Improving and Optimizing your Viable Environmental Monitoring Program Jason Willett Midwest Manager
Where did this Specification Come From? U.S. Railroad tracks are 4 8.5 in width. That s a strange number Where did that specification come from? Because that was the specific width of the English railroad tracks and the U.S. Railroads were built by English settlers (expatriates). But, why is that so? Because, in England, the railroads were built by the same people who made the original tramcars. They had used the same gauges as the tramcars, which had that specific spacing. But, why is that so? Because, the people who built those tramcars used the same tools as they did for building wagons. And the wagon axles had that specific spacing. But, why is that so? Because, the the old, ragged, roads in England had wheel ruts on the sides of the road. This specific spacing was needed to keep the wagon wheels from maneuvering out of the ruts and breaking on the rough surfaces. But, why is that so? Because, the roads where originally built by Imperial Rome and those ruts were dug over many years by their chariots. That was the specific width of the chariot wheels. But, why is that so? The Roman chariots were specifically spaced just wide enough to accommodate the rear ends of two horses. 2
Where did this Specification Come From? So, the next time you are handed a specification and you are forced to wonder, What horse s ass came up with this?, you may be right! The response, We ve always done it this way, would be correct as well. 3
Let s Start From the Beginning
System Components Contamination Control is a proven system that prevents external contamination from entering controlled environments Environmental Monitoring tests that the system you designed is keeping your areas under control Cleaning and Disinfection corrects the inappropriate introduction of contamination 5
Areas of Control Within a Facility Personnel Equipment Disinfectants and cleaners Water HVAC Manufacturing Processes 6
What is Environmental Monitoring? It Is: Demonstrates Control of Aseptic Operations Identifies problems and trends in process and facilities Cleaning and Disinfection corrects the inappropriate introduction of contamination Data collection to support root cause analysis of excursions Is is Not: A test for final product release An analytical test Highly reproducible or recoverable Always constant Always linked to the smoking gun (cause and effect) Total picture of control 7
Global Regulations, Guidelines and Literature
Current Documentation United States United States Pharmacopeia 39, <1116> Microbiological Control and Monitoring of Aseptic Processing Environments FDA Guidance for Industry, Aseptic Processing, cgmp (October 2004) Europe GMP, Manufacture of Sterile Medicinal Products, Annex 1 (October 2009) World Health Organization Technical Report 961, Annex 6: GMPs for sterile pharmaceutical products (2011) Environmental Monitoring of Clean Rooms in Vaccine Manufacturing Facilities - Points to consider for manufacturers of human vaccines (November 2012) Japan Japanese Pharmacopoeia 16 (October 2011) - Microbiological Evaluation of Processing Areas for Sterile Pharmaceutical Products PDMA - Guidance on the Manufacture of Sterile Pharmaceutical Products by Aseptic Processing (October 2006) PIC/S Annex PE009-11: Guide GMP for Medicinal Products (March 2014) PDA Technical Report #13 (Revised) Fundamentals of an Environmental Monitoring Program (2014) 9
Determination of Sample Locations Document How to Determine Locations EU, WHO Based on formal risk analysis study and the results obtained during room classification and/or clean air devices PIC/S Based on a formal risk assessment. Consider data obtained during room classification and previous monitoring data Critical locations should be covered FDA Guidance Representative locations Locations posing the most microbiological risk to the product Locations where significant activity or product exposure occurs during production USP Locations considered should include those in proximity of the exposed product, containers, closures, and product contact surfaces Best selected with consideration of human activity during operations Evaluate impact of personnel movement and work within the area Entry points and areas within and around doors and airlocks The most likely route of contamination is airborne, so the samples most critical to risk assessment are those that relate to airborne contamination near exposed sterile materials. Japanese Guidance Japanese Pharmacopoeia Size of working area, scope of operations and process flow of materials and products Potential product contamination Not Specified 10
