cleanroom control and best practices€¦ · cleaning and disinfection best practices •how often...

Copyright © 2018 STERIS Corporation. All Rights Reserved.

13 Aug 2020

Cleanroom Control and Best Practices

Beth Kroeger

Technical Service Senior Manager

[email protected]

STERIS Life Sciences

Contamination Control Solutions

mailto:[email protected]


• Definition of Cleanroom and how classified

• Cleanroom behavior

• Cleanroom startup

• Cleaning and Disinfection Best Practices• Cleaning and disinfection techniques/development of Cleaning SOP

• Rotation of disinfectants/sporicidal agents

• Rinsing

Agenda


Origin of the Cleanroom

• Invented in 60’s by Willis Whitfield “Mr. Clean”

– Pioneer of Unidirectional airflow.

• Invented room 1000 x cleaner

• “Particle counters broken”


Why do we need Cleanrooms?

• 21 CFR 211.46(b) “Equipment for adequate control over air pressure, micro-organisms, dust, humidity and temperature shall be provided when appropriate for the manufacture, processing, packaging or holding of a drug product.”


Clean-room Classifications

• Clean-room users concerned with 2 types of particulates:

: lint, soil, skin flakes, etc.

: bacteria, viruses, molds and spore forming organisms


FS209 Cleanroom

Classification

ISO 14644-1 Cleanroom

Classifications

EU GMP Annex 1 Grades

≥ 0.5µm particles/m3

Viable Microbes(cfu/m3)

Mean Airflow Velocity

(fpm)

Air-changes/hr

100,000 8 D 3,520,000 100 5 – 10 5 – 48

10,000 7 C 352,000 10 10 – 15 60 – 90

1,000 6 - 35,200 7 25 – 40 150 – 240

100 5 A & B* 3,520 1 40 – 80 240 – 480

*at rest. Grade B in operation is equivalent to a Grade C at rest.

Cleanroom Classifications/requirements


Don’t forget, in case of questions….

Contamination Sources

People• Gowning rooms

• Attire (clothing, shoe covers,

hoods, face masks, goggles, etc)

• Conduct

• Standard Operating Procedures

Facility• Poor design

• Aging facility

• Maintenance

• Cleaning and

disinfection

Materials• Pass through sterilizers

(autoclaves, dry heat ovens,

depyrogenation tunnels, etc.)

• Decontamination chambers (EO,

VHP, UV, etc.)

• Material handling airlocks

Genus A/B (6729) C/D (2500)

Micrococci (and related) 38% 40%

Staphylococci 21% 11%

Bacillus (and related) 13% 10%

Pseudomonas (and related) <1% 8%

Corynebacterium (and related) 3% 5%

Rhodococci <1% N/A

Fungi N/A 3%


• Failure to have sanitation program/not following procedures

• Poor aseptic technique

Procedures to prevent microbial growth

Disinfectant contact time not specified

Disinfectant used in ISO 5 area not sterile

No sanitization of items into hood

Failure to consistently sanitize gloved hands

Failure to work slow and deliberate


Personnel Practices:Cleanroom Behaviors

• Manipulations• Keep body/hands out of the path of unidirectional flow

• Contact sterile materials only with sterile instruments

• Always walk and move slowly: • Prevents turbulence to laminar airflow in the room.

• Can increase particles in area.

• No excessive talking


• Limit movement

• Do not lean against walls or equipment.

• Do not touch floors. • If process requires work on floor, put down barrier or leave after task is

complete.

• If you drop something, pick it up, spray it off and IPA/EtOH item and hands.

• May have to exit

• Do not touch face, masks, or Safety glasses once gowned.

• NO GUM and NO Reaching for phones inside of gown!

Personnel Practices:Cleanroom behaviors


• Personnel transmission:

• Inspectors are paying attention to operator’s gloves

• Integrity (packaged and while worn)

• Frequency of sanitization

• Compatibility

• Residue removal

• Proper gowning

• Adulteration

• Expect written procedures to detail methods used to don

• Expect micro surface sampling after manipulations

Personnel PracticesHealth and Hygiene


• Your gown has been compromised/soiled.

• If gown or inner glove is torn.

• Your gown contacts the floor due to kneeling, etc.

• After plating

• Any doubt your suit is compromised.

