MICROBIOLOGICAL ANANLYSIS OF PHARMACEUITCAL WATER SAMPLES AND ENVIORMENTAL MONITORING

Under the Guidance : Presented By: MR. Ajeet Kumar Anand Amrit

rajMicrobiologist “(QC lab) M.Sc Industrial Biotechnology

Intas pharmaceutical Ltd D.I.B.N.S College Dehradun

AIMS AND OBJECTIVES

MICROBIOLOGICAL ANANLYSIS OF PHARMACEUITCAL WATER SAMPLES AND ENVIORMENTAL MONITORING

Objectives: Microbial Limit Test: This test is performed to estimate

the number of viable aerobic microorganism present in water sample.

Environmental monitoring: To estimate the number of viable aerobic microorganisms present in environment and to lay down procedure of microbial monitoring in micro lab.

INTRODUCTION

o What is the need of microbial analysis of water and Environmental monitoring?

Introduction about :

Microbial analysis of water Microbial limiting test

Environmental monitoring

LIST OF MEDIAS AND INSTRUMENTS USED IN MICROBIOLOGY LAB

Medias used in Micro-Lab: Soybean casein digest agar Sabouraud dextrose agar R2A Cetramide agar Macconkey broth Macconkey agar Brilliant green agar Xylose lysine deoxycholate agar Bismuth sulphate agar Triple sugar iron agar Violet red bile agar Maintol salt agar Baired parker agar Soybean cacine digest medium Buffer peptone water Peptone water Tetrathionate bile brilliant broth

•Instruments:•Autoclave•Hot air oven•Microwave oven•Laminar air flow(laf)•Bacterial incubator•ph meter•Electronic balance•Refrigerator•Water bath•Vortex machine•Colony counter•Magnetic stirrer with hot plate•Air sampler•Heating block

ANALYSIS OF PHARMACEUTICAL WATER:

Water is one of the most widely and abundantly used substances in pharmaceutical manufacturing. It is required for a variety of purpose ranging from manufacturing processes to the preparation of the final dosage forms.

TYPES OF WATER

Raw Water

Potable Water

Purified Water

ANALYSIS OF PHARMACEUTICAL WATER: Microbial limiting test - Estimation of the number of viable

aerobic microorganism present and for detecting the presence of designated microbial species in purified water.

Two methods:

Total microbial count  Pour plate method Membrane filtration

Pathogen testing

Test for Escherichia coli Test for Salmonella Test for Pseudomonas aeruginosa Test for Staphylococcus aureus

ANALYSIS OF PHARMACEUTICAL WATER: MICROBIOLOGY LAB TESTING Sample Microbiology lab testing 

Total bacterial count Pathogen testing Total fungal count

Pre -enrichment

broth E.coli Enrichment broth

P. aeruginosa Salmonella S. aureus

ENVIRONMENTAL MONITORING -

To estimate the number of viable aerobic microorganisms present in environment and to lay down procedure of microbial monitoring in production and warehouse.Methods of E.M

By Settle plate. By Active Air sampler. By Swab. By Contact plate.

RESULTS AND DISCUSSION :

Microbial Limit Test: shows the total viable aerobic count in a water.

Table 1: Showing total aerobic bacterial count on soyabean caesin digest Agar.

Table 2 :Showing total aerobic fungal count on Sabouraud Dextrose Agar.

Pictures : Identification of selective bacteria.

Samples Number of colony forming

units(CFU)

Test for E.coli, S.aureus, P.aeruginosa, Salmonella,

Coliforms

Sample A <10 CFU Negative( E. coli, S. aureus, P.aeruginosa,

Salmonella, Coliforms absent)

Sample B <10 CFU Negative( E. coli, S. aureus, P.aeruginosa,

Salmonella, Coliforms absent)

Sample C <10 CFU Negative( E. coli, S. aureus, P.aeruginosa,

Salmonella, Coliforms absent)

Sample D <10 CFU Negative( E. coli, S. aureus, P.aeruginosa,

Salmonella, Coliforms absent)

Sample E <10 CFU Negative( E. coli, S. aureus, P.aeruginosa,

Salmonella, Coliforms absent)

Samples Number of colony forming units

(CFU

Test for Candida albicans, Aspergillius niger

Sample A <10 CFU Negative(Candida albicans, Aspergillius niger)

Sample B <10 CFU Negative(Candida albicans, Aspergillius niger)

Sample C <10 CFU Negative(Candida albicans, Aspergillius niger)

Sample D <10 CFU Negative(Candida albicans, Aspergillius niger)

Sample E <10 CFU Negative(Candida albicans, Aspergillius niger)

RESULTS AND DISCUSSION

Environmental monitoring: Table 1- showing results of environmental monitoring by settle

plate technique. Table 2- showing results of environmental monitoring by air

sampling technique. Table 3- showing results of environmental monitoring by

contact plate technique. Table 4- showing results of environmental monitoring by swab

method.

Area 1 Total bacterial count ( TBC )

Total fungal count ( TFC )

Air lock 1 60 CFU/Plate NIL

Air lock 1 75 CFU/Plate NIL

Passage 56 CFU/Plate NIL

Media preparation room

40 CFU/Plate NIL

Incubation room 28 CFU/Plate NIL

Buffer zone 20 CFU/Plate NIL

MLT room 05 CFU/Plate NIL

Washing Area 45 CFU/Plate NIL

Laminar air flow 01 CFU/Plate NIL

Area 2 Total bacterial count ( TBC )

Total fungal count ( TFC )

Air lock 1 50 CFU/m³ NIL

Air lock 1 40 CFU/m³ NIL

Passage 35 CFU/m³ NIL

Media preparation room

20 CFU/m³ NIL

Incubation room 05 CFU/m³ NIL

Buffer zone 18 CFU/m³ NIL

MLT room 02 CFU/m³ NIL

Washing Area 25 CFU/m³ NIL

NO OF PERSONS 3

Total bacterial count ( TBC )

Total fungal count ( TFC )

Person 1 65 CFU/Plate NIL

Person 2 47 CFU/Plate NIL

Area 4 Total bacterial count ( TBC )

Total fungal count ( TFC )

Refrigerator surface 45 CFU/Plate NIL

Floor of MPR room 55 CFU/Plate NIL

Near weighing balance 30 CFU/Plate NIL

Working table 43 CFU/Plate NIL

Buffer zone floor 25 CFU/Plate NIL

Laminar air flow surface

00 CFU/Plate NIL

Incubator door 03 CFU/Plate NIL

CONCLUSION

Microbial limiting test is very important for pharmaceutical water because there are several microorganisms which interfere with these water samples so their tests are necessary, there are several Water samples which cannot be detected by chemical methods hence microbiological assay are very useful for resolving doubts regarding possible change in potency of water used in pharmacy industry, environmental monitoring microbiological quality of different locations in pharmaceutical facility are far very critical measures in controlling clean area and environment.

ANY QUESTIONS ?