II.
Electrons from water are moved through PSII to a mobile, hydrophobic molecule,
plastaquinone (PQ) to form its reduced form, PQH2. PSII is a complicated structure with
many polypeptide chains, lots of chlorophylls, and Mn, Ca, and Fe ions. A Mn cluster,
called the oxygen evolving complex, OEC, is directly involved in the oxidation of wate.
137
Two key homologous 32 KD protein subunits, D1 and D2, in PSII are transmembrane
proteins and are at the heart of the PSII complex. Another photosystem, PS1, is also
found further "downstream" in the electron transport pathway. It takes electrons from
another reduced mobile carrier of electrons, plastocyanin (PCred) to ferredoxin, which
becomes a strong reducing agent. Ferrodoxin is a protein with an Fe-S cluster (Fe-S-Fe-S
in a 4 membered ring, with 2 additions Cys residues coordinating each Fe). It ultimately
passes its electrons along to NADP+ to form NADPH. A summary of the light reaction in
plants and standard reduction potentials of the participants, are shown below. Note that
many of the complexes produce a transmembrane proton gradient. In contrast to
mitochondria, the lumen (as compared to the mitochondrial matrix) becomes more acidic
that the other stroma. Protons then can move down a concentration gradient through the
C0C1ATPase to produce ATP required for reductive biosythesis of glucose.
Figure: THE LIGHT REACTIONS OF PHOTOSYNTHESIS
Photosystem II:
The net reaction carried out by PS2 is the oxidation of water and reduction of
plastoquinone.
PQ + H2O --> PQH2 + O2 (g)
Note that water is not converted to 2H2 + O2 , as in the electrolysis of water. Rather the
Hs are removed from water as protons in the lumen of the cholorplast, since the part of
PSII which oxides water is near the lumenal end of the transmembrane complex.
Protons required to protonated the reduced (anionic) form of plastaquinone to form
PQH2, an activity of PSII found closer to the stroma, derive from the stroma. which then
can be used to protonated the "anionic" form of reduced PQ to form PQH2.
A quick look at standard reduction potentials shows that the passing of electrons from
water (dioxygen SRP = +0.816 V) to plastoquinone (approx SRP of 0.11 ) is not
thermodynamically favored. The process is driven thermodynamically by the energy of
the absorbed photons.
Recently the crystal structure of PSII from a photosynthetic cyanobacterium was
determined. It consists of 17 polypeptide subunits with metal and pigment cofactors and
over 45,000 atoms. (Zouni, Nature, 409, 739, 2001). Of particular interest is the P680
chlorophyll reaction center, which consists of four monomeric chlorphylls adjacent to a
cationic Tyr-D side chain which destabilizes the chlorophyll molecules. When H2O gets
oxidized to form dioxygen, 4 electrons must be remove by photoactivated P680. In PSII,
this process occurs in 4, single electron steps, with the electrons first being transferred to
140
a Mn4 cluster cofactor (of composition Mn4Ca1Cl1-2(HCO3)y. This inorganic Mn cluster
is often called OEC (oxygen evolving complex) or WOC (water oxidizing complex). The
electrons passed through the Mn complex are delivered to P680 by a photoactive Tyr free
radical (Tyr Z). The actual structure of the OEC could not be resolved, but other
structural and spectroscopic data support the structure below (Chem. Rev., 2001, 101, 21-
35), which also shows a possible mechanism for electron and proton transfer from water
to form dioxygen. 5 discrete intermediates of the OEC, S0-S4, are suggested from the
experimental data (Kok cycle). These were postulated from experiments in which
spinach chloroplast were illuminated with short light pulses. A pattern of dioxygen
release was noted that repeated after 4 flashes. Ultimately, light absorption by P680
forms excited state P680* which donates an electron to pheophytin (which passes them to
quinones) to form P680+, which receives electrons from the OEC, specifically the TyrZ
radical.
