Randy Hutt, Ph.D. 2016Associate Director Sterility Assurance and MicrobiologyRhutt@luitpold.com 631-924-4000 Ext. 632

Part IHow to Handle Excursions in Environmental Monitoring

(EM) and Personnel Monitoring (PM) in an

Aseptic Processing Plant

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The information provided in this presentation only represents my personal views and not the views of LuitpoldPharmaceutical, Inc.

› Randy Hutt, Ph.D.

› Associate Director Sterility Assurance and Microbiology

› Luitpold Pharmaceuticals, Inc.

› AUG2016

Company Disclaimer

› EM excursions are environmental results that exceed established alert or action levels.

› EM excursions should not be the sole determining factor used to release or reject batches without further evaluation.

› Excursions leading to trends should be used to help determine what corrective/preventive actions are needed for microbial control.

› EM excursions are referred to as being “Out-of-Limit” results not “Out-of-Specification” results. Examples of OOSs would be bioburden, endotoxin, and sterility.

Introduction

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REFERENCES

1. USP General Chapter (current Version 38) <1116> Microbiological Evaluation of Clean Rooms and Other Controlled Environments

2. Volume 4- EU Guidelines to Good Manufacturing Practice Medicinal Products for Human and Veterinary Use- Annex 1 Manufacture of Sterile Medicinal Products (corrected version)-Effective March 1, 2009

3. Sterile Drug Products Produced by Aseptic Processing- Current Good Manufacturing Practice 2004

4. PDA Technical Report No. 13 (Revised) Fundamentals of a Microbiological Environmental Monitoring Program

5. International Organization for Standardization (ISO) 14644 Cleanrooms and Associated Controlled Environments

6. WHO Technical Report Series, No. 961, 2011- Annex 6 WHO good manufacturing practices for sterile pharmaceutical products

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› Batches rejected due to microbial contamination Gram-negative organisms on the floor:– Microorganism identified from a contact plate.– An investigation was performed.

› Since the organism is a Gram-negative, it was most likely a water borne organism.

› Inspection of the aseptic area was performed and pooling water was identified as the most likely source of contamination.

› The suspected source was sampled using RODAC plates and swabs.

› Results confirmed suspicions.

– Root cause was confirmed and as a follow up a CAPA was created. › The CAPA included a fix to the slope of piping of jacket water towards non-sterile

manufacturing area and corrected lack of air-breaks to drain.

No more Gram-negative organisms were isolated.

War Stories - Gram-negative organisms

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› Found Gram-positive spore-formers around aseptic area in manufacturing plant situated in a rural area.– An investigation was performed.

› The organism was identified as a spore former.

› Usual sources are likely found in soil, cardboard boxes, wooden pallets, and raw materials.

› Due to the rural location of the plant, many employees lived near/on farms.

› Root cause identified the most probable source to be personnel tracking into the aseptic area.

– CAPA was initiated to create procedural controls to provide dedicated shoes for employees entering the aseptic processing area.

This greatly reduced finding these organisms in the aseptic area.

War Stories - Gram-positive spore-formers

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› Is the testing of 20 or even 40 samples as required in a non-destructive sterility test sufficient to provide assurance of sterility?

› This test is simply a probability function, because a heavy amount of contamination in a batch of 100,000 units would need to be present to detect it in only 20-40 units.

Therefore, it is critical to control the environment with Engineering Controls (# of air changes per hour/Laminar Air Flow, etc.) and to test the Environment and the Personnel for possible contamination and provide assurance of sterility.

Sterility Testing

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› Types of Environmental Monitoring include

• As built / commissioning EMPQ

• At rest testing (Static)

• In-Operation testing (Dynamic)

Environmental Monitoring

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› Types of sampling include:

• Viable – evaluating for living organisms • Active

• Passive

• Surface

• Non-viable – Total airborne particulates• Continuous

• Total particulate

Environmental Monitoring

Performance of Non-Viable Monitoring:- There are 2 types of non-viable methods that are used in

an aseptic filling facility and monitor both 0.5 µm and 5.0 µm.- Continuous real-time monitoring is performed using

fixed probes, for example, Particle Monitoring System (PMS).

- Total particulate monitoring is performed using portable Lasair units. Portable Lasair units are placed at specified locations where sampling is needed.

It is helpful to use hand-held units to discern where particulates are being generated in the case of any alarms due to high particulate levels.

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› The purpose for Non-Viable sampling is to:

• Verify room classification when an area is new.

• Evaluate air-cleanliness, ensuring compliance, during normal operations.

