Risk Based Study of Sample Spot Fixing for Conducting Environmental Monitoring Test in Classified Areas of Pharmaceutical Industry.

Environmental monitoring is a study Programme designed to study load of Viable Microorganism as well as Non-Viable Particles present in (controlled and classified area) environment. The use of risk assessment approaches is an important current Good Manufacturing Practice (cGMP) tool in microbiological environmental monitoring Programme (1). The prime goal of environmental monitoring of cleanrooms are to regulate the numbers of airborne viable and non-viable particles within defined limits, predict the risk to the environment and regularly estimate the efficacy of cleaning and disinfecting processes (2). Pharmaceutical manufacturing comprises a complicated, multi-phase processing system associated with major risks of microbial contamination through diverse sources. The product quality is greatly influenced by microbial contamination in several processing steps. There are two types of pharmaceutical products currently being manufactured are sterile and non-sterile products (3) Environment plays key role in microbial access in Pharmaceutical and biological products. The quality of product depends on environmental controls in manufacturing area. To obtain a pharmaceutical product free of contamination, you need an adequate environmental monitoring system. The system includes identification, testing and removal of bioburden to ensure the quality of product (4). The frequency of monitoring ensures contamination free environment maintenance. Environmental monitoring is a programed scheme to explain the regulation of viable and non-viable particles in critical sites (5).

Clean Rooms

Clean Area Separation is a segregation area which is required between classified area and external environment to prevent cross contamination. According to WHO classification, it as classified as Class A, Class B,Class C and Class D and according to ISO Classification ,it is classified as ISO 5,ISO 6, ISO 7 and ISO 8 (6).

Risk Assessment approach

These risk assessment approaches are not only concerned with selecting environmental monitoring locations. They integrate the environmental monitoring system with a complete review of operations within the cleanroom to ensure those facilities, operations, and practices are also satisfactory (12,13,14).

Risk Assessment Tools

A risk assessment procedure is required to determine locations for environmental monitoring. Such risk-based approaches are recommended in ISO 14698 and regulatory authorities are increasingly asking drug manufacturers about this subject. Risk-based approaches include Failure Mode and Effects Analysis (FMEA), Fault Tree Analysis (FTA), and Hazard Analysis and Critical Control Points (HACCP), Quantitative Microbiological Risk Assessment (QMRA). (10,11).

Objective of Study

1.The scope of this study, is fixing the sampling spot based on risk assessment for performing environmental monitoring.

2.To correlate the result of Finger dab test and non-viable count report with the sanitization of area and cleanliness of Personnel working in the area.

MATERIAL AND METHODOLOGY

The research was conducted in Quest Pharmaceuticals Pvt., Ltd. The company has been manufacturing non sterile dosages forms (Tablet, Capsules, Semisolids and Soft gel capsules) and distributing its products throughout Nepal.

The research was designed on the basis of statistical sampling model as ISO 14644-1 for airborne particles quantification in cleanroom (Class D) and data was taken from third party report, Qual Tech Services, Total Aerobic Microbial Count based on risk based approach and Finger dab test to understand the cleanliness standard of personnel working in the areas. Clean rooms were divided into five groups as Dispensing Room-I, Granulation Room-I, Compression room-IV, Tablet Inspection room and Blister Packing room based on different Production activities done in the plant and further the individual rooms were divided in to five different sample spot as

a. Near machine where production activity is done

b. Area where the man movement is maximum

c. Difficult to clean area

d. Near Return Air Loop

e. Near Drainage area

 based on risk of contamination of products during activities to find the Hot spot areas for higher microbial load.

Study Module for Total Aerobic Microbial Count(TAMC)

Study Period: 22/06/2022- 24/06/2022 (3 days)

Media: Soyabean Casein Digest Agar and Sabouraud Dextrose Agar

Method: Settle Plate Method (90 mm Petriplates)

Limit: Not more than 100 cfu/plate (4 hours Exposure) 25 plates of Soyabean Casein Digest Agar and 25 plates of Sabouraud Dextrose Agar was exposed for 4 hours in all sampling points. The study was conducted for three successive days. Sterile Culture media plates were exposed on petriplates stand on respective sampling sites in each room for four hours. After completion of exposure time, the petriplates were aseptically transported to Microbiology laboratory. Soyabean Casein Digest Agar plates were incubated in incubator for 72 hours and Sabouraud Dextrose Agar petriplates were incubated in BOD Incubator for 5-7 days. After the completion of incubation period, colony on Petriplates were counted using Colony Counter and result were interpreted.

Finger Dab Test

Finger Dab test was performed in the Dispensing Room-I, Granulation Room-I, Compression room-IV, Tablet Inspection room and Blister Packing room from the personnel working in the respective areas. In-house Limit of Finger Dab test was set to 100 cfu as the production area was operating in class D area and non-sterile solid dosages was formulated in the area. Personnel working in areas were selected randomly and each of them was evaluated by finger DAB test. All the fingers including thumb of personnel’s gloves were gently imprinted and impression of all these workers were obtained on labeled Perti plates containing Soyabean casein digest agar. All the plates were incubated for at 35°C for 72 hours and results were recorded.

Non-Viable Count

Report of Third party validation is taken and is performed by Qual Tech Services as approved vendor for Clean room validation in the plant.

RESULT AND DISCUSSION

1. Result of Viable Microbial Count

1.Dispensing Room-I

2.Granulation Room-I

3.Compression Room-IV

4.Tablet Inspection Room

5. Blister Packing Room-II

After reviewing the results obtained for different locations of five different rooms, it can be concluded that the results are under to the defined limits. The worst case location is “Near Return Air Loop”. The TAMC and TYMC, was observed high at sampling Location “Near Return Air Loop” in comparison to the other sampling Locations for three consecutive days.

2.Result of Finger Dab Test

In House Limit: Not more than 100 cfu/5 finger

Altogether tested 10 samples in 5 different rooms in the working areas, the cleanliness status of personnel was found satisfactory according to In-house limit. The result of right hand (five finger) and Left hand (five finger) was in the specified in house limit for finger dab test.

Result of Non-Viable Count (Rest Condition).