141
Figure: OEC - Mechanism of Water Oxidation
Investigators have made non-peptide mimetics of superoxide dismutase to facilitate
therapeutic removal of excess superoxide formed in brain and heart tissue. These may
142
arise after an oxidative burst from reperfusion of these tissues after heart or brain
attacks. Likewise, scientist are trying to build synthetic PSII-OEC complexes which
could be used to form dioxygen or hydrogen gas for fuels.
In summary, for PSII in plants
1. a pair of chlorophylls (P680) in the D subunits absorb light (maximum absorbance
around 680 nm) and reach an excited state
2. electron transfer from P680 to a nearby chlorophyll with a lower energy level for
the excited state electron occurs. This chlorophyll has 2 H+
ions in the
chlorophyll instead of Mg2+
occurs. The P680 now becomes cationic, P680+.
3. This "anionic" chlorophyll transfers an electron to oxidized plastoquinone.
4. The P680+, a strong oxidizing agent, removes one electron from H2O-OEC
complex.
Steps 1-4 repeat three more times, each requiring another photon and each cycle
producing another electron which passes on through the system. Remember that when O2
acts as an oxiding agent, it gains four electrons. The first produces superoxide, the next
peroxide, and two more produce oxide which when protonated is water. Hence two
waters and four cycles are required to remove the four electrons required to produce
dioxygen.
PSII
PSII from the PDB
143
A similar mechanism is found in PSI, except plastacyanin, not dioxygen is oxidized, with
electrons moved to ferrodoxin. This is likewise a difficult process since the reduction
potential for oxidized plastocyanin (the form that can act as a reducing agent) is +0.37
while for ferrodoxin it is -0.75. This transfer of electrons is an uphill thermodynamic
battle since the more positive the standard reduction potential, the better the oxidizing
agent and the more likely the agent becomes reduced. What drives this uphill flow of
electrons. Of course, it is the energy input from the photon.
Chime: Light Harvesting Complex from Spinach 1RWT Jmol: Light
Harvesting Complex from Spinach 1RWT
Chime: Detailed Photosystem II from S. elongatus (1S5L) Jmol:
Detailed Photosystem II from S. elongatus
Chime: Photosystem I: A Photosynthetic Reaction Center and Core Antenna
System From Cyanobacteria 1JBO
Jmol: Photosystem I: A Photosynthetic Reaction Center and Core Antenna
System From Cyanobacteria 1JBO
144
Plant Protection
Plants have evolved a great ability to absorb light over the entire visible range of the
spectra. Can they absorb to much energy. The answer is yes, so plants have developed
many ways to protect themselves. IF too much light is absorbed, the pH gradient
developed across the thylacoid membranes becomes greater. This is sensed by a protein,
PsbS, and through subsequent conformational changes transmitted through the light-
harvesting antennae, the excess light energy is dissipated as thermal energy. Mutants
lacking PsbS showed decreased seed yield, a sign that it became less adaptable under
conditions of stress (such as exposure to rapidly fluctuating light levels. Molecules called
xanthophylls (synthesized from carotenes - vit A precursors) such as zeaxanthin are also
important in excess energy dissipation. These molecules appear to cause excited state
chlorophyll (a singlet like excited state dioxygen) to become deexcited. Without the
xanthophylls, the excited state chlorophyll could deexcite by transfer of energy to ground
state triplet dioxygen, promoting it to the singlet, reactive state, which through electron
acquisition, could also be converted to superoxide. These reactive oxygen species (ROS)
can lead to oxidative damage to proteins, lipids and nucleic acids, alteration in gene
transcription, and even programmed cell death. Carotenoids can also acts as ROS
scavengers. Hence both heat dissipation and inhibition of formation of ROS (by such
molecules as vitamin E) are both mechanism of defense of excessive solar energy
Given that both plants and animals must be protected from ROS, antioxidant molecules
made by plants may prove to protect humans from diseases such as cancer,
cardiovascular disease, and general inflammatory diseases, all of which have been shown
145
to involve oxidative damage to biological molecules. Humans, who can't synthesize the
variety and amounts of antioxidants that are found in plants, appear to be healthy when
they consume large amounts of plant products. These phytomolecule also have other
properties, including regulation of gene transcription which can also have a significant
effect on disease propensity.