Viable Monitoring Frequency Document EU, WHO & PIC/S FDA Guidance USP Frequency The Grade A zone should be monitored at a frequency and sample size such that all interventions, transient events and any system deterioration would be captured and alarms triggered if alert limits are exceeded. Based upon the relationship to the operation performed Sample sizes should be sufficient to optimize detection of environmental contaminants at levels that might be expected in a given clean area Suggested Frequency of Sampling for Aseptic Processing Areas Sampling Area/Location Frequency of Sampling Clean Room/RABS Critical zone (ISO 5 or better) Active air sampling Each operational shift Surface monitoring At the end of the operation Aseptic area adjacent critical zone All sampling Each operating shift Other nonadjacent aseptic areas All sampling Once per day Isolators Critical zone (ISO 5 or better) Active air sampling Once per day Surface monitoring At the end of the operation Non-aseptic areas surrounding the isolator All sampling Once per Month 11
Viable Monitoring Frequency, continued Document Japanese Guidance Frequency To be determined in accordance to the air cleanliness level required for individual working areas. For example: Grade Microorganisms in Surfaces Particles air A & B Every working shift After completion of processing Continuously C & D Twice a week Twice a week Twice a week Japanese Pharmacopoeia Processing area Critical area (Grade A) Frequency of monitoring Each Shift Clean area adjacent to critical area (Grade B) Other clean areas (Grade C, D) Potential product/container contact areas Non-product/container contact areas Each Shift Twice a week Once a week 12
Viable Sampling Methods Document How to Determine Locations EU, WHO, PIC/S Air: Settle plates and volumetric air Surfaces: (Examples) swabs and contact plates FDA Guidance Air: Active device, optional settling plates Surfaces: Touch plates, swabs and contact plates USP Air: Choose from a variety of active samplers, settle plates Surface: contact plates or swabbing Japanese Guidance Air: Settle plates, impact and filtration methods Surfaces: contact plate or swabbing Japanese Pharmacopoeia Air: Settle plates, impact and filtration methods Surfaces: contact plates or swabs 13
Viable Limits EU, WHO, PIC/S & Japan Recommended Limits for Microbial Contamination* a Grade Air Sample - 90mm Settle Plates - CFU/m 3 CFU/4 Hours * b 55mm Contact Plates - CFU/Plate 5 Fingers Glove Print - CFU/Glove A <1 <1 <1 <1 B 10 5 5 5 C 100 50 25 - D 200 100 50 - a. These are average values. b. Individual settle plates may be exposed for less than 4 hours. 14
Viable Limits FDA (2004 GMP) TABLE 1 - Air Classifications a Clean Area Classification (0.5 μm particles/ft 3 ) ISO Designation b > 0.5 μm particles/m 3 Microbiological Active Air Action Levels c (CFU/m 3 ) Microbiological Settling Plates Action Levels c,d (diam. 90mm; CFU/4 hours) 100 5 3,520 1 e 1 e 1,000 6 35,200 7 3 10,000 7 352,000 10 5 100,000 8 3,520,000 100 50 a. All classifications based on data measured in the vicinity of exposed materials/articles during periods of activity. b. ISO 14644-1 designations provide uniform particle concentration values for cleanrooms in multiple industries. An ISO 5 particle concentration is equal to Class 100 and approximately equals EU Grade A. c. Values represent recommended levels of environmental quality. You may find it appropriate to establish alternate microbiological action levels due to the nature of the operation or method of analysis. d. The additional use of settling plates is optional. e. Samples from Class 100 (ISO 5) environments should normally yield no microbiological contaminants. 15
Viable Limits Japanese Pharmacopoeia Grade Table 4: Recommended limits for environmental microorganisms* 1 Airborne microorganisms* 2 (CFU/m 3 ) Minimum air sample (m 3 ) CFU on a surface Instruments/Facilities (CFU/24-30 cm 2 )* 3 Gloves A <1 0.5 <1 <1 B 10 0.5 5 5 C 100 0.2 25 - D 200 0.2 50 - * 1 Maximum acceptable average numbers of microorganisms under each condition. * 2 These values are by using a slit sampler or equivalent. * 3 Viable microbe cell number per contact plate (5.4-6.2 cm in diameter). When swabbing is used in sampling, the number of microorganisms is calculated per 25 cm 2. For gloves, usually, put their all fingers on the plate. 16
Viable Limits - United States Pharmacopoeia Microbial Control Parameters (Incident Rate): Suggested Initial Contamination Recovery Rates in Aseptic Environments* Room Classification Isolator/Closed RABS (ISO 5 or better) Active Air Sample (%) Settle Plate (9 cm) 4 h Exposure (%) Contact Plate or Swab (%) Glove or Garment (%) <0.1 <0.1 <0.1 <0.1 ISO 5 <1 <1 <1 <1 ISO 6 <3 <3 <3 <3 ISO 7 <5 <5 <5 <5 ISO 8 <10 <10 <10 <10 * All operators are aseptically gowned in these environments (with the exception of background environments for isolators). These recommendations do not apply to production areas for nonsterile products or other classified environments in which fully aseptic gowns are not donned. 17
