KEEPING CLEANROOMS CLEAN: SELECTION AND USE OF DISINFECTANTS AND DETERGENTSTim Sandlehttp://www.pharmamicroresources.com/

INTRODUCTION The importance of cleaning and disinfection in cleanrooms

Criteria for the selection of disinfectants

In use requirements (disinfectant rotation)

The basics of disinfectant validation Cleaning techniques Reviewing microflora Environmental monitoring.

IMPORTANCE OF CLEANING AND DISINFECTION IN CLEANROOMS

• Cleanrooms– Design

• Air– Filtration (HEPA)– Pressure differentials

– Air changes– Clean-up times– UDAF: air velocity

– Personnel– Gowning– Behaviours

– Cleaning and disinfection

IMPORTANCE OF CLEANING AND DISINFECTION IN CLEANROOMS Regulatory requirements

EU GMP Annex 1 FDA Guide for Aseptic Filling (2004) FDA Code of Federal Regulations: 21 CFR 211.56b, 21 CFR 211.56c and CFR 211.113b

USP Chapter <797> USP Chapter <1072> ‘Selection of a Disinfectant for Use in a Pharmaceutical Manufacturing Environment’

WHAT ARE DETERGENTS? Detergents are cleaning agents Detergents work by penetrating soil and reducing the surface tension

Cleaning, in the context of cleanrooms, is the process to remove residues and ‘soil’ from surfaces to the extent that they are visually clean.

WHAT ARE DISINFECTANTS? "[a] chemical or physical agent that inactivates vegetative micro-organisms but not necessarily highly resistant spores“

ISO standard on Aseptic Processing (ISO 13408–11),

HOW DO DISINFECTANTS DIFFER? Disinfectants vary in:

Spectrum of activity Modes of action Efficacy Whether they are bacteriostatic or bactericidal

Factors influenced by the active ingredient

TYPES OF DISINFECTANTS #1• Different disinfectants target different sites within the microbial cell:– Cell wall, – Cytoplasmic membrane – Cytoplasm.

• Different approaches to the categorisation of disinfectants:– Chemical nature, – Mode of activity – Bacteristatic and bactericidal effects on micro-organisms.

TYPES OF DISINFECTANTS #2 Non-oxidising

Many commercially available types Oxidising

Fewer commercially available types Widest spectrum of activity

TYPES OF DISINFECTANTS #3 Non-oxidising disinfectants

Alcohols Aldehydes Amphoterics (anionic and cationic character),

Phenolics Quaternary ammonium compounds

TYPES OF DISINFECTANTS #4 Oxidising disinfectants

Halogens like iodine, chlorine and chlorine dioxoide

Oxidising agents like peracetic acid and hydrogen peroxide.

SELECTION OF DISINFECTANTS• Selection:

– Are the disinfectants used appropriate?– Is the effectiveness periodically assessed?

– What is the scientific rationale for selection?

• Normal:– Select two disinfectants (ideally with different modes of action)• Ideally one should be sporicidal

– Some elect to keep a third disinfectant in reserve

A RATIONALE FOR SELECTION # 1• Spectrum of activity

– What types of different microorganisms will the disinfectant destroy?

– How effective is the disinfectant against microorganisms in different physiological states?

– Ideal – spectrum should be as wide as possible.• Sporicidial

– Does disinfectant x need to be sporicidal?– Not all types need to be– Health and safety– Surface corrosion

• Contact time– How rapid is the disinfection action?– How long does a surface need to remain wet?

A RATIONALE FOR SELECTION # 2• When selecting the two disinfectants, they should have different modes of action. – This is also tied to the regulatory expectation that disinfectants are rotated.

• Does the disinfectant require a certain temperature and pH range to function correctly?– E.g. Does it need to work in a coldroom? If so, is additional validation required?

• Is the disinfectant compatible with the in-use detergent?– Sometimes detergent residues can neutralise the active ingredient in the disinfectant.

A RATIONALE FOR SELECTION # 3• Does the disinfectant leave surface residues?

– What is the extent?– Does this matter? Will it be wiped away with WFI?

• Is the disinfectant compatible with all types of surfaces.

• Will the disinfectant damage the surface material? – Is a surface rinse required (WFI or alcohol)?– Is the disinfectant likely to absorbed into the material over time?

• Will the disinfectant meet the requirements of the validation standards to measure bactericidal, fungicidal and, if appropriate, sporicidal activity?

A RATIONALE FOR SELECTION # 4• What presentation is required for the disinfectant?– Prediluted preparation in a trigger spray? – Ready to use concentrate?– Impregnated wipe?