Production of Hydrogen: Hydrogenases (repeated from 8B)
Our world desperately needs an energy efficient way to produce H2 for energy production
without producing waste pollutants. Catalytic cracking of molecules and newly
developed fuel cells offer two possibilities. Wouldn't it be great if a reactant like water
could be used for H2 production (without the use of electrolysis) or expensive metal
catalysts? Nature may show the way. Bacteria (even E. Coli found in our GI system) can
use simple metals like iron to produce H2 from H+ with electrons for the reduction of H
+
coming from a donor (such as a reduced heme in proteins):
Dred+ H+ <=> Dox + H2
The reaction is also reversible in the presence of an acceptor of electrons from H2 as it
gets oxidized:
Aox+ H2 <=> Ared + H
+
The enzymes that catalyze hydrogen production are hydrogenases (not
dehydrogenases). Note that the name hydrogenases best reflects the reverse reaction
when a molecule (P) in an oxidized state gets reduced (to S) and H2 gets oxided to H+.
146
Crystal structures of hydrogenases show them to be unique among metal-containing
enzymes. They contain two metals bonded to each other. The metal centers can either be
both iron or one each of iron and nickel. The ligands interacting with the metals are two
classical metabolic poisons, carbon monoxide and cyanide. Passages for flow of
electrons and H2 connect the buried metals and the remaining enzymes. The metals are
also bound to sulfhydryl groups of cysteine side chains. It appears that two electrons are
added to a single proton making a hydride anion which accepts a proton to form H2. In
the two Fe hydrogenases, the geometry of the coordinating ligands distorts the bond
between the two iron centers, leading to irons with different oxidation numbers.
Electrons appear to flow from one center to the other, as does carbon monoxide as well.
Ultimately, hydrogenases or small inorganic mimetics of the active site could be coated
on electrodes and used to general H2 when placed in water in electrolytic experiments.
147
APPENDIX F
A Murder Mystery Utilizing Paper Chromatography
Adapted from:
This is the html version of the file http://www.liverpool.gov.uk/Images/tcm21-65147.pdf.
Google automatically generates html versions of documents as we crawl the web.
Contents of the Unit
Overview Sheet to outline all the activities
Resource list
Newspaper report to set the scene of the murder mystery
Police report sheet which give the pupils further information about the victim and each of
the suspects
Pupil running record sheet, so they can record results and eliminate suspects as each test
is completed.
Lesson plans for the four activities
Pupil worksheets for the four activities
148
Resources/Equipment
test tubes
beakers
chromatography paper
methanol
acetone
scissors
leaves of spinach, Oregon grape, purple flowers, and any dark green leaf
tooth picks or capillary tubes
hot plate
hot water bath
sand
ruler
tape
plastic pipettes
Note: Use the same laboratory procedure given in the main body of this paper.
149
Overview of the Unit
The murder has taken place in a Liverpool school. The victim is a 55 year-old
headteacher who has worked at the school for over 25 years. The police have closed the
school and forensic scientists have concluded that the following testing should be carried
out while police questioning continues.
• Chromatography tests on the leaf remains where shoe prints indicating a struggle had
taken place in the garden near the front of the school. It is autumn in Liverpool, and leaf
litter is all over the yards and streets of the town. The plants in the garden of the school
are unique to this community, and will definitely establish a cause and effect
circumstantial connection between the victim and the murder suspect.
Who did it?
The school (Honey Lane High) had been under scrutiny by the LEA for a while and the
headmaster had been under a lot of pressure to meet targets and succeed on a minimal
budget. Word had broken out amongst gossip mongering staff that the school was under
threat of staff
numbers being cut and / or complete closure of the school. The Head through no fault of
his own had become an obvious source of blame for the problems the school was facing.
