msq plus north america sales training chapter 1 – basic ms beta training version b 20-dec-11
TRANSCRIPT
MSQ Plus North America Sales Training
Chapter 1 – Basic MS
Beta Training
Version B
20-Dec-11
2 Proprietary & Confidential
Agenda for the NA MSQPlus training program
• Comparison of Detectors: General, MS only, MS/MS
• Functional modules common to all MS systems
• MS Features and their importance to customers
• MSQPlus Strengths, Weaknesses, Opportunity, and Threats
• Decision Tree to better understand when to pursue an MS only order or an MS/MS order
3 Proprietary & Confidential
MSQPlus beta Sales Training
Attending
Sales Rep Region Manager
Alex Tagol North West Ken Larkey
Zee Nadjie West Ken Larkey
Natalie Odneal Mid West Ken Larkey
Matt Dilts Ohio Valley Ken Larkey
Tucker Lincoln North East Andy Rodriguez
Chuck Costanza South West Mike Hey
Rita Clement South East Mike Hey
Mark Netsch CoE Matt Neely
Charles Ontiveros CoE Matt Neely
4 Proprietary & Confidential
MSQPlus beta Sales Training
Program Goals
•Transfer general knowledge of the MSQPlus, and the competitive landscape, our market position in order to better sell the product.
•Provide feedback on beta training modules to optimize them prior to launch to the NA Sales Team for Chromatography/Advanced Detection.
•Impact on MSQPlus order receipts within the 1st quarter of 2012
•Resource within your region for MSQPlus sales
5 Proprietary & Confidential
MSQPlus beta Sales Training
By the end of this module you will know….
•The key components of Mass Spectral Analysis
•Advantages and limitations of MS in the determinations of unknowns
•Advantages of using Collision Induced Dissociation with MS and MS/MS instruments.
•Advantage of MS over general detection.
•Advantage of MS/MS over MS only detection
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Today’s Agenda - Comparison of Detectors
• Detection and Mass Spectrometry Overview
• Analyte Identification by Mass & MS Patterns
• Analyte screening: Retention Time vs. Mass
• Improved MS Specificity through Fragmentation
• What is MS/MS?
• Differentiating Factors: General, MS, & MS/MS Detection
Detection and MS Overview
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What does Mass Spectrometry Do?
• Confirm the identity of pesticides and herbicides – Contract Labs
• Determine toxic substances in Catfish - USDA
• Determine Low level ClO4 Analysis – EPA, Universities, Municipalities, Contract Labs
• Determine explosives residues – RCMP, ATF, Fire depts., MRI
• As III / As V Speciation - EPA
• Determine Haloacetic Acids & Bromate – EPA, Municipalities
• Determine the molecular weight of small molecules with biological activity – Pharmaceutical Labs
• Fraction collection of compounds based upon molecular ion - Pharmaceutical Labs
• Determine plating bath contaminants – Semiconductor Labs
Did you know that mass spectrometry is used to...
Mass Spectrometers have a broad range of applications: ICMS & LCMS.ThermoFisher is the ICMS market leader.
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Attributes of the Ideal Detector
•High sensitivity and the same predictable response
•Response to all solutes, or has a predictable specificity
•Linear response over a wide range of solute amounts
•Unaffected by changes in temperature and mobile phase
•Does not contribute to extra-column band broadening
•Reliable and convenient to use
•Fast response
•Provides qualitative information on the detected peak
•Does not destroy the solute
A combination of different detectors can help customers achieve attributes of the Ideal Detector
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The Power of Ultra High Performance Liquid Chromatography
0.0 0.5 1.0 1.5 2.0 2.5 3.0Time (min)
UltiMate 3000UHPLC+
AccelaSystems
ViperFittings
Columnsand
Consumables
World-Class Thermo Scientific Chromatography PortfolioUltra High Performance Liquid
Chromatography with UV/VIS detection is the primary workhorse in most analytical labs around the world
HPLC with UV/VIS is a very valuable analytical tool because it offers:
1. High analytical data quality for routine analysis
2. Robust and easy-to-maintain operation
3. Straightforward method building
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What if You Want More Information?
Absorbance trace shows a number of components and their possible identity if tR standards are available.
Full-scan MS gives additional/alternate components, possible chemical formulae but can still leave co-eluters unresolved.
Tandem mass spectrometry, MSn, can offer differentiation of co-eluting species and information about structure.
