Microbiological Control

EXCELLENCE IN MICROBIOLOGYMicrobiological Control for Non-Sterile Pharmaceuticals Pharmig Monograph No. 2 Pharmaceutical Quality Group Monograph No. 12  Pharmig Pharmaceutical Quality Group of The Chartered Quality Institute Pharmig Monograph No. 2 PQG Monograph No. 12 MICROBIOLOGICAL CONTROL FOR NON-STERILE PHARMACEUTICALS November 2008 EXCELLENCE IN MICROBIOLOGY  Contents Contents Page General Introduction ii Foreword v From the Chairmen vi Acknowledgements vii Chapter 1 Introduction 1 Chapter 2 Roles and Responsibilities 5 2.1. Introduction 2.2. Microbial Governance Chapter 3 Personnel 7 3.1. Introduction 3.2. Training 3.3. Hygiene 3.4. Dress Code/Changing Chapter 4 Facilities 11 4.1. Introduction 4.2. Area Classification 4.3. Access to Areas 4.4. Building Requirements 4.5. Facility Qualification Chapter 5 Cleaning and Disinfection 23 5.1. Introduction 5.2. Facilities 5.3. Equipment 5.4. Cleaning Agents 5.5. Validation of Cleaning Chapter 6 Microbiology Laboratories 31 6.1. Introduction 6.2. Relationship to the Quality Unit 6.3. Microbiology Laboratory Facility Design 6.4. Autoclaves 6.5. Disinfectants 6.6. Media 6.7. Micro-organisms 6.8. Identification 6.9. Microbiological Testing of Product, Starting Materials and Intermediates Chapter 7 Risk Assessment and Management 37 © 2008 Pharmig and The Chartered Quality Institute 7.1. Introduction 7.2. Risk Assessment Associated with Non-Sterile Products All rights reserved. No part of this publication may be reproduced, stored 7.3. Water Activity in a retrieval system, or transmitted in any form or by any means, without Chapter 8 Microbiological Monitoring 43 the written permission of Pharmig and The Chartered Quality Institute. 8.1. Introduction 8.2. Environmental Monitoring Policy 8.3. Designing the Microbiological Monitoring Programme Published by Pharmig and The Chartered Quality Institute Chapter 9 Reporting and Trending of Microbiological Data 61 ISBN 978-0-9560804-0-0 9.1. Introduction 9.2. Batch-Specific Data 9.3. Periodic Summary Reports This monograph is available from: 9.4. Out of Specification/Out of Trend Handling Pharmig, T5 The Maltings, Roydon Road, Stanstead Abbotts, Hertfordshire SG12 8HG, United Kingdom. Appendix Mind Maps 65 www.pharmig.org.uk Glossary 69 Bibliography 73 The Chartered Quality Institute, 12 Grosvenor Crescent, London SW1X 7EE, United Kingdom. www.thecqi.org Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals i  General Introduction General Introduction The Pharmaceutical Quality Group Pharmig EXCELLENCE IN MICROBIOLOGY The Pharmaceutical Quality Group in the United Kingdom developed from a small group of The Pharmaceutical Microbiology Interest Group (Pharmig) is a non-profit making pharmaceutical quality executives who met initially in 1977 to assist in the preparation of professional organisation, established in 1991, that represents the interests of individuals a supplier questionnaire for discussion at the 1978 seminar of the European Organisation who work in, have responsibility for, or work alongside microbiology. for Quality Control. The group has expanded since that time and is now incorporated with the Chartered Quality Institute. Pharmig is the influential forum for microbiology in the pharmaceutical, healthcare and allied industries. It provides a focus for continuing professional development and serves as The objectives of the group are: a unique network for the exchange of microbiological information through training courses, 1. The open exchange of information and experience concerning pharmaceutical quality conferences, publications and its website forum. matters. Initial meetings were held at members’ places of work and included site tours. The first Annual 2. The development of a unified approach to pharmaceutical quality assurance and good Conference was held in 1992. This has now become a well-established annual diary event. manufacturing practices. 3. To promote and represent the status of pharmaceutical quality assurance professionals. The Group has grown significantly since 1991 expanding the portfolio of products it now offers 4. To promote education and training in the achievement of pharmaceutical quality. to the Membership whilst remaining true to the initial needs of microbiologists which include: Organising meetings, training courses and conferences that provide topical information and views on microbiologically related topics The Pharmaceutical Quality Group initiated a project in 1983 with the intention of writing a series of monographs, the prime objective of which was to augment the published, Advancing the science of microbiology and its practical application official Codes of Good Manufacturing Practice by providing practical, comprehensive Influencing the development of regulations and guidelines surrounding microbiology and non-mandatory guidelines. The target audience was identified as management and Acting as a confidential forum for the dissemination of information concerning all supervisory personnel engaged in the manufacture and supply of medicines, with particular aspects of microbiology emphasis on their future training and education. The series of monographs currently contains the following titles: Publications 1. Pharmaceutical Premises and Environment Pharmig launched its first publication – ‘A Guide to Disinfectants and their use in the Pharmaceutical Industry’ in 2006 (Monograph No.1). 2. Pharmaceutical Manufacturing (Processing and Packaging) 3. Elements and Philosophy of Pharmaceutical Quality Assurance A series of Limulus Amoebocyte Lysate (LAL) Fact Sheets on pyrogen/endotoxin testing have also been produced by the LAL Action Group and range from ‘What is LAL/BET?’ 4. Pharmaceutical Distribution through to ‘Product Validations – Quantitative Methods’. 5. Pharmaceutical Auditing Pharmig produces a quarterly technical newsletter for its members and has conducted, 6. Bulk Pharmaceutical Chemicals – (withdrawn) in the past, various surveys that include Non-sterile manufacturing, Aseptic Manufacturing, 7. Pharmaceutical Contract Manufacture and Analysis Environmental Monitoring, Water Systems and Disinfectants. 8. Pharmaceutical Documentation This monograph on ‘Microbiological Control for Non-Sterile Pharmaceuticals’ is Pharmig’s 9. Pharmaceutical Packaging Validation (produced jointly with The Packaging Society) first publication in collaboration with PQG. 10. Cleaning Validation 11. Good Quality Control Laboratory Practice ii Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals iii  Foreword Pharmaceutical Microbiology Advanced Training (PMAT): Following a collaborative effort by Pharmig and the University of Manchester, a Master’s Foreword Degree in Pharmaceutical Microbiology Advanced Training (PMAT) was launched in September 2007. This is a university accredited qualification in pharmaceutical “Dead flies cause the ointment of the apothecary to send forth a stinking savour” was the microbiology for the medical and healthcare industry. It is a modular, distance-learning ancient observation of King Solomon as recorded in the Bible (Ecclesiastes Chapter 10, programme designed for scientists and managers in the field of Pharmaceutical verse 1). This early reference to a defective medicine was clearly the result of microbial Microbiology, the first of its kind in Europe. spoilage of a non-sterile pharmaceutical arising from inadequate control. Whilst this historical reference might be expected through lack of scientific understanding, pharmaceutical manufacturers today have no such excuse! The focus of microbiological control of pharmaceuticals has been largely, and rightly, directed to sterile products. However, there is a clear expectation from the MHRA and other Regulators that the microbiological contamination risks associated with non-sterile products are fully understood and that appropriate measures are taken to avoid contamination during manufacture. I welcome this monograph from Pharmig and PQG as the existing guidance on the microbiological control of non-sterile pharmaceuticals is limited. This is a comprehensive guide on all aspects of microbiological control from facility design to staff training which taps into a great deal of useful experience from the industry. Whilst this guidance may need to be interpreted for particular applications, the principles should be applicable across the whole range of non-sterile manufacture. It is good to see the emphasis on risk analysis to justify levels of microbiological control. The reporting and investigation of microbiological excursions and the need for results trending are also helpfully discussed. I am delighted to commend this monograph which I feel sure will further strengthen the endeavours of both Industry and Regulators to protect the public through the provision of safe and effective medicines which are free from undesirable micro-organisms. Gerald W Heddell Director Inspection, Enforcement and Standards Division Medicines and Healthcare products Regulatory Agency iv Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals v  Acknowledgements From the Chairmen Acknowledgements This monograph is the result of a collaborative effort between members of the PQG and The concept of a monograph for Microbiological Control for Non-Sterile Pharmaceuticals Pharmig. It represents the recommendations and opinions of a number of individuals was first discussed at the joint PQG/Pharmig meeting in 2001 and raised by PQG’s then experienced in different aspects of microbiological control for non-sterile pharmaceuticals, chairman Peter Gough. The monograph was initiated by Kerry Hawitt and overseen by a which have been integrated by our joint editors. steering team comprising Philip Butson, Stewart Green, Kerry Hawitt, Bob Johnson and Ashley McCraight. The PQG and Pharmig both strive to provide guidance on subjects where there is minimal regulatory guidance or legislation available, as is the case with this subject. In the absence The monograph was edited by Philip Butson and Kerry Hawitt. of guidance, it is easy to either not do enough, leading to a risk of patient harm and/or regulatory censure, or to do too much, thus adding costs without benefit. This is especially Original chapter drafts were prepared by Richard Burt, Derek Davey, Rob De Raaij, Kerry the case with non-sterile pharmaceuticals due to the wide range of products that fall Hawitt, Gail Henry, Julia Mottishaw and Trevor Munton with additional authoring by Philip under this banner and the consequent wide range of risk profiles. The information provided Butson and Kerry Hawitt. is intended to be applicable to all non-sterile dosage forms, including liquids, creams, ointments, tablets, capsules, powder inhalers, aerosols etc. It is applicable to herbal and Thanks are due to the following additional individuals for their contributions as reviewers: over-the-counter products as well as prescription pharmaceuticals. Ian Anderson, Kathy Armstrong, David Begg, Elaine Doyle, Wanda Jay, Bob Johnson, David Keen, Ashley McCraight, Colin Newbould, Wendy Pimblett, Tim Sandle, Kirit Sanghani. Any responsible manufacturer of non-sterile dosage forms should ensure that: A thorough assessment is conducted of the risks that may be posed to patients by Additional thanks are due to the following who provided feedback, either individually or on their particular product portfolio. This should include the raw materials used, the behalf of their representative organisations, during the consultative review process: process and the premises in which manufacture takes place Mike Murray – Association of the British Pharmaceutical Industry Based on this risk assessment document, actions are implemented to control the risks Penny Viner – Herbal Forum to an acceptable level and appropriate monitoring is carried out to demonstrate this Mr R S Iyer – Indian Pharmacopoeia control Richard Funnell – UK Medicines and Healthcare products Regulatory Agency (MHRA) All personnel involved in the manufacture are aware of the potential risks, their John Harwood – UK National Health Service (NHS) Pharmaceutical Production Committee sources and what steps they need to take to ensure that control is maintained Liz Allanson Tony Cundell Using these principles and the detailed information that follows, manufacturers should be Peter Gough able to select and apply the appropriate control and monitoring measures to protect the William Miele well-being of their patient customers without excessive costs and use of resources. Erica Notman Consequently, this monograph should contribute to the success of the organisation. Norman Randall Donald Singer Scott Sutton Ashley McCraight Stewart Green Chairman, Pharmaceutical Quality Group Chairman, Pharmig Not surprisingly given the lack of clear regulatory expectations and the range of individuals consulted, a number of different opinions were expressed by the various reviewers. The final decisions regarding content were made by the editors and the naming of an individual above should not be taken as their agreement with the whole content of the final monograph. vi Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals vii  Chapter 1 – Introduction Chapter 1 Introduction 1.1. Microbiological Control for Non-Sterile Pharmaceuticals The pharmaceutical industry is highly regulated by the application of the principles of Good Manufacturing Practice (GMP). In most countries, government agencies provide guidance to pharmaceutical manufacturers, which is intended to facilitate the manufacture of safe, unadulterated and efficacious drug products. The sterile pharmaceutical sector has a well defined set of expectations and regulations which provide clear statements relating to microbiological controls and monitoring. In contrast, the expectations for non-sterile pharmaceuticals are poorly defined, with few specifics written in either legislation or guidance publications. However the expectations are implied in various places within the GMP guidance, for example: EU GMP 1.4 ‘Quality Control’ requires “procedures [to be] available... where appropriate for monitoring environmental conditions for GMP purposes” EU GMP Section 5 ‘Production’ states that “at every stage of processing, products and materials should be protected from microbial and other contamination” (5.10); “cross- contamination should be avoided by appropriate technical or organisational measures” (5.19) and “measures to prevent cross-contamination and their effectiveness should be checked periodically” (5.20) EU GMP Annex 7 ‘Manufacture of Herbal Medicinal Products’ states that “effective measures should be taken to prevent the spread of... micro-organisms brought in with the crude plant and to prevent cross-contamination” EU GMP Annex 9 ‘Manufacture of Liquids, Creams and Ointments‘ notes that “Liquids, creams and ointments may be particularly susceptible to microbial and other contamination during manufacture. Therefore special measures must be taken to prevent any contamination” CFR Part 211.113 ‘Control of microbiological contamination’ states that “appropriate written procedures, designed to prevent objectionable micro-organisms in drug product not required to be sterile, shall be established and followed” USP 31-NF26 <1111> ‘Microbiological Attributes of Non-sterile Pharmaceutical Products’ includes the following: “Strict adherence to effective environmental control and sanitation, equipment cleaning practices, and good personal hygiene practices in pharmaceutical manufacture is vital in minimising both the type and the number of micro-organisms” “Monitoring, in the form of regular surveillance, should include an examination of the microbiological attributes of Pharmacopeial articles” viii Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 1  Chapter 1 – Introduction “It is essential that ingredients and components be stored under conditions of this expectation in the industry; all responders had a programme in place. There was designed to deter microbial proliferation” still significant variation in the scope and execution of monitoring programmes, and whilst this could be partly due to variation in the facilities and operations, it was clear that it was The Therapeutic Goods Association of Australia (TGA) states that “it is not a not always driven by a risk assessment. requirement that every batch of product be tested for microbial content at release. Instead, for each product manufacturers should aim to develop confidence that the This monograph has been created with the input of experts from across the industry manufacturing procedures do not permit contamination by excessive numbers of based in the UK, Europe, US and India. The principal objective is to provide the microbes or by pathogenic organisms”. The TGA also require absence of all pharmaceutical and associated industries with an international bench mark and guidance pseudomonads not just Pseudomonas aeruginosa. on technical best practices for microbiological control and monitoring in manufacturing facilities for non-sterile pharmaceuticals. The regulatory agencies therefore expect the industry to take a risk based approach to microbiological control and apply appropriately justified monitoring in the manufacture of The monograph emphasises a risk based approach rather than providing a ‘one size fits non-sterile pharmaceuticals. all’ solution and seeks to encourage organisations to understand their premises, products and processes prior to deciding on their approach. In this way, manufacturers should be Generally, companies understand and embrace the need for delivering microbiological able to select and apply appropriate control and monitoring measures to protect the well- control. However, there is a broad spectrum of interpretation on how to deliver being of their patient customers without excessive costs and use of resources. In the appropriate control across industry. majority of cases, the microbiological risk associated with non-sterile products is low and extensive monitoring should not be required. For some companies, the establishment of initiatives to deliver microbiological control and monitoring has required extensive efforts from Microbiology and QA staff to define and Through the review process of this monograph, it was identified that there are currently implement procedures from basic principles. This effort is being repeated many-fold across significant differences in approaches, largely driven by differing regulatory pressures, the industry; each company ‘reinventing the wheel’ in different forms and to different between the US and EU. This is an important consideration for facilities that are part of a extents owing to the lack of defined published criteria. global supply strategy