Specifications, Methods and precedence of source with particular reference to microbiological requirements in TGOs.

Transcription

1 Specifications, Methods and precedence of source with particular reference to microbiological requirements in TGOs. Fergus O Connell RACI Pharmaceutical Science Group (NSW) 10/08/2015

2 Currently there are 22 current TGOs (as of 04/08/15) 6 address microbiology in some respect: TGO No General Standard for Pyrogen and Endotoxin Content of Therapeutic Goods (28/02/1995) To be ceased 01 Apr 2018 Reason for Ceasing Sunsetting TGO No Standard for Disinfectants and Sterilants (13/11/1996) - As amended To be ceased - No date listed. TGO No Standard for Sterile Therapeutic Goods (27/11/1998) To be ceased 01 Oct 2017 Reason for Ceasing Sunsetting

3 TGO No Microbiological Standards for Medicines (22/09/2008) To be ceased 01 Oct 2024 Reason for Ceasing - Sunsetting TGO No Standard for Tablets and Capsules (29/10/2008) To be ceased 01 Apr 2019 Reason for Ceasing Sunsetting Note sets fiducial limits for antibiotics where the microbiological method of assay is used in the test. TGO No Standard for WFI for Parenteral Medicines (18/10/2011) To be ceased 01 Apr 2022 Reason for Ceasing - Sunsetting

4 TGO 50 Goods. shall comply with Test for Pyrogens specified in the BP Bacterial Endotoxin Test (BET) of the USP. Medical devices primarily & Parenterals (>15 ml injected) Interestingly, Appendix XIV C. Test for Bacterial Endotoxins (LAL Test) of the BP is not listed. Note: Since December 2012, the endotoxin test methods have been harmonized. The 1987 guideline (withdrawn in 2011) on validating the Limulus Amebocyte Lysate test is still referenced.

5 TGO 63 - Standard for Sterile Therapeutic Goods Appendix XVI Test for Sterility shall constitute the standard for therapeutic devices that are labelled as sterile or sterilised or otherwise purport to be sterile or sterilised.

6 TGO 77 Microbiological Standards for Medicines Addresses the requirements of both Sterile & Nonsterile medicines. Section 7 Sterility and Bacterial Endotoxin Testing Section 8 PET of a multidose medicine. Section 9 Non-sterile medications. The order or pharmacopoeias referenced are not comprehensive microbial limit specifications. Sponsors must determine risks from other objectionable organisms.

7 Section 7 Sterility & Endotoxins Sterile Products: BP Appendix XVI A. Sterility Endotoxins: - BP Appendix XIV C. Test for Bacterial Endotoxins (LAL Test) Note these appendices are not referenced in the order. The medicine must comply with. a default standard.

8 Media for Sterility Testing Fluid thioglycollate: primarily for anaerobic bacteria; Note - it will also detect aerobic bacteria. Soya-bean casein digest both fungi and aerobic bacteria.

9 Growth Promotion & Method Suitability Media: Separate containers Inoculated with <100CFU Fluid thioglycollate: Clostridium sporogenes, Pseudomonas aeruginosa, Staphylococcus aureus. Soya-bean casein digest Aspergillus brasiliensis, Bacillus subtilis, Candida albicans.

10 Test for Sterility Membrane Filtration is performed wherever possible. Direct Inoculation also available e.g. medical devices. The media is again inoculated with <100CFU as per the growth promotion step. The primary function of a Method Suitability test (Validation) is to demonstrate that: The product has no antimicrobial properties or that these have been neutralized, The test can recover small numbers of microorganisms in the presence of the product (<100 CFU).

11 Sterility Specifications Simple Growth of each organism in the Growth Promotion Studies and Method Suitability tests. No growth in routine testing. Confirmatory steps are required e.g. streaking out on plates to confirm viability, where a clear Growth/No Growth result is not obtained.

12 Endotoxins Validity of testing requires 2 aspects to be met: The suitability of the material to be used The absence of endotoxins in the water & reagents the sensitivity of the amoebocyte lysate must be checked Absence of product interference with the test: the sensitivity of the amoebocyte lysate is determined in the presence and in the absence of the product under examination. There must be no significant difference between the 2 sensitivity values.

13 Limit = K/M K = M = Endotoxin Limit threshold pyrogenic dose of endotoxin per kilogram of body mass; maximum recommended bolus dose of product per kilogram of body mass. Note: products to be injected at frequent intervals or infused continuously, M is the maximum total dose administered in a single hour period. Table of the BP suggests values for K.