Viable Monitoring of Compressed Gases Document EU, WHO & PIC/S How to Determine Locations Not Specified FDA Guidance USP Japanese Guidance Japanese Pharmacopoeia A compressed gas should be of appropriate purity (e.g., free from oil) and its microbiological and particle quality after filtration should be equal to or better than that of the air in the environment into which the gas is introduced. Microbial monitoring of manufacturing clean rooms, RABS, and isolators should include compressed gases, surfaces, room or enclosure air, and any other materials and equipment that might produce a risk of contamination. Environmental monitoring targets should also include compressed air or gas that comes in contact with the environment and equipment. Not Specified 18
ISO Regulations Misinterpretation? ISO 14644 Covers the classification of air cleanliness in cleanrooms and associated controlled environments exclusively in terms of concentration of airborne particles (has nothing to do with viable) ISO 14698 Establishes the principles and basic methodology of a formal system of biocontamination control (Formal System) for assessing and controlling biocontamination when cleanroom technology is applied for that purpose. Annex B: This annex describes a technique for determining the collection efficiency of samplers used for counting airborne microbes. Manufacturers or third-party testing organizations will usually perform this evaluation (this has no bearing on compressed gas/air). This is the unit s VALIDATION. ISO 8573-7 Specifies a test method for distinguishing viable, colony-forming, microbiological organisms from other solid particles which may be present in compressed air. A quantitative assessment MAY be made using the method given in Annex B (Slit to Agar) 19
Qualification of Viable Air Samplers An analysis provided by the manufacturer of the device (according to ISO 14698) An on-site comparative analysis (side by side) of the current device against the new device being implemented (e.g. Performance Qualification) Trending/performance of the newly implemented device (e.g. Performance Qualification) A study assuring that desiccation of the nutrient media does not occur during the length of the sample and subsequent incubation period. An Installation Qualification and Operational Qualification need to be completed prior to implementation. 20
Types of Viable Air Samplers
Portable Viable Air Samplers Sieve Impaction (many devices) Surface Air Sampler (SAS) Centrifugal Air Sampler (RCS) Slit to Agar Sampler (many devices) Sterilizable Microbial Atrium (SMA) Gelatin Filter Air Sampler (MD-80) What makes them all different? 22
Settle (Passive) Plates This method is still widely used as a simple and inexpensive way to qualitatively assess the environments over prolonged exposure times. Published data indicate that settling plates, when exposed for 4 5 hour periods, can provide a limit of detection for a suitable evaluation of the aseptic environment. Settling plates may be particularly useful in critical areas where active sampling could be intrusive and a hazard to the aseptic operation. One of the major drawbacks of most mechanical air samplers is the limited sample size of air being tested. 23
Isolator/RABS/Fixed Location Air Samplers Similar to their portable versions, however, these are designed so that the moving/active components (pumps, vacuums, electronics, etc.) are physically located outside of the barrier or room. It is generally NOT recommended to use a portable device inside an ISO 5 area for the following reasons: Portable devices are NOT able to be fully sterilized Portable devices generate particles Portable devices disrupt unidirectional airflow 24
Viable Compressed Gas Samplers 25
Rapid Technologies for Viable Air Sampling 26
Examples of Installed HMI/TouchScreens 27
Examples of Fixed Sample Locations 28
Ability to Monitor/Control from Outside ISO 5 29
Document Microorganism Incubation Conditions EU, WHO, PIC/S Not Described Not Described FDA Guidance USP Japanese Guidance Japanese Pharmacopoeia Media Selection & Incubation Fungi (yeast & molds) as well as bacteria Bacteria, yeast, and molds. Strict anaerobes not performed, but micro-aerophilic organisms may be sampled for, if warranted. Aerobic Bacteria, fungi (yeast & molds) & anaerobic bacteria Aerobes, Yeast, fungi & Anaerobes (not typically targeted) TABC: 30-35 C for 48-72 Hours TCYM: 20-25 C for 5 7 Days Time and incubation are set once appropriate media is determined. SCDM (TSA): 20-35 C for not less than 72 hours (longer for slow growers) The incubation condition of the medium should be suitable for growth of the target microorganisms (Huh?) Aerobes: 30-35 C for more than 5 days Fungi: 20-25 C for more than 5 days Anaerobes: 30-35 C for more than 5 days If SCDM is used for TVC, then 25-30 C for more than 5 days First incubate at 20-25 C temperature for 72 hours for fungal growth and then same plates transferred to 30-35 C for further 48 hours for bacterial growth. Favors Yeast & Molds First incubate at 30-35 C temperature for 48 hours for bacterial growth and then same plates transferred to 20-25 C for further 72 hours for fungal growth. Favors Gram Positive Cocci 30