and therefore need to comply with the minimum expectations of all countries. However, the effective use of a risk based approach should result in decisions The original Parenteral Drug Association (PDA) Technical Guideline 13 indicated that the regarding control and monitoring which have global application. approach used for the monitoring of sterile manufacturing environments could be applied to non-steriles. If a company interprets this as applying the full remit of environmental The considerations in this monograph may be applied to handling any non-sterile material monitoring for sterile product manufacture to non-sterile manufacturing, then the result including starting materials, radio-pharmaceuticals, Investigational medical products will be costly and in excess of what is warranted by a risk based approach. Such an (IMPs), drug substances and all formulations of non-sterile drug products, in both the approach is not recommended in this monograph. private and public sectors. The Pharmaceutical Research and Manufacturers of America (PhRMA) 1997 industry The fundamental requirements for good microbiological control in non-sterile manufacturing review stated that routine monitoring of non-sterile manufacturing facilities should not be and quality assurance are established through cross functional management oversight, mandatory, but dictated by the circumstances and required microbial quality of the drug qualified and trained personnel, and well designed and maintained facilities, supported by a product manufactured. This is the forerunner of the application of risk based assessment suitable microbiological laboratory function. This infrastructure is addressed in Chapters 2 to determine and justify appropriate monitoring and controls, which is recommended by to 6 respectively. this monograph. It should be recognised that microbiological monitoring is a mechanism for understanding Cundell (2005) summarises the results of three other surveys in the US dating from microbiological control and the potential risk to the product and patient, not a control 1994, 1998 and 2002 in which most of industry was shown to conduct some form of measure in itself. It is important that manufacturers understand and control the microbial viable monitoring, albeit at varying frequencies and using various techniques. hazards within their facility. There should be an understanding of the ongoing performance of the facility. The risk based assessment may be used to minimise and target any In 2002, Pharmig conducted a survey of its membership and found that 81% of monitoring activity to check periodically the effectiveness of the measures to prevent respondent companies had a programme in place for their non-sterile manufacturing cross-contamination required by EU GMP 5.20. facilities. However, there was considerable variation in the scope and execution of such programmes. The survey was repeated in 2007 and showed an increase in the percentage of companies surveyed who had adopted a programme, reflecting the evolution 2 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 3  Chapter 2 – Roles and Responsibilities Once the infrastructure has been evaluated a documented risk assessment can be carried Chapter 2 out to develop an appropriate monitoring programme which should be subject to periodic review, this is considered in Chapters 7 and 8. Roles and Responsibilities Data derived from such programmes should be analysed and interpreted to provide a basis for the on-going risk assessment process. The reporting and trending of data is considered in Chapter 9. 2.1. Introduction Some mind maps are included in an Appendix. These may be useful as memory joggers Historically, the need for microbiological monitoring and control in non-sterile manufacture has or for audit purposes. been challenged. Recent surveys suggest that monitoring is now generally practiced, but this does not necessarily mean that the importance of comprehensive microbiological control has been recognised. Just as quality is embedded as part of everybody’s responsibility, safeguarding against microbial contamination should also be seen as everybody’s responsibility. Quality Assurance’s role is to set standards, policy and provide guidance and oversight with the site Microbiologist leading programmes of good microbiological control, in cooperation with the Qualified Person (in Europe). This is equally applicable and adaptable to manufacturing facilities of all sizes and types, e.g. classic pharmaceutical manufacturers or specialist hospital units. 2.2. Microbial Governance Top level microbial oversight ‘governance’ should be driven by senior level site management and not limited to the senior Microbiologist or QA manager. Clear direction needs to be given to the site emphasising that microbiological contamination control is a key factor in Good Manufacturing Practice. Multi-functional involvement of representatives from manufacturing (technical and operations), engineering, QA management and QC Microbiology should have a collective responsibility to ensure the appropriate quality systems are in place to ensure microbiological control (Figure 2.1). Within Europe the Qualified Person (QP) should also be included since The Code of Practice for Qualified Persons (QP COP) includes requirements for QPs to consider all relevant factors “including any not specifically associated with the output batch directly under review (e.g. calibration and maintenance records, environmental monitoring)” (QP COP 7.9). The Code of Practice for Qualified Persons also emphasises the need for the QP to consult with other relevant experts on matters such as “potential environmental or microbiological risks” (QP COP 7.11). The QP is dependant upon the knowledge and expertise of his/her colleagues to enable them to fulfil their legal obligations (QP COP 7.14). 4 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 5  Chapter 3 – Personnel There should be governance of every aspect of site functionality that could impact Chapter 3 microbial contamination and control. Direction, prioritisation and appropriate review and action are required relating to: Change control Personnel Facility and engineering projects New product introductions 3.1. Introduction Risk assessments A potential source of microbiological contamination for pharmaceutical products comes Monitoring data from the personnel. This includes not only the production operations staff but also Environmental monitoring performance, as determined by the risk assessment personnel from other disciplines including engineers, QA, QC sampling, management, security personnel, cleaning staff and contractors; in short, anyone who may enter the Product, intermediate, starting material and water microbiological testing production areas. Deviation handling All individuals who enter the production areas at any time therefore need training on matters relating to microbiological control, in particular hygiene. Access to production In addition to the normal roles and responsibilities in the quality management system, areas must be controlled to ensure entry is restricted to trained personnel and specific microbiological expertise should be used in the following: accompanied visitors. Risk assessment (Chapter 7) Determination of appropriate monitoring as per the risk assessment (Chapter 8) Production personnel are in a position to directly impact the microbiological quality of Setting of appropriate action and alert levels (Chapter 8) products and should be given a basic microbiological awareness to underpin procedural requirements for hygiene and good working practices. Trending and Interpretation of testing and environmental monitoring data (Chapter 9) In situ support to operational areas Design and delivery of training (section 3.2) 3.2. Training Review of/consultation on equipment, system and facility designs to ensure they are There is a general requirement for manufacturers to provide training in accordance with hygienic and thus minimise future contamination risks GMP for all personnel whose activities may affect the quality of the product. Ensure appropriate product/starting material specifications are set from a microbiological perspective (Ph Eur 5.1.4) Pharmaceutical manufacturing sites have a broad spectrum of personnel in terms of education and experience. Typically only a minority of staff will have microbiological Investigation of Microbiological Deviations, Out of Specification (OOS)/Out of Trend background knowledge. It is therefore important to provide appropriate levels of training to (OOT) data (section 9.4) all relevant functions as part of the overall microbiological control arrangements. The training of staff, including any contract staff, should emphasise the importance of their A site should have access to someone with appropriate training and background to make, particular role in safeguarding the products made in the facility from contamination. communicate and influence good assessments and decisions from a microbiological perspective. This will typically, but not necessarily, be a graduate microbiologist but must In relation to the manufacture of non-sterile pharmaceutical products it is important to be someone with documented experience and appropriate qualifications. It may be cover the following from a microbiological perspective: necessary to contract this resource from a reputable third party supplier/consultant. All staff: Introduction to micro-organisms and microbiological contamination control Entry and exit of production facilities (including gowning) Personal hygiene training 6 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 7  Chapter 3 – Personnel More specialised training, dependant on role: The prohibition of eating, drinking, chewing or smoking, or the storage of food, drinks, Environmental sampling, monitoring and control smoking materials or personal medication in the production or storage areas Microbiological risks associated with specific production tasks A requirement for personnel to wash their hands before entering the manufacturing facility Risk assessment and design of microbiological control features Classified area/Clean room practice (if required by facility/formulation) Statistical/data analysis 3.4. Dress Code/Changing Training must be documented and regularly reviewed. It is important that training is Every person entering the manufacturing area should wear protective garments effective and competency should be assessed to demonstrate this (EU GMP 2.9). appropriate to the operations to be carried out (EU GMP 2.16). Non dedicated clothing should not be allowed in the production areas. Training should be tailored on a facility by facility basis with an appreciation of the specific risks associated with the product formulations and facility design. In general, a non-sterile manufacturing environment will require the wearing of suitable protective over-garments, hair cover (e.g. mob cap), beard or moustache cover (where Trainers should be appropriately communicative, knowledgeable and experienced in the applicable) and overshoes. Dedicated shoes are recommended for personnel working in subject to provide adequate training and answers to technical queries. production areas for long periods. Records/documents e.g curriculum vitae, training records etc, demonstrating the trainer’s competency should be reviewed/archived. Although it is only an explicit GMP requirement to remove jewellery and make up for sterile manufacture (EU GMP Annex 1), consideration should be given to requiring the removal of jewellery, in particular rings with stones and earrings, and make up by staff involved in 3.3. Hygiene non-sterile manufacture. There is a general requirement for manufacturers to ensure hygiene programmes are One piece over-garments are often used as they provide a better level of protection to established, understood and implemented. The programmes should be adapted to the both the operator and the product. different needs of the facility and cover the following: Health Visitors not entering production rooms may only require a laboratory coat, rather than a one piece over-garment. Hygiene Clothing of personnel (section 3.4) Over-garments may be disposable or washable. Procedures should be in place to manage dirty garments and safeguard against cross-contamination. Additional procedural controls Typically, for non-sterile manufacture the hygiene programme will include: should be in place to provide assurance over any laundry process. Consideration should be Medical examination upon recruitment given to: The need for qualification of the laundry process A procedure for the notification of health conditions that may affect product quality and subsequent actions to prevent contact with starting materials, primary packaging Repair and retirement/replacement of garments materials and manufactured products by those personnel with: Third party (i.e. outsourced laundry operations) Infectious disease Quality assessment Open lesions on any exposed part of body Technical agreement Shedding skin conditions, such as eczema or psoriasis (skin scales may harbour objectionable micro-organisms that may impact pharmaceutical products and Gloves must be worn when performing production activities (including the handling of patients) components) to ensure there is no contact between operators’ hands and exposed Gastric upsets products or any part of equipment that comes into contact with product (EU GMP 2.18). Frequently, the wearing of gloves is required as part of company procedures for entry into Steps to be taken to prevent direct contact between the operator’s skin and any production areas to provide a greater level of contamination control. pharmaceutical product/product contact equipment 8 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 9  Chapter 4 – Facilities Typically, the requirement for wearing of masks is driven by personnel protection Chapter 4 considerations. However, masks may be required for product protection when manufacturing certain higher risk products such as aqueous inhaled products and those formulations with a higher availability of water (Chapter 7). Facilities 4.1. Introduction The design and operation of pharmaceutical facilities are critical to microbiological control. Whereas there are well defined criteria for the design and operation of sterile manufacturing facilities, there are few specific regulatory requirements provided for the manufacture of non- sterile products. Whilst this provides for greater flexibility in interpretation, it also leads to a greater need to assess risk and justify facility design and working practices. There are a number of potential sources of microbial contamination within a pharmaceutical facility, including: Personnel Water Air/compressed gases Product (active, starting materials, packaging components) Equipment Consumables and tools Pests and vermin Facility design and layout Process design Personnel and material flows should be considered when planning the facility and processes. For example, when working with herbal products the risk of microbial contamination from the starting materials may be significant and additional care should be taken with staff movement and production scheduling. Facility and equipment design cannot eliminate microbial contamination, but good hygienic design can mitigate several of these risks. For example, ergonomic design can help people to work more hygienically; well designed water and HVAC systems minimise microbial ingress and aid monitoring; equipment designs with smooth surfaces of appropriate materials can facilitate effective cleaning; points of access for pests and vermin should be minimised. 10 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 11  Chapter 4 – Facilities 4.2. Area Classification capability of the facility design but does not require the application of a formal area classification. See Chapter 7 for further consideration of risk assessment. This is a complex subject and this chapter can only summarise the overall position. Typically for non-sterile manufacture, formal area classifications are not required. The The design of an appropriate facility will consider not only the air supply quality, but also practical achievement of appropriate microbiological control should be a focus in addition aspects such as air change rates and pressure differentials which should reflect the to chemical cross-contamination control and environmental health and safety concerns. potential risks. However, facilities may be designed to achieve a given specification which will be assessed during the commissioning. For example, a tablet manufacturing facility may be designed to There are no regulatory specifications for pressure differential or air change rates for non- achieve an equivalent of EU GMP Grade D (at rest), if tested, but would not be formally sterile products. Current industry practice is to have pressure differentials of 10–20 Pa classified as such. between areas of different standards. For lower risk activities, air change rates are typically 10–15 per hour. Higher risk activities have higher air change rates, typically The engineering standards for area classifications are based on the number and size of greater than 20 per hour. particles in the air. Within the pharmaceutical industry it is the viable particles that are of greater concern. This is recognised by the EU GMP Guide (Annex 1), for example, which The facility design also has a bearing on appropriate operational practices. For example, incorporates recommended microbial limits within the area grades. The setting and where a processing room is at negative pressure to a corridor for the containment of monitoring of appropriate microbial action and alert levels for non-sterile manufacturing is active ingredients, there is an increased risk of microbial ingress from the corridor. discussed in Chapter 8. Consequently, an effective cleaning regime for the corridor is required to ensure that microbial levels are kept low. A number of different standards and nomenclature have been used across the pharmaceutical industry. Table 4.1 summarises these and provides approximate Many pharmaceutical companies and engineering design houses have evolved their own equivalencies. It should be noted that the US Federal Standard 209E has been withdrawn guidelines and engineering specifications based on experience. These are of value, but the but it is given for reference because its terminology is still commonly used. importance of specifying and designing a facility based on an assessment of risk to the product and patient is emphasised. The use of specific risk assessments as the basis for the standards applied will result in different