› “In Grade A and B zones, the monitoring of the ≥ 5.0 µm particle concentration count takes on a particular significance as it is an important diagnostic tool for early detection of failure. The occasional indication of ≥ 5.0 µm particle counts may be false counts due to electronic noise, stray light, coincidence, etc. However, consecutive or regular counting of low levels is an indicator of a possible contamination event and should be investigated. Such events may indicate early failure of the HVAC system, filling equipment failure or may also be diagnostic of poor practices during machine set up and routine operations.”

Non-Viable Testing

Viable Monitoring:

a) Active air sampling - Using SAS air sampler - QuantitativeAssesses air quality in the controlled environment specified volume > 1 m3. Active air sampling is performed by using SAS air samplers, RCS, or other sampler which pulls in a specific quantity of air for a specific time.

b) Passive air sampling - Using Settle plates (90mm) - QuantitativeAssesses air quality in the controlled environment over prolonged exposure times. Settle plates are placed on the designated surface for a specific length of time. (e.g., Filling machine inside a fill room or on settle plate stands). Per Annex 1, individual settle plates may be exposed for < 4 hours.

c) Surface sampling - Using RODAC plates (55mm) - Quantitative or Swabs - Qualitative Checks cleaning effectiveness and environmental control.

Personnel Monitoring (PM) sampling (Using RODAC plates) - QuantitativeEvaluates aseptic technique and possible impact to aseptic controls. RODAC plates are touched/pressed against specified locations on aseptically gowned personnel (e.g., gloved fingerprints, forearms, and chest).

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Room Classif.-VP

AA SP CP or Swab

Glove/Garment

ISO/Rabs < 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%

US Requirements/Limits-NVP for classification; VP for EM and PM- Monthly Contamination Recovery Rate

ISO 14644-1Class-NVP

>=0.5micron/m3

>=5 micron/m3

ISO 5 3,520 29

ISO 6 35,200 293

ISO 7 352,000 2,930

ISO 8 3,520,000 29,300

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NVP Max. permitted # particles/m³ = or greater than tabulated size

At Rest In Operation

Grade 0.5 µm 5.0µm 0.5 µm 5.0 µm

A 3,520 20 3,520 20

B 3,520 29 352,000 29

C 352,000 2,900 3,520,000 29,000

D 3,520,000 29,000 Not Defined

Not Defined

EU Requirements

Grade- VP AA SP CP Glove

A <1 <1 <1 <1

B 10 5 5 5

C 100 50 25 -

D 200 100 50 -

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Alert Levels:

Microbiological levels, specified in the SOPs, which when exceeded should result in an investigation to ensure that the process is still within control. Alert levels are specific for a given facility and are established on the basis of a baseline developed under an EMP. These alert levels can be modified depending on the trend or statistical analysis performed in the monitoring program.

Note that Alert levels are always lower than action levels.

Action Levels:

Microbiological levels in the controlled environment specified in the SOPs , which when exceeded should trigger an investigation and a corrective action based on the investigation.

Alert and Action Levels/Limits for Viables

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Are statistically determined by history and trend analysis…usually mean + 2 SD;

Lower than action level and determined by 100 data points (or a year of data sometimes).

An alert may or may not trigger an investigation; define in SOPs.

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How to determine Alert levels:

Alert Levels

• The Contamination Recovery rate is the rate at which environmental samples are found to contain any level of contamination. For example an incident rate of 1% would mean that only 1% of the samples taken have any contamination regardless of colony number. • This rate should be based on actual monitoring data by room,

sample type (e.g., AA, CP, SP or Personnel) and should be retabulated monthly.

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Contamination Recovery Rates per USP

Percent Recovery Rate =

Total number of Samples with recovery(100)

Total number of samples

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• Most probable number method, otherwise known as the method of Poisson distribution with many zeroes, and is a method of getting quantitative data from negative incidents. “Recommendation from USP <1116> on Contamination Recovery Rates”, is an article by Scott Sutton, where he discusses the MPN approach as a method given the Poisson distribution of the data and very low numbers of samples.

Most Probable Number Method (MPN)

(MPN) = 2.303 * Log(10)Total number of sampling

Number of sampling yielding zeroes

For Viable Recoveries-Note: Limit of detection on a plate= 1 CFU (EU requirement for investigation in Grade A)

Limit of quantification = 25 CFU per plate (e.g., 25-250 CFU/plate)

• Therefore, EM Alert and Action Levels of 1-10 CFU are of questionable accuracy

(background noise). A significant excursion is 15 CFU (per USP <1116>). How to handle 10-15 CFU??

