This standard operating procedure outlines the process for analyzing water used in drug manufacturing, including collecting initial samples, conducting chemical tests to measure parameters like hardness, total suspended solids, total dissolved solids and total organic carbon, and performing bacterial and microbial tests to identify microbes like Escherichia coli and Staphylococcus aureus using various agar mediums and notifying supervisors if any results exceed limits.

1. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 1 of 4
Confidential
PURPOSE
To provide the procedure to be used in analyzing water for use in drug manufacturing.
SCOPE
Applicable to all sampling points of the water system and includes chemical and microbial tests.
APPLICABLE TO
Microbiologist
Head of Department
SOP Coordinator
All Staff (as assigned on a case by case basis)
RELATED PROCEDURES
SOP WA 01 for Water Sampling and Analysis for Chemical and Microbial Parameters
SOP WA 02 How to use TOC Analyzer
SOP WA 03 OC Verifying and Recording TOC Results
SOP WA 04 Interpreting Chemical Analysis Results on Water
SOP WA 05 Identifying Microbes in Water Used for Manufacturing
Author: Nadine Sookhoo
Date: April 30, 2016
Approval: Jason Chu
Date: May 6, 2016
Quality Assurance

2. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 2 of 4
Confidential
Definitions:
Acronyms
SOP Standard Operating Procedure(s)
GMP Good Manufacturing Practices
mg Milligram
TSS Total Suspended Solids: is the dry-weight of particles trapped by a filter
TDS Total Dissolved Solids: refer to any minerals, salts, metals, cations or anions dissolved in
water.
TOC Total Organic Carbon: is the amount of carbon found in an organic compound and is
often used as a non-specific indicator of water quality.
PAF Pseudomonas agar for Fluorescein: used for the detection and differentiation of
Pseudomonas aeruginosa by enhancement of fluorescein production.
PAP Pseudomonas agar for Pyocyanin: is a blue, secondary metabolite with the ability to
oxidize and reduce other moleculesand therefore can kill microbes competing against P.
aeruginosa.
G.C. Giolitti Cantoni broth: An anaerobic enrichment broth for Staphylococcus aureus
TAMC Total Aerobic Microbial Count: number of aerobic bacteria.
Author: Nadine Sookhoo
Approval: Jason Chu

3. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 3 of 4
Confidential
(Microbiology Department Head and all Microbiology Scientists)
PROCEDURE
Responsibility Actions
Microbiologist
Microbiologist
1.0
1.1
2.0
Checking
Initial Sample Testing:
1.1.1 Collect samples as per SOP WA 01 for Water Sampling and
Analysis for Chemical and Microbial Parameters
Chemical Analysis
2.1 Tests:
2.1.1 Hardness:
-Take 100 ml sample, add 2 ml of ammonia buffer pH 10,
50 mg of mordant black 11 mixture and add 1 ml of 0.01M
NaCl
- Mix until pure blue colour is produced
- Measure volume of NaCl used and calculate hardness by
following formula:
Hardness mg/L = ml NaCl/Sample Volume x 1000
- Record results in Log Book

4. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 4 of 4
Confidential
2.1.2 Total Suspended Solids (TSS):
- Place the gouch crucible clean and dry in oven for one
hour at 105C
- Cool crucible in desiccator and take the empty weight of
crucible then filter 30 ml water sample from crucible with
vacuum pump and calculate TSS with the following
formula:
TSS = W2-W1 x 1000/ ml of solution taken
- Record results in Log Book
2.1.3
2.1.4
2.1.5
Total Dissolved Solids (TDS):
- Measure the conductivity at 25C with a calibrated conductivity
meter and convert the value in TDS with the following formula:
TDS in mg/L = conductivity in mS x 0.667
- Record results in Log Book
Total Organic Carbon (TOC):
- Analyze the sample for TOC in a calibrated TOC analyzer as per
SOP WA 02 How to use TOC Analyzer
- If the TOC results are above alert limit and action limit, follow
SOP WA 03 OC Verifying and Recording TOC Results, Limits are
outlined in Table 1 Alert and Action Limit for TOC of Water
System
- Record results in Log Book
- If any chemical tests are out of the acceptable ranges, as
outlined in SOP WA 04 Interpreting Chemical Analysis Results
on Water, immediately notify the Microbiology Head of
Department

5. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 5 of 4
Confidential
3.0
3.1
3.1.1
3.1.2
3.1.3
Bacterial and Microbial Tests:
Tests:
Test for Escherichia coli:
- If the inoculated MacConkey’s broth tube shows acid and gas
formation, inoculate the following selective media plates and
incubate at 30-35C for 24-48 hours for presumptive
identification of the pathogen, as outlined in Table 2 Agar
Mediums used and Resulting Colonies Formed for Escherichia
coli
- Record results in Log Book
Test for Staphylococcus aureus:
- If growth is present in Giolitti Cantoni (G.C.) broth, usually
characterized by black settled growth at the bottom of the
broth under anaerobic conditions, inoculate the following
selective media plates and incubate at 30-35C for 24-48 hours
for presumptive identification of pathogen, as outlined in
Table 3 Agar Mediums used and Resulting Colonies Formed
for Staphylococcus aureus
- Record results in Log Book
- After colonies have formed, identify and record in log books
all strains as outlined in SOP WA 05 Identifying Microbes in
Water Used for Manufacturing
- If any strains are above the cited limits in SOP , alert
Microbiology Head of Department immediately

6. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 6 of 4
Confidential
4.0 Appendix
Table 1 Alert and Action Limit for TOC of Water System:
S.No Type of Water Alert Limit (ppb) Action Limit (ppb)
1 Purified water 300.0 500
2 Water for injection 250.0 500
3 Pure steam 250.0 500
Table 2 Agar Mediums used and Resulting Colonies Formed for Escherichia coli
Medium Description of Colony
MacConkey’s Agar Brick red may have surrounding zone of
precipitated bile.
Eosin Methylene Blue Agar Metallic sheen with dark grey colonies
Table 3 Mediums used and Resulting Colonies Formed for Staphylococcus aureus
Medium Description of Colony
Mannitol Salt Agar Medium Yellow colonies with yellow zones
Vogel Johnson Agar Medium Black surrounded by yellow zone
Baird Parker Agar Medium Black, shiny, surrounded by clear zones of 2-
5mm

7. AAPS Labs SOP
Standard Operating Procedure
Document Number
Version Number
Effective Date
:
:
:
SOP WA23
07
May 6, 2016
STANDARD OPERATING PROCEDURE FOR
THE ANAYLSIS OF WATER FOR USE IN DRUG
MANUFACTURING
Page 7 of 4
Confidential
References
http://www.pharmaguideline.com/2010/01/sop-for-procedure-for-analysis-of-water.html, SOP for
Analysis of Water, Accessed Saturday April 30, 2016