facilities applying different standards to the EU GMP ISO 14644 US Federal 0.5µm 5.0µm manufacture of non-sterile products. Annex 1 2008 1999 Pt 1 Standard 209E particles/m3 particles/m3 ‘At Rest’ (withdrawn) Where higher levels of product protection are required, e.g., for inhaled products, a graduated control standard with additional garment changes may be built into the design A-B 5 100 3,520 Grade A =20 and incorporated within operational procedures. Grade B = 29 C 7 10,000 352,000 2,900 Older facilities may have inherent design issues which are sub-optimal. For example some D 8 100,000 3,520,000 29,000 manufacturing facilities may have been constructed from materials no longer considered appropriate to the manufacturing environment, such as high and irregular ceilings or unprotected light fittings. Where older facilities have such deficiencies compared to Table 4.1: Different Standards for Air Classification current standards, these should be captured within a risk assessment and plans created to address them through refurbishment, or justifications written to accept them in association with appropriate operational procedures. The EU GMP Guide Annex 1 also provides ‘in operation’ particle standards for each grade (except class D), which are relevant for sterile manufacture, but typically not required for 4.3. Access to Areas non-sterile production areas. It does not matter how well a facility is designed unless it is operated appropriately and well In the absence of specific requirements for non-sterile products, the design and monitoring maintained, and an important consideration is access to the area by personnel and materials. requirements of the facility should reflect the risk profile of the products manufactured. So, for example, the level of control applied might increase moving from tablets to creams/liquids on the basis of available water and the level of control applied to inhaled products enhanced due to route of administration. This may require an increase in the 12 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 13  Chapter 4 – Facilities 4.3.1. Personnel Access Wooden pallets and cardboard cartons should not be taken into manufacturing or packaging areas. Whenever possible, carts, pallet jacks, trolleys, etc. should be dedicated Access to the facility must be restricted to personnel that are fully trained and assessed to the production area. as competent to work in that area, who are appropriately attired (see Sections 3.2 – 3.4). All visitors should be accompanied at all times by fully trained personnel. Access is Active pharmaceutical ingredients, starting materials and packaging components will have normally restricted by the use of an electronic access control system, or similar. been dispensed for a specific job and should be provided in appropriate clean, sealed impermeable (e.g. plastic/stainless steel) containers. Personnel access must only be made via changing rooms. The changing room design contributes to the assurance of appropriate personnel access and microbial contamination In-process materials and consumables should also be supplied in sealed, clean plastic bags control. and, if these are delivered in cardboard boxes, the plastic bags should be removed from the boxes before being taken into the area. Hand washing facilities should be included in the design of changing rooms and used prior to access to the production areas (EU GMP 2.19). It is useful to have a ’step over’ line or All containers taken into processing areas should be clean and dry. bench to define the boundaries of clothing requirements. An interlock may be appropriate in some facilities. Tools (Chapter 5.3.1) have the potential to contaminate the equipment and product contact parts and their introduction to the facility needs to be appropriately controlled with The requirements for area entry must be written into procedures and it is usual to have consideration given to cleaning and sanitisation requirements. Similarly, the introduction of pictures of correctly clothed personnel and mirrors to enable staff to check them before new equipment and tooling into a production environment should be assessed and entry. A copy of the gowning procedure should be present in the changing room. appropriate actions taken to ensure that contamination is not brought into the facility. The changing room should be provided with filtered air. Intermediate (bag) filters will typically be suitable for this purpose, though High Efficiency Particulate Air (HEPA) filtration 4.4. Building Requirements may be used. The air pressure should be negative with regards to the manufacturing area corridor, but positive relative to external adjacent areas. The design of the building and level of product protection provided by the facility should be determined by the nature of the operations intended to be carried out. 4.3.2. Materials Access 4.4.1. Surfaces Materials include active pharmaceutical ingredients, starting materials, in process materials, packaging components, consumables and tools, which may be brought into the Materials of construction, surface finish and general design also need to be reflective of facility. These are all potential sources of microbial contamination and it is essential that the different operating and cleaning regimes. suppliers and delivery process of these items are assured to ensure that quality is maintained. This is especially the case for materials of biological origin. Non-sterile manufacturing areas need to have smooth surfaces that will not harbour micro-organisms and can withstand the routine cleaning regimes required. Warehouses should be well ventilated, cool and low in humidity; materials should be checked regularly and used as quickly as possible especially in the case of herbal materials. Floors should be constructed of durable cleanable materials with minimal joins and will typically be epoxy resin* or sheet vinyl, not vinyl tiles. Terrazzo is used less in new facilities, Dispensing operations and traffic within a facility should minimise dust carryover and hence but may still be used for areas with potentially very high traffic and heavy moving loads. risk of cross-contamination of the production areas. *Note: Epoxy resin may have grit included to improve traction. However there can be Materials access must be via specified routes – generally via air locks, although certain microbiological and general GMP implications arising from this – over time, this can items when justified may be introduced via the personnel changing rooms, e.g. equipment wear-out of the resin leading to potential for contamination and poorly sealed floors. for environmental monitoring. At points of entry to the facility or to rooms, “sticky” mats or polymeric flooring areas can be beneficial in the prevention of cross-contamination by removing particulate material from the soles of shoes or wheels. However, they should be frequently maintained (e.g. daily) so that a fresh surface is presented and they do not themselves become a source of contamination. 14 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 15  Chapter 4 – Facilities In older facilities, walls may be made of plaster and simply emulsioned. Typically, Water from any process system must have an ‘air break’ into a tundish before it is piped elastomeric or flexible paints are now being used instead of emulsion. The latter may be away – conventionally to a floor drain. The floor drain will be trapped but there is still suitable for low risk applications only. Increasingly, areas are being fitted out with walls potential for the water in the drain to be contaminated. The floor drain should be made from proprietary pre-finished coated panels. These are more expensive but have the connected via a break tank (as minimum) to the foul drain system. Although it does allow advantages of quick construction, greater flexibility if room layouts need to be changed and for the containment of contaminated spillage (preventing it reaching the local foul system), greater hygienic control. the primary purpose of the break tank is to prevent any backflow of contaminated waste from the foul drain into the process drainage system. Windows/viewing panels should be non-opening, encased, flush with the wall panels and appropriately sealed to prevent collection of dust and microbial matter. Because drains are the primary link between the room and the external drainage system, where present they require disinfection on a regular basis based upon a known Ceilings can be a simple silicone sealed lay-in grid, but these are difficult to clean properly understanding of the microbial contamination risk presented. and cannot withstand more than a minimal overpressure. It is therefore conventional to have a solid surface in a similar finish to the walls. Wash bays, which tend to remain wet for long periods, are major potential sources of microbial contamination and particular attention needs to be paid to their design to ensure Lights should be encased, flush with the ceiling or wall panels and appropriately sealed ‘puddling’ and stagnant water does not occur. It is important that air changes, robust with silicone to prevent collection of dust and microbial matter. construction, excellent floor seals and good working practices are in place to ensure that the wash bay does not encourage a residual microbial population or affect the surrounding facility. Coving or moulding should be used at the junctions between floor and wall and between ceiling and wall to eliminate right-angle joints and aid cleaning. The joints should be sealed Sinks and safety showers should be minimised to meet functional and safety requirements with silicone and the coving maintained in a good condition to prevent collection of dust and appropriately located. They present a potential risk as they are sources of water, often and microbial matter. potable water, and should be assessed and monitored appropriately. Doors for a new facility should not be made of wood. In an older facility where wooden doors still exist, they should be sealed and well maintained. A risk assessment should be 4.4.3. Heating, Ventilation and Air Conditioning (HVAC) System conducted regarding their potential replacement. The detailing around doors should be sealed with silicone and well maintained to prevent collection of dust and microbial matter. The HVAC system is crucial as it supplies air at the correct cleanliness and in the correct volumes to help keep particles entrained within the airflow so they are passed out of the Sinks within the production areas should be made of stainless steel and designed to be room. The HVAC is also key to the correct maintenance of pressure differentials, self-draining to minimise any pooling or trapping of water and to maximise cleanability. temperature and in most cases humidity throughout the facility including changing rooms. All of these aspects play a part in microbial contamination control. Benches and trolleys should be made of cleanable, impermeable materials and have smooth surfaces. The use of mobile benches can facilitate cleaning. However, where Humidity control is key for some storage areas if materials are hygroscopic, increased mobile units are used, consideration must be given to maintaining cleanliness of the moisture levels may present a risk to microbial proliferation. wheels, e.g., through the use of sticky mats between rooms. Pressure differentials and air change rates are covered in Section 4.2. 4.4.2. Drainage Air ‘quality’ is improved by filtering the air through progressively more efficient filter media. Absolute filtration, using High Efficiency Particulate Air (HEPA) filters removing 99.97% of Any waste water from sinks, wash-in-place systems or wash bays must be disposed of particles sized greater than 0.3µm, is commonly used throughout the industry and is and, if not correctly designed and maintained, there is the potential issue of back recommended for higher risk activities. However, less efficient filters, e.g. bag filters contamination from the ‘foul’ waste pipework. removing 90% of particles sized greater than 0.5µm, with turbulent air flow are sufficient for most lower risk activities, e.g. tablet manufacture. As drains are a potential source of contamination, they should be minimised. Where used, they should be hygienically designed to reduce the risk of aerosols, standing water and Other factors to be considered in the design and maintenance of air systems is where the splash back. Drain covers, surfaces and joints should be smooth and appropriate filters are positioned, whether extract filters are required and how much ‘fresh air make containment features should be incorporated within the design. up’ is used. This chapter will only consider filter positioning as the other two areas tend to be driven by the safety aspects of the product rather than microbial aspects. 16 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 17  Chapter 4 – Facilities It is cheaper to install and easier to maintain filters housed centrally, with the air handling 4.4.5.1. Purified Water plant, where the air is then distributed to all the rooms. From a microbial aspect it can however be a totally false economy as the ductwork then becomes part of the controlled Purified water is typically used as process water for non-sterile pharmaceutical products environment and it can be virtually impossible to seal, clean and monitor the ductwork. and for the final wash/rinse while cleaning product contact surfaces. In addition, any changes of components or instrumentation, within the ductwork, are in contact with ‘product contact air’ and need to be subjected to much more rigorous change The specifications for purified water are given in the pharmacopoeias and are not covered control. Depending on design it can also be much more difficult to spot problems in a in this monograph. Piped purified water systems are typically continually monitored on-line centrally filtered system and any problems will affect all areas served from that filter bank. for conductivity and Total Organic Carbon (TOC) and routinely monitored for chemical and microbial quality (section 8.3.3.6). Terminal filters, which are actually situated above the production rooms, are more expensive and are slightly more difficult to change but return the additional investment in Purified water is generated from potable water, e.g. by reverse osmosis or distillation, simplicity of operation and greater levels of quality assurance. though the latter is typically only used for the generation of Water for Injections. Deflectors are often incorporated between the filter/air duct and the room and are The problems with purified water systems usually involve bacteria (Hutcheson 2008). Most important for air distribution and flow. However, they also create opportunities for the microbial problems arise from the storage and distribution of the water rather than its collection of dust and micro-organisms. Consideration should be given to the positioning of generation – the development of biofilms can be extremely difficult to resolve. However, the air blades, to ensure that the flow of air is not directly onto product and contact improperly maintained generation components, e.g. carbon beds, softeners, reverse surfaces, and to cleaning, so that they do not become a source of contamination in osmosis membranes, can also contribute to subsequent contamination downstream within themselves. the distribution system. The design and installation of the components of the storage and distribution system 4.4.4. Compressed Gases should enable validated cleaning and sanitisation. Compressed gases and their distribution systems will have a microbiological impact if not Appropriate placement of outlets should be built into the design to assist operational specified, installed and maintained to the appropriate standards. logistics, to ensure representative sampling and the minimisation of dead legs. See section 8.3.3.8 for further consideration. Various design features may be utilised to prevent the development of biofilms and control contamination. The design of the system should include continual circulation with adequate 4.4.5. Water flow rate to aid the prevention of biofilm formation (typically 1–3 m/s). Other common design features are the capacity to heat the water to elevated temperatures (typically Water and its distribution systems will have a microbiological impact if not specified, 65–85°C) for sanitisation purposes and the inclusion of high intensity UV lamps. The installed and maintained to the appropriate standards. inclusion of UV lamps downstream of potential microbial reservoirs, e.g. carbon beds, softeners, has the added advantage of enabling ozone to be used for sanitisation purposes, There are various qualities of water used within non-sterile manufacturing facilities, some of if required. Where UV lamps are used they should be regularly checked and maintained to which are detailed in pharmacopoeia and all of which need to be suitable for their intended ensure they are clean and provide the correct wavelength and energy output. The inclusion use. Regulatory requirements are specifically mentioned in the EMEA CPMP ‘Note for of filters within the distribution loop is difficult to justify and is not advisable. guidance on quality of water for pharmaceutical use’ and the EU GMP Guide, e.g. Annex 9 relating to the manufacture of liquids, creams and ointments; Chapter 3 regarding Older systems, without such design features may require regular disinfection using an sanitisation of pipe work; Chapter 1 regarding utility qualification; Part II, 4.3 regarding oxidising agent to control biofilm (for example, ozone, hydrogen peroxide, hypochlorite). water for API manufacture. Each of these methods has disadvantages, e.g. additional design considerations (UV light to destroy ozone) or Health and Safety risks, and none is fully effective. Extensive flushing Incoming water should be sampled at point of entry on a routine basis (section 8.3.3.6). is required to remove chemical residues if hypochlorite is used, which is expensive and Where process water is treated by the manufacturer to create the appropriate quality, the disruptive. treatment process should be validated and routinely monitored. Typically, stainless steel pipework will be used, though systems with plastic piping have also been used. Sanitary design for valves is an expectation. The design must minimise opportunities for stagnation, e.g. dead legs, or sites for residue accumulation, which can 18 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 19  Chapter 4 – Facilities promote microbial proliferation. Good control over welding processes is required to ensure 4.5. Facility Qualification that these do not introduce surfaces which provide opportunities for biofilm formation. The commissioning, qualification and validation of a facility is very important. There is a The system should also be suitably sized in relation to the anticipated consumption of the GMP requirement to identify the validation required to prove control over critical water; general guidance is for a total water replacement every 24 hours. Where purified operations and a risk-based approach, with a multi-functional team, should be used to water usage volumes are low, bottled purified water may be considered as an option. determine the scope and extent of this work. Microbiological considerations should be included within this risk assessment. This topic is not covered in detail in this monograph; Procedures should be in place to cover the maintenance of the water system, including the reader is advised to refer to more specialised guides, such as those produced by ISPE, details of shut down and start up mechanisms. 