Mass Spectrometry Offers
1.Sensitivity
2.Selectivity
3.Information
LC AbsorbanceUV-VIS
0
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LC Full Scan MSXIC
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MS2 XIC 167.9
MS2 XIC 181.9
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135.96181.89 300.83107.89
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107.96 301.01
181.9
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+ +++
Mass Spectrometer: Basic Schematic
LC SampleInlet
DataSystem
IonizationSource
MassAnalyzer(s) Detector
++
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+
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Mass Spectra
Creation of Ions
Mixture of Solvent and Compounds
Detection of Ions
Separation of Ions
“A device to measure the mass to charge ratio of individual molecules that have been converted ions”
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Key Advantages of using MS Detection
•Selectivity – Co-eluting peaks can be isolated by mass selectivity and are not constrained by chromatographic resolution.
•Peak assignment – A unique mass for the compound of interest is generated, ensuring correct peak assignment in the presence of complex
matrices.
•Molecular weight information – Confirmation and identification of known and unknown compounds.
•Structural information – In-source Collision Induced Dissociation (CID) enables molecular ion fragmentation providing structural determination.
•Sample matrix adaptability – Decreases sample preparation time.
•Quantitation – Quantitative and qualitative data can be obtained easily with limited instrument optimization and simultaneous scan modes.
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MSQPlus Mass Detector: Advanced LC Detection
For cost chemists who require increased
productivity through dependable simplicity,
the ThermoFisher MSQPlus is a turnkey
HPLC/MS solution providing superior
chromatographic efficiency and enhanced
mass detection with unsurpassed ease of
use, performance, and ruggedness.
MSQPlus Value Statement
Analyte ID by Mass - Specificity
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Identification/Confirmation by Mass
Mass Spectrum of Methomyl - LCMS
Molecular Ion
Base Peak
Adduct
MW = 162
Formula: C5H10N2O2S
m/z 163
m/z 231
m/z 100 m/z 300
The mass spectrum and its interpretation require experience and knowledge in order to identify an analyte. Software can help but does not provide the answer.
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Identification/Confirmation by Mass
Mass Spectrum
Chromatogram
Unique pattern can identify compound
m/z 79 m/z 81
Target Analyte
Isotopic patterns can aid in compound identification
Isotope ratios are “gifts of nature” that give patterns to help indentify analytes
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Mass vs. Chromatography Selectivity
Analytes can be distinguished by mass even of they co-elute with other compounds or matrix
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Mass Detection Enhances Chromatography
Compound B(RT: 1.43min)
Compound A(RT: 1.40min) 205 m/z285 m/z
Which is which?
Compound A or Compound B
Co-eluting peaks can be distinguished by mass – SIM or Extracted Ion
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Chromatogram Overlays help Indentify Analytes
(1) Scan Masses
(2) Sum All Ions
(3) Plot
“Total” Ion Chromatogram (TIC)
“Extracted” Ion Chromatogram“Extracted” Ion Chromatogram (EIC)
Mass Extactions and the resulting Mass Chromatogram help to correlate a specific mass to a chromatographic peak
(4) Mass Spectrum
Analyte screening: Mass Spectrometric Detection complements General Detection
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Screen Detection Comparison: Conductivity vs MS
Extracted ion chromatograms:1. Sodium, m/z 232. Ammonium, m/z 183. Hydrazine, m/z 334. Potassium, m/z 395. 2-(2-Aminoethoxy)ethanol,
m/z 1066. 5-Amino-1-pentanol, m/z 1047. Morpholine, m/z 888. 2-Diethylaminoethanol, m/z 1189. Cyclohexylamine, m/z 10010. Magnesium, 2411. Calcium, m/z 20
0 2 4 6 8 10 12 14 16
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Extracting Ions from the Full Scan and then overlaying the resulting Mass Chromatograms, Correlate Mass with Peaks from a General Detector
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Screen Detection Comparison: Conductivity vs MS
Column: IonPac® CS12A-MS, 100 x 2 mm I.D.EG40 Gradient: 8-24 mM MSA in 4 min, then hold, using EG40Flow Rate: 0.25 mL/minSuppressor: CAES™, 21 mADetection: 1. Conductivity
2. Mass specMS Conditions: MSQ™, +ESI,
monovalents @ 50 V, divalents @100 V,
350 ºC, SIM channels as indicatedPeaks:
1. Lithium 500 µg/L2. Sodium 500 3. Ammonium 5004. Potassium 5005. Magnesium 2506. Calcium 250
NH4+, 18 m/z
Na+, 23 m/z
Li+.H2O, 25 m/z
K+, 39 m/z
Mg2+, 24 m/z
½ Ca2+, 20 m/z
Cond.