There are 5 suspects:
An Irate Parent
A Dinner Lady
The Caretaker
150
Head of Science
Lab technician
Who did it?
Due to the school‘s financial problems the head had had to ask all staff to ‗tighten their
departmental belts‘ which obviously did not meet approval of everyone. The Lab
Technician, feeling the strain of being overworked and underpaid as well as having to
hold back spending an already minuscule budget, could not take the pressure anymore
and felt something had to be done. In his mind the only thing to do was ‗get rid‘ of the
Headmaster and replace him with someone who going to run the school efficiently and
prevent closure and subsequent loss of his job. He hoped it would also reduce the amount
of nagging he was getting from the Head of Science who was also feeling the pressure of
the School‘s downfall. He had noticed how the headmaster usually left work at 5.30pm
everyday and figured the best way to do it was to strangle him. Obviously the head would
put up a fight so he thought the best way was to use some kind of liquid to blur his vision
and co-ordination. He had a look around the prep-
room and chose some dilute hydrochloric acid (he was on a budget remember). At 5.20
he went over to the head‘s office and demanded the head do something to save his job.
When the head told him it was out of his control the lab technician couldn‘t take anymore
of what he thought
was just excuses. He got up and grabbed the head by the neck and of course he fought to
get away. Once free, the head went out the door and into the school garden but the lab
technician threw the acid in his face and he shrieked out in chronic pain as the acid
151
burned. The lab technician pushed him to the ground and again grabbed his neck tight
until eventually the headmaster‘s struggle stopped and he was dead.
Additional Information
The activities in the unit of work represent the bare skeleton of the project. It is entirely
up to individual teachers what additional tasks are delivered. The unit could be extended
to include:
- Drama role-plays
- Concept Cartoon constructions
- Concept Maps
- Project/Research work (into individual topics)
- Creative Writing (Newspaper Articles)
- Presentation work
Murder Mystery
Learning Objectives
Activity
Be able to make predictions, introduce the new murder mystery, and give suggestions
about how to test the pupils, the newspaper report, and the evidence found at the crime
scene.
police report. Pupils to make predictions and suggestions on how to identify the evidence.
152
Chromatography
Learning Objectives
The pupils will learn:
• Learn that leaves are made up of mixtures of different pigments or colours that can be
separated using chromatography.
Activity
Pupils to experiment with a range of leaves.
Pupil Outcomes
Completed worksheet
Pupil explanation of what they did and what they found out.
• Carry out a simple experiment to separate a mixture of plant pigments into separate
colours.
• Understand the practical uses of chromatography in everyday life. (E.g. in a crime
scene investigation)
Lesson 1
Introducing the Murder Mystery
Learning Objectives
The pupils will:
• Be able to make predictions and suggestions about how to test the evidence found at the
crime scene.
153
Activity
Introduce the project. Introduce the newspaper report and the police report and ask pupils
to work in pairs to highlight the text identifying all the key points. Ask pupils to suggest
who they think the murderer is, giving reasons. Pupils to record their ideas on the pupil
running record. Discuss their findings and talk about what evidence they will need to
collect.
Discuss how the case will proceed and talk about the science tests the pupils will need to
complete to draw a conclusion.
Differentiation
All pupils will: have made a prediction and recorded it on the pupil running record.
Most pupils will: will be able to give reasons for their predictions
Some pupils will: be able to suggest appropriate science tests to examine the evidence
further.
Organisation
Pupils to work in small groups
Newspaper report
Police report
Pupil running record
Assessment strategies
Q + A Session
Completed crime scene reports
154
Evaluation
LIVERPOOL NEWS
Murder of Liverpool Headteacher
By Fred Snooper
A Liverpool Headteacher was found dead in his office last night.
Victim
Henry Headley, the headteacher of Honey Lane High in Liverpool was found in the
school garden at approximately 6pm by the school caretaker who was doing a routine
check of the school before locking up for the night. It is thought the headteacher had
stayed behind after school to do work and catch up on paperwork. Mr. Headley had been
the headteacher of Honey Lane High for 25 years and it is a mystery as to why a well-
respected member of the Honey Lane community would meet such an untimely end.