2001, for further information on this topic. Operating procedures should require outlets to be flushed before usage to ensure use of the circulating water and to remove possible stagnant water or contamination from the surface of the outlet. The flushing of outlets prior to sampling for monitoring purposes should be equivalent to that applied in operational use. The use of hoses and temporary piping is a major source of contamination to product in non-sterile facilities and therefore their use should be minimised. Where used they should be subject to appropriate controls to minimise the risk of contamination from this source. For example, they should not be left on the outlets; they should be dried after use, hung vertically in appropriate locations to ensure free drainage, monitored and regularly cleaned, sanitised and replaced. 4.4.5.2. Hot and Cold Potable Water Potable water may be used for hand washing and initial cleaning or wash-in-place systems. These are very similar to domestic supplies (except that hot water is often in a re- circulating loop) and do not conventionally cause many microbial issues. It is wise to ensure the hot water does not get ‘too hot’ or staff will avoid using it for critical hand washing activities. In some cases potable water may be softened before use, which adds complexity to the water system. 4.4.6. Steam Clean steam is used for cleaning and sanitisation of production tools and equipment, supply for autoclaves and humidification of air where required. The microbial risk from the steam itself is low due to the physical characteristics of steam and its production from purified water using a heat exchange. Care should be taken with respect to condensation generated upon cooling of the steam on surfaces, which may be sources of contamination that may then be spread by the condensate (Section 8.3.3.7). 20 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 21  Chapter 5 – Cleaning and Disinfection Chapter 5 Cleaning and Disinfection 5.1. Introduction Cleaning and disinfection are vital parts of the maintenance of facilities and equipment in order to ensure that contamination risk is minimised. Facilities and equipment need to be designed so that they are fit for the intended purpose and cleanability is an important factor to consider in the design/technical specification. The design should aid the cleaning and disinfection process rather than cleaning and disinfection procedures being used to compensate for poor facility or equipment design. The assurance of appropriate cleaning and selection of appropriate disinfection agents through validation are also important. For a more detailed consideration of cleaning and cleaning validation, including chemical considerations, refer to the PQG monograph on ‘Cleaning Validation’, 1999. For a more detailed consideration of disinfectants, refer to Pharmig’s ‘A Guide to Disinfectants and their use in the Pharmaceutical Industry’, 2006. 5.2. Facilities Areas must be regularly cleaned and, where necessary, disinfected to a written procedure and programme. A cleaning log is required which records the areas cleaned, agents used, type of cleaning (as appropriate) and the identity initials of the operator. Sufficient time and resource should be allocated for cleaning activities. Frequently, some or all GMP facility cleaning activities are carried out by contract staff. Where contract GMP cleaners are used, a technical agreement should be in place with the contract agency. Only named staff should have access to GMP areas and be involved in GMP cleaning activities. A permanent member of staff should have oversight of the contract cleaning operations to ensure that they are being conducted satisfactorily. Whether contract or in-house staff are used for cleaning activities, they must have documented training and competency (Chapter 3) and the responsibility for cleaning needs to be clearly defined and traceable to accountable management. Standard Operating Procedures (SOPs) are required and need to be part of the document control system. SOPs need to state the areas to be cleaned and their frequency, together with materials, equipment and methods. 22 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 23  Chapter 5 – Cleaning and Disinfection Risk assessment and hazard analysis tools should be used to formulate the cleaning Disinfectants should only be applied to relatively clean surfaces since they can be programme (Chapter 7). inactivated by residues of dirt and other materials, e.g. product. Chemical disinfectants need sufficient contact time and this can be aided by using repeated applications rather The frequency of different cleaning and disinfectant activities will vary depending on the risk than a brief wipe or mop application of the solution. Knowledge of the properties of the assessment (which takes account of room usage and formulation). For example, floors, disinfectant used is essential to ensure its optimal usage. It is important to use detergent fittings and benches may have a routine daily clean, while higher level walls and ceilings and disinfectants which are compatible with each other. This also applies to the may be cleaned less frequently, e.g. weekly or monthly. compatibility of disinfectants that are rotated with each other. The SOP needs to contain sufficient detail to allow the correct application of the cleaning Disinfection programmes should be supported by laboratory studies with representative and disinfecting agents. Steps for preparing the correct concentrations are particularly micro-organisms (USP <1072> Disinfectants and Antiseptics). relevant. The manufacturer’s recommendations for preparation and use should be followed. Floor and sink drains, when present, are a potential source of microbial contamination. The method and sequences of cleaning is important; contact time, application Their design needs to include traps and screens where appropriate (see 4.4.2) and temperature, mechanical action and the chemistry of the cleaning agents should all be consideration should be given to the risks of aerosols and splash back as part of the risk considered during the design of the cleaning process. assessment. A treatment with alkali may be used to eliminate blockages and a compatible disinfectant should be used to reduce the microbial load. The effectiveness of the Loose dirt and spillages of material require removal first to ensure that they are not disinfection should be determined and documented. The frequency of the disinfection spread around the facility. This is typically done using a vacuum cleaner of appropriate should be identified through risk assessment. design, i.e. one that has a filtered exhaust to ensure that it does not present a source of recontamination. Utensils used for all aspects of cleaning must be managed so they do not present a source of contamination. They should be selected to reflect the requirements of the area Fittings and fixtures including door handles, light switches and telephone receivers should and may be dedicated to specific areas of the facility. Utensils used in the preparation of be included in the cleaning regime. cleaning and disinfectant agents must be dedicated for this purpose. Whilst mop and buckets remain a widely used method, consideration should be given to disposable mop Hot water or hot water with detergent followed by a hot water rinse may be all that is heads and wipes. Buckets and mop heads need to be of a design which allows them to be necessary in some areas, whereas other facilities will require detergent clean followed by stored clean and dry. It is important that mop heads are stored dry, frequently sanitised a disinfectant. This should be determined and documented by the risk assessment. Where and replaced on a regular basis. Water should never be left to stand in the buckets but used, all residual cleaning and sanitising agents must be removed from product contact used and immediately discarded to ensure that a potentially significant source of surfaces to avoid product contamination. microbiological contamination is removed as soon as possible. Disinfectant rotation is a contentious issue attracting considerable debate within industry A bucket system which allows the separation of ‘used’ solutions of detergent/disinfectant and regulatory agencies. Annex 1 of EU GMP states that “where disinfectants are used, from fresh solutions is an aid to effective cleaning. The mop is wrung out in a separate more than one type should be employed”. However, this is specific to sterile manufacture bucket from that used to contain the fresh cleaning agent. This reduces the rate at which and not necessarily applicable or justifiable to non-sterile manufacture. Whilst this the disinfectant solution becomes dirty and diluted during use, and prevents the reapplication requirement has lead to widespread application in non-steriles, there are arguments of dirt to the cleaned surface, thus reducing the risk of recontamination and inactivation. against applying it. The principle scientific argument for rotation is to ensure activity against the full spectrum of flora within the facility; one category of agent may not work Mechanical cleaning equipment may be necessary to provide efficient, thorough cleaning. against all types of organisms (e.g. sporicides for use against spore forming organisms). Such items include cloths and brushes to enhance manual processes and scrubbing However, the counter argument is that the periodic use of an alternative and potentially machines and vacuum cleaners, wet or dry. less effective disinfectant may have an adverse effect on the efficacy of the disinfection procedure (Pharmig, 2006). One approach could be to use routinely a single disinfectant Machines used for mechanical cleaning need to be carefully maintained and operated in only introducing a different disinfectant or sporicide in the event of an adverse trend in accordance with SOPs and cleaned themselves on a routine basis, including any internal environmental monitoring data. Another approach is to use a different disinfectant or filters or surfaces where they can be accessed. This cleaning and maintenance needs to sporicide at a defined extended interval. be documented. 24 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 25  Chapter 5 – Cleaning and Disinfection All items, including machines used for mechanical cleaning, need to be stored clean and Biofilm development is a risk with some items of equipment and must be considered as dry. Residues of liquid and moisture encourage microbial growth and the development of part of risk assessment both in relation to design and cleaning processes. Where biofilms resistant strains. There should be a defined drying method/area and storage area, room do occur, remediation can be difficult and contingency plans for dealing with them should or cupboard for dried cleaning utensils and materials. be considered. Status labels should indicate the clean/dirty or quarantine/released status of the equipment. 5.3. Equipment Methods for equipment cleaning vary from manual cleaning to automated systems, which 5.3.1. Tools and Tooling may be integral to the equipment. Automated systems, such as integral Clean-in-Place (CIP) systems and stand alone automated washers, when fully validated, are preferable Tools and tooling should not be forgotten: due to their demonstrated reproducibility. Tools – items used to maintain or adjust equipment Tooling – change parts for equipment (often product contact) Equipment components (e.g. tooling) may require dismantling to enable effective Cleaning- out-of-Place (COP). Items which prove difficult to clean (e.g. valves) may benefit from additional steps such as sonication. Tooling should be dealt with as part of the equipment as detailed in section 5.2. Hosing and tubing that is not single use must be appropriately cleaned, dried and stored Tools generally fall into two categories: to prevent pooling of water, i.e. hung with no loops or kinks to allow free flow drainage of Tools that stay within the facility internal water by gravity and replaced at defined intervals. Specialist tools that are brought into the area, often by external service personnel It is important that all cleaning regimes (automated and manual) are fully validated or Tools that stay with the equipment should be cleaned on a regular basis and following use. subject to verification (i.e. demonstrated efficacy) to ensure that the equipment used for pharmaceutical production is free from residues of product or cleaning material and Specialist tools should be checked for appropriate cleanliness before entry to the area and microbial contamination. appropriately cleaned before (as required) and after use. Cleaning process hold times must be defined and validated in accordance with various global GMP guidance and requirements (FDA guide to inspections validation of cleaning processes; EU Annex 15 (38); PIC/S guidance 7.7.2; Health Canada – Cleaning Validation 5.4. Cleaning Agents guidelines (part of 3.2)). Cleaning provides an important mechanism for improving microbiological control by removing physical contamination from surfaces which would otherwise reduce the ability of The dirty equipment hold time (DEHT) is the time between the end of manufacture and disinfectants to remove micro-organisms. When selecting a cleaning process it is cleaning. The DEHT should be kept to a minimum because over time product residues may important to consider both the chemical and microbiological cleaning capabilities of the become harder to remove and the risk of microbial cross-contamination increases. process to be adopted. Maximum times should be defined and supported by validation. Cleaning efficiency is derived from a number of parameters; time, temperature, The clean equipment hold time (CEHT) is the time from cleaning to next use and is mechanical action and cleaning chemistry. The relative efficiencies of different important to assure equipment does not become re-contaminated during storage either by combinations of these parameters are illustrated by Figure 5.1. microbial contamination or dust. The CEHT should be a defined and documented period up to which the cleanliness of the equipment has been demonstrated. This may be achieved It is preferable not to use chemical cleaning agents, thus avoiding the need for additional through a hold time study, i.e. validated. assurance of the removal of chemical residue. However, some kinds of chemical cleaning agent may be required to ensure a consistent and effective clean. Equipment design and material of construction can significantly impact the ability to achieve an effective clean. The Microbiologist should be involved in review of equipment design to facilitate hygienic design and the inclusion and positioning of appropriate and accessible sampling points where required. 26 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 27  Chapter 5 – Cleaning and Disinfection Cleaning agents must be prepared in accordance with manufacturer’s instructions (unless Water, chemical, mechanical and thermal action otherwise justified and documented) to ensure appropriate concentrations are always Water and thermal action used, otherwise this may negate the validation. Water and chemical action Water and 5.5. Validation of Cleaning mechanical action Trials should be carried out to provide evidence that the process will remove any residue Cleanliness adhering to the surface of the facility or equipment and leave the surface free from Water only residues and microbial contamination. Validation should demonstrate the consistent removal of cleaning agents and meeting of pre-defined limits for microbial and chemical contamination. Ongoing assurance of cleaning practices should be gained from the routine monitoring programme. For further details relating to cleaning validation and disinfectant validation refer to PQG (1999) and Pharmig (2006). Time Figure 5.1: Relative Cleaning Efficiencies (After Wilkinson, 2008) If a cleaning agent is required, the agent should be selected based upon the following considerations: Maximise Active Pharmaceutical Ingredient (API), starting material, intermediate and product removal Compatibility with equipment surfaces Supplier assurance Environmental, toxicological, health and safety impact Cost effectiveness Rinsability It is important to understand the chemistry of your cleaning agents and the mechanism of residue removal. Detergents are primarily used for the removal of soil/dirt residues whilst disinfectants are primarily targeted at microbiological control. For further details relating to disinfectants refer to Pharmig (2006). All cleaning agents should be freshly prepared with water of a suitable quality (typically purified water, although potable water may be acceptable). Cleaning agents and prepared solutions should state the dates of preparation and expiry clearly on the container. 28 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 29  Chapter 6 – Microbiology Laboratories Chapter 6 Microbiology Laboratories 6.1. Introduction The microbiology laboratory and the microbiologists who support the site are the focal point of the site’s mechanism for understanding microbiological control. An important objective of the quality control microbiology laboratory is to provide information in the form of interpreted microbiological data to the site. These data need to be generated and managed in accordance with GMP. The following information provides a standard that is considered appropriate to support the activities in a non-sterile facility. Further details on microbiological best laboratory practices may be found in the USP general chapter <1117>. 