Personnel Practices:Re-gown if…..


• Adequacy of design and qualification studies to ensure facility is meeting expectations for air balance and air flow

• Practices

Inadequate Facility/studies

Failure to perform filter leak tests

No control over opening and closing doors to different classifications

Issues with room pressure differentials


• After HEPA’s installation, protect return vents with filter prior to Construction clean

• Cleaning of debris, dirt, soiling, particles

• Removing cardboard, boxes, equipment, hoses

• Single or Double Cleaning with a formulated cleaner

• Acidic, Neutral, or Basic Cleaner can be utilized

• Neutral cleaner is often used to prevent any substrate compatibility issues

• Goal: Clean Pristine Surfaces

• Source: [International Standard 14644-5, Cleanroons and associated controlled environments – Part 5: Operations: 2004]

New Cleanroom Facility

Information

Chapter 19, p. 213 - 218Start-up of Cleanrooms, Initially and After a Worst-Case EventBy Jim Polarine and Beth Kroeger

Chapter 4, p. 97 – 120 How Issues Related to Utilities, Surfaces, and Practices Impact Cleanroom EnvironmentsBy Jim Polarine and Beth Kroeger


• After the initial cleaning and large debris removal is complete• Clean construction work may still occur (no invasive work)

• Frequency in ISO 146441-2• ~ every 12 months, not exceeding 366 days in use or 400 days when not

in use

• ISO 5 every 6 months

• May damage filter or seals w/ install – buy extra

• The room should meet all the ISO-14644 (including ISO-14644 part 3) specs for particles and air control

• If Aseptic Manufacturing is taking place, then the requirements for the Aseptic Processing Guide (2004) should be met for EM

Certification


• Particle Concentration Test

• Airflow Test• volume of air per unit of time (m3/hr)

• Determines air exchange rate

• Air Pressure Differential Test

• Temperature Test

• Humidity Test

• Recovery Test• Ability to eliminate airborne particles

• Evaluates 100:1 recovery time

• Installed filter system Leak Test

ISO 14644 (part 2 – specs; part 3 – test methods)


• Once the cleaning program is established, it is important to follow the recommendations below to maintain the environment during routine operations.

• No cleaning should take place in an area during open operations and/or environmental monitoring.

• Rooms divided by lines of demarcation, separating two classes in the same room, should be cleaned and monitored per the stricter classification.

• Solutions and equipment from a less-strict classification should not be used to clean a stricter room classification.

Cleaning around the facility


Environmental Control:Buildings and Facilities

• HVAC function• Temperature and RH control is vital

• 30 – 60% RH

• 16 - 24°C

• Maintain 24 hours a day

• A velocity of 0.45 m/s has generally been established, with a range of 20% around the set point, when measured 15 – 30 cm from the filter face

• Airflow: AC/HR depends on room and system• Class 100,000: > of 20 airchanges/hr

• Class 10,000: > of 50 airchanges/hr

• Class 100: > of 100 airchanges/hr

• Blockage and placement of feed and return vents

References: • FDA Guidance for Industry, Sterile Drug Products

Produced by Aseptic Manufacturing –Current Good Manufacturing Practice. Sep. 2004. p 4

• Parenteral Drug Association (2015) PDA Points to Consider for Aseptic Processing, Part 1, January 2015, Parenteral Drug Association.

• ISO 14644-4 Design, Construction, and Start-up



• HVAC function • HVAC systems maintain pressure between spaces

• Dependent on nature of operation

• Pressure differential: 0.04 – 0.06” WC (10 – 15 Pascals)

• “Close the door policy”

• Monitor continuously, alarms documented, and deviations investigated.

• Procedures in place for excursions• CONFIRM



• Airlocks, Passthroughs & Gowning rooms• Restricted access.

• Doors should be interlocked.• Locked, alarmed (audible/visual) or procedures

• DRAFT Annex 1 now requires interlocks, also describes a time delay

• Divided into “Dirty” and “Clean” sides.• Room cleaned to the higher level of ISO level

• Cart with wipes and disinfectants available.

