principles of aseptic processing

37
PRINCIPLES OF ASEPTIC PROCESSING Jim Hardy GHGBioSciences, Inc. f [email protected] [email protected]

Upload: nsf-health-sciences

Post on 16-Apr-2017

792 views

Category:

Documents


1 download

TRANSCRIPT

Page 1: Principles of Aseptic Processing

PRINCIPLES OF ASEPTIC PROCESSING

Jim HardyGHGBioSciences, [email protected]@gmail.com

Page 2: Principles of Aseptic Processing

Agenda Background Microbiology Basics Contamination Control

Sources of Contamination

Cleaning & Disinfection 4 Pillars of Aseptic

Techniques

2

Page 3: Principles of Aseptic Processing

What are Aseptic Practices?Definitions

Aseptic = without microorganismsA methodology that prevents the introduction

of unwanted organisms into an environment.A practice that removes or kills

microorganisms from hands and objects.

Sterile = the complete destruction of all forms of microbial life, including bacterial spores.

The meaning of this word is absolute; there is no such thing as "partially sterile“

Something is either sterile or non-sterile

3

Page 4: Principles of Aseptic Processing

Aseptic TechniquesHistory & Milestones

4

1600 1700 1800 1850 1900 2000

Microscope

Iodine

Carbolic Acid

HEPA

19th Century SurgeryIgnaz Semmelweis

Page 5: Principles of Aseptic Processing

Useful:-E.coli for production of rDNA/insulin

-Aspergillus for production of penicillin

-Production of foods, for example:

-Dairy industry: Streptococcus thermophilus to make cheese

-Yeast in beer / wine making

-Bakers yeast

Harmful:-contamination

-food decay

-contamination of medication

-germs

Page 6: Principles of Aseptic Processing

When are Aseptic Practices Used?

Sampling Raw Materials for Qualification Release Sampling of Utilities (WFI, pure steam,

compressed air, and specialty gases) Bioburden & Sterility Testing Environmental Monitoring Manufacturing in classified areas (i.e. cleanrooms) Compounding/Formulation Fill/Finish Operations

6

Page 7: Principles of Aseptic Processing

Good Aseptic PracticesRegulatory Aspects

21 CFR 211 211.28 (b) Personnel shall practice good sanitation and health habits. 211.113 (b) Appropriate written procedures, designed to prevent microbiological

contamination of drug products purporting to be sterile shall be established and followed.

211.84 (c) Sterile equipment and aseptic sampling techniques shall be used when necessary.

7

To ensure our drug products are safe, pure, and effective

Eudralex Vol. 4 Part 1, Ch. 2 Personnel working in areas where contamination is a hazard, e.g. clean areas or areas

where highly active, toxic, infectious or sensitizing materials are handled, should be given specific training. (2.10)

EC Guide to Good Manufacturing Practice Revision to Annex 1: Manufacture of Sterile Medicinal Products The manufacture of sterile products should be carried out in clean areas entry to which

should be through airlocks for personnel and/or equipment and materials. Clean areas should be maintained to an appropriate cleanliness standard and supplied with air which is passed through filters of an appropriate efficiency.

Page 8: Principles of Aseptic Processing

Microbiology and Microorganisms

8

Page 9: Principles of Aseptic Processing

Why is Microbiology Important?

Defined as “the study and science of microorganisms” Most microbial contaminants are not pathogens;

however, their presence in drug product and raw materials can affect the safety, purity, and efficacy by: Causing turbidity Causing product degradation Shifting the pH Introducing endotoxins and other toxins

Microbial testing (bioburden) is performed to ensure that an adulterated product does not reach the patient

9

Page 10: Principles of Aseptic Processing

Types of Microorganisms Bacteria Fungi

YeastMold

Mycoplasma Microscopic algae Viruses

10

T4 VirusMycoplasma Bacteria

Saprolegnia Mold E Coli Bacteria

Page 11: Principles of Aseptic Processing

Bacteria Prokaryotes: Unicellular microorganisms with no

nucleus & rarely have membrane-bound organelles Ubiquitous to every habitat on Earth 0.5 -1.0 microns in length : 3 million can fit

into an area the size of a pin head Wide range of shapes, spheres, rods, spiral Approximately 10X as many bacterial cells as human

cells in the human bodyLarge populations on the skin and in digestive tract

11

Page 12: Principles of Aseptic Processing

Bacterial Reproduction

Binary FissionOne cell divides into

two identical cellsA single bacterial cell can

multiply to 9 x1030 in a 24-hour period in ideal conditions

Refrigeration slows bacterial growth, but does not stop it completely

12

Bacterial Binary Fission

E.coli can double their number every 20 minutes!!