6.2. Relationship to the Quality Unit The microbiology unit is most commonly part of the quality control function owing to the nature of the work being performed. However, the microbiology laboratory capability may be managed through a third party contract arrangement overseen by the quality assurance organisation. Good quality control laboratory practice within the microbiology laboratory is essential to the production of reliable and reproducible microbiological data. It should be remembered that non-sterile environmental monitoring data will typically provide an overall picture of the microbiological control within the facility, rather than data relating to specific product batches. 6.3. Microbiology Laboratory Facility Design The microbiology laboratory facility design should be hygienic, ergonomic and fit for purpose. Its function requires a design that considers the product, people and process flows so as to minimise the risk of cross-contamination. Thought needs to be given to the receipt, storage and handling of the different types of material with respect to the level of contamination. Statutory requirements to operate at Containment level 2 should be adhered to for health and safety reasons and to facilitate good quality control laboratory practice (Advisory Committee on Dangerous Pathogens, HSE Books, 2001). 30 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 31  Chapter 6 – Microbiology Laboratories 6.4. Autoclaves 6.6.2. Incoming Inspection Autoclaves should be validated and subject to revalidation depending upon the use. GMP Received media should undergo a pre-defined incoming approval process before it is critical usage such as the sterilisation of in-house prepared media should be revalidated released for use by the laboratory. annually. Non-GMP critical tasks, such as decontamination of lab waste prior to disposal, may only require revalidation on a two yearly basis. The validation should cover the Media receipt from vendors should include a check that transport conditions have been different cycles and load patterns that may be required. appropriate. Each lot of ready made media should undergo a defined set of growth promotion, sterility 6.5. Disinfectants and chemical testing or be covered by validated skip lot testing. Disinfectants qualified for use in the production area should be suitable for use in the laboratory. 6.7. Micro-organisms Validation includes assessing activity against pharmacopoeial organisms and local isolates 6.7.1. Local Isolates and checks to ensure that all detergents and disinfectants are mutually compatible. Each laboratory should maintain a collection of organisms representative of the local flora Application of disinfectants as part of a cleaning/sanitisation programme in the typically isolated across the site for use in testing and validation activities. Since changes microbiology laboratory should be followed by a purified water rinse to ensure residue is in local flora may occur, it is recommended to review and, if appropriate, replace the removed effectively. strains in the collection with fresh isolates regularly, e.g. annually. 6.6. Media 6.7.2. Objectionable Organisms The quality of the work generated by a microbiology laboratory depends on the quality of Regulations require industry to prevent release of finished product that contain the culture media employed and so safeguarding the quality of this media is critical. Media objectionable organisms (21 CFR211.165(b), Ph Eur. 5.1.4). Some objectionable preparation, storage and quality control all need to be defined to assure the quality. It is organisms are specified in the pharmacopoeias but these are not exclusive and other important to ensure that the manufacturer’s instructions are followed for the preparation organisms may be objectionable depending on the nature of the product, route of of media and that activities are recorded. administration and intended patient population. There is an expectation that the significance of other micro-organisms is evaluated (e.g. Ph Eur. 5.1.4, USP <1111>). Special care should be taken to ensure that media used for environmental monitoring do not introduce contamination into the production environment. In preference, commercially A risk based assessment should be conducted, including personnel with specialised manufactured irradiated and double bagged plates confer a greater level of assurance. training in microbiology and data interpretation. In addition to dealing with isolates as they arise, it is advisable that an assessment is done proactively to generate a documented list of objectionable micro-organisms, which should be incorporated into procedures and 6.6.1. Vendor Assurance of Media Suppliers internal specifications as appropriate. Suppliers of media should be approved prior to use following an assessment of their In certain production environments, such as for the production of herbal medicines, Quality Management System (QMS). Data supporting media shelf lives and storage by-products of objectionable organisms, for example, aflatoxins, should be considered. conditions should be available and each batch of media should be supplied with an Certain materials may need to be sampled if toxins are known potential contaminants appropriate Certificate of Analysis (CoA). (Ph.Eur. monograph 1433). Ready to use media supplied by a vendor with a good supply history may undergo a pre- defined validation exercise leading to routine acceptance on the basis of the vendor’s CoA and only periodic testing (e.g., every 10 lots or 6 months). Hence, the vendor’s data can be used for growth promotion, selectivity and sterility. 32 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 33  Chapter 6 – Microbiology Laboratories 6.8. Identification Maximum hold times of aqueous solutions should typically not exceed 24 hours. For dried intermediates longer hold times may be established. In order to provide a good level of For non-sterile manufacturing an assessment should be made and procedures written confidence, it is recommended that the validation period exceeds the defined maximum defining the extent of the identification required. A simple morphological description and hold time. Gram stain may be justified for samples taken from low risk environmental isolates. A more detailed identification, to genus or species level, may be required for higher risk isolates such as those from starting materials or product, or for investigational purposes. 6.9.3. Product It is less likely that a level of identification beyond phenotypic identification will be required. genotypic identification may be helpful when investigating adverse trends or product failure There are general pharmacopoeial chapters giving expectations of microbiological purity of but the benefit may not justify the additional cost. non-sterile products (Ph Eur 5.1.4) and if tested the product should meet these minimum requirements whether there is a registered specification or not. The registered Whichever method is chosen, the laboratory must show its ability to use the system and specification for non-sterile products will typically be based on these pharmacopoeial identify known organisms, including local isolates, to ensure that any identifications requirements. Depending on development data, it may be possible to justify not performing performed in routine analyses are valid. end product testing or to perform skip lot testing. For low risk non-sterile products, annual or 6 monthly testing of finished product may be acceptable. Any such reduced testing must be reflected in the Marketing Authorisation and specification for the product. 6.9. Microbiological Testing of Product, Starting Materials and Intermediates Where product is held in bulk form, hold times should be established and validated as for intermediates. 6.9.1. Starting Materials All testing must be documented to controlled procedures and recorded Where a starting material or packaging component is liable to microbiological contemporaneously (in real time) following the verified method. contamination testing will be required to ensure the absence of objectionable organisms (21 CFR211.84(d)(6)). Consideration should also be given to the need for neutralisation agents in the media to remove any inherent antimicrobial properties in the starting materials. Microbiological testing of starting materials may be conducted on a skip lot basis as determined and justified by documented risk assessment. 6.9.2. Intermediates Intermediates include, for example, granulation solutions, coating solutions and suspensions for spray drying. A risk assessment should be made to determine the risk of microbial contamination; typically aqueous intermediates should have validated hold times and be subject to periodic microbiological monitoring. Where it is decided that there will not be any microbiological testing of intermediates the justification of the risk assessment should be documented. An appropriate in-house specification should be established for intermediates with the input of a microbiologist. 34 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 35  Chapter 7 – Risk Assessment and Management Chapter 7 Risk Assessment and Management 7.1. Introduction A full consideration of pharmaceutical quality risk assessment and management is beyond the scope of this monograph and the reader is encouraged to consult ICH Q9 (now Annex 20 of the EU GMP Guide) and more specialist publications to gain a greater understanding of this important topic. Key definitions that need to be understood are: Risk: “The combination of the probability of occurrence of harm and the severity of that harm” (ICH Q9; ISO/IEC Guide 51) Harm: “Damage to health, including the damage that can occur from loss of product quality or availability” (ICH Q9) Hazard: “The potential source of harm” (ICH Q9; ISO/IEC Guide 51) Risk management is fundamentally about understanding what is most important for the control of product quality and then focussing resources on managing and controlling these things to ensure that risks are reduced and contained. Before risks can be managed, or controlled, they need to be assessed. Two important points to remember for any risk assessment are: 1. There is no such thing as ‘zero risk’ and therefore a decision is required as to what is ‘acceptable risk’. 2. Risk assessment is not an exact science – different people will have a different perspective on the same hazard. Consequently, whichever tool is used, it is important that a team approach is taken utilising an agreed set of definitions/scores. Whichever tool is used, risk assessment involves: Identifying hazards Analysing the risk associated with each hazard Evaluating how significant the risks are 36 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 37  Chapter 7 – Risk Assessment and Management The outcome of the assessment can then be used to determine appropriate control Processes requiring heavy physical labour or increased numbers of personnel might strategies to reduce the risk to a level that is deemed acceptable. These strategies are increase the risk of contamination from personnel through enhanced sweating or typically focussed on reducing the probability of a risk occurring and/or increasing the shedding of skin particles probability of it being detected. Whether the processing step has the potential to increase or reduce bioburden Risk assessments and the actions arising from them need to be documented and subject to periodic review to ensure that the assessment reflects the actions taken and the most Fundamental characteristics of the formulation, such as hygroscopicity, should also be recent data. considered as part of the risk assessment. Where possible, data should be used to support the risk assessment. Microbiological 7.2. Risk Assessment Associated with Non-Sterile Products process validation data can be a valuable aid in estimating the risk of microbial contamination or proliferation in each stage of the process. A science-based microbiological risk assessment should be carried out for non-sterile manufacturing activities to establish what microbial risks are involved with the facility, The growth of micro-organisms on manufacturing equipment and tools will be influenced by equipment and processes used. In many cases a generic risk assessment can cover a the cleaning and sanitisation routine used and their design. Microbiological cleaning number of products, but product-specific considerations should not be forgotten. The validation data can be useful in estimating this risk. outcome of the risk assessment will help determine appropriate controls associated with the running of the facility or specific processes and an appropriate monitoring programme. Knowledge of the starting materials and their potential contribution to bioburden is essential. A description of the full manufacturing process is a useful starting point in assessing the Water is a frequently used material and there needs to be assurance of the quality of risks involved. This may be simply done as a process map or flow diagram, with activities, water used. Typically, pharmacopoeial grade purified water will be used for non-sterile equipment used and process parameters added on. manufacture. Modern purified water systems generate water of very high microbiological quality and data should be available to demonstrate this. Important factors to consider in the manufacturing process include: Solvents used Product-contact utilities, such as compressed air and Nitrogen, must not be forgotten. These will typically be filtered close to the point of use, but reliance should not be placed Water-based processes provide a more favourable environment for micro- on filtration alone. Monitoring data should be available. organisms; the use of other solvents might decrease the risk of microbial growth pH The manufacturing environment itself, including the design and classification (when applied) of the facilities, contributes to the risk of microbiological contamination. Environmental Values above 10 or below 2 generally being detrimental to micro-organisms monitoring data should be obtained and used as part of the risk assessment. Osmolarity of solutions High osmolarity typically being detrimental to micro-organisms GMP and personal hygiene routines are also important considerations as part of the risk assessment. Temperatures used 25–35°C promoting microbial growth; significantly higher or lower temperatures Having gathered the information required, various tools may be used to conduct the risk being detrimental to micro-organisms assessment. Methods commonly used for microbiological risk assessments include HACCP (Hazard Analysis and Critical Control Points) and FMEA (Failure Mode and Effects Analysis). Drying If the water activity of the product is reduced below 0.6, then microbial growth will Some tools deliver a numerical assessment of the risks from different sources. The risks be suppressed (section 7.3) can then either be accepted or action taken to mitigate them. Where high risks of Hold times and overall campaign length microbiological contamination remain after mitigation, controls must be introduced at these points with an appropriate level of monitoring. The Microbiologist and QP should be Longer processing times may increase the opportunity for microbial proliferation involved in the risk assessment, the decisions on which risks to mitigate and the unless the conditions are detrimental to microbial growth appropriate mitigation actions. 38 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 39  Chapter 7 – Risk Assessment and Management 7.3. Water Activity Dosage Form aw Nasal inhalant (aqueous based) 0.99 A fundamental component of assessing the risk for microbiological control in non-sterile manufacturing is an understanding of whether the product or intermediates during the Antacid 0.99 production process are able to support growth or sustain viability of micro-organisms. Rectal cream 0.97 Oral liquid 0.90 Water activity (aw), is a measure of the free water in a material and is therefore a useful measure to aid the determination of microbiological risk. The details on how to determine Oral suspension 0.87 water activity are outside the scope of this monograph. Topical ointment 0.55 Water activity ranges from 0.0 (completely dry) to 1.0 (pure water). It is a more accurate Liquid filled capsule 0.45 index for microbial growth than water content as micro-organisms have a limiting aw below Tablets 0.36 which they cannot grow (typically ~ aw 0.6) (Pharmig Water Activity – Technical Guidance Suppositories 0.30 Document 1, 2008). Typically, products with water activities less than 0.6 are not susceptible to microbial Table 7.1: Water Activity of Pharmaceutical Dosage Forms growth of organisms found with compendial microbiological culture media. Friedel and Cundell, 1998 USP 31-NF26 <1112> ‘Application of Water Activity Determination to Non-Sterile Pharmaceutical Products’ provides examples of the application of water activity determination to non-sterile pharmaceutical products which can facilitate a risk based approach and may be used to: Develop product formulations Set microbiological release specifications Establish microbial testing programmes Determine potential shelf life stability from microbial growth It should be noted that water activity is temperature dependent and a lack of temperature control or temperature stability may lead to variable water activity in your product. Whilst the water activity provides a useful indicator of the ability of the formulation to support growth it should be remembered that some organisms present may remain viable and be pathogenic at low levels (e.g. Salmonella spp), therefore, good control during manufacturing is still essential. Table 7.1 shows the typical Water Activity of representative non-sterile pharmaceutical drug products. 