Standard test procedure for Sterilized water for Injection BP

This article consists of standard test procedure for Sterilized water for injection as per BP (British Pharmacopeia)

standard test procedure for Sterilized water for Injection BP

1. Description:

Procedure : Take clean and dry colourless, transparent, neutral glass with a flat base test tube.

Rinse the test tube with sample twice. Take about 50 ml sample in the test tube.

Check visually appearance of sample.

Specification: Clear and colourless liquid.

 

2. Length:

         Istrument:Vernier Caliper

Procedure : Measure the Length in mm and Measure crease to crease by Vernier Caliper.

Note down the results in test data sheet.

 

3. Width:

         Istrument: Vernier Caliper

Procedure : Measure the Width in mm and Measure crease to crease by Vernier Caliper.

Note down the results in test data sheet.

 

4. Height:

         Istrument: Vernier Caliper

Procedure : Measure the Height in mm and Measure crease to crease by Vernier Caliper.

Note down the results in test data sheet.

 

5. pH (at 25ºC):

        Istrument: PH Meter

Procedure : Take clean and dry 100 ml glass beaker. Rinse the glass beaker with sample twice.

Take about 50 ml sample in the glass beaker.  Adjust the temperature of the sample to 25 ºC.

Measure the pH with the help of pH meter.

(Specification: 5.0 to 7.0)

 

 

6. Conductivity (at 25⁰c):

        Istrument: Conductivity Meter.

Procedure : Take clean and dry 100 ml glass beaker. Rinse the glass beaker with sample twice.

Take about 50 ml sample in the glass beaker. Adjust the temperature of the sample to 25 °C

Measure the conductivity with the help of conductivity meter.

Specification: Not more than 5.0 µs.cm^-1.

7. Heavy metals:

        Equipment: Measuring Cylinder.

Procedure : Note: Prepare blank, standard and sample solutions at same time and in same manner.

Concentrated aqueous solution:

Take a clean and dry glass evaporating dish. Take 200 ml of sample using measuring cylinder, and transfer in glass evaporating dish. Add 0.15 ml of 0.1 M nitric acid in glass evaporating dish using graduated pipette. Place glass evaporating dish on water bath till the volume is reduced up to 20 ml.

Test solution:

Take clean and dry colourless, transparent, neutral glass with a flat base test tube.

Rinse the test tube with sample twice. Mark the test tube as heavy metals sample. Take about 12 ml concentrated aqueous solution in the test tube using graduated pipette. Add 2 ml of buffer solution pH 3.5 in the test tube using bulb pipette and mix well.  Add 1.2 ml of thioacetamide reagent using graduated pipette and Mix immediately.

Lead Standard Solution (0.1% Pb): Dissolve 0.400 g of lead (II) nitrate in sufficient water to produce 250.0 ml.

Lead Standard Solution (100 ppm Pb): Dilute 1 volume of lead standard solution (0.1% Pb) to 10 volumes with water   immediately before use.

Lead Standard Solution (10 ppm Pb) : Dilute 1 volume of  lead standard solution (100 ppm Pb) to 10 volumes with water   immediately before use.

Lead Standard Solution (1 ppm Pb) : Dilute 1 volume of  lead standard solution (10 ppm Pb) to 10 volumes with water   immediately before use.

 

Reference solution (standard) :

Take clean and dry colourless, transparent, neutral glass with a flat base test tube. Rinse the test tube with sample twice. Mark the test tube as heavy metals standard Pipette 10 ml of lead standard solution (1ppm Pb) using bulb pipette and transfer in test tube. Add 2 ml concentrated aqueous solution in the test tube using bulb pipette. Add 2 ml of buffer solution pH 3.5 in the test tube using bulb pipette and mix well.  Add 1.2 ml of thioacetamide reagent using graduated pipette and Mix immediately.

 

Blank solution : Take clean and dry colourless, transparent, neutral glass with a flat base test tube.

Mark the test tube as heavy metals blank. Pipette 10 ml of water using bulb pipette and transfer in test tube. Add 2 ml concentrated aqueous solution in the test tube using bulb pipette. Add 2 ml of buffer solution pH 3.5 in the test tube using bulb pipette and mix well.  Add 1.2 ml of thioacetamide reagent using graduated pipette and Mix immediately

Note: If the result is difficult to judge, filter the solutions through a membrane filter (pore size 3 µm, without the prefilter). Carry out the filtration slowly and uniformly, applying moderate and constant pressure to the piston. Compare the spots on the filters obtained with the different solutions.

 

Specification: Any Brown colour in the sample should not be more intense than that of the standard.

Specification: Not more than 0.1 ppm.

 

8. Acidity or alkalinity:

 Acidity:

Take a clean and dry 100 ml glass beaker. Take 20 mL of sample using measuring cylinder in glass beaker. Add 0.05 mL of phenol red solution . Add 0.15 mL of 0.01 M hydrochloric acid.

 

Specification: The solution is becomes yellow.

 

Alkalinity:

Take a clean and dry 100 ml glass beaker. Take 20 mL of sample using measuring cylinder in glass beaker. Add 0.05 mL of phenol red solution . Add 0.1 mL of 0.01 M sodium hydroxide

 

Specification:   .

 

 

 

 

9. Oxidisable substances:

        Instrument :  Water Bath

Procedure : Take a clean and dry 250 ml glass beaker. Take 100 ml of sample using measuring cylinder in glass beaker. Add 10 ml of dilute sulphuric acid using bulb pipette in glass beaker. Add 0.4 ml of 0.02 M potassium permanganate using graduated pipette in glass beaker. Place glass beaker on water bath till the sample is boiled for 5 min.