Page 13: Principles of Aseptic Processing

Fungi

Eukaryotic organismA cell that has a complex structure

enclosed within a membraneContains a nucleus Ubiquitous to every habitat on Earth

TypesYeastMold

Can be multi-cellular (mold) or single-celled (yeast)

Reproduction is sexual or asexualCommonly via spores

13

Mold

Yeast

Page 14: Principles of Aseptic Processing

Yeast Unicellular (single-celled) fungi Average size = 3 - 4 µm in diameter Both aerobic and anaerobic respiration Ideal growth conditions:

Neutral pHTemperature 10˚ – 37˚ C

Part of normal flora of human body Reproduce asexually by budding or

sexually by spores like other fungi Converts sugar into alcohol = alcohol

tolerant

14

Budding Yeast Cells

Yeast Cells with Spores

Page 15: Principles of Aseptic Processing

Molds Multi-cellular filaments called “hyphae”

A colony of hyphae is called a mycelium Functions as a decomposer Ability to survive extreme temperature and

pH Visible as a downy or furry coating on food

or surfaces Reproduce through small spores

Spores can be asexual (mitosis) or sexual (meiosis)

15

Aspergillus niger

Penicillium

Page 16: Principles of Aseptic Processing

Microbes and the Human Body

Where are microbes found on humans?SkinEyesNoseMouthUpper throatIntestines

16

Hands10,000 -100,000 cm2

Groin1-20 million / cm2

Feet1 million / cm2

Scalp~ 1 million / cm2Forehead

100 -1,000 / cm

Saliva~ 10 million / gm

Nasal Fluid~ 10 million / gm

Armpit1-10 million / cm2

Feces>100 million / gm

Page 17: Principles of Aseptic Processing

Identification of Microorganisms Why?

Identify sources of contamination Identify trends in the environment Monitor disinfectant effectiveness

How?1. Appearance or morphology2. Gram staining to identify Gram-negative or Gram-positive bacteria3. Secondary tests to identify families4. Biochemical tests to further speciate (Vitek and API)5. Genetic sequencing analysis (MicroSeq)

17

Fewer than 1% of the world’s microbes have been identified!!

Morphology: Size, ShapeTexture, Color

Page 18: Principles of Aseptic Processing

Bacterial Identification Gram Positive Microorganisms

18

Gram Positive Cocci Staphylococcus aureus

Gram Positive Rods Bacillus sp.

Page 19: Principles of Aseptic Processing

Bacterial Identification Gram Negative Microorganisms

A source of endotoxin

A single E Coli can contribute 2 million Endotoxin Molecules!

19

Gram Negative Rods Escherichia coli

Gram Negative Cocci Neisseria gonorrhoeae

Endotoxin

Cell Wall

Page 20: Principles of Aseptic Processing

Contamination Control

20

Page 21: Principles of Aseptic Processing

What is Contamination?The introduction of undesirable impurities

into an environment Viable (living) Contamination:

Examples: bacteria, yeasts, and molds

Non-Viable Contamination: Examples: lint, dust particles, skin flakes, hair, pollen, smoke, chemical

substances, etc.

21

Page 22: Principles of Aseptic Processing

ContaminantsCommon Particulates

22

Fungi

Water Vapor Smoke

Bacteria

Skin Flakes

Influenza Virus

Page 23: Principles of Aseptic Processing

Contamination Control

Contamination – any effect or action that has a negative impact on a product's integrity making it unfit for useChemical composition pH Sterility & Pyrogenicity Biological or Therapeutic Potency Physical appearance Particulate Matter (e.g. dust, glass or precipitation)