40 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 41  Chapter 8 – Microbiological Monitoring Table 7.2 shows the water activity required by certain organisms for growth Chapter 8 (or the threshold at which they can no longer grow). Organism aw Microbiological Monitoring Most Bacteria >0.91 Pseudomonas spp 8.1. Introduction Listeria monocytogenes Mucor plumbeus 0.97 This chapter covers microbiological monitoring in non-sterile manufacturing. Other forms of environmental monitoring, as listed below, are relevant to non-sterile manufacturing, but Escherichia coli 0.95 are not detailed in this monograph: Serratia marcesens Particulates Shigella spp Temperature Bacillus subtilis 0.94 Humidity Many yeasts 0.88 Differential pressures Staphylococcus aureus (reduced oxygen conditions) 0.86 Room air changes Penicillium chrysogenum 0.83 Air flow patterns Many moulds 0.80 Microbiological monitoring should deliver a monitoring programme that provides value Halobacterium halobium added information enabling successful microbiological control. However, the methods are (halophilic bacterium) 0.75 very insensitive; increasing the number of samples taken increases the risk of diverse Zygosachharomyces rouxii results and there is no direct correlation between number of organisms found and product (osmophilic yeast) 0.60 contamination risk. Xeromyces bisporus (xerophilic fungi) 0.65 In general, the monitoring programme should demonstrate the effectiveness of production and maintenance activities, operator discipline and housekeeping activities and compliance No Microbial Proliferation <0.60 to defined standards. Table 7.2: Minimum Water Activity required for Growth Monitoring should be targeted to gain detailed information which can help build a picture Kabara and Orth, 1997 of the facility and enable appropriate risk assessment, thus helping trouble-shooting and the identification of any ‘hot spots’. It should be remembered that monitoring does not represent microbiological control in itself but is a mechanism to understand the effectiveness of the various controls in place and identify areas for improvement. 8.2. Environmental Monitoring Policy It is recommended that an environmental monitoring policy be drafted, which once effected should result in a state of control within the facility. All sampling techniques, sample locations, and sample frequencies should be based on this policy. Sampling that does not add to the understanding of the facility's state of control is a waste of resources and should not be performed. 42 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 43  Chapter 8 – Microbiological Monitoring The degree of environmental monitoring applied to non-sterile manufacturing activities Specific instructions regarding interpretation of results (e.g. in between read-outs to should be aligned with the risks associated with the product, process and patient, which prevent problems with overgrowth, how to handle too-numerous-to-count results, should be documented and defined in an environmental monitoring policy and standard whether scoring per colony type is performed) operating procedures (section 8.2.1) following a robust risk analysis. How alert and action limits are set and revised Surveys conducted over the last 14 years (PhRMA 1994, Pharmig 2001, Pharmig 2007) How a trend is defined all indicate that there is no clear consensus on microbiological monitoring programmes. How to handle excursions of the alert or action limits, or when a trend is observed However, regulatory guidelines and regulatory inspectors do expect microbiological (standardised investigation procedure) monitoring in non-sterile manufacturing to be in place. EU GMP 5.20 requires periodic assessment of measures to prevent cross-contamination and USP <1111> describes Records of the microbiological monitoring programme data should be generated. The monitoring in the form of regular surveillance. However, the nature of such a programme records must allow full traceability allowing for information retrieval and data manipulation, should be driven by circumstances, supported by risk assessment (Chapter 7) rather than detailing: a mandated set protocol as defined for sterile manufacturing. How, when, where and by whom a sample was taken and further processed If an area is determined as low risk, such as the manufacturing of tablets, the policy Which media and sampling methods were used: should focus on the bioburden control measures and on targeted monitoring, for example, Type and lot numbers the monitoring of relatively higher risk aqueous coating solutions. Routine monitoring can be performed at a lower frequency in such cases. There is little value in generating large Equipment identification (where applicable) amounts of data when the resource could be more effectively applied to preventive Room/Equipment status: measures. For the highest risk activities, monitoring on a batch by batch basis may be Clean status deemed appropriate in addition to bioburden control measures. Which activities were ongoing in the area at the moment of sampling: Routine monitoring is recommended to provide qualitative information with regards to the flora Dynamic present on site but should be supported through the risk assessment. A periodic species level identification should be applied to routine monitoring to build up a general knowledge of the site At rest flora to species level and enable identification of any changes in the facility. 8.3. Designing the Microbiological Monitoring Programme 8.2.1. Procedures and Documentation In order to design an appropriate microbiological monitoring programme several aspects The microbiological monitoring programme should be documented in standard operating should be considered which include but are not limited to: procedures which should include the following topics: Location For each sample (where applicable): Frequency Type of sample, media and disposables used Technique type Moment of sampling (at rest or in operation, but also related to operations, Time of monitoring (in operation/at rest) e.g. at the end of operations, prior to cleaning) Clean status Pre-treatment of the sampling location (disinfection at point of use, flushing of media) should mimic actual use Each aspect contributes different information regarding the efficiency of the control measures Frequency and locations of sampling and should be tailored to suit an individual site to provide a value added ‘complete’ picture. Length of time of monitoring Sample treatment, incubation and interpretation of the measurement Post-treatment of the sampling location, e.g. removal of residual media from the sampling event 44 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 45  Chapter 8 – Microbiological Monitoring 8.3.1. What to Monitor Suitable locations for monitoring should be identified, and if selected as part of the final monitoring programme, mapped and diagrammatically or photographically represented in The risk assessment process should highlight worst case areas and activities in the procedures to help accurate identification of sampling points. facility; these may include areas and activities with proximity to product and product contact, high human activity and/or traffic (e.g. door handles), areas with poor air flow, where water is in use and where drains are present. 8.3.2. When to Monitor: Frequency Special consideration should be given where contamination may be transferred and include: If the areas within the manufacturing facility have been formally classified, then regulatory Splash back inspectors will expect the frequency and approach to monitoring defined by EU GMP Annex 1 to be followed. Generation of aerosols Pooling of water For manufacturing facilities and areas which have not been formally classified, the Exposure to incoming contamination sources (e.g. starting materials, air, people etc.) frequency of monitoring should be based upon the risk assessment. Cleaning materials and equipment Again, there is no clear consensus approach across the non-sterile pharmaceutical The act of cleaning and storage industry and frequencies range from weekly to annual monitoring. Typically, a monitoring programme will set different frequencies depending on the particular aspect being Transfer by equipment wheels or by foot monitored. A non-specific example illustrating the approach may assign a weekly monitor Access to voids to final fill and wash bays, a monthly monitor to dispensary, unclassified production rooms, Tools microbiology laboratory, preparation rooms and equipment and a quarterly monitor for change areas, corridors and equipment storage rooms. The areas covered by the environmental monitoring programme should be tailored based Once the frequency has been set and implemented, data review and trending should be on risk. Areas that should be considered for monitoring include the following: used to assess the ongoing appropriateness of the frequencies chosen, which may be Dispensary increased or decreased depending on data. Data should be presented in a form that will Final fill (primary packaging) enable interpretation and identification of trends, e.g. graphical representation. In response to an Out of Trend or Out of Specification result, the planned action may involve Unclassified production areas special cause monitoring or an increased frequency for a period of time; thus providing Equipment more information to aid the resolution of the root cause and for the assessment of Equipment storage product quality. The routine frequency may be reinstated once results have returned to normal levels. Wash bays Packaging 8.3.3. How to Monitor: Techniques Change areas Corridors 8.3.3.1. Choice of Media Preparation rooms Pharmacopoeias stipulate media required for certain monitoring activities and techniques, Storage vessels for example, water monitoring or absence of pathogens testing using selection and Microbiology laboratory enrichment media. These media must be used unless supported by regulatory commitments and validation to demonstrate equivalence to the stipulated media. The baseline flora within the microbiology laboratory should be understood to provide For bioburden determinations, it is an expectation to ensure monitoring can capture information in the event of an investigation. bacteria, yeasts and moulds. Common media and incubation conditions for monitoring are given in table 8.1. 46 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 47  Chapter 8 – Microbiological Monitoring Each particulate may be associated with thousands of micro-organisms and the final Media Temperature Duration Comments dispersion of the particulates and therefore the resulting number of colony forming units Tryptone Soy Agar 30–35°C up to 3 days predominantly for (CFUs) is influenced by the sampling method. The number of CFUs in a sample equals the (TSA) (soybean-casein (48 hours USP the detection number of CFUs sampled from the environment. However, a limitation of these types of digest agar) <1116>) of bacteria microbiological testing is that number of CFUs is not equivalent to the exact number of micro-organisms that are present. For these purposes, the correlation is sufficient to provide an indication of microbiological control. Sabouraud 20–25°C up to 5 days predominantly for dextrose agar (SDA) (72 hours USP the detection There are two principle mechanisms for monitoring air, active and passive capture. <1116>) of yeasts and moulds As discussed throughout this monograph, there are no stipulated requirements for microbiological monitoring of unclassified areas and classification is only required in certain Table 8.1: Common Media and Conditions for Monitoring specified conditions for non-sterile manufacturing. Monitoring during operation should be considered as part of the risk assessment. If separate selective media are used for bacteria and yeast/mould, then each sample location requires sampling with each media to ensure representative data is obtained. 8.3.3.2.1. Passive Air Monitoring Alternatively, if validation supports, bacteria, yeasts and moulds can all be monitored using The passive air monitoring captures micro-organisms, which are typically attached to a a single medium (e.g. TSA) with dual incubation (e.g., 20–25°C first, then 30–35°C). If substrate particle, settling from the air by gravitational sedimentation onto ‘settle plates’ this approach is taken, recovery of normal flora needs to be demonstrated in the validation containing growth medium. Settle plates are typically Petri-dishes containing a nutrient after the prolonged incubation period. medium which are set out in predetermined locations for periods up to 4 hours. Longer manufacturing periods may be monitored by sequential settle plates. Data obtained should Scientifically, the use of low nutrient media with longer incubation times may be justified be corrected for the actual exposure time. because environmental isolates are, by nature of the environment they are isolated from, damaged. Nutrient rich media can shock damaged micro-organisms sufficiently to hinder The advantages of passive air sampling include the ability to sample throughout the or prevent growth on that medium, rendering some isolates viable, non-culturable and production period with minimal intervention, hence minimal turbulence. As such, settle hence resulting in an unrepresentative low recovery rate. Validation data should always plates are the method of choice for measuring air borne contamination in laminar air flows. demonstrate improved or equivalent recovery capability to more established media. The disadvantages of passive air sampling include poor sensitivity which is unable to detect Neutralising agents added to media should be considered where sampling locations may rapid changes, no determination of the volume of air sampled and is semi-quantitative at expose the samples to antimicrobial activity, for example, disinfectant residues on surfaces. best. Settle plates may, however, be used to assess concentration of micro-organisms in the environment and hence, as a tool for calculating contamination risk. 8.3.3.2. Air A quantitative value based upon the settling rate at which contaminants sediment from the air could be applied. However, since the sedimentation rate is dependent on the size of Airborne contamination includes bacteria (typically Gram positives), yeasts and moulds and aggregates and airflow patterns and is sensitive to disturbances in the airflow, this may arise from several sources in the manufacturing environment, which may include: technique should be used as a qualitative or semi-quantitative measure of the environment. Shedding skin cells from operators Aerosols from contaminated liquids An exposure time of four hours is considered to be the maximum due to the potential for agar media to lose moisture and hence alter the properties of the plates to support Dust generated from building or engineering works growth. The maximum exposure time should be validated to demonstrate suitable recovery Dust generated from the manufacturing process, for example, starting materials or of representative micro-organisms. dry blends Contaminated air 48 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 49  Chapter 8 – Microbiological Monitoring 8.3.3.2.2. Active Air Sampling representative micro-organisms from representative surfaces should be incorporated into the validation of this method. Active air sampling devices are designed to actively draw in air and capture the micro- organisms contained within the sample. A defined volume of air is drawn into the device; It is advisable to consider addition of neutralising agents to the media of RODAC plates to and the micro-organisms are captured either by impaction on the surface of media, counteract any residual antimicrobial activity of product, disinfectant or cleaning agents left trapped in liquid or onto a filter which may then be transferred to growth media. on the surface to be sampled. Factors that may influence the rate of capture include the velocity of air sucked in and RODAC plates are suitable for sampling flat and smooth surfaces only. deflection of the flow; where strong air flows are present, attention must be given to the exact placement of the equipment in the flow. The act of sampling using active methods Application of the RODAC media to a surface leaves a residue of media on that surface can cause disruption to laminar air flows. The equipment used should be covered by a and sampling by this method must incorporate a subsequent cleaning of the surface to preventive maintenance and calibration programme. remove traces of the residual media. 8.3.3.3. Surfaces 8.3.3.3.2. Swabs Surfaces in the manufacturing environment are typically contaminated by micro-organisms Swabs are more suitable for sampling surfaces that are uneven or difficult to access. settling from the air, by direct touch of an operator, product or starting materials, tools or ineffective cleaning. Swabs should be moistened before sampling. Typically swabs are moistened in a sterile isotonic buffered salt solution to help maintain viability of micro-organisms. The preferred method for sampling surfaces is determined by the properties of the surface. Regular, flat and smooth surfaces with low levels of soil, moisture and contamination may Several types of swabs are available for use, for example, alginate or cotton buds. Swabs be sampled using contact plates. Irregular surfaces, liquids, or surfaces with higher levels are wiped over a standardised surface area in a defined manner (templates may be of soil or contamination may be sampled more effectively by swabs. advantageous) and unlike RODAC sampling, may require further manipulation in the laboratory to transfer the micro-organisms from the swab into the growth environment. Surface monitoring forms a main mechanism for the microbiological assessment of cleaning, by cleaning validation or monitoring and determination of clean equipment hold The viability data obtained from swabs may be impacted (proliferation or reduction) by the times and dirty equipment hold times (section 5.3). lag time between sampling and processing in the laboratory or by the transportation process to the laboratory, so both should be defined procedurally and incorporated into Surface sampling should be considered as part of the risk based assessment; the location method validation. of sampling sites and frequency of sampling should be defined accordingly. Consideration should be given to the benefits and limitations of the various techniques available when Neutralisation of the growth media should be considered and the sampled surface should choosing the most appropriate for each location. be cleaned following sampling to ensure no contaminating residues are left. Swabs have poor transfer efficiency and recovery rates of micro-organisms from the swab 8.3.3.3.1. Contact Plates material are highly variable and depend on the technique used by the operator. Although it is possible to validate swabs as semi-quantitative, swab sampling may be used more Contact plates, also called RODAC (Replicate Organism Detection and Counting), are effectively as a qualitative measure. typically 55mm diameter plates where the media is slightly over filled and the surface, a raised pad of agar, allows for direct application to a surface for sampling. In theory, micro- Note that the correlation between contact plates and swab samples can be very poor. In organisms present on the surface in the area in contact with the agar, transfer to the addition to the variations in sampling efficacy, the mixing of swab solution can disrupt clumps agar surface and develop in to CFUs when incubated. It is known, however, that the of bacteria leading to an increase in the number of colonies observed (section 8.3.7). pressure of application can impact recovery and that micro-organisms adhere to surfaces differentially depending on multiple factors which include the surface type, organism type and length of time it has resided on the surface. Although a better recovery rate and efficacy is expected from RODAC plates, in relation to the recovery from swabs, the transfer is not complete (in the region of 10–50%) and recovery capability of 50 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 51  Chapter 8 – Microbiological Monitoring 8.3.3.4. Personnel provide information on the organisms that are circulating in the water system but may also be readily contaminated through the sampling process. A site should consider monitoring of personnel in the risk assessment. Monitoring of personnel is not mandatory for non-sterile manufacturing and is unlikely to be performed A possible source of microbial contamination in the system is the feed water. Feed water, in routinely, but may be useful in investigations and training programmes. most cases from the municipal supply, should be included in the monitoring programme. Action limits may be derived from the national regulations for potable water and alert limits from If assessed as a risk, gloved fingers are typically sampled by applying finger dabs (including historical performance; the suggested minimal frequency for sampling and testing is monthly. the thumb print) to nutrient agar plates, for example, Tryptone Soy Agar and incubating for 3 to 5 days at 30–35°C. The method for testing of water and recommended action limits are provided in the applicable monographs of the European Pharmacopoeia, however, it is expected to have Sampling of un-gloved hands can provide information on whether an individual carries a internally derived alert levels based upon the historical performance of the system. particular micro-organism on the skin which may be of use for investigational purposes if required. The performance qualification of the water system requires frequent monitoring and typically takes place over one year; in practice the performance qualification continues over the life of the system through the ongoing monitoring programme. 