More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
SOP for Microbiological analysis of
Raw water, Potable water & Purified
water
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
1.0 OBJECTIVE:
To lay down a procedure for Microbiological analysis of Raw Water, Potable Water & Purified Water.
2.0 SCOPE:
This SOP is applicable for Microbiological analysis of Raw Water, Potable Water & Purified Water in
Microbiological Lab of Quality Control Area.
3.0 RESPONSIBILITY:
Officer / Executive – Microbiologist
4.0 ACCOUNTABILITY:
Head – QC
6.0 PROCEDURE:
6.1 TOTAL AEROBIC MICROBIAL COUNT:
6.1.1 Sample the Raw water, Potable water & Purified water as per SOP, Titled “Sampling of Water for
Microbiological Analysis”. Prepare the R-2Amedia as per SOP, Titled “Preparation of Culture
Media” (Note: Sample and Result observation due on holiday shall be done next working day).
6.1.2 For Total Aerobic Microbial Count filter 1 ml of raw and potable water through 0.45µ Membrane
Filter and wash the membrane with 100 ml sterile water and place the Membrane Filter to pre-
incubated R-2A media plate for bacterial count. Label the plates with Sampling Point, Date of
Sampling, Date of Testing, and Date of release and Media Reference No.
6.1.3 Incubate the R-2A media plates at 30 to 350C for 5 (five) days for total microbial count.
6.1.4 For Total Aerobic Microbial Count filter 1 ml of Purified water and Process Potable water through
0.45µ Membrane Filter and wash the membrane with 100 ml sterile water and place the Membrane
Filter to pre-incubated R-2A media plate for bacterial count. Label the plates with Sampling Point,
Date of Sampling, Date of Testing, Date of release and Media Reference No.
6.1.5 Incubate the R-2A plates at 30 to 350C for 5 (five) days for total microbial count.
6.1.6 Observations and Results:
5.1.6.1 Raw Water and Potable Water: Examine the plates for the growth and count the number of colonies with the help of
colony counter. Express the count in term of the number of microorganisms per ml of raw water or potable water.
5.1.6.2 Purified Water and Process Potable Water: Examine the plates for the growth and count the
number of colonies with the help of colony counter. Express the count in term of the number of
microorganisms per ml of purified water.
6.3 TEST FOR SPECIFIED MICROORGANISMS:
6.3.5 Pretreatment of Sample:
6.3.5.4 Filter 100 ml of water sample through membrane filter of nominal pore size NMT 0.45µm and
47 mm diameter.
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
6.3.5.5 Transfer the filter to 100 ml Soyabean Casein Digest Medium.
6.3.5.6 Incubate the medium at 30-350C for 18-24 hrs.
6.3.5.7 Examine the medium for turbidity.
6.3.6 Test for Escherichia coli:
6.3.6.4 Shake the tube and transfer 1 ml of pretreated sample (SCM) to 100 ml of Mac Conkey Broth
and incubate 42 to 440C for 24 to 48 hrs.
6.3.6.5 Streak a portion from MacConkey broth on the surface of MacConkey Agar media and
incubate 30 to 350C for 18 to 72 hrs.
6.3.6.6 Upon examination, if none of the colonies confirm to the description given in Table-1, the
sample meets the requirements for the absence of the E. coli.
6.3.6.7 Run Positive and Negative Control with test.
6.3.6.8 If colonies show characteristic growth, carry out gram staining as per SOP, Titled
“Techniques for Microbial Culture Staining”.
6.3.6.9 If colonies show characteristic growth as per Table-1, carry out the identification by BBL
Crystal ID System in plant-III.
6.3.7 Test for Salmonella spp.
6.3.7.4 Shake the tube and transfer 0.1 ml of pretreated sample to 10 ml of Rappaport Vassiliadis
Salmonella Enrichment Broth and incubate at 30 to 350C for 18 to 24hrs.
6.3.7.5 Streak a portion from the Rappaport Vassiliadis Salmonella Enrichment Broth on surface of
Xylose Lysine Deoxycholate Agar Medium and incubate 30 to 350C for 18 to 48 hrs.
6.3.7.6 Upon examination, if none of the colonies confirm to the description given in Table-1, the
sample meets the requirements for the absence of the Salmonella spp.
6.3.6.1 Run Positive and Negative Control with test.
6.3.6.2 If colonies show characteristic growth, carry out gram staining as per SOP, Titled
“Techniques for Microbial Culture Staining”.
6.3.6.3 If colonies show characteristic growth as per Table-1, carry out the identification by BBL
Crystal ID System.
6.3.7 Test for Pseudomonas aeruginosa:
• Shake the tube and streak one loop full pretreated sample (SCM) on to the plate of Cetrimide
Agar Medium and incubate 30 to 350C for 18 to 72 hrs.
• Upon examination, if none of the colonies confirm to the description given in Table-1, the
sample meets the requirements for the absence of the Pseudomonas aeruginosa.
• Oxidase Test: If growth of suspected colonies occurs, place suspected colony on the oxidase
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
disc or paper strips or discs that previously had been impregnated with N, N-dimethyl-p-
phenylenediamine dihydrochloride. If there is no development of a pink color, changing to purple
color, the sample meets the requirements of the test for absence of Pseudomonasaeruginosa.
