ALS Ltd. · ASX:ALQ
ALS — Technical Presentation (March 2016)
54 slides from ALS's technical presentation, March 2016
Slide 1: Microbiology for Oral and Topical Products - The basics
Microbiology for Oral and Topical Products - The basics Scott Colbourne Business Manager NSW – ALS Food & Pharmaceutical R IG H T S O L U T I O N S | R I G H T PA RT N E R
Slide 2: Contents
Contents • TGO 77 - Introduction • Tests Performed – Common Tests – Preservative Efficacy – Endotoxin – Bioburden • Method Turn Around Times (TAT) • The Confirmation Process • Method Validation • Causes of counts being high initially, then low when retested • Topical Products Microbiology for Oral a
Slide 3: TGO 77 - General
TGO 77 - General • Therapeutic Goods Order No. 77 – Microbiological Standards for Medicines (TGO 77) • This Order specifies the minimum microbiological requirements with which a medicine must comply throughout its shelf life in Australia. • On and from 1 January 2010, each medicine to which this Ord
Slide 4: TGO 77 - General
TGO 77 - General • Acceptance criteria is the same as used in the BP, Ph.Eur. and the USP: • A medicine is considered to be non-compliant with a microbial count acceptance criterion if the CFU (colony forming unit) count exceeds two (2) times the stated limit. • When the Order establishes a limit of
Slide 5: TGO 77
TGO 77 • Section 7: Sterility and Bacterial Endotoxin testing – A medicine must comply when these tests are required by an individual or general monograph – Where a sponsor labels or packages a medicine in a way that states or implies that the medicine is sterile Microbiology for Oral and Topical 5
Slide 6: TGO 77
TGO 77 • Section 8: Efficacy of antimicrobial preservation of multidose medicines – Applies to multidose sterile medicines (e.g. eye drops) – Applies to multidose non-sterile medicines (e.g. oral liquids) Microbiology for Oral and Topical 6 Products - The Basics
Slide 7: TGO 77
TGO 77 • Subsection 9(1): – Applies to prescription, OTC and many complementary medicines – Acceptance Criteria • BP Appendix XVI (Ph. Eur. 5.1.4): Limits defined by route of administration • USP chapter <1111>: Almost identical to the BP criteria – Testing consists of some/all of the following: Tot
Slide 8: TGO 77
TGO 77 • Subsection 9(2): – Applies to a complementary medicine oral dosage form containing raw material of natural origin Microbiological Quality Units Schedule 1: Schedule 2: Containing Solely herbal substances * Total aerobic microbial count CFU per g or mL ≤ 104 ≤ 107 Total yeast and mould count
Slide 9: Common Tests – what and why?
Common Tests – what and why? • Total Aerobic Microbial Count (TAMC) What? A generic count of micro-organisms present. The results are non-specific Why? Gives a general indication of the level of micro-organisms present in the sample tested Can be referred to as… Total Microbial Count (TMC), Standard
Slide 10: Common Tests – what and why?
Common Tests – what and why? • Yeast and Mould What? Generic test for the levels of these present. Non-specific. Why? • They cause biodegradation of natural materials • Some yeasts are useful in fermentation (e.g. Bread and Beer) • However some (e.g. Candida albicans) are opportunistic pathogens and
Slide 11: Common Tests – what and why?
Common Tests – what and why? • Bile Tolerant gram negative bacteria What? • The name comes from their gram staining method of bacterial differentiation. Basically they do not retain crystal violet. • Large family of bacteria including Enterobacteriaceae, Salmonella and E.Coli Why? • In humans, disea
Slide 12: Common Tests – what and why?
Common Tests – what and why? • Salmonella What? Pathogen found in cold/warm blooded animals and the environment Why? • It causes food poisoning, such as gastrointestinal issues • To ensure there is no presence of pathogens Microbiology for Oral and Topical 12 Products - The Basics
Slide 13: Common Tests – what and why?
Common Tests – what and why? • E. coli What? A coliform almost exclusively of faecal origin Why? • E. coli presence is an effective confirmation of faecal contamination. • Most strains of E. coli are harmless, but some can cause serious illness in humans. Microbiology for Oral and Topical 13 Product
Slide 14: Common Tests – what and why?