8.3.3.5. Drains The performance qualification may be divided into 3 phases: Drains present the potential for back-contamination within the manufacturing environment Phase 1: for 2–4 weeks to: with respect to splash back and aerosols and are a source of standing contaminated Establish the operating ranges water (section 4.4.2). Hygienic design mitigates associated risks, but an effective sanitisation programme should be incorporated as part of microbiological control. The Develop operating practices effectiveness of the microbiological control should be monitored. Support cleaning and maintenance procedures Demonstrate production and delivery of water of the required quality The efficacy of disinfectants, method of application and contact time should be determined and documented in a study to support microbiological control practices. Phase 2: for 2–4 weeks to demonstrate consistent operations producing and delivering water A qualitative monitor is typically achieved using swab sampling of the drain surfaces and or Phase 3: for one year to demonstrate extended performance over seasonal fluctuations liquid from the drain. Swabs provide easier access to difficult to reach locations and are the method of choice for wet areas. Depending on the outcome of the risk assessment with regards to the positioning of the drain in relation to manufacturing operations, areas Sampling should be representative of point of use and all individual points-of-use should be around the drain may also be sampled, for example, splash back areas. included in the monitoring programme. Points-of-use may be sampled according to a rotating scheme, whereby each point-of-use is, for example, tested at least monthly with Acceptance criteria should be based on objectionable organisms for the site (section representative samples obtained on a weekly basis. 6.7.2) and new isolates should be identified to species level. If organisms defined as objectionable to the product are identified there should be an assessment of the potential Typical sampling frequencies are: risk to product and a decision made with regards to appropriate actions, e.g. additional Potable feed Monthly/Quarterly disinfection of the drain. Product use Weekly Washing Weekly The frequency of monitoring drains should be justified through the risk assessment but typically a quarterly monitor is advised once a baseline flora is understood. Sample points will depend on the design specification of the system and should ensure coverage of the: 8.3.3.6. Water Start and end of the distribution system and generation plant Feed water into the system Micro-organisms in water systems are usually present in two forms: those attached to the surfaces of the system in biofilms and micro-organisms circulating in the water which are Intermediate points within the generation plant may also be considered to monitor the generally derived from organisms shed from the source biofilms. Water samples will performance of the different components. 52 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 53  Chapter 8 – Microbiological Monitoring 8.3.3.7. Steam 8.3.4. Acceptance Criteria Refer to section 4.4.6. Data generated by monitoring should be assessed against predetermined acceptance criteria in order to understand if the area remains under microbiological control. The microbiological risk of steam usage should be considered in the risk assessment and included in the monitoring programme if appropriate and at a suitable frequency determined by the risk. 8.3.4.1. Setting Limits Extreme care should be taken if sampling steam and a condensate of the steam should be For contamination levels, only upper limits are set. Typically, two limits are set for each generated for sampling. The sample should then be subjected to testing similar to that aspect tested; these are normally termed as “alert” for the tightest limit and “action” for used for water. the widest limit. Unlike the situation for sterile product manufacture, limits for non-sterile product manufacture are not typically set by regulations; it is for the manufacturer to set Limits should be established by historical evaluation and re-evaluated on a periodic basis. appropriate limits. However, there is a regulatory expectation that limits will be based not only on risk assessment but also on historical data, such that deviations from the norm are readily detectable. 8.3.3.8. Compressed Gases Excursions above the action limit will indicate a decline in the effectiveness of one or more Compressed gases which come into direct contact with product are a potential source of of the control measures; the excursion must initiate an investigation to determine the contamination and should be considered in the risk assessment. However, they typically cause and impact. represent a low risk due to the pressurised environment, low water availability and in line filtration. Consequently, frequent monitoring is unlikely to be required and it may be possible to Excursion of the alert limit acts as an early warning signal for this decline, and may initiate demonstrate satisfactory control with a documented study as part of the initial qualification. an investigation or an analysis of earlier data to detect a trend. Microbiological sampling apparatus may be bespoke or commercially obtained and should The limits set need to be a balance between taking unnecessary action or insufficient include a flow meter and mechanism for obtaining the sample securely; this may require a action when needed. A brief outline is provided below, but it is recommended that a flow rate controller to ensure integrity of the sample. statistician is consulted to ensure an appropriate data analysis is performed. The decision regarding the approach to setting the appropriate limits should be captured in a written For anaerobic gases due attention should be paid to ensure maintenance of anaerobic justification document. conditions at all stages including sampling, transportation to the laboratory and incubation. When very low counts are found, with a majority of the measurements revealing 0 colony A controlled amount of gas is directed through the equipment, on to the sampling surface; forming units, the results approach a Poisson distribution (but with tailing due to random the surface may be a filter membrane which will require transfer to suitable medium for effects); this is normally the case for areas classified as ISO 6 or lower and Laminar Air growth or impaction on to growth media. Flow cabinets. Microbiological monitoring of compressed gases should complement physical monitoring of When higher counts are found, the results may approach a normal distribution; this may be particulates, moisture and hydrocarbons. the case for ISO classes 7 and up where more counts are typical. The data should be plotted graphically to ensure the correct distribution is chosen for modelling. If the distribution is The frequency of microbiological monitoring should be determined by the risk assessment unknown, then the use of cumulative probability will allow limits to be calculated. and take into account filter performance etc. An approach for tabletting manufacture may include microbiological sampling of the source each month with annual sampling of outlets. For Poisson-distributed data, the alert limit may be set at the 95th percentile of the data- Consideration should be given to the accessibility of sample outlets that are integral to set, and the action limit at the 99th percentile. manufacturing equipment. For normally distributed data, the limits may be set at the average plus 2x standard Limits should be established by historical evaluation and re-evaluated on a periodic basis. deviation (equivalent to 95% probability) and 3x standard deviation (equivalent to 99.7% probability) respectively. 54 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 55  Chapter 8 – Microbiological Monitoring The figures calculated from the distributions should be critically assessed to ensure they 8.3.6. Investigations are commensurate with risk. For example, action limits significantly tighter than EU grade C are unlikely to be required for most non-sterile products. In an established facility the majority of data collated from environmental monitoring should confirm microbiological control. When excursions occur (see section 9.4), investigations The data set used for determination of limits must include all seasonal variations; ideally a are typically led by microbiologically trained personnel. However, consideration should be period of 12 months or a multiple thereof. It is advisable to remove outliers from the data given to assigning ownership and accountability of the investigation to personnel from the set, since these may lead to limits being set which are too high. The method for removal area where the excursion occurred, with support from microbiology staff, as this often of outliers depends on the distribution, but must be described in the procedure for helps achieve a greater commitment to resolution. establishment of limits. Individual values that are removed, e.g. due to assignable but irrelevant contamination (special cause variation), must be justified. For microbiological environmental monitoring the subsequent investigation typically includes: Identification of the micro-organisms and comparison with the defined objectionable In the beginning of execution of the monitoring plan, or after modifications to the organisms list for that site environment, equipment or processes, the data-set may be limited. It is recommended Review of activities in the area, functioning and suitability of equipment and interview of that a period of intensive monitoring is performed at this stage. The number of data points personnel collected should allow for the detection of trends (to ensure a baseline is reached) and to determine the correct statistical distribution. As guidance a minimum of 10 data points is Review of cleaning records and review of data and trends of related environmental recommended for each aspect. monitoring samples If it is necessary to gain a better understanding of the extent of a problem, the The limits should be reviewed and, if necessary, adapted periodically; typical review periods monitoring programme can be increased with more sample locations and/or a higher are annually or bi-annually. sampling frequency Adjustment to lower (tighter) limits is normally easy to justify; adjustment to higher limits Typical corrective actions upon excursion may include: may be necessary due to, e.g. increased activity in the area, and can be acceptable if Communication with the area management from where the excursion occurred investigations have revealed that the environment and processes are under control. Increase in monitoring More frequent or more rigorous cleaning and/or disinfection 8.3.5. Performing Monitoring Retraining or communication with personnel Before monitoring commences, the following should be defined and in place: Modification of procedures for operations or cleaning An approved protocol or procedure Maintenance or modification of the air handling system Approved and validated methods Approved and validated equipment The type of organism isolated may provide an indication of the contaminating source and Floor plan identifying sample points thus aid the determination of appropriate investigations and actions. For example: isolation of airborne organisms (e.g. Moulds or Bacillus spp.) Trained personnel to execute sampling (this may include production personnel) Investigation of Trained personnel to execute testing The air supply Defined sample transit criteria to ensure timely and careful transfer to the laboratory Maintenance of pressure differentials and air flow patterns Containment of building work Remedial actions may include use of a sporocide or disinfectant 56 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 57  Chapter 8 – Microbiological Monitoring Isolation of water borne organisms (e.g. Pseudomonas spp.) Specificity Investigation of Detection level/limit Water data Quantification (limit, linearity and range – where appropriate) Leaks, spills, opportunities for aerosol production and pooling water in the area Ruggedness Drains Robustness Humidity (e.g. washroom areas and equipment) Methods used to recover micro-organisms from the environment are difficult to validate Damaged surfaces or unsealed surfaces where water may accumulate and reproducibly and accurately and are fraught with technical issues, for example: harbour micro-organisms Contaminants may not be completely removed during sampling Remedial action may include Contaminants may not be completely transferred from the sampling tool to the growth Sanitisation of the water system medium Disinfection of surfaces or drains Transit time from sampling to the laboratory for processing may proliferate or Repair to damaged surfaces or seals deteriorate viability Dehumidification Organisms may be sufficiently damaged by the environment to render them non- culturable The growth media may shock the organisms, hindering their growth Isolation of personnel borne organisms (e.g. Staphylococcus spp.) may include: Organisms are frequently isolated in clumps, which may or may not be broken up by Investigation of the analytical method, hence, a single clump of cells may be a colony forming unit but Dressing and hygiene practices if agitated sufficiently during analysis multiple colony forming units may result in much Personnel monitoring higher perceived recovery Remedial action may include Consequently, these methods provide an indication of the microbiological status of the area Training in basic microbiology and hygiene sampled and validation should provide sufficient confidence of reproducibility to enable Personnel counselling trends to be established, but an absolute value of the inherent population of micro- organisms is unlikely to be obtained. The validation may include in vitro style testing using spiked environments to provide a 8.3.7. Validation of Monitoring Techniques measure of the method’s recovery capability. For example, coupons of various surface types found in the manufacturing environment, may be spiked with known numbers of Analytical methods should be validated (EU GMP Chapter 6). The variables and how the viable cells and the method used to recover those cells from the surface. However, the variables are controlled should be understood and accounted for in the design of the micro-organisms’ differential ability to adhere to different surfaces, impacted by age of the validation. This applies equally to both traditional and rapid microbiological methods. culture, surface condition and type of organism, can all add further complexity to the validation effort. A risk based assessment should be used to determine the approach to Critical considerations for validation are: method validation, ensuring validity of data generated. Is the method appropriate? Is the method reproducible? It is recommended that challenge organisms used during validation of monitoring methods Is the method comparable (if the method replaces a current method)? includes a range of local isolates to demonstrate the ability to recover such isolates. The validation should include a rationale for the approach taken including the choice of representative local isolates. Consideration should also be given to how long local isolates Recommended validation parameters include: are retained before there is a need to select new isolates from the local environment. Accuracy Precision 58 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 59  Chapter 9 – Reporting and Trending of Microbiological Data When a method, medium or equipment is changed, a comparative analysis between the Chapter 9 methods must be made. Independent validation of both methods is insufficient. Preferably both methods are used simultaneously for a certain period. Based on the obtained results, new action and alert levels may be set, and a new baseline for trend evaluation may be Reporting and Trending of determined. Microbiological Data 9.1. Introduction Raw monitoring data by itself is of little value and it needs to be appropriately analysed and presented in order to provide trends and appropriate focus for microbiological control. The reporting and trending of data provides an opportunity for the effectiveness of microbiological control and the appropriateness of the monitoring programme to be reviewed and modified. Where good control is demonstrated, there may be opportunities to reduce the level of monitoring, thus reducing cost without compromise to product or patient. 