MICROBIOLOGICAL ANANLYSIS OF PHARMACEUITCAL

WATER SAMPLES AND ENVIORMENTAL MONITORING

Under the Guidance : Presented By:
MR. Ajeet Kumar Anand Amrit raj
Microbiologist (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/m
NIL
Air lock 1

40 CFU/m

NIL

Passage

35 CFU/m

NIL

Media preparation room

20 CFU/m

NIL

Incubation room

05 CFU/m

NIL

Buffer zone

18 CFU/m

NIL

MLT room 02 CFU/m

NIL

Washing Area

25 CFU/m

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 ?