Common Tests – what and why? • Staphylococcus Aureus What? There are over 30 Staphylococcus types of bacteria, but Staphylococcus aureus causes most staph infections Why? • Can cause skin infections (most common), Pneumonia, food poisoning, toxic shock syndrome and blood poisoning (bacteremia) • A s
Slide 15: Common Tests – what and why?
Common Tests – what and why? • Pseudomonas aeruginosa What? It is found in soil, water and most man-made environments Why? • An opportunistic human pathogen of immunocompromised individuals • Because it thrives on moist surfaces, this bacterium is also found on and in medical equipment, including ca
Slide 16: Common Tests – what and why?
Common Tests – what and why? • Candida Albicans What? • A fungus that is a constituent of the normal gut flora that live in the human mouth and gastrointestinal tract. • Present in 80% of the human population without causing harmful effects Why? • Causes opportunistic oral and genital infections in
Slide 17: Preservative Efficacy Testing (PET)
Preservative Efficacy Testing (PET) • What is preservative efficacy? • The test consists of challenging the preparation by inoculating with suitable micro-organisms, at a prescribed temperature, withdrawing samples at specified intervals and counting the organisms present. • The preservative propert
Slide 18: Preservative Efficacy Testing (PET)
Preservative Efficacy Testing (PET) • What is preservative efficacy? • The test consists of challenging the preparation by inoculating with suitable micro-organisms, at a prescribed temperature, withdrawing samples at specified intervals and counting the organisms present. • The preservative propert
Slide 19: Preservative Efficacy Testing (PET)
Preservative Efficacy Testing (PET) • What is preservative efficacy? • The test consists of challenging the preparation by inoculating with suitable micro-organisms, at a prescribed temperature, withdrawing samples at specified intervals and counting the organisms present. • The preservative propert
Slide 20: Preservative Efficacy Testing (PET)
Preservative Efficacy Testing (PET) • What is preservative efficacy? • The test consists of challenging the preparation by inoculating with suitable micro-organisms, at a prescribed temperature, withdrawing samples at specified intervals and counting the organisms present. • The preservative propert
Slide 21: Endotoxin (LAL)
Endotoxin (LAL) • What is endotoxin testing? • The outer membrane of a Gram-negative bacterium is composed of lipopolysaccharide (an endotoxin). • The qualitative determination of bacterial endotoxin concentration is performed by the Limulus Amebocyte Lysate (LAL) gel clot test Microbiology for Oral
Slide 22: Endotoxin (LAL)
Endotoxin (LAL) • What is endotoxin testing? • The outer membrane of a Gram-negative bacterium is composed of lipopolysaccharide (an endotoxin). • The qualitative determination of bacterial endotoxin concentration is performed by the Limulus Amebocyte Lysate (LAL) gel clot test • What samples are te
Slide 23: Endotoxin (LAL)
Endotoxin (LAL) • What is endotoxin testing? • The outer membrane of a Gram-negative bacterium is composed of lipopolysaccharide (an endotoxin). • The qualitative determination of bacterial endotoxin concentration is performed by the Limulus Amebocyte Lysate (LAL) gel clot test • What samples are te
Slide 24: Bioburden
Bioburden • What is Bioburden testing? • This testing does not constitute sterility testing, but is a method used to determine the microbial bioburden of the sample. • Bioburden is normally defined as the number of bacteria living on a surface that has not been sterilised Microbiology for Oral and T
Slide 25: Bioburden
Bioburden • What is Bioburden testing? • This testing does not constitute sterility testing, but is a method used to determine the microbial bioburden of the sample. • Bioburden is normally defined as the number of bacteria living on a surface that has not been sterilised • What samples are tested?
Slide 26: Bioburden
Bioburden • What is Bioburden testing? • This testing does not constitute sterility testing, but is a method used to determine the microbial bioburden of the sample. • Bioburden is normally defined as the number of bacteria living on a surface that has not been sterilised • What samples are tested?
Slide 27: Bioburden
Bioburden • What is Bioburden testing? • This testing does not constitute sterility testing, but is a method used to determine the microbial bioburden of the sample. • Bioburden is normally defined as the number of bacteria living on a surface that has not been sterilised • What samples are tested?