9.2. Batch-Specific Data In contrast to the situation with sterile product manufacture, environmental monitoring data is typically not produced specifically to support batch disposition in non-sterile manufacturing. Where batch-specific environmental monitoring is established as necessary to support the quality assurance of the product this forms part of the batch disposition assessments. In addition, if general monitoring action limits are exceeded they must be reported to the Quality Unit and a boundary for investigation should be drawn taking into consideration the impact on products manufactured in the facility since the previous satisfactory monitoring. It is important that Qualified Persons are aware of any potential impact on the products they are responsible for releasing. 9.3. Periodic Summary Reports Data should be assessed and trended as it is generated. Data outside the set specification or limits must be reported to the Quality Unit (9.2). If the facility is under good control, the majority of data will be reported via periodic summary reports. Periodic summary reports should be generated and reviewed by the applicable cross functional team (Chapter 2) including functional management and the Quality Unit. The frequency of generating a formal periodic summary report will depend on a number of factors such as the sampling frequency, the facility history and the products manufactured. 60 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 61  Chapter 9 – Reporting and Trending of Microbiological Data The summary report should be used to determine appropriate actions relating to the Laboratory environmental data facility and its operations and reassess the testing frequencies and limits (Chapter 8). Data entry/calculations Consideration should be given to the most appropriate means of presenting the summary data to other site functions. A combination of graphical and tabulated formats providing a If no attributable laboratory error is determined, then a wider investigation, involving a visual representation with clear supporting summary text should be the aim. Data should be cross-functional team will be required to determine the root cause. presented covering an extended period of time, for example, over a rolling year, to determine whether trends are present. It may also be useful to compare data with previous Upon excursion of the alert limit (OOT) a limited investigation may follow; as a minimum an rolling 12 months to establish understanding of successive seasonal variations. OOS and analysis for trends of the applicable sample and related samples is expected. OOT data should be specifically highlighted and addressed within the summary reports (section 9.4). For excursions outside of the specification or action limits, or when an adverse trend is confirmed there should be a more extensive investigation. It is recommended to standardise Often microbiological data will contain many zero data points which can present issues in the investigation procedure which will be specific to the type of OOS or action limit breached. statistical analysis of the data. It is important to select appropriate analysis tools which do not lead to the masking of significant events or trends (Chapter 8). If an OOS result is deemed to be valid, then further investigation is required to establish the root cause(s) and appropriate corrective and preventive actions need to be determined. Numerical methods (e.g. CUSUM calculations) or trend analysis software may be used for the detection of trends; this ensures objectivity and speed in trend detection. Any software tool Predetermined corrective action steps may be detailed in procedures. For example, in the used to generate graphical representations or analyse data must be qualified appropriately. situation where an environmental monitoring sample shows an OOS (action level) or OOT (alert level), particular steps to clean and retest may be taken and if the resulting retests are satisfactory, no further corrective actions may need to be taken. 9.4. Out of Specification/Out of Trend Handling Even when procedure-driven corrective actions are taken, consideration should be given to Out of specification (OOS) results include all suspect results that fall outside the whether any actions are required to prevent recurrence of the root cause. specifications or acceptance criteria established in new drug applications, official compendia, or by the manufacturer within their internal procedures, e.g. Action levels for The information from the OOS and OOT investigations should be analysed and shared with environmental monitoring data. the quality unit and other relevant business functions to aid determination of root cause, impact on product disposition and agreement of any further corrective and preventative Out of Trend (OOT), with respect to environmental monitoring data, may be defined as the actions that may be appropriate. OOS and OOT data should also be specifically highlighted exceeding of an alert level or a defined statistical signal. and addressed within periodic summary reports. The management of microbiological OOS and OOT results follows the same principles applicable to chemical OOS and OOT investigations (see PQG, 2005) but requires additional considerations, for example, microbiological contamination may not be homogeneous, and may include specific requirements detailed in the compendial tests. These requirements should be documented in procedures. For OOS investigations, it is critical to perform an initial investigation to determine whether the OOS result is due to laboratory error. A checklist may be useful to aid such investigations and might include: Sample handling/storage Media/components Equipment Methods Analyst technique 62 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 63  Appendix – Mind Maps Appendix – Mind Maps Figure A.1: Overall Microbiological Control Mind Map Figure A.2: Personnel Microbiological Control Mind Map 64 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 65  Appendix – Mind Maps Figure A.3: Facilities Microbiological Control Mind Map Figure A.5: Laboratory Operations Microbiological Control Mind Map Figure A.4: Cleaning Microbiological Control Mind Map Figure A.6: Formulation Microbiological Control Mind Map 66 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 67  Glossary Glossary Aflatoxin Naturally occurring mycotoxins that are produced by many species of Aspergillus. May be associated with herbal starting materials. At rest A term for the static state with all services operating, but without personnel present. Baseline Observations or data used for comparison or as a control. Bioburden Figure A.7: Monitoring Microbiological Control Mind Map The total microbial load with which an object or sample is contaminated. Biofilm An organised microbial system consisting of layers of microbial cells associated with surfaces, often with complex structural and functional characteristics. Biofilms have physical/chemical gradients that influence microbial metabolic processes. They can form on inanimate devices and also cause fouling (Prescott, Harley & Klein, 2005). CFR (Code of Federal Regulations) US regulations. Parts 210 and 211 relate to pharmaceutical Good Manufacturing Practice. CFU (Colony Forming Unit) Viable micro-organisms (bacteria, yeasts and mould) capable of growth under the prescribed conditions (medium, atmosphere, time and temperature) develop into visible colonies (colony forming units) which are counted. The term colony forming unit (CFU) is used because a colony may result from a single micro-organism or from a clump/cluster of micro-organisms. It is normally expressed as CFU per g or mL. Cleaning The removal of soil from a surface. Clean room A room in which the concentration of airborne particles is controlled to a defined standard. This is achieved by controlling the introduction, formation and retention of particles. Contact time The total time an organism is exposed to the antimicrobial action of a disinfectant. 68 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 69  Glossary Containment level 2 Intermediate/in-process material The facility requirements for the containment of Hazard Group 2 biological agents as A partly manufactured material which requires further processing before it becomes a defined by the Advisory Committee on Dangerous Pathogens (Advisory Committee on bulk or filled product. Dangerous Pathogens, 2001). Local isolate Contaminant An organism isolated from the local environment, e.g. production, laboratory. A foreign agent not introduced as part of processing, such as airborne particulates or adventitious micro-organisms. Medium (plural, media) Any liquid or solid material prepared for the growth, maintenance, or storage of micro- Dead leg organsims. Any length of pipework that does not allow effective circulation of its contents, resulting in the potential for contamination. Microbiological Control The sum total of activities undertaken to manage and mitigate risks from microbiological Disinfectant sources. An agent that reduces the level of micro-organisms to one that is safe for the relevant purpose. Usually a chemical agent but sometimes may be a physical one such as X-rays Microbiological Monitoring or ultraviolet light (Pharmig 2006). Sampling, testing, trending and reporting activities undertaken to assess and understand the state of microbiological control. Dynamic state Refers to environmental or particle monitoring when a room is occupied by personnel. Objectionable organism An organism that is risk assessed to be objectionable with respect to its potential impact FMEA (Failure Mode and Effects Analysis) to patients. Some objectionable organisms are specified in the pharmacopoeias but these A risk assessment and risk mitigation tool. are not exclusive and other organisms may be objectionable depending on the nature of the product, route of administration and intended patient population. Genotype The precise genetic constitution of an organism. Outlets User points in a water system. HACCP (Hazard Analysis and Critical Control Points) A formalised process by which a manufacturer ensures that all steps critical to product safety Out-of-Specification (OOS) are assessed and that adequate safety procedures are identified, implemented and reviewed. An OOS result is one which falls outside the specifications or acceptance criteria established in new drug applications, official compendia, or by the manufacturer within HEPA (High Efficiency Particulate Air) their internal procedures, e.g. Action levels for environmental monitoring data. Describes the system for filtering (diluting) air into clean-rooms. Standard HEPA filters remove 99.97% of 0.3µm particles. Out-of-Trend (OOT) With respect to environmental monitoring data, the exceeding of an alert level or a defined Herbal drugs statistical signal. Herbal drugs are mainly whole, fragmented, or cut plants, parts of plants, algae, fungi or lichen, in an unprocessed state, usually in dried form but sometimes fresh. Certain exudates Phenotype that have not been subjected to a specific treatment are also considered to be herbal drugs. The observable properties of an organism. Herbal drugs are precisely defined by the botanical scientific name according to the binominal system (genus, species, variety and author). (European Pharmacopoeia, 6th edition, 2008). Potable water Water which complies with local drinking water standards. Typically this is WHO (World Incubation Health Organisation) drinking water quality. The process of maintaining appropriate conditions (temperature, humidity, time) to allow microbial cells to replicate. RODAC (Replicate Organism Detection And Counting) The international acronym for a contact plate (that is an agar plate, with a raised surface, applied to a surface for the enumeration of micro-organsims). 70 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 71  Bibliography Risk assessment A systematic process of organising information to support a risk decision to be made Bibliography within a risk management process. It consists of the identification of hazard and the analysis and evaluation of risks associated with the exposure to those hazards (ICH Q9). ADVISORY COMMITTEE ON DANGEROUS PATHOGENS ‘The management, design and operation of microbiological containment laboratories’ Settle plate ISBN 0 7176 2034 4 (HSE Books, Her Majesty’s Stationery Office, 2001) An agar plate used for passive air-sampling. The plate is exposed for a fixed duration, after which it is incubated, and the number of micro-organisms which have settled on it and BAIRD, R.M.& BLOOMFIELD, S.F. , Microbial Quality Assurance in Cosmetics, Toiletries and Non-Sterile have produced colonies are counted. Pharmaceuticals. 2nd Ed. Taylor & Francis. Bristol, PA. 1996. p 51 & 115 Sporicide CFR Part 211.113 Control of microbiological contamination An agent that destroys microbial spores, especially a chemical substance that kills bacterial spores (Pharmig 2006). CUNDELL, A. M. Environmental Monitoring in Non-Sterile Product Manufacturing. In Environment Monitoring. Edited J. Moldenhauser Davis Horwood/PDA 2005 Starting material CUNDELL, A. M. ‘Managing the Microbiological Quality of Pharmaceutical Excipients’, PDA Journal of Any substance used in the production of a medicinal product but excluding packaging Pharmaceutical Science and Technology, Vol 59, No 6, November – December 2005, p 381–395 materials (EU GMP Glossary). EMEA CPMP ‘Note for guidance on quality of water for pharmaceutical use’ CPMP/QWP/158/01, Tundish May 2002 A receptacle within a drain forming a reservoir for water thus acting as a buffer, smoothing out flow, between processing areas and the drainage system. EUROPEAN COMMISSION ‘The Rules Governing Medicinal Products in the European Union’ Volume 4 ‘Good manufacturing practice (GMP) Guidelines’ http://ec.europa.eu/enterprise/pharmaceuticals/eudralex/vol4_en.htm Water activity (aw) A measurement of the energy status of the water in a system. It is defined as the vapour EUROPEAN PHARMACOPOEIA pressure of water above a sample divided by that of pure water at the same temperature; therefore, pure distilled water has a water activity of exactly one. FRIEDEL, RR & CUNDELL, AM; Pharmacopoeial forum vol 24 number 2 March/April 1998 p 6087-6090 HUTCHESON, J. ‘Validation of Purified Water Systems’ presentation at Pharmig conference on non- sterile monitoring (2008) INTERNATIONAL CONFERENCE ON HARMONISATION (ICH) ‘Q9 – Quality Risk Management’ (2005) www.ich.org (also now incorporated as Annex 20 of European Commission GMP Guide and available via Eudralex) INTERNATIONAL CONFERENCE ON HARMONISATION (ICH) ‘Q4B ANNEX 4A Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions on Microbiological Examination of Non-Sterile Products: Microbial Enumeration Tests General Chapter’ INTERNATIONAL CONFERENCE ON HARMONISATION (ICH) ‘Q4B ANNEX 4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions on Microbiological Examination of Non-Sterile Products: Tests for Specified Micro-organisms General Chapter’ INTERNATIONAL CONFERENCE ON HARMONISATION (ICH) ‘Q4B ANNEX 4C Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions on Microbiological Examination of Non-Sterile Products: Acceptance Criteria for Pharmaceutical Preparations and Substances for Pharmaceutical Use General Chapter’ 72 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 73  Bibliography ISO/IEC Guide 51:1999 ‘Safety Aspects – Guideline for their inclusion in standards’ WILKINSON, S: ‘Cleaning Validation in Non-Sterile Manufacturing’ Presentation to Pharmig non-steriles conference, February 2008 ISPE Baseline pharmaceutical engineering guides for new and renovated facilities: Volume 2 Oral solid dosage. 1998 ISPE Baseline pharmaceutical engineering guides for new and renovated facilities: Volume 5 Commissioning and Qualification. 2001 KABARA, JJ AND ORTH, DS (ed.) Preservative-free and self preserving cosmetics and Drugs: Principles and Practise, Marcel Dekker, p 1–14 (1997) MEDICINES AND HEALTHCARE PRODUCTS REGULATORY AGENCY (MHRA) Inspection and Standards Division ‘Rules and Guidance for Pharmaceutical Manufacturers and Distributors’ Pharmaceutical Press (2007) (Also referred to as ‘The Orange Guide’) MONTGOMERY, E. & TAWIAH, W. M., Microbiological Considerations When Selecting Excipients During Product Development, American Pharmaceutical Review. Vol. 7, Issue No. 4. July, 2004. PDA TECHNICAL REPORT 13 ‘Fundamentals of an Environmental Monitoring Program’, 2001. PHARMACEUTICAL QUALITY GROUP (PQG) ‘Monograph 10 – Cleaning Validation’ ISBN 0 906810 63 9 (Institute of Quality Assurance, 1999) Available from www.thecqi.org PHARMACEUTICAL QUALITY GROUP (PQG) ‘Monograph 11 ‘Good Quality Control Laboratory Practice’ ISBN 0 906810 79 5 (Institute of Quality Assurance, 2005) Available from www.thecqi.org PHARMIG ‘A Guide to Disinfectants and their use in the Pharmaceutical Industry’ ISBN 0 9551030 0 3 (The Pharmaceutical Microbiology Interest Group, 2006) Available from www.pharmig.org.uk PHARMIG ‘Technical Guidance Document 1 – Water Activity’ (2008) PHRMA ENVIRONMENTAL MONITORING WORK GROUP ‘Microbiological monitoring of environmental conditions for Non-Sterile Pharmaceutical Manufacturing’. Pharmaceutical Technology March 1997; PRESCOTT, HARLEY & KLEIN: ‘Microbiology’, 6th edition (2005) THE CODE OF PRACTICE FOR QUALIFIED PERSONS: Institute of Biology, Royal Pharmaceutical Society of Great Britain, Royal Society of Chemistry (Also included in ‘The Orange Guide’) RANDALL, C. ‘Environmental monitoring of Non-Sterile Manufacturing’ presentation at Pharmig Microbiological monitoring and control of Non-Sterile manufacturing meeting (2005) UNITED STATES PHARMACOPOEIA USP <1072> Disinfectants and Antiseptics USP <1112> Microbiological Attributes of Non-Sterile Pharmaceutical Products – Application of Water Activity Determinations 74 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 75  Notes 76 Microbiological Control for Non-Sterile Pharmaceuticals Microbiological Control for Non-Sterile Pharmaceuticals 77  EXCELLENCE IN MICROBIOLOGY Pharmig T5 The Maltings, Roydon Road, Stanstead Abbotts Hertfordshire, SG12 8HG, United Kingdom T: + 44 (0)1920 871 999 F: + 44 (0)1920 871 156 E: [email protected] www.pharmig.org.uk The Chartered Quality Institute 12 Grosvenor Crescent, London, SW1X 7EE T: +44 (0)20 7245 6722 F: +44 (0)20 7245 6788 E: [email protected] www.thecqi.org 9 780956 080400