STP for borewell generation point (raw water storage tank)

PROCEDURE
1. Description : Take the sample and observe the colour and clarity visually
Open the bottle and smell the odour. Take few ml of sample
from the bottle and taste. Record the observations in the
test protocol.
2. pH : Take 100 ml of sample and record the on a calibrated pH meter at 25 ± 2 °c
Limit: 6.5 to 8.5
3. Total Hardness: Take 100ml Sample add 10ml ammonia ammonium chloride
as CaC03 buffer Use Erichrome Black T Indicator and titrate with 0.05M
E.DTA Each ml 0.05M E.D.T.A =0.005mg
Limit NMT 200 mg / Liters
4.Total Dissolve Solid : Take 20 ml Sample and Water Bath Evaporate and dry weight
NMT 500 mg/Liters

5. Total Alkalinity : Take 100 ml Sample and titrate with 0.02N HCL and use
methyl orange Indicator end point red colour.

Each 0.02N HCL =0.05mg
Limit 200 mg /Liters
6. Heavy Metals:
Test Sample:
Solution is prepared as per the monograph and 25 ml is transferred in Nessler’s
cylinder.Adjust the pH between 3 to 4 by adding dilute acetic acid or dilute ammonia
solution.Dilute with water to 35 ml.Add freshly prepared 10 ml of hydrogen sulphide
solution.Dilute with water to 50 ml.Allow to stand for five minutes.View downwards
over a white surface

Standard Sample
Take 2 ml of standard lead solution and dilute to 25 ml with water.Adjust the pH
between 3 to 4 by adding dilute acetic acid or dilute ammonia solution. Dilute with
water to 35 ml. Add freshly prepared 10 ml of hydrogen sulphide solution.Dilute
with water to 50 ml.Allow to stand for five minutes.View downwards over a white
surface.

Observation:
The color produce in sample solution should be greater than standard solution. If
color produces in sample solution is less than the standard solution, the sample will
pass the limit test of heavy metals and vice versa.
LIMIT: NMT 0.1 ppm.

7. Nitrates :
Reagents : (a)Potassium chloride solution (b)Diphenylamine solution

(c) Nitrate standard solution (d) Conc. Sulphuric acid

Procedure: Place 5 ml of the sample in a test tube immersed in iced water, add 0.4 ml of a 100 gm/l solution of

Potassium chloride solution R, 0.1 ml of Diphenylamine solution R, add drop wise with shaking, 5 ml of nitrogen

free Sulphuric acid R. Transfer the tube to a water bath at 50º C. After 15 minutes, any blue colour in the solution is

not more intense than that in a reference standard prepared at the same time in same manner using a mixture of 4.5 ml

of nitrate free water R, and 0.5 ml of nitrate standard solution (2 ppm NO3).
LIMIT: NMT 45 ppm.

8. Aluminium : Not more than 10 ppb, determined using the following solutions. Test solution. To 400
ml of the water under examination, add 10 ml of acetate buffer solution pH 6.0 and 100
ml of distilled water. Reference solution. Mix 2 ml of aluminium standard solution
(2 ppm Al), 10 ml of acetate buffer solution pH 6.0 and 98 ml of distilled water. Blank
solution. Mix 10 ml of acetate buffer solution pH 6.0 and 100 ml of distilled water.
LIMIT: NMT 0.03mg/Liters.

9. Ammonium:
Reagents: (a) 1 Ml of alkaline potassium solution
(b) 1 ppm Ammonium standard solution

Procedure: To 20 Ml add 1 Ml of alkaline potassium terraiodomercurate solution R. after 5 min,
examine the solution down the vertical axis of the tube. The solution is not more
intensely coloured than a standard prepared at the same time by adding 1 ml of alkaline
potassium terraiodomercurate solution R to a mixture of 4 ml of ammonium standard
solution (1 ppm NH4) R and 16 Ml of ammonium free water R.
Limit NMT 0.05 mg/Liters

10. Chloride: Take 100ml sample add potassium chromate Indicator Titrate with 0.1 N Silver
Nitrate end point
Each ml 0.1 N Sliver Nitrate =0.0035
Limit 250mg/Liters

11. Sulphate:
Reagents: (a) Barium chloride solution
(b) 0.1 M Hydrochloric acid

Procedure: To 10 ml add 0.1 ml of dilute hydrochloric acid R and 0.1 ml of Barium Chloride solution
R1. The solution shows no change in appearance for at least 1 h.

LIMIT: 200 mg/Liters

12. Calcium and Magnesium:
Reagents:(a) 0.01 M Sodium Edetate
(b) Mordant black II mixture
(c) Ammonia Chloride Buffer (pH 10.0).