Importance of Aseptic TechniqueParenteral (needle injection) administration bypasses the skin

and gastrointestinal tract, the body’s natural barriers to infectionGiving a patient a contaminated product can cause serious

adverse events including DEATH

23

Page 24: Principles of Aseptic Processing

ContaminationSources

Personnel (most common)○ Touch Contamination○ Skin / Hair / Mucous

Membranes○ Clothing

Equipment Improper Cleaning

/Contaminated SuppliesAir

○ HVAC / HEPA Failure○ Infiltration○ Internal Generation

24

Page 25: Principles of Aseptic Processing

Particle GenerationPersonnel

25

NOTE: Men shed more particles than women

Activity Particles Generated

Standing or Sitting 100,000

Slight head/hand movement

500,000

Body, arm movement w/ toe tapping

1 million

Changing from sitting to standing

2.5 million

Slow walk 5 million

Running 30 million

Page 26: Principles of Aseptic Processing

Contamination ControlPreventative Practices

Gowning Labcoats in the labs Scrubs/Cleanroom suits for manufacturing areas Face masks, beard covers, hair nets, etc.

Good hygiene practices Personnel/material/waste flow in critical areas HEPA filters Sanitization of equipment/materials Slow, deliberate movements in controlled areas Cleaning techniques

26

Page 27: Principles of Aseptic Processing

The HEPA Filter

High Efficiency Particulate Attenuation

Page 28: Principles of Aseptic Processing

Unidirectional Air FlowAir Flow Patterns•Air moving devices displace the air in a unidirectional (one direction) pattern to reduce turbulence of the air and displace contaminants.

•Air is directed using walls, curtains, or panels.

•Smoke studies are done to allow the airflow to be visualized.

•Machinery and large objects can affect airflow by creating local zones of air turbulence

Page 29: Principles of Aseptic Processing

Room Air Changes

Air is circulated through the Cleanroom and carries contaminants as air passes through in unidirectional flow.

Recirculated air passes through HEPA filters.

To maintain ISO 5 (Grade A, Class 100) conditions requires the air volume to be replaced a minimum of 25 air changes per hour (ACH) ; air velocity should at least be 0.45 m/s.

20 ACH are typically required for ISO 7 areas (Grade C areas; Class 10,000) and ISO 8 (Grades D areas; Class 100,000.

29

AHU1:Recirculated Air

AHU2: Outdoor Air

HEPA Filters

Page 30: Principles of Aseptic Processing

Types of Cleaners/Disinfectants

30

Spor Klenz Active Ingredient 22% Acetic Acid 4.5% Peracetic Acid 10% Hydrogen Peroxide

70% IPA

LpHsePhenolic Disinfectant Acidic, (pH 2.6 – 3.0)

Vesphene IIse Alkaline pH 10.4-10.6

Bleach Sodium Hypochlorite

Page 31: Principles of Aseptic Processing

Disinfectant Action

31

UV Light

SurfaceProteins

Protein Denaturation by Base

Disruption of Cell Membrane

Protein Hydrolysis by Acid

Page 32: Principles of Aseptic Processing

Microbial Killing Efficiency Antimicrobial Agents Compared

32

Page 33: Principles of Aseptic Processing

Contamination Control Personal Resposibility

Personal Hygiene Regular Bathing/showering (includes washing

hair) Practice good oral hygiene Clean clothes and shoes

Illness Notify your management of illness or open

wounds Restrict access to controlled areas

When working in Controlled Areas No make-up, jewelry, or cologne No food, drinks, or gum/candy Avoid getting sunburned (flaking skin)

33

Page 34: Principles of Aseptic Processing

Contamination Control Personal Responsibility

Frequent hand washing If you smoke, drink water

after to reduce the introduction of smoke particles into controlled areas

Maintain a clean, organized workspace

34

Page 35: Principles of Aseptic Processing

4 Pillars of Aseptic Techniques

Personnel training & monitoringEnvironmental monitoringFacilities design & HVAC validationProcess simulation (media fills)

Page 36: Principles of Aseptic Processing

Let’s Review! What does “aseptic” mean? Name 3 scenarios that require aseptic technique Name 3 types of microorganisms List the different techniques that QC employs for microbial

identification What are the two categories of particulate contamination? Name 3 sources of contamination List 3 ways to control contamination Describe YOUR personal responsibilities toward contamination

control

36

Page 37: Principles of Aseptic Processing

Q & A

37