Slide 28: Method Turn Around Times
Method Turn Around Times Turnaround Time (Days) Method If Confirmation is If Clean Required Total aerobic microbial count 5 n/a Yeast and Mould 5-7 n/a Bile tolerant Gram negative bacteria 2-3 n/a Salmonella 3-4 5-6 E. coli 3-6 4-8 Staphylococcus Aureus 2-4 4-6 Extras not listed in TGO 77 section 9(
Slide 29: Confirmation Process
Confirmation Process • Have you ever received an email like this? Hi John, Please note that the below sample is suspect for E.Coli: Lot: 12345 sample A, DOM 15/05/2015 The sample is under confirmation and the final result will be updated once complete Thank you and best regards Jenny Bloggs Microbio
Slide 30: Confirmation Process
Confirmation Process • Have you ever received an email like this? Hi John, Please note that the below sample is suspect for E.Coli: Lot: 12345 sample A, DOM 15/05/2015 The sample is under confirmation and the final result will be updated once complete Thank you and best regards Jenny Bloggs Microbio
Slide 31: Confirmation Process
Confirmation Process • Have you ever received an email like this? Hi John, Please note that the below sample is suspect for E.Coli: Lot: 12345 sample A, DOM 15/05/2015 The sample is under confirmation and the final result will be updated once complete Thank you and best regards Jenny Bloggs Microbio
Slide 32: Confirmation Process
Confirmation Process • Have you ever received an email like this? Hi John, Please note that the below sample is suspect for E.Coli: Lot: 12345 sample A, DOM 15/05/2015 The sample is under confirmation and the final result will be updated once complete Thank you and best regards Jenny Bloggs Microbio
Slide 33: Confirmation Process
Confirmation Process • What tests can require confirmation? Turnaround Time (Days) Method If Confirmation is If Clean Required Total aerobic microbial count 5 n/a Yeast and Mould 5 n/a Bile tolerant Gram negative bacteria 3 n/a Salmonella 4 2-3 E. coli 5 7 Staphylococcus Aureus 3 6 Extras not listed
Slide 34: Confirmation Process
Confirmation Process • What tests can require confirmation? Turnaround Time (Days) Method If Confirmation is If Clean Required Total aerobic microbial count 5 n/a Yeast and Mould 5 n/a Bile tolerant Gram negative bacteria 3 n/a Salmonella 4 2-3 E. coli 5 7 Staphylococcus Aureus 3 6 Extras not listed
Slide 35: Method Validation
Method Validation • What is microbiological method validation? • Verifying that the method used including media and sample dilution are suitable to recover the micro organism Microbiology for Oral and Topical 35 Products - The Basics
Slide 36: Method Validation
Method Validation • What is microbiological method validation? • Verifying that the method used including media and sample dilution are suitable to recover the micro organism • How is it done? • By inoculating the sample with the bacteria, incubating and recovering >70% of the micro organism Microbi
Slide 37: Method Validation
Method Validation • What is microbiological method validation? • Verifying that the method used including media and sample dilution are suitable to recover the micro organism • How is it done? • By inoculating the sample with the bacteria, incubating and recovering >70% of the micro organism • How l
Slide 38: Method Validation
Method Validation • What is microbiological method validation? • Verifying that the method used including media and sample dilution are suitable to recover the micro organism • How is it done? • By inoculating the sample with the bacteria, incubating and recovering >70% of the micro organism • How l
Slide 39: Method Validation
Method Validation • Why do they fail? • Due to antimicrobial efficacy of the product • Presence of preservatives • <70% recovery of the validation test Microbiology for Oral and Topical 39 Products - The Basics
Slide 40: Method Validation
Method Validation • Why do they fail? • Due to antimicrobial efficacy of the product • Presence of preservatives • <70% recovery of the validation test • How can I avoid validation delays on new products? • By sending in samples for validation in advance, e.g. process validation or pilot scale batch
Slide 41: High to Low Counts