Motive
A murderer has not yet been identified but police are still continuing with their
questioning. It is thought that the murderer may have worked at the school and perhaps
had a personal vendetta against the head. The motive for such a vendetta has not yet been
established. The school has been closed until further notice and will remain closed until
the police have identified a suspect or suspects. Police are asking people if they have any
information about the crime or saw anything suspicious near or around the school on the
day in question to come forward and tell them what you know. A phone line has been set
up to phone in with any relevant information.
155
Lesson 2
Chromatography
Time Allocation
50 mins - 1 hr
Learning Objectives
The pupils will:
• Understand that chromatography is a process used to separate particles in a liquid or
mixture of liquids.
• Understand that chromatography has many applications and that one of them is in a
crime scene.
• Carry out an experiment to separate a mixture of pigments in leaves to identify the
source of the pigments.
Activity
Introduction
Recap the last lesson and recall what evidence the last lesson had concluded. Introduce
this
lesson by asking pupils what other evidence was worth looking at from the initial police
report.
Explain that Chromatography is used to separate the particles in a mixture of liquids.
Explain
some of its applications in everyday life.
Development
156
Explain the practical and what they are going to do to the pupils. The pupils will take the
leaves found on the victims shoes and match them with leaves of plants found on the
suspects shoes or in their home…… and place them on the chromatography paper and
they will be labelled with the suspects‘ names. At the end none will match the victims.
The rest of the class will carry out chromatography .
Teacher-led discussion of the results and what this evidence suggests. Class should think
why
this evidence is important even though it has turned out to be irrelevant to the case.
Differentiation
All pupils will: carry out an experiment to separate a mixture of pigments unique to the
rare plant to identify the source.
Most pupils will: understand that chromatography is used to separate liquids.
Most pupils will: understand that chromatography has many applications and that one
of them is in a crime scene.
Some pupils will: understand that chromatography is used to separate the
particles/molecules in a mixture of liquids.
Organisation
Pupils to work in small groups
Risk Assessment
Discuss safety issues of working with substances that the pupils are unaware of – talk
about
157
the need to wash hands, not to taste or smell the liquids.
Assessment strategies
Q + A Session
Completed worksheets
Evaluation
Merseyside Police
Crime Investigation
Chromatography Testing
• Carry out the chromatography experiment as shown.
• Record the colours that appear when the pigments separate
• Tick or cross the third column if any sequence of colours are the same as the victims.
Name
Colours present
Match
Victim
Doreen Bridgeley
Irving Stinkley
Sam Givens
Harriet Dustier
Mr. Cosworth
158
Conclusion:
Are any of the colours the same as those from the victims ?
Are any of the colour sequences the same as those from the victim?
Is this evidence relevant to the case?
Why do you think it was necessary to investigate this evidence?
Merseyside Police Department
Fazakerley Forensics Department
Date:__________________
Name:_______________
Chromatography Results Sheet
Colours Found
Red, Yellow, and/or Green
Victim
A
B
C
D
E
Chromatogram:
Merseyside Police
159
Crime Investigation
Chromatography Testing
• Carry out the chromatography experiment as shown.
• Record the colours that appear when the leaf pigments separate
• Tick or cross the third column if any sequence of colours are the
same as the victims.
Name
Colours present
Match Victim
Doreen Bridgeley
Irving Stinkley
Sam Givens
Harriet Dustier
Mr. Cosworth
160
Conclusion:
Are any of the colours the same as those from the victims pen?
Are any of the colour sequences the same as those from the victim?
Is this evidence relevant to the case?
Why do you think it was necessary to investigate this evidence?
Chromatography:
Leaf pigments from the suspect matched that on the victim‘s shoes.What might this
mean? Does this mean the ink stains are relevant to the murderer‘s identity
Who at this point do you think killed Headley?