• Cleaning often referred to as 2-step cleaning.• Disinfectant/Sterilant followed by IPA or EtOH as a rinse agent or where residue is a

concern


• Federal Insecticide, Fungicide and Rodenticide Act (FIFRA)

• All germicidal products fall under the FIFRA as amended (1988) and administered by EPA

• FDA regulation as medical device per Food Quality Protection Act of 1996 if used to reprocess other medical devices or if used as a sterilantfor medical devices

Disinfectant regulation


EPA classifications

• Sanitizer

• Disinfectant

• Sterilant


• Proper use results in bacteria reduction of >99.9% • Used on precleaned surfaces

• How many bacteria can be found on the head of a pin?

• Log reduction 1,000,000

• 1-Log Reduction would reduce the number of bacteria 90%• Example: 100 bacteria would be reduced to 10, or 10 reduced to 1

Sanitizer defined


Disinfectant defined

• Proper use results in 100% kill of vegetative bacteria, target viruses and target fungi

• May or may not require precleaning• Serum efficacy

• Staphylococcus aureus

• Salmonella enterica

• Pseudomonas aeruginosa


• Proper use results in 100% kill of all microorganisms, including bacterial spores

• Demonstrated 6-7 Log reduction

• Always requires precleaning

Sterilant defined


Cleaning and Disinfection: Techniques

• Grid

• Overlapping strokes

• Pull and lift

• Figure 8 for floors or unidirectional mopping strokes

• Modified Figure 8 for walls

Anne Marie Dixon, Ch. 11, Cleaning of Non-Product Contact Surfaces, p 226, in Cleaning and Cleaning Validation for the Pharmaceutical and Medical Device Industries, Vol. 1 Basics, Expectations, and Principles. Paul L. Pluta, Ed., PDA, Bethesda, MD, and DHI Publishing, LLC, River Grove, IL. 2009.


Cleaning and Disinfection:Techniques

• Two Bucket routine:– Disinfectant in front or both

– Bucket under wringer is rinse & waste

• Three Bucket routine:– Third bucket used for catching wrung

out solutions

• Changing out the use dilutions– Every 600 ft2/ 55.74 m2 (ISO 5,6)

– Every 1000 ft2/ 92.90 m2 (ISO 7,8) Bucket #1 contains

disinfectant solution

Bucket #2 contains rinse

solution

Bucket #3 contains wrung

out waste

Link to the YouTube video for the 3 bucket mop technique: http://youtu.be/ajZ5_QHErqU

Videos/Three bucket mop technique_Walls.wmv

http://youtu.be/ajZ5_QHErqU


Visible Particulate on Surfaces

excessive soil is easy to see


Result of typical technique using wetted wipers


Single pass stroke to remove particulate


Importance of overlapping

strokes


• Cleanest to dirtiest area

• Top to bottom and back to front

• Most critical to least critical surface

• Clean the farthest corner of the room towards the exit that will be used to leave the room.

• Rooms divided by demarcation with two “classifications”

• Clean to the stricter classification

• Mops and buckets: ISO level dedication

• Different Mop for Walls, Floors, and Ceilings

• Double glove when cleaning floor drains

Cleaning and Disinfection: Techniques

34


• Don’t complicate procedures.

• Give yourself some leeway when starting up a cleaning program.

• Define daily, weekly, monthly, etc. Consider weekends and holidays.• Daily can mean “business day”

• Weekly – 7 ± 3 days

• Monthly – 30 days ± 10 days

• Triple clean definition• Different for everyone

• Satisfies monthly, weekly and daily requirements.

Cleaning SOP development



• Draft procedures to handle unplanned events• Non-routine cleaning – spills, excursions, disturbances

• Address room pressure excursion cleaning• If excursion occurs: “room will be two-stepped….if less

than 5 min. with traffic or less than one-hour without..”• If excursion exceeds above: “room will be two stepped….tagged

OOS, Management and Quality notified….”

• Room is monitored and placed back into service.


• Alternation of antimicrobial actives• Two disinfectants in sequence,

regular rotation, with sporicidal agent as needed

• One disinfectant daily, with sporicidalweekly or monthly

Key issue - Rotation


• Why do facilities rotate?• Good practice to ensure all microorganisms eliminated

• Inspectors from FDA, MHRA, etc. expect it

• Recommended by USP 37 <1072>, Aseptic Processing Guide, Annex 1, Orange Guide etc.