• Health and safety standards. – Operator welfare. – Impact upon the environment.

• Cost– Use over a large surface area.

• If the disinfectant is required for use in an aseptic filling area then it will need

• to be sterile filtered or supplied sterile in a suitably wrapped container. – Can the disinfectant be readily filtered?

A RATIONALE FOR SELECTION # 5 Hand sanitizers are also required (for cleanroom staff to apply either to skin or to gloved hands) as part of a comprehensive disinfection programme.

SELECTING DETERGENTS There are two key considerations when selecting detergents: The chemical composition of the detergent (ideally neutral and non-ionic solutions)

The compatibility of the detergent with the disinfectant.

IN-USE RULES #1 Any disinfectant will only be effective if it is used: At the correct concentration (minimum inhibitory concentration),

If applied to relatively clean surfaces using appropriate cleanroom grade mops or cloths

If left for the correct contact time Its effectiveness will also depend upon the numbers and types of microorganisms present

IN-USE RULES #2• Rotation

– Typically two disinfectants are rotated. – The theoretical argument for rotating two disinfectants is to reduce the possibility of resistant strains of micro-organisms developing.• Concern for antibiotics, but little evidence for biocides used on surfaces

– A requirement of some regulatory bodies• EU GMP Guide states that “where disinfectants are used, more than one type should be employed” (Annex 1).

• USP (<1072>) is less exacting and poses some questions about the scientific need for rotation.

IN-USE RULES #3 Establishing the Disinfectant Expiration Date

WHAT ELSE INFLUENCES HOW DISINFECTANTS WORK?Different types of microorganisms have varying levels of resistance to broad spectrum disinfectants

Most Resistant Prions/TSEs Coccidia Bacterial Endospores Mycobacteria Yeast-like/Mold-like fungi

Gram-Negative Bacteria

Gram-Positive Bacteria

Least Resistant

WHAT ELSE INFLUENCES HOW DISINFECTANTS WORK? The location of microorganisms. Microorganisms in suspension are easier to kill than those affixed to surfaces.

Temperature and pH Interfering substances (‘soil’).

THE BASICS OF DISINFECTANT VALIDATION # 1 Each disinfectant must be validated to ensure its efficacy

Regulatory agencies expect the pharmaceutical manufacturer to evaluate the efficacy of their disinfectants

There are many different approaches for disinfectant validation, such as: ASTM (American Society for Testing Materials)

AOAC (Association of Official Analytical Chemists International)

BSI (British Standards Institution) TGA (Australian Therapeutic Goods Administration)

USP (United States Pharmacopeia)

THE BASICS OF DISINFECTANT VALIDATION # 2 The standard European approach for disinfectant validation consists of: A basic suspension test A two-part simulated-use surface test.

North American surface test.

In addition, field trials or a type of on-going assessment is common

THE BASICS OF DISINFECTANT VALIDATION # 3 Basic Suspension Test

Designed to measure the efficacy of a disinfectant against selected microorganisms after a predetermined contact time

The selection of a suitable sterile neutralizer is required

After challenging a disinfectant solution with a microbial population the mixture is plated after the required contact time, and the surviving microorganisms enumerated

THE BASICS OF DISINFECTANT VALIDATION # 4 Quantitative suspension test

The purpose of the quantitative suspension test is to evaluate the activity of a disinfectant against a range of microorganisms under conditions which more closely simulate practical use.

The test consists of inoculating a prepared sample of the disinfectant under test in simulated ‘clean’ and ‘dirty’ conditions using a challenge suspension of bacteria or fungi.

THE BASICS OF DISINFECTANT VALIDATION # 5 Surface Test

More relevant than the suspension test Representative manufacturing surface samples are inoculated with a selection of microbial challenge organisms.

A disinfectant is applied to the inoculated surfaces and exposed for a predetermined contact time. Then the surviving organisms are recovered using a qualified disinfectant-neutralizing broth and test method (such as a contact plate or swab)

THE BASICS OF DISINFECTANT VALIDATION # 6

Test variables: Different surfaces Optimal contact times

Types of microorganisms (reflective of the natural microflora of the facility)

THE BASICS OF DISINFECTANT VALIDATION # 7 Field Trials

Review appropriateness of cleaning frequencies for: Variation in microbial counts The most frequently isolated organisms Atypical microbial isolates Isolates associated with continued adverse trends

Isolates associated with both environmental/personnel and product bioburden

CLEANING AND DISINFECTION TECHNIQUES #1

The techniques of cleaning and disinfection are important. It does not matter how effective the purchased detergents and disinfectants are unless they are used in the correct way, otherwise areas will not be cleaned effectively and unduly high levels of microbial contamination will remain.