Let s Apply What We Now Know You currently manufacture a single batch, spanning an 8 hour operating shift Air Sampler A - operates at 180 L/Min and samples a cubic meter in 5.5 minutes. Air Sampler B - operates at 100 L/Min and can utilize an interval sampling mode. Air Sampler C - operates at 28.3 L/Min and can actively sample for 3-4 hours on a single plate. How many plates do you need to sufficiently cover your production, how many will you bring into the APA at a time? How many interventions have you planned for plate changes (do you stop)? How many plates will you utilize (and subsequently have to review)? What did you potentially miss between samples/sampling? 31
What We May Have Missed Multiple air samples can lead to false positives (operator/analyst contamination), mishandled plates & more laboratory work. Non-sampling time between plate changes (or during passive state of of interval sampling) can result in missed contamination events. 32
Establishing and Executing Alert & Action Levels
Establishing Alert & Action Levels Document EU, WHO & PIC/S How to Determine Locations Appropriate alert and action limits should be set for the results of particulate and microbiological monitoring. If these limits are exceeded operating procedures should prescribe corrective action. FDA Guidance Based on : The relationship of the sampled location to the operation The need to maintain adequate microbiological control throughout the entire sterile manufacturing facility Historical databases, media fills, cleanroom qualification and sanitization studies USP Alert Levels: Not Specified Action levels: Should be based on empirical process capability Japanese Guidance Action level by referring to data contained in monitoring frequency table. Alert level based on results of a PQ test Japanese Pharmacopoeia An environmental control program document is prepared for each area used for the processing of sterile pharmaceutical products. The procedures in the document include: 5) alert and action levels, and 6) actions to be taken when specified levels are exceeded. 34
Establishing Alert & Action Levels 100 80 Number 60 40 20 0 Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec G+C G+R G-R YST Mold 35
Executing Alert & Action Levels Regulations require: Immediate follow-up Identification of microorganisms Corrective actions/preventive actions (CAPAS) if appropriate Indicates potential deviation from normal operational conditions Document and follow-up Additional or modified sample plan Designed to allow appropriate capability of reaction before reaching Action level No actions required 36
Identification of Microorganisms
Identification of Microorganisms Document Microorganism EU, WHO, PIC/S, Japanese GMP & Japanese Pharmacopoeia Not Described FDA Guidance Monitoring of critical and immediately surrounding clean areas as well as personnel should include routine identification of microorganisms to the species (or, where appropriate, genus) level Establishing an adequate program for differentiating microorganisms in the lesser-controlled environments, such as Class 100,000 (ISO 8), can often be instrumental in detecting such trends (migration of microorganisms into clean areas) At minimum, the program should require species (or, where appropriate, genus) identification of microorganisms in these ancillary environments at frequent intervals to establish a valid, current database of contaminants present in the facility during processing (and to demonstrate that cleaning and sanitization procedures continue to be effective) USP A successful environmental control program includes an appropriate level of identification of the flora obtained by sampling. A knowledge of the flora in controlled environments aids in determining the usual microbial flora anticipated for the facility and in evaluating the effectiveness of the cleaning and sanitization procedures, methods, agents and recovery methods. The information gathered by an identification program can be useful in the investigation of the source of contamination, especially when recommended detection frequencies are exceeded. 38
Identification of Microorganisms Corynebacterium tuberculostearicum 2.9% Bacillus cereus, 4.3% Staphylococcus hominis, 5.6% Staphylococcus epidermidis, 8.7% Staphylococcus luteus, 9.6% Staphylococcus capitis, 2.8% Bacillus pumilus, 2.7% Staphylococcus warneri, 2.5% Bacillus subtilis, 2.4% Bacillus licheniformis, 0.9% Staphylococcus haemolyticus, 2.2% Ralstonia pikettii, 1.7% Paenubacillus glucanolyticus, 1.5% Bacillus megaterium, 1.5% Proprionibacterium acnes, 1.0% Source: Bacteria Most Often Submitted for Identification Testing During 2010, Barry A. Friedman, posted May 17, 2011 39
Distribution of Microorganisms People Air/Soil Water/Liquid 40
Identification of Microorganisms Biochemical (Selective Assays) Gram Stain Catalase Test Coagulase Test Oxidase Test Indole Test 41
Identification of Microorganisms Phenotypic Identification (incorporates reactions to different chemicals or different biochemical markers) BD BBL Enterotube II BD BBL Crystal ID Panels 42 BioMerieux API Strips 42