14 Important points to remember MVD Maximum Valid Dilution maximal assurance ve result means endotoxin conc. of the product < endotoxin limit. Depends on: endotoxin limit sensitivity of the lysate MVD Calculation λ = the labelled lysate sensitivity in the gel-clot technique (IU/mL) or the lowest concentration used in the standard curve of the turbidimetric or chromogenic techniques.

15 λ: the labelled lysate sensitivity in the gel-clot technique (IU/mL) or the lowest concentration used in the standard curve (turbidimetric or chromogenic techniques). The endotoxin limit for active substances administered parenterally is specified in monographs. See reference - USP Endotoxin Limits for common injectables. Lysate sensitivity down to EU/ml typical.

16 Interference: Products are diluted to eliminate interference. NOTE: amoebocyte lysate reacts with some β-glucans in addition to endotoxins. Dilutions reduce the sensitivity of the test. BP - test for bacterial endotoxins is preferred over the test for pyrogens: usually considered to provide equal or better protection to the patient.

17 Endotoxins Substitution of an LAL test for the rabbit pyrogen test required in a monograph: constitutes the use of an alternative method of analysis and hence requires validation.

18 Section 8 Preservative Efficacy Multidose medicines must comply with BP Appendix XVI C. Liquid oral antacid medicine may comply with USP chapter <51> Note Only the BP and Equivalent EP Chapters are referenced other than USP <51> for liquid oral antacids. Reason different acceptance criteria. BP - some antacids.. are unlikely to achieve target criteria.

19 PET - Method The preparation is challenged, wherever possible in its final container, with a prescribed inoculum of suitable micro-organisms Storing the inoculated preparation at a prescribed temperature. Withdrawing samples from the container at specified intervals of time and counting the organisms in the samples so removed. Recovery of each microorganism must be validated Also neutralizers.

20 PET Organisms Organisms Description Use Pseudomonas aeruginosa Gram -ve rod Standard Organism Staphylococcus aureus Gram +ve cocci Standard Organism Candida albicans Yeast Standard Organism Aspergillus brasiliensis Mould Standard Organism Escherichia coli Gram -ve rod Recommended for oral preparations Zygosaccharomyces rouxii Osmotolerant fructophilic yeast Recommended for oral preparations with a high concentration of sugar.

21 PET - Acceptance Criteria. The A criteria express the recommended efficacy to be achieved. In justified cases where the A criteria cannot be attained, the B criteria must be satisfied. E.g. - an increased risk of adverse reactions,

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23 Antacids BP Requirements 3 log reduction for bacteria in 14 days USP Requirements No Increase

24 Section 9 Non-Sterile Medicines Non-sterile medicines must comply with the relevant acceptance criteria of: Appendix XVI B. Microbiological Examination of Nonsterile Products Section 1 - Test for Specified Micro-organisms (Ph. Eur. method , USP <62>) Section 2 - Microbial Enumeration Tests (Ph. Eur. method , USP <61>) Note: these are listed separately in TGO77 (issued pre-harmonisation).

25 Microbial Limits Tests A collection of tests and specifications. Products are tested determine the absence or limited occurrence of specified micro-organisms Title BP EP USP Microbial Enumeration Tests Appendix XVI B <61> Tests for specified organisms Appendix XVI B <62> Alternative microbiological procedures - equivalence to the Pharmacopoeia method has to be demonstrated.

26 Microbial Enumeration Tests Bioburden determination quantitative. Harmonisation added significantly to the USP chapters Inoculum of < 100 CFU Growth is described in greater detail Harvesting and resuspension of cells detailed Media growth promotion requirements et-cetera. Two separate tests Total Aerobic Microbial Count (TAMC) Total Yeast and Mould Count (TYMC)

27 Method Validation The suitability of the counting method must be demonstrated in the presence of the product. Sample prep depends on the physical characteristics of the product: Water-soluble Non-fatty products Fatty products Fluids or solids in aerosol form Transdermal patches. Recovery of <100 CFU from the lowest possible dilution required. Neutralisation/removal of antimicrobial activity.

28 Membrane Filtration Plate-count methods Pour-plate Spread-plate Methods of recovery Most-probable-number (MPN) NOTE: This is the least precise or accurate method. Not suitable for enumeration of moulds. Reserved for TAMC where no other method is available.