0 84
Selected Ion Monitoring (SIM) can help Screen for Known Compounds and also, Correlate Mass with Peaks from a General Detector
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Chromatogram Overlays help Identify Analytes
Extracted Mass Chromatogram m/z 81
Mass spectrum
Conductivity & Mass Chromatogram overlays – Link peaks time
m/z 79 m/z 81
General Detector Chromatogram
Chromatograms
Compound Identification - Chromeleon makes the process of Extracting Ions from the Full Scan and then overlaying the applicable Chromatograms - Easy
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Quantitation using MS
ClO4-QuanY = 0.0122265+0.0012984*X R 2̂ = 0.9996 W: 1/X
0 5000 10000ppt
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ClO4-QuanY = 0.0122265+0.0012984*X R^2 = 0.9996 W: 1/X
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Selected Ion Monitoring (SIM – MS) or Selected Reaction Monitoring (SRM – MS/MS) are used for Quantitation
Improved MS Specificity through Fragmentation
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MS/MS on MSQ Plus? In-Source CID
A: MorphineThere is no significant fragmentation when cone voltage is set at 70 V.
B: HydromorphoneFragmentation dominates the spectrum when cone voltage is set at 125 V.
The MSQPlus via Collision Induced Dissociation can produce fragments of the molecular ion creating “MS/MS like” Spectra
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CID – More Energy Increases Fragmentation
Mass Spectra - Anatoxin-a
Cone voltage 50 V
Cone voltage 30 V
Cone voltage 70 V
HN CH3
O
IncreasingEnergy
IncreasesFragmentation
Potential
Increasing Collision Energy (CID – MSQPlus Cone Voltage), will cause fragmentation of the molecular ion, adducts, etc. This can help in analyte ID.
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CID can help Elucidate Structure
Functional Group has m/z 44
Mass spectral peaks separated by 44 Daltons
Mass Chromatogram of Triton-X 100 (surfactant)
By increasing Collision Energy (CID), functional groups can fragment away from the molecular ion, helping to elucidate molecular structure
MS/MS – General Overview
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MS/MS – Basic Description
Q1
Product Ions
Precursor Ions
Q2Q3
● MS/MS or MSn enable determination of product ions. ● Compared to a single quads, MS/MS instruments have greater specificity due to the uniqueness
product ions. ● Compared to a single quads, MS/MS instruments have greater sensitivity due to reduced chemical
noise.
Step 1: Isolate Target Precursor Ion from the mass spectrum
Step 2: CID
Step 3a: MRM - Analyze MS/MS mass spectrum
Step 3b: SRM - Monitor Selected Ions
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MS/MS – The Advantage: Low Chemical Noise
Inte
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cou
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S]
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35,000
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ClO4 - 99 - 11.97
0
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ClO4 - 101 - 11.98
9.00 10.00 11.25 12.50 14.000
12,500
20,000
Retention Time [min]
4 ClO4 - IStd - 11.95
Conductivity
SIM_01: 99 m/z
SIM_02: 101 m/z
SIM_03: 107 m/z
RT: 0.00 - 14.02 SM: 7G
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RT: 11.60AA: 740997SN: 5299
RT: 11.58AA: 235803SN: 6226
RT: 11.59AA: 227857SN: 21147
NL: 3.76E4TIC F: - c ESI SRM ms2 99.102 [83.012-83.312] MS Genesis Sample_E_3
NL: 1.18E4TIC F: - c ESI SRM ms2 101.120 [84.988-85.288] MS Genesis Sample_E_3
NL: 1.15E4TIC F: - c ESI SRM ms2 107.120 [89.020-89.320] MS Genesis Sample_E_3
MS/MS SRM – No Chemical noise MS SIM – Chemical noise present
ClO4 in Infant Formula ClO4 in Urine
Differentiating Factors: General, MS & MS/MS Detection
34 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Sensitivity Less SensitiveDetection levels in the PPM levels.
More SensitiveSelected Ion Monitoring (SIM) Detection levels in the low to mid PPB levels.MSQPlus Optimized for Low Mass, Low Level Detection.
More Sensitive in MatrixProduct Ions are isolated from high chemical background lowering detection levels.
Specificity Less SpecificAnalyte determination by retention time only. Confirmation may be available from 3D spectrum
More SpecificMS detectors differentiate by mass/charge
More SpecificProduct Ion mass provides more specific analyte information. Requirement is sample dependent.