• Run Positive and Negative Control with test.
• If colonies show characteristic growth, carry out gram staining as per SOP, Titled
“Techniques for Microbial Culture Staining”.
• If colonies show characteristic growth as per Table-1, carry out the identification by BBL
Crystal ID System.
• Test for Staphylococcus aureus:
• Shake the tube and streak one loop full pretreated sample (SCM) on to the plate of Mannitol
Salt Agar Medium and incubate at 30 to 350C for 18 to 72 hrs.
• Upon examination, if none of the colonies confirm to the description given in Table-1, the
sample meets the requirements for the absence of the Staphylococcus aureus.
• Run Positive and Negative Control with test.
• If colonies show characteristic growth, carry out gram staining as per SOP, Titled
“Techniques for Microbial Culture Staining”.
5.2.5.5 If colonies show characteristic growth as per Table-1, carry out the identification by BBL
Crystal ID System.
Test for Bile-Tolerant Gram-Negative Bacteria (Enterobacteria) for Raw Water Potable
Water :
• Transfer 1 ml of sample to 100 ml Enterobacteria Enrichment Broth Mossel. Incubate the
medium at 30 to 350C for 24 to 48 hrs.
• After Incubation, Subculture on the plate of Violet Red Bile Glucose Agar and incubate at 30
to 350C for 18 to 24 hrs.
• Upon examination, if none of the colonies confirm to the description given in Table-1, the
sample meets the requirements for the absence of Enterobacteria.
• Run Positive and Negative Control with test.
• If colonies show characteristic growth, carry out gram staining as per SOP, Titled
“Techniques for Microbial Culture Staining”.
• If colonies show characteristic growth as per Table-1, carry out the identification by BBL
Crystal ID System.
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
• ACCEPTANCE CRITERIA:
• Purified Water:
• Total Aerobic Microbial Count
Limit : 100 cfu/ml
Action Limit : 80 cfu/ml
Alert Limit : 60 cfu/ml
• Specified Microorganisms – Escherichia coli, Salmonella spp., Pseudomonas aeruginosa,
Staphylococcus aureus should be absent.
• Raw Water / Potable Water:
• Total Aerobic Microbial Count
Limit : 500 cfu/ml
Action Limit : 400 cfu/ml
Alert Limit : 300 cfu/ml
• Specified Microorganisms– Escherichia coli, Salmonella spp., Pseudomonas aeruginosa,
Staphylococcus aureus, Bile-Tolerant Gram-Negative Bacteria (Enterobacteria) should be
absent
6.0 REFERENCES:
United State Pharmacopoeia 37
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
Indian Pharmacopoeia 2012
British Pharmacopoeia 2012
7.0 ANNEXURES:
ENCLOSURES: SOP Training Record
8.0 DISTRIBUTION:
• Controlled Copy No. 01 Quality Assurance Department
• Controlled Copy No. 02 Quality Control Department
Master Copy Quality Assurance Department
9.0 ABBREVIATIONS:
cfu Colony Forming Unit
Hrs. Hours
IP Indian Pharmacopoeia
ml Milliliter
No. Number
NMT Not More Than
QA Quality Assurance
QC Quality Control
SOP Standard Operating Procedure
SCM Soyabean Casein Digest Medium
spp. Species
UV Ultra Violet
USP United State Pharmacopoeia
10.0 REVISION HISTORY:
CHANGE HISTORY LOG
Revision No. Details of Changes Reason for Change Effective Date Updated By
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
ANNEXURE-I
MICROBIOLOGICAL ANALYSIS RECORD OF PURIFIED WATER
e of Sampling
R. No
mpling points
ume sampled
Incubator ID No.
e of Testing
Test : Total aerobic Microbial Count
Test Method : Membrane Filtration
Name of media : R-2A Media Reference No.: ………………
Volume of Sample :1 ml
Incubation Temp. : 30-350C for five days
Date of Incubation:…………………………… Date of Observation:.………………..….
Observations:
Sampling Point No.
Date
Average cfu.
Results : Total Aerobic Microbial Count / ml of purified water
-ve Control
Remarks:………………………………………………………………………………………………………………………
……………..……………………………………………………………………………………………………………………
LIMIT: Total Aerobic Microbial Count
Alert Limit: 60 CFU/ml Action Limit : 80 CFU/ml Limit : 100 CFU/ml
Test : Test for Specified Microorganisms Date of Test:
Enrichment of Sample:
Test Method : Membrane Filtration
Name of media : Soyabean Casein Digest Medium
Volume of Sample : 100 ml Media Reference No. :…………………
Incubation Temp. : 30-350C for 18 - 24 hrs
Date of Incubation :………………………................ Date of Observation :………………..…………
Sampling Positive Negative
Points control control
Observation
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
Remarks:……………………………………………………………………………………………….…
…………………..
……………………………………………………………………………………………………………
……………..……..
1.0 Test for Escherichia coli:
(I) Date :……………………………………….…
Name of media : MacConkey Broth Media Reference No.:
………………
Incubation Temp. : 42-440Cfor 24 -48 hrs
Date of Incubation :…………………………… Date of Observation:
.………………..……..
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
More than 2000 document related to pharmaceutical Industry in Pharmatalks Telegram
community👇👇👇👇👇👇👇👇 t.me/Pharmatalks