• Why is it important to rotate like or compatible chemicals?• Some disinfectants contain incompatible ingredients

• For example, cationic (Quats) and anionic (Phenolics) surfactants cannot be mixed




• Recent scientific discussion on need for rotating disinfectants

– Bacteria resistance?

• Chemical disinfectants

– have a higher biocidal activity and bacterial populations are generally very low in cleanrooms.

• Persistent bacterial populations due to:

– Ineffective cleaning methods

– Disinfectant cannot reach the bacteria

– Misuse of disinfectant


Antibiotic Action Sites


• Disinfectants have three mechanisms of action or ways that they affect or kill an organism• Cross-linking, coagulating, clumping

• Structure and function disruption

• Oxidation

• Chlorine – Oxidizing proteins, lipids, and carbohydrates

• Peroxide compounds – Oxidizes cell membrane and protein coat on spore causing them to collapse exposing core to lethal disinfectant

• Phenols – penetrates and disrupts proper functioning of the cell wall

How Disinfectants Work


• Alcohols cause cell proteins to clump and lose their function

Alcohol

Bacteria’s lipid membrane is held together because the hydrophobic part of the lipid sticks closely in the core

Reference: https://www.youtube.com/watch?v=j5jaHwuSuzo


• Quaternary Ammonium Compounds denatures proteins of the bacterial or fungal cell, causes substances to leak out of cell causing death

Quaternary Ammonium Compounds

Images reference: https://teamaquafix.com/biocides-food-processing/


“The antimicrobial agents typically employed in cleanrooms continue to be

effective because they have numerous effects on a number of aspects of

cellular physiology. That means multiple mutations would be required in a

short period of time (ex. 5 minutes) with exposure to low numbers of cells

typically found in a cleanroom to overcome their detrimental effects. As

such, resistance of a cell to agents used in a disinfection process would be

highly unlikely given the environmental conditions and low cell number.”

Microbial Resistance?

“…. Routinely used disinfectants should be effective against the

normal microbial vegetative flora recovered from the

facility.….” and “…Therefore, a sound disinfectant program

also includes a sporicidal agent, used according to a written

schedule and when environmental data suggest the presence of

sporeforming organisms.”

FDA guidance on aseptic manufacturing (2004)

Eudralex annex 1 (2008)

Given this knowledge, the pharmaceutical and biotechnology industries

have moved away from the rotation of two disinfecting agents. This

formerly common practice led to high residue levels and subordinate

efficacy performance. Today most firms use a system whereby a

disinfectant is rotated with a sporicide to more effectively reduce the

bioburden levels. The rotation of a disinfectant with a sporicide is

superior to the use of rotations of multiple disinfectants.”

JP guidance on aseptic manufacturing (2006)

“Cleaning and disinfection of facilities and

equipment A program for cleaning and

disinfecting facilities and equipment should be

established taking into account the potential

development of drug-resistant microorganisms.

The program should contain procedures for

screening and classifying bacterial isolates in each

manufacturing environment. »

(3) If selected disinfectants might have inferior

efficacy against microorganisms isolated from the

environment, the efficacy should be reevaluated and

the replacement with or alternate use of different

disinfectants should be considered and implemented, as

appropriate.

(4) If environmental monitoring data indicate or

suggest the presence of spore-forming bacteria or

fungi, suitable sporicides or fungicides should be

selected for disinfection.”

PIC/S

WHO

“61. The sanitation of clean areas is particularly important. They

should be cleaned thoroughly in accordance with a written

programme. Where disinfectants are used, more than one

type should be employed. Monitoring should be undertaken

regularly in order to detect the development of resistant strains.»

PDA TR70 cleaning and disinfection (2015)



• CURRENT USP <1072> DISINFECTANTS AND ANTISEPTICS

• “The development of microbial resistance to disinfectants is less likely to occur…, as disinfectants are more powerful biocidal agents than antibiotics”.

• “…they are normally applied in high concentrations against low populations of microorganisms usually not growing actively, so the selective pressure for the development of resistance is less profound”.

• “…frequently isolated microorganisms ….may be periodically subjected to use-dilution testing with the agents used in the disinfection program to confirm their susceptibility, as there are real differences among different species in resistance to the lethal effects of different sanitizers”.