Selectivity Not SelectiveTarget analytes that co-elute or elute with matrix cannot be differentiated. Different wavelengths may separate coeluters.
More Selective Co-eluting analytes can be differentiated by mass. A target compound’s mass can differentiate it from matrix.
More SelectiveProduct Ion mass provides more selective analyte information. Requirement is sample dependent.
Ease of Use Easy ModerateSimple walk-up Mass Spec. The MSQPlus is one of easiest to use MS detectors available.
ComplexRequires skilled and experienced operator.
35 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Sensitivity Less SensitiveDetection levels in the PPM levels.
More SensitiveSelected Ion Monitoring (SIM) Detection levels in the low to mid PPB levels.MSQPlus Optimized for Low Mass, Low Level Detection.
More Sensitive in MatrixProduct Ions are isolated from high chemical background lowering detection levels.
Specificity Less SpecificAnalyte determination by retention time only. Confirmation may be available from 3D spectrum
More SpecificMS detectors differentiate by mass/charge
More SpecificProduct Ion mass provides more specific analyte information. Requirement is sample dependent.
Selectivity Not SelectiveTarget analytes that co-elute or elute with matrix cannot be differentiated. Different wavelengths may separate coeluters.
More Selective Co-eluting analytes can be differentiated by mass. A target compound’s mass can differentiate it from matrix.
More SelectiveProduct Ion mass provides more selective analyte information. Requirement is sample dependent.
Ease of Use Easy ModerateSimple walk-up Mass Spec. The MSQPlus is one of easiest to use MS detectors available.
ComplexRequires skilled and experienced operator.
36 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Sensitivity Less SensitiveDetection levels in the PPM levels.
More SensitiveSelected Ion Monitoring (SIM) Detection levels in the low to mid PPB levels.MSQPlus Optimized for Low Mass, Low Level Detection.
More Sensitive in MatrixProduct Ions are isolated from high chemical background lowering detection levels.
Specificity Less SpecificAnalyte determination by retention time only. Confirmation may be available from 3D spectrum
More SpecificMS detectors differentiate by mass/charge
More SpecificProduct Ion mass provides more specific analyte information. Requirement is sample dependent.
Selectivity Not SelectiveTarget analytes that co-elute or elute with matrix cannot be differentiated. Different wavelengths may separate coeluters.
More Selective Co-eluting analytes can be differentiated by mass. A target compound’s mass can differentiate it from matrix.
More SelectiveProduct Ion mass provides more selective analyte information. Requirement is sample dependent.
Ease of Use Easy ModerateSimple walk-up Mass Spec. The MSQPlus is one of easiest to use MS detectors available.
ComplexRequires skilled and experienced operator.
37 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Sensitivity Less SensitiveDetection levels in the PPM levels.
More SensitiveSelected Ion Monitoring (SIM) Detection levels in the low to mid PPB levels.MSQPlus Optimized for Low Mass, Low Level Detection.
More Sensitive in MatrixProduct Ions are isolated from high chemical background lowering detection levels.
Specificity Less SpecificAnalyte determination by retention time only. Confirmation may be available from 3D spectrum
More SpecificMS detectors differentiate by mass/charge
More SpecificProduct Ion mass provides more specific analyte information. Requirement is sample dependent.
Selectivity Not SelectiveTarget analytes that co-elute or elute with matrix cannot be differentiated. Different wavelengths may separate coeluters.
More Selective Co-eluting analytes can be differentiated by mass. A target compound’s mass can differentiate it from matrix.
More SelectiveProduct Ion mass provides more selective analyte information. Requirement is sample dependent.
Ease of Use Easy ModerateSimple walk-up Mass Spec. The MSQPlus is one of easiest to use MS detectors available.
ComplexRequires skilled and experienced operator.
38 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Speed of Analysis
SlowerChromatographic separation required.
FasterDue to analyte ID by mass, complete chromatographic separation may not be necessary. Lower requirement for sample pre-treatment.
Faster in MatrixProduct Ions are isolated from high chemical background lowering requirement for sample pre-treatment.
Cost of Operation
LessFewer consumables
More Higher cost in terms of operator skill and consumables. The MSQPlus is one of the lowest cost and easiest to use MS detectors available.
MoreHigher than Single Stage MS but may be justified depending on sample requirements.
Purchase Price
Less MoreThe benefit in terms of superior Sensitivity, Specificity, Selectivity, and Speed of Analysis, justifies higher cost.
MoreThe benefit in terms of superior analysis in matrix relative Sensitivity, Specificity, Selectivity, and Speed of Analysis, justifies higher cost.