Identification of Microorganisms Phenotypic Identification - continued Biolog GEN III OmniLog ID System BioMerieux Vitek 2 System 43
Identification of Microorganisms Genotypic Identification (based on gene sequencing, 16S rrna gene) Thermo Fisher Scientific MicroSeq DuPont RiboPrinter 44
Identification of Microorganisms Proteotypic (MALDI-TOF): Spectral analysis based on ribosomal protein expression BioMerieux Vitek MS System Bruker MALDI Biotyper 45
Identification of Microorganisms Fatty Acid Analysis/GC Optical/Raman Spectroscopy Midi Sherlock Q-FAME Batelle REBS 46
Viable Monitoring of Surfaces
Different Surface Types Irregular Surfaces Residues Particulate and Viable particulates All may complicate monitoring and disinfection 48
Sampling Methods Contact plates (RODAC) and swabs Contact Plate and Swab recovery studies Contact plates can offer better recovery than swabs and utilized more often (where surface and location permits) Flocked swabs offer better recovery than spun/cotton swabs due to physical composition Rinse samples can be performed on interior surfaces of kettles and tanks using membrane filtration and sterile water (or Ringer s, etc). Sampling done on equipment, work surfaces, floors, walls and product contact surfaces after production is complete 49
Viable Monitoring of Personnel
Viable Monitoring of Personnel Personnel Monitoring: Routine during production Gloved fingers: underside where contaminants are most likely Critical gown sites for aseptic operations Contact plate testing of people leaving/exiting area Routine failures on a person requires a corrective action (re-training and/or re-qualification) Gown Training Certification Surface sampling of gown at key garment locations Personnel MUST participate in a media challenge at least once annually if they will be filling aseptic product 51
Gowning Qualification Set strict limits for qualification Remove those who fail and retrain them Testing needs to be done by Quality. They should not qualify themselves Determine Qualification Test locations (8) Hood, goggles, mask, zipper, sleeves, hands (both), thighs Re-qualify on an annual basis 52
Viable Monitoring of Water Systems
Water Sampler Collection & Hold Time Sample Collection and Testing After validation of controlled state Collect samples consistent with manufacturing practices Sample locations Test as soon as possible after collection Samples should be taken from holding and distribution system to assess microbiological quality for its intended use If manufacturing flushes use points prior to use, collect samples using same flush cycle Sample through hoses (not directly from the tap) if manufacturing requires use of hoses Chosen to demonstrate microbiological quality throughout the distribution system If not possible, refrigerate samples at 2-8 C upon receipt in the laboratory Time between collection and examination 24 hours. 54
Microbial Monitoring/Testing of Water Azbil BioVigilant IMD-W Manual Filtration EMD Millipore Milliflex Mettler-Toledo 7000RMS EMD Millipore Milliflex Quantum BioMerieux ScanRDI 55
Cleaning & Disinfection
Chemical Selection Sanitizers Reduce some level of microbial contamination; least effective agents Examples: Isopropyl alcohol (70% IPA), and Ethyl alcohol (62% EtOH) Disinfectants Reduce higher levels of vegetative microorganisms than sanitizers depending on the strength and contact time Examples: Quaternary Ammonium compounds, Phenolics Sporicides Effective against all microorganisms provided the wetted contact time is achieved Examples: Sodium hypochlorite, Hydrogen peroxide/peracetic acid, Hydrogen peroxide (6% or greater) 57
Cleaning & Disinfection Program Cleaning and Disinfection address mistakes in the Contamination Control System If done appropriately, it cleans particulates and microbes that have been allowed to enter the controlled area It does not have any preventive capabilities. It cleans or kills what exists but does not prevent the introduction of particulates and microbes, nor their growth. Too many times disinfection is considered a preventive measure. It is not. 58
Cleaning versus Disinfection Proper cleaning is critical for effective disinfection Would you disinfect without brushing or cleaning the surface? 59
The System is a Cycle 4. Assure appropriate application and test again 1. Control What Enters the Environment 3. Validate disinfectants and disinfection system in conjunction with cleaning and disinfection SOP s 2. Test the environment and address the contamination 60
In Conclusion Understand Environmental Monitoring because it can be a very unique and daunting task The guidelines are just that, guidelines. Establish a well-justified and validated program that works for you and meets the various global recommendations A good EM program also controls the introduction of contamination Cleaning and disinfection are critical aspects to contamination control Monitoring systems should not affect final product 61
Any Questions? 62