Rotation

UPIP VAPI Working group on the Annex 1 revision


Draft Annex 1: Disinfection – 2007 versus 20XX

• More than one type of disinfecting agent should be employed (if Regs require) and should include the periodic use of a sporicidal agent

• Disinfectants should be shown to be effective for the duration of their in-use shelf-life taking into consideration appropriate contact time and the manner in and surfaces in which they are utilized

• Monitoring should be undertaken regularly in order to show the effectiveness of the disinfection program and to detect the development of resistant and/or spore forming strains.

• Cleaning programs should be effective in the removal of disinfectant residues.

Future Requirements for Disinfectant Rotation


•Do I need to rinse?

• 483 observations • Your firm does not always keep laminar flow hoods visually clean of

residue on HEPA filter surfaces and covering grates……I observed white and yellow residue on the HEPA filters…..and in areas up to approx. eight inches square on the filter…..

• I observed white particles on the floor of the clean room…approximately two to three millimeters square.

Cleaning and Disinfection:Rinsing


• Efficacy concerns:• Residues inhibiting Biocides

• Do residues support harbor microbial growth?

• Do residues inhibit preceding actives (rotational chemistries)?

• Functional Issues: • Sticky or opaque surfaces due to residues

• Aesthetic Issues: • Surfaces do not look “clean”

• Safety Issues:• Transfer (direct and indirect) from surfaces to manufactured drug

products or medical devices

• Personnel safety (slippery, tacky, sticky, etc.)

Cleaning and Disinfection:Rinsing


Typical Components in Disinfectants and what they look like

Component Residue Appearance

Water Water spots

SurfactantVery faint yellow to clear, gelatinous and sticky

White solid beads.

Chelants White or pinkish grey solid

Solvent No residue visible

Bases White to tan crystals

Acids Clear glaze with points of precipitates

Builders White powder

AntimicrobialsWhite to yellow

Brown, pinkish tanClear, gelatinous, sticky glaze

OxidantWater spots

No residue visible to white to tan powder or crystals from additives

Dyes Red, yellow, blue, color residues

Where does the residue come from?

• Compatibility issues• Different agents in rotation program

• Surface substrates

• Corrosion – attack of the impurities in the metal by chemical agent

• Deterioration by melting, softening, discoloration

• Drying – soft substrates such as Plexiglas, Kydex, Mipolam, epoxy.

• Staining

• Poor cleaning practices

• Drug product/process spills


Residue on Substrates


Residues on Substrates:

Desco Quartz Epoxy flooring system, 1.5 years, floors daily with phenolicsand 1/month application of sporicidalagent


Residue removal trial

Desco Quartz Epoxy flooring system in an ISO 8 gowning room marked off in a grid pattern for in-situ testing. Two-year-old floors are disinfected daily with phenolics and once per month application of sporicidal agent one time per month without rinsing.

Desco flooring after residue removal trial using:• WFI (lane 1),• Sterile oxidative detergent (lane 2)• Sterile neutral detergent (lane 3)


Residue removal trial

Application of sterile detergent Desco flooring after residue removal trial using sterile neutral detergent and rinse

Up-close before image

Up-close after image after cleaning with sterile neutral detergent


Residue removal – Microbial removal


Cleaning and Disinfection Best Practices

• How often to clean??? • Environmental cleaning frequency determined by:

• ISO Classification of area

• Activity level in area or use

• Environmental monitoring feedback

• Type of process being performed & equipment used

• Substrates

• Visual Observations

• Incorporate rinse/cleaning into cleaning and disinfection program

• Nondestructive mechanical action to remove contaminants and residue

• In addition to sanitizers, disinfectants or sporicides


• For clean rooms we still recommend• Disinfectant Rotation

• Phenols or Quats on monthly basis (never a phenol and a quat)

• Sterilant use routinely based on environmental data

• Rinsing on a routine basis• Not necessarily daily, but SOPs should

include weekly/monthly rinsing with IPA or water

• Evaluate cleanability of heavy residue

Contamination Control Program Recommendation

ContaminationControl Program

RinsabilityResidue

Compatibility

Efficacy

Disinfectant

Stability


13 Aug 2020

Cleanroom Control and Best Practices

Beth Kroeger

Technical Service Senior Manager

[email protected]

STERIS Life Sciences

Contamination Control Solutions

mailto:[email protected]

cleanroom control and best practices€¦ · cleaning and disinfection best practices •how often...

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