CLEANING AND DISINFECTION TECHNIQUES #2Important features:

Order of cleaning and disinfection In order for a disinfectant to work effectively, ‘soil’ (such as grease and dust particles) must be removed first using a suitable cleanroom grade detergent.

Preparation of detergents and disinfectants Detergents should be made in hot purified water (>25°C). Disinfectants are normally prepared in cold water.

For aseptic processing areas, detergents and disinfectants are often prepared using Water-for-Injections and sterile filtered into the filling area.

The correct amount of detergent must be used.

Note expiry time

CLEANING AND DISINFECTION TECHNIQUES #3 Important features continued:

Cleaning materials Wipes used must be effective at removing non-viable particulates, capable of applying a disinfectant, and capable of the subsequent removal of viable contamination.

For use in an Aseptic Filling Area, detergents and disinfectants must be applied using non-particle shedding, clean room certified mop heads, cloths and wipes. These should be of a non-woven, non-particle shedding and lint-free variety.

CLEANING AND DISINFECTION TECHNIQUES #4 Cleaning steps

The cleaning and disinfection process consists of: Sweeping away dust and debris (if applicable) Application of a detergent solution through wiping or mopping.

Application of a disinfectant solution through wiping or mopping.

Removal of disinfectant residue through wiping or mopping with Water for Injections or 70% IPA.

CLEANING AND DISINFECTION TECHNIQUES #5Cleaning techniques

The cleaning technique should be defined and standardised.

Poor cleaning technique = problems. The physical treatment of a surface or item of equipment is as important as the chemical.

CLEANING AND DISINFECTION TECHNIQUES #6 Appropriate records should be kept of cleaning activities in process areas.

Detergents and disinfectants for use on surfaces (walls, floors) must be applied using the double or triple-bucket system to avoid cross contamination.

CLEANING AND DISINFECTION TECHNIQUES #7 Two-bucket Technique Dip the mop in the disinfectant

Mop the floor (preferably a specified number of square metres per “dipping”)

Dip the mop in the bucket of water

Rinse the excess water off the mop head back into the bucket of water (this is to prevent carrying too much water back into the disinfectant and diluting it)

Dip the mop in disinfectant Repeat

Three-bucket Technique

Dip the mop in disinfectant

Mop the floor Dip the mop in the bucket of water

Rinse the excess water off the mop head into the third (empty) bucket

Dip the mop in disinfectant

Repeat

CLEANING AND DISINFECTION TECHNIQUES #8 Cleaning and disinfection

using cloths and mop heads = saturating the cleaning item and wiping the area using a series of parallel overlapping strokes (with an approximate one quarter overlap) and never in circular motions.

The direction of the cleaning is towards the operator (from top to bottom, from back to front).

Only one application of the disinfectant or detergent should be applied to avoid over concentration.

Cleaning and disinfection should begin with the visually ‘cleanest’ area first and towards the ‘dirtiest’ area last (typically the application of detergents or disinfectants involves starting from the far side of a room and towards the exit in a series of overlapping parallel strokes).

CLEANING AND DISINFECTION TECHNIQUES #9 Cleaning and disinfection frequencies

At different frequencies for different areas

Based on the criticality of the area and this will vary between facilities

Floors and working surfaces will often be cleaned at a higher frequency than walls or ceilings

HAND SANITISATION Hands, whether gloved or ungloved, are one of the main ways of spreading infection or for transferring microbial contamination

Therefore, regular glove sanitization should be undertaken in aseptic processing areas

There are many commercially available hand sanitisers with the most commonly used types being alcohol-based liquids or gels.

ENVIRONMENTAL MONITORING: EVIDENCE Effectiveness of cleaning and disinfection is shown by environmental monitoring, especially microbiological surface samples Swabs Contact plates (RODAC)

Environmental monitoring data reviewed for trends - allows the frequency of cleaning and disinfection to be based on risk.

ENVIRONMENTAL MONITORING: EVIDENCEFrequency of disinfectant rotation is based on the environmental monitoring data.

Microflora recovered can also be revealing:Knowing the potential origins of different micro-organisms can indicate the potential source of contamination.

Knowing the species of a microorganism can provide invaluable information as to its origins.

For example, many Gram-positive cocci are part of human skin flora, Gram-positive rods can be transferred into clean areas via equipment and on footwear, and Gram-negative rods are often linked to water sources.

Endospore forming bacteria may indicate that cleaning and disinfection practices are not effective.

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