Procedure: To 100 ml sample add 2 ml of Ammonium chloride buffer solution pH 10.0 R, 5
mordant black II triturate R and 0.5 ml of 0.01 M Sodium Edetate; a pure blue colour
is produced.

LIMIT: 75 mg/Liters.
13. Microbial Contamination:
Total Viable Aerobic Microbial Count
Total Viable Aerobic Microbial Count (TVAC) values provide density of aerobic and facultative
bacteria in the water sample, which can grow at 37ºC. The TVAC values, are useful in warning
about excessive growth in any water.

(A) Apparatus and materials:

• Incubator with arrangement at stable 37ºc.
• Petri dishes, diameter 90 mm.
• Conical flasks, 150ml, for dilution.
• Pipettes, bacteriological
• Gas burner
• Colony counter

(B) PROCEDURE:

Pour plate method

• Prepare SCDA as per SOP and sterilize all the required glassware as per SOP.
• Collect 250 ml of sample in sterilized sampling bottles and transfer to microbiology lab for analysis.
• Transfer sterilized media, Petri plates micropipette and microtips to LAF-2 of assay room and start LAF as per SOP.

• Pipette 1 ml of sample to two Petri plates and promptly add approx. 15-20 ml of SCDA that previously sterilized and

cooled to about 450C in two Petri plates and mix the sample by rotating the petriplates.
• Incubate SCDA plates at 300C to 350C for 48 hrs for bacterial count and after 48 hrs transfer the plates at 200C to 250C for 72 hrs for fungal count.
• After incubation count the numbers of colonies with colony counter and operates colony counter as per SOP.

LIMIT: Total Bacterial Count: 500 cfu/ml, Total Fungal Count: Nil

C.TESTS FOR SPECIFIED MICRO-ORGANISMS (PATHOGENS)
(A) APPARATUS AND MATERIAL
• Selective media
• Inoculating loop
• Colony counter
• Compound microscope
• Pure culture of E. coli, Salmonella abony, P. aeruginosa, S. aureus

PROCEDURE
(i) E. coli:
• Transfer 10 ml of sample to SCDM and make it 100 ml and incubate at 30°C to 35°C for 18
to 24 hrs.
• After incubation transfer 1 ml of SCDM to 100 ml of Mac-conkey broth and incubate at 42°C
to 44°C for 24 to 48 hrs.
• After incubation streak a loopful of SCDM to surface of preincubated Mac-conkey agar plates
and incubate at 30°C to 35°C for 18 to 72 hrs.
• After incubation if none of the colonies are brick red in colour and have a surrounding zone of precipitated bile, the sample

meets the requirement for the test of absence of E. coli.Run positive and negative control with test.

(II) Salmonella:

• Transfer 10 ml of sample to SCDM and make it 100 ml and incubate at 30°C to 35°C for
24hrs.

• After incubation transfer 0.1 ml of SCDM to 10 ml of Rappaport Vassilliadis Salmonella enrichment broth and incubate at 30°C to 35°C for 18 to 24 hrs.

• After incubation streak a loopful of SCDM to surface of preincubated Xylose lysine deoxycholate agar plates and incubate at 30°C to 35°C for 18 to 48 hrs.
• After incubation if none of colonies are red with or without black centres, the sample meets the requirement for the test of absence of Salmonella.
• Run positive and negative control with test.

(iii) Pseudomonas aeruginosa :
• Transfer 10 ml of Spl. to SCDM and make it 100 ml and incubate at 30°C to 35°C for 18 to 24 hrs.
• After incubation streak a loopful of SCDM to surface of preincubated Cetrimide agar plates and
incubate at 30°C to 35°C for 18 to 72 hrs.
• After incubation none of the plate contains green colour colonies the sample meets the requirement for the test of absence of P.aeruginosa.
• Run positive and negative control with test

(IV) Staphylococcus aureus:

• Transfer 10 ml of sample to SCDM and make it 100 ml and incubate at 30°C to 35°C
for 18 to 24 hrs.
• After incubation streak a loopful of SCDM to surface of preincubated Mannitol salt agar plates and incubate at 30°C to 35°C for 18 to 72 hrs.
• After incubation none of the plate contains yellow or white colonies surrounded by a
yellow zone the sample meets the requirement for the test of absence of P. aeruginosa.
• Run positive and negative control with test.

SOP for testing of ETP water

sop for Calibration and Maintenance of Laboratory Instruments and Equipment

Disposal of Residual Sample or Left Over Material

sop for for Laboratory Incident

standard operating procedure temperature monitoring

sop for drying of silica gel

sop for operation of infrared moisture balance

sop for preparation of mobile phase

sop for treated RO water

sop for Preparation and Issuance of Analysis protocol standard

sop of placebo and impurity stock solutions

sop for disposal of residual sample