Mr. Cosworth – The Head of Science
Mrs. Doreen Bridgeley – The Dinner Lady
Mr. Irving Stinkley – The Lab Technician
Mrs. Harriet Dustier – The School Cleaner
Mr. Sam Givens – The School Caretaker
I think ________________ killed Headley because ___________________
161
1. Looking at the evidence on this page: Who must have been involved in the murder?
2. Make up a story to say why these people may have wanted the Headmaster dead!
Investigator’s Running Record of Evidence
Name:
Date:
First Impressions
From the evidence I have seen so far I think____________________________ is the
murderer. My reasons for this are ________________________________________
Results from the victim:
Evidence
Which suspects can you eliminate?
Why?
162
References:
Hess, Amber & Olson, Andrew. Retrieved June 21, 2008 from
http://www.sciencebuddies.org/science-fair-
projects/project_ideas/Chem_p008.shtml?fave=no&isb=c2lkOjEsaWE6Q2hlbSx
wOjEscmlkOjMxMzgzMDg&from=TSW
Schmidtke, Sabrina. Paper Chromatography. Retrieved June 27, 2009 from
http://peer.tamu.edu/podium_poster_presentations/Paper%20Chromatography%2
0Handout.doc
Candy Chromatography, Retrieved July 1, 2009, from
http://scifun.chem.wisc.edu/HOMEEXPTS/candy.htm
Candy Chromatography: What Makes Those Colors? Retrieved June 27, 2009
from http://www.sciencebuddies.org/science-fair-
projects/project_ideas/Chem_p042.shtml
Do Chromatography with Candy and Coffee Filters, Retrieved June 27, 2009 from
http://chemistry.about.com/od/chemistryexperiments/ht/candychroma.htm
Paper Chromatography Experiment. Retrieved June 26, 2009 from
http://www.scienceprojectlab.com/paper-chromatography-experiment.html
Chromatography. From Wikipedia. Retrieved June 24 2009 from
http://en.wikipedia.org/wiki/Chromatography
Make Your Own TLC Plates for Chromatography Science Project. Retrieved June
26, 2009 from http://www.scienceprojectlab.com/plates-for-cromatography-
science-project.html
Separations: Chromatography of M&M and Ink Dyes. Retrieved July 6, 2009 from
163
http://chemistry.bd.psu.edu/jircitano/Chromatography05.pdf
Separating Leaf Pigments Using Paper Chromatography. Retrieved July 1, 2009 from
www.sciencebuddies.org/science-fair-projects
What Color Are the Leaves Really Turning? Retrieved July 1, 2009 from
www.sciencebuddies.org/science-fair-projects
Autumn Leaf Color Why Do Leaves Change Color in the Fall? Retrieved June 29, 2009
form www.sciencemadesimple.com/leaves
Helmenstine, A.M. (2007). How To Do Paper Chromatography With Leaves. About.com:
Chemistry. The New York Times Company. Retrieved December 6, 2007 from
http://chemistry.about.com/cs/howtos/ht/paperchroma.htm
History of Chromatography. Retrieved July 6, 2009 from
http://en.wikipedia.org/wiki/History_of_chromatography
Dr. Jakubowski. Ch 8 Oxidation/Phosporylation Photosynthesis: The Light Reactions.
Retrieved July 10 2009 from
http://www.google.com/imgres?imgurl=http://employees.csbsju.edu/hjakubowski/
classes/ch331/oxphos/photosynth.gif&imgrefurl=http://employees.csbsju.edu/hjak
ubowski/classes/ch331/oxphos/olphotsynthesis.html&h=850&w=705&sz=39&tb
nid=YzmOhpbia04REM:&tbnh=145&tbnw=120&prev=/images%3Fq%3Dphotos
ynthesis%2Blight%2Breactions&hl=en&usg=__HXPAbIHGPaYnB8nYGaR-
uTz3dEw=&ei=kFpvSr32F4jIsQOfvJj7Ag&sa=X&oi=image_result&resnum=2&
ct=image
164