• The USP monthly recovery rate was developed and is appropriate to use to look for trends both prior to a specific recovery in investigations and monthly for management Quarterly Trend Reports or Quality Council meetings. Discuss trend definitions…

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When to write an investigation- USP vs. EU

Investigation Methods:

Fishbone method: Method, Materials, Equipment, Environment, Personnel, Process

5 Whys: Keep asking “Why” 5 times to drill down to details

Kepner-Tragoe Analytical Trouble-Shooting: Compare differences of where the problem is and where it is not

Internal Investigation for Viable Recoveries

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Use of Risk Assessment Methods to determine Severity of Microbial contamination should it have occurred, the Likelihoodof it occurring, the ability to Detect it.

Of course, severity for sterile production is defined as product contamination, so it is deemed as serious.

Your table for RA can include Risk Factors (and a risk rate of High, Medium or Low or a Quantitative Assessment) and Mitigating factors for each risk factor. At the end, draw a conclusion of risk to SISPQ of the product.

Risk Assessment as part of an EM/PM Investigation

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Example Fishbone Diagram of Cause and Effect for Potential Causes of Contaminated Product-http://www.microbiologynetwork.com/contamination_control.asp

› Identify non-conforming sample (with excursion) by location, type of sample (EM- AA/SP/CP vs. PM), count, date and time taken.

› Identify contaminating organism to allow for speculation of possible source; Review history of same organism in facility. Is there a trend?

› Ensure media is not contaminated and Growth Promotion Tests were acceptable.

› Ensure incubators within calibration.

› Ensure EM personnel were adequately trained.

QC Microbiology Data for Investigation

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› Can be led by QA or performed by Production personnel.

› Details to be evaluated and documented for EM/PM excursions:– Check Sanitization just prior to the event; proper use-dilutions/approved

disinfectants.

– Check Training up to date for Mechanics/Operators; gown-qualified

– Verify Pressure differentials.

– Verify Temperature/% Relative Humidity within limits

– Ensure no Non-Viable particulate alarms

– Check filter maintenance/certification.

– Evaluate if any recent construction or equipment maintenance in area.

– Ask: Are other batches affected?

Manufacturing Investigation

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› Issues to evaluate and document:

– Disinfectant preparation-dilutions, expiration date.

– Sanitization procedures for materials passing in to Aseptic Area and for disinfection of rooms.

– Disinfectant Efficacy Studies.

– Actions taken in the room during filling of batch (if after filling completion)

Contact Plates - Static monitoring or taken after completion of filling

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Contact plates resulting in Personnel Excursions:

• Type of organism is critical.

• Are people sanitizing their hands with alcohol sufficiently? May wish to retrain staff if they have repeated “hits” and 3 consecutive hits may warrant disqualification until retrained in aseptic technique, if video reviews show unacceptable aseptic techniques.

Note that organisms pass through pores in gowns- Need time limit in aseptic area.

Contact Plates- Continued

› Root Cause Assessment- based on all the facts provided by QC and Production– Evaluate people, material flow, training, sanitization, and etc.– Determine most probable root cause, if possible. It may not be for 1 CFU.

› Risk and Impact Assessment-– Could the organism have contaminated a batch of product? – Is the product aseptically filled or terminally-sterilized?– If the organism is Gram-negative or a mold, is there concern for endotoxin

or allergens, respectively?

Determine Corrective Actions/Preventive Actions (CAPAs) (recommendations can be obtained from QC and Production)

Investigation Summary- May be a QA role

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› The severity of the risk is highest when product, containers, closures or product contact surfaces can be contaminated.

› Once the Severity is established, this is considered in relation to the likelihood (Probability) of occurrence and the ability to detect (Detection) the occurrence. If these issues result in a high risk, it may lead to batch rejection, even if root cause cannot be determined.

› Controls that serve to reduce the likelihood of the occurrence or increase the ability to detect the occurrence are also considered as follow-up action.

› High risk activities require corrective (remedial) actions.

Risk Assessment

› Corrective Action: Immediate actions taken after identification of root cause

› Preventive Actions: Prediction of problem and actions taken to avoid the occurrence through self-initiated actions and analysis related with processes/products.

› CAPA Effectiveness should be checked by QA or Compliance

CAPAs

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› QA Batch Disposition- Acceptable for further processing or Reject?

› Be sure to evaluate all affected batches.

Batch Disposition

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› People are the greatest source of contamination in an aseptic area…so in designing/re-designing a facility, keep that in mind.

› Isolators in production are very helpful or Restricted Area Barriers (RABs).

› Blow/Fill/Seal is another way to go.

› Any questions?

Conclusion