39 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Speed of Analysis
SlowerChromatographic separation required.
FasterDue to analyte ID by mass, complete chromatographic separation may not be necessary. Lower requirement for sample pre-treatment.
Faster in MatrixProduct Ions are isolated from high chemical background lowering requirement for sample pre-treatment.
Cost of Operation
LessFewer consumables
More Higher cost in terms of operator skill and consumables. The MSQPlus is one of the lowest cost and easiest to use MS detectors available.
MoreHigher than Single Stage MS but may be justified depending on sample requirements.
Purchase Price
Less MoreThe benefit in terms of superior Sensitivity, Specificity, Selectivity, and Speed of Analysis, justifies higher cost.
MoreThe benefit in terms of superior analysis in matrix relative Sensitivity, Specificity, Selectivity, and Speed of Analysis, justifies higher cost.
40 Proprietary & Confidential
Differentiating Factors of LC/IC Detection
Factor General Detection Single Stage MS Multi Stage MS
Speed of Analysis
SlowerChromatographic separation required.
FasterDue to analyte ID by mass, complete chromatographic separation may not be necessary. Lower requirement for sample pre-treatment.
Faster in MatrixProduct Ions are isolated from high chemical background lowering requirement for sample pre-treatment.
Cost of Operation
LessFewer consumables
More Higher cost in terms of operator skill and consumables. The MSQPlus is one of the lowest cost and easiest to use MS detectors available.
MoreHigher than Single Stage MS but may be justified depending on sample requirements.
Purchase Price
Less MoreThe benefit in terms of superior Sensitivity, Specificity, Selectivity, and Speed of Analysis, justifies higher cost.
MoreThe benefit in terms of superior analysis in matrix relative Sensitivity, Specificity, Selectivity, and Speed of Analysis, may justify higher cost.
Quiz
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Quiz - Comparison of Detection
1. The Goal of this beta Sales Training exercise is.a.Obtain general knowledge of the MSQPlus and the competitive
landscape
b.Better understand the MSQPlus market position and how to sell the product
c.Impact on MSQPlus order receipts within the 1st quarter of 2012
d.Resource within your region for MSQPlus sales
43 Proprietary & Confidential
Quiz - Comparison of Detection
2. An ideal detector has the following attributes.a.Linear response over a wide range of analyte amounts
b.Utilizes mass to identify the analyte
c.Makes use of software to provide the correct answers
d.Is made in China
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Quiz - Comparison of Detection
3. Mass Spectral Analysis requires which one of the following.a.Physical Separation
b.Ionization
c.Fragmentation
d.Nuclear Reaction to change into a stable isotope
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Quiz - Comparison of Detection
4. Name 2 ways and LCMS can be used to identify an unknown.a.A mass spectral library purchased from NIST
b.Unique isotope ratios
c.Mass of the molecular ion
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Quiz - Comparison of Detection
5. Mass Chromatograms from a Mass Spectrometer can be used to Correlate Mass to a Specific Peak from a General Detector.
a.True
b.False
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Quiz - Comparison of Detection
6. Collision Induced Dissociation (CID) is an MS feature that is used to….
a.Cause fragmentation of the Molecular Ion creating “MS/MS like” mass spectra
b.Elucidate molecular structure
c.Add clusters and adducts to the molecular ion
d.Is only available on MS/MS instruments
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Quiz - Comparison of Detection
7. Advantages of MS/MS over MS only instruments include ….a.Lower detection levels due to lower chemical background
b.Less sample preparation for high matrix samples
c.Improved specificity
d.Superior detection at sales price equivalent to MS only instruments
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Quiz - Comparison of Detection
8. Advantages of MS over general detection include ….a.Determinations of co-eluting analytes
b.Lower detection levels via Selected Ion Monitoring
c.Simultaneous confirmation of knowns and determination of unknowns
d.Cost and Ease of Operation
50 Proprietary & Confidential
Future beta training sessions
# Training Module Date
1 Comparison of Detectors 15-Dec-11
2 Functional modules common to all MS systems
5-Jan-12
3 MS Features and their importance to customers
10-Jan-12
4 MSQPlus Strengths, Weaknesses, Opportunity, and Threats
12-Jan-12
5 Decision Tree to better understand when to pursue an MS only order or an MS/MS order
17-Jan-12
Launch of Training Modules to the entire NA Sales Team will be in the form of Adobe Connect Self Paced Sessions and occur within 2 wks beta training.
51 Proprietary & Confidential
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