29 Acceptance Criteria If acceptance criteria has been prescribed 10 1 CFU: Max acceptable count = CFU: Max acceptable count = CFU: Max acceptable count = 2000, and so forth. Note on Bacterial Counts USP/FDA Countable Range: CFU/plate First proposed in 1916 after a study of Milk Analysis. American Society for Testing and Materials (ASTM) CFU/membrane, CFU/pour plate

30 Tests for Specified Indicator Organisms Staphylococcus aureus Pseudomonas aeruginosa Salmonella spp Escherichia coli Clostridia* Candida Albicans* Bile Tolerant Gram-Negative Bacteria (BTGN)* * newly specified in the USP upon harmonization in 2009

31 The first step enrichment to encourage growth of low numbers of the specified organism. Followed by subculture on selective and/or differential media. Samples are prepped as per the enumeration methods. Specified Organism BTGN Dilution Media Temp & Time 1:10 dilution with >1g of product Volume = 1g of product Subculture Casein soya bean digest broth Enterobacteria enrichment broth-mossel Violet Red Bile Glucose Agar C for 2-5 hours C for hours As above Function Resuscitate bacteria without encouraging multiplication. Test for Absence As above

32 section 9 of this order or the pharmacopoeias mentioned. Should not be regarded as comprehensive microbial limit specifications. Ref - Section 3 of TGO 77 The sponsor must determine the risk to the medicine from other objectionable microorganisms. What is an objectionable organism?

33 Objectionable Organisms TGA: The significance of, and risk from these other microorganisms should be evaluated in terms of the formulation of the medicine, its route of administration, method of application, the population for which the medicine is intended. underlying illness/disease in the population. the possible use of immunosuppressive agents or corticosteroids by the user. Risk may differ for neonates, infants, the debilitated.* *USP <1111>

34 Burkholderia cepacia as an Objectionable Organism The primary hazard - those with cystic fibrosis (or predisposed to pneumonia) and the use of inhaled medications. It is a clear pathogen when introduced into the air passages of a susceptible population. Objectionable in most non-sterile product presentations for general use? - remains unclear. US FDA has issued 483 findings for a wide range of product types where it was detected.

35 Schedules 1 of TGO77 Microbial attributes for a complementary medicine oral dosage form containing raw material of natural (animal, vegetal or mineral) origin. Microbiological Quality Acceptance Criteria Total aerobic microbial count Total yeast and mould count Bile-tolerant Gram negative bacteria Salmonella Escherichia coli Staphylococcus aureus Less than or equal to 10 4 CFU per g or per ml Less than or equal to 10 2 CFU per g or per ml Less than or equal to 10 2 CFU per g or per ml absent in 10 g or 10 ml absent in 1 g or 1 ml absent in 1 g or 1 ml

36 Schedules 2 of TGO77 Applies to: complementary medicine oral dosage form containing raw material of natural (vegetal) origin that is a herbal medicinal product consisting solely of one or more herbal substances (whole, reduced or powdered) to which boiling water is added before use. Microbiological Quality Total aerobic microbial count Total yeast and mould count Bile-tolerant Gram negative bacteria Escherichia coli Salmonella Acceptance Criteria Less than or equal to 10 7 CFU per g Less than or equal to 10 5 CFU per g Less than or equal to10 2 CFU per g absent in 1 g absent in 10 g

37 The TGA reminder to sponsors of listed herbal teas the pharmacopoeial test methods are not optimal for the detection of low numbers of enteric pathogens in the presence of high numbers of competing microorganisms. See Reference: Guidance on Therapeutic Goods Order No. 77

38 Thank you

39 References: Guidance on Therapeutic Goods Order No microbiological-standards-medicines List of Therapeutic Goods Orders FDA - Bacterial Endotoxins/Pyrogens calguides/ucm htm ICH guideline Q4B - Annex 14 to Note for Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions on Bacterial Endotoxins Tests General Chapter deline/2010/09/wc pdf

40 USP Endotoxin Limits for common injectables British Pharmacopoeia - SC IV P. Guidelines for Using the Test for Sterility What is an Objectionable Organism? American Pharmaceutical Review Posted October 12, Articles/ What-is-an-Objectionable-Organism-Objectionable- Organisms-The-Shifting-Perspective/ ARGOM Appendix 2: Guidelines on quality aspects of OTC applications See table 10.2 for limits.