High to Low Counts • How can a result be high in one test and then low when retested? • This can be caused by a number of factors – The sample may not be completely homogenous – Lab error – The sample has anti-bacterial properties – Preservative taking effect Microbiology for Oral and Topical 41 Pro
Slide 42: High to Low Counts: Sample Not Homogeneous
High to Low Counts: Sample Not Homogeneous • The laboratory generally uses 10g of the sample in the required media, e.g. 90 mL Microbiology for Oral and Topical 42 Products - The Basics
Slide 43: High to Low Counts: Sample Not Homogeneous
High to Low Counts: Sample Not Homogeneous • The laboratory generally uses 10g of the sample in the required media, e.g. 90 mL • Of that 90 mL, 0.1ml to 1ml is removed for each test Microbiology for Oral and Topical 43 Products - The Basics
Slide 44: High to Low Counts: Sample Not Homogeneous
High to Low Counts: Sample Not Homogeneous • The laboratory generally uses 10g of the sample in the required media, e.g. 90 mL • Of that 90 mL, 0.1ml to 1ml is removed for each test • A reported result of 500 cfu/mL, may only be 5 colonies on an agar plate. This is a low number and with repeat testi
Slide 45: High to Low Counts: Sample Not Homogeneous
High to Low Counts: Sample Not Homogeneous • The laboratory generally uses 10g of the sample in the required media, e.g. 90 mL • Of that 90 mL, 0.1ml to 1ml is removed for each test • A reported result of 500 cfu/mL, may only be 5 colonies on an agar plate. This is a low number and with repeat testi
Slide 46: High to Low Counts: Lab Error
High to Low Counts: Lab Error • This is always a possibility and our quality procedures and controls minimise this risk. However it can never be 100% removed. Microbiology for Oral and Topical 46 Products - The Basics
Slide 47: High to Low Counts: Lab Error
High to Low Counts: Lab Error • This is always a possibility and our quality procedures and controls minimise this risk. However it can never be 100% removed. • If we believe lab error is a possibility we will try to look for a pattern, e.g. • Do all the similar samples have a high count? • Does the
Slide 48: High to Low Counts: Lab Error
High to Low Counts: Lab Error • This is always a possibility and our quality procedures and controls minimise this risk. However it can never be 100% removed. • If we believe lab error is a possibility we will try to look for a pattern, e.g. • Do all the similar samples have a high count? • Does the
Slide 49: High to Low Counts: Anti-Bacterial Properties
High to Low Counts: Anti-Bacterial Properties • The sample may contain substances that are anti-bacterial (e.g. preservative systems) • Samples are stored for up to 5 days before a retest can occur and substances in the sample itself may lower the bacterial count • Therefore possibly high levels upo
Slide 50: Topical Products
Topical Products • What should I test? Microbiology for Oral and Topical 50 Products - The Basics
Slide 51: Topical Products
Topical Products • What should I test? • Based on Cutaneous acceptance criteria in the Pharmacopoeias (BP, USP, Ph. Eur) Microbiological Quality Units Pass Criteria Total aerobic microbial count CFU per g or mL ≤ 102 Total yeast and mould count CFU per g or mL ≤ 101 Staphylococcus aureus in 1g or 1m
Slide 52: Topical Products
Topical Products • What should I test? • Based on Cutaneous acceptance criteria in the Pharmacopoeias (BP, USP, Ph. Eur) Microbiological Quality Units Pass Criteria Total aerobic microbial count CFU per g or mL ≤ 102 Total yeast and mould count CFU per g or mL ≤ 101 Staphylococcus aureus in 1g or 1m
Slide 53: Topical Products
Topical Products • What should I test? • Based on Cutaneous acceptance criteria in the Pharmacopoeias (BP, USP, Ph. Eur) Microbiological Quality Units Pass Criteria Total aerobic microbial count CFU per g or mL ≤ 102 Total yeast and mould count CFU per g or mL ≤ 101 Staphylococcus aureus in 1g or 1m
Slide 54: Further Reading
Further Reading British Pharmacopoeia (2015): Appendix XVI D. Microbiological Quality of Non-sterile Pharmaceutical Preparations and Substances for Pharmaceutical Use United States Pharmacopoeia – National Formulary (38): <1111> MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: ACCEPTANCE CRITERIA