Specification: The solution remains faintly pink.

 

10. Chlorides:

Procedure : Take clean and dry colourless, transparent, neutral glass with a flat base test tube. Mark the test tube as chloride sample. Take 10 ml sample using graduated pipette in test tube. Add 1 ml of dilute nitric acid using graduated pipette in test tube. Add 0.2 ml of silver nitrate solution using graduated pipette in test tube.

Specification: The solution shows no change in appearance for at least 15 minutes.

 

11. Nitrates:

        Instrument :  Water Bath

Procedure :

Note: Prepare standard and sample solutions at same time and in same manner.

Test solution: Take clean and dry 1000 ml glass beaker. Fill the beaker with ice cubes about ¾ of its capacity. Take clean and dry colourless, transparent, neutral glass with a flat base test tube. Mark the test tube as nitrate sample and immersed in ice containing beaker. Pipette 5 ml sample using bulb pipette and transfer in test tube. Add 0.4 ml of a 100 g/l solution of potassium chloride in the test tube using graduated pipette. Add 0.1 ml of diphenylamine solution in the test tube using graduated pipette. Add 5 ml nitrogen free sulphuric acid dropwise with shaking in the test tube using bulb pipette. Transfer the test tube to a water-bath at 50 °C.

 

Nitrate standard solution: Take clean and dry colourless, transparent, neutral glass with a flat base test tube. Mark the test tube as nitrate standard. Pipette 4.5 ml nitrate-free water using graduated pipette and transfer in test tube. Add 0.5 ml of nitrate standard solution (2 ppm NO₃) using graduated pipette.

Immerse this test tube in ice containing beaker. Add 0.4 ml of a 100 g/l solution of potassium chloride in the test tube using graduated pipette. Add 0.1 ml of diphenylamine solution in the test tube using graduated pipette. Add 5 ml nitrogen free sulphuric acid dropwise with shaking in the test tube using bulb pipette. Transfer the test tube to a water-bath at 50 °C.

Nitrate Standard Solution (2 ppm NO₃): Dilute 0.2 ml of a 0.163% w/v solution of potassium nitrate to 100 volumes with water.  Immediately before use.

 

Specification: After 15 min, any blue colour in the solution is not more intense than that in a reference solution.

Specification: Not more than 0.2 ppm

 

12. Sulphates:

Procedure : Take clean and dry colourless, transparent, neutral glass with a flat base test tube. Mark the test tube as sulphates sample.  Take 10 ml sample using bulb pipette in test tube. Add 0.1 ml of dilute hydrochloric acid using graduated pipette in test tube. Add 0.1 ml of barium chloride solution using graduated pipette in test tube.

Specification: The solution shows no change in appearance at least for 1 hour.

 

13. Ammonium:

 Note: Prepare standard and sample solutions at same time and in same manner.

 Procedure :

Test solution:

Take clean and dry colourless, transparent, neutral glass with a flat base test tube.

Mark the test tube as ammonium sample.  Take 20 ml sample using bulb pipette in test tube.

Add 1ml of alkaline potassium tetraiodomercurate solution using bulb pipette in test tube.

Ammonium Standard Solution (1 ppm NH₄) : Dilute 1  volume of ammonium standard solution (3 ppm NH₄) to 3  volumes with water  immediately before use.

 

Standard solution: Take clean and dry colourless, transparent, neutral glass with a flat base test tube.

Mark the test tube as ammonium standard.  Take 4 ml Ammonium standard solution (1ppm ammonia) using bulb pipette in test tube. Add 16 ml of water using graduated pipette in test tube and mix well. Add 1ml of alkaline potassium tetraiodomercurate solution using bulb pipette in test tube.

 

 Reagents:

Alkaline potassium tetraiodomercurate solution: Dissolve 15gm of Mercuric iodide in 50ml of water. add 11gm of Potassium iodide and volume made to 100 ml with water. Immediately before use, mix 1 volume of this solution with an equal volume of a 250 gm / lit solution of Sodium Hydroxide.

Ammonium Standard Solution (3 ppm NH₄): Dilute 1  volume of a 0.0741% w/v solution of ammonium chloride to  100 volumes with water .

Specification: The solution is not more intense coloured than standard.

Specification: After 5 minutes examine the solution down  the vertical axis of the tube; the solution is       not more  intensely coloured than a  standard prepared.

14. Calcium and magnesium:

Procedure : Take a clean and dry 250 ml glass beaker. Take 100 ml of sample using measuring cylinder in glass beaker. Add 2 ml of ammonium chloride buffer solution pH 10.0 using bulb pipette in glass beaker.

Add 50 mg of mordant black 11 triturate in glass beaker. Add 0.5 ml of 0.01 M sodium edetate using graduated pipette in glass beaker.

Specification: A pure blue colour is produced.

 

15. Residue on evaporation:

        Instrument : Analytical Balance,Water Bath,Vaccume  Oven.

Procedure : Take a clean and dry glass evaporating dish that has been previously dried at

105 ⁰ C ± 2 °C. For at least ½ hours. Weigh the empty glass evaporating dish on analytical balance. Note down the weight into test data sheet (W1). Take 100 ml of sample using measuring cylinder, and transfer in glass evaporating dish. Put glass evaporating dish on water bath till the sample evaporate completely only residue is remain. Remove the evaporating dish from water bath, dry the residue in oven. Place evaporating dish containing residue in oven at 105 ⁰ C ± 2 °C for 1 hour. After drying is completed, open the oven, and allow it to cool to room temperature in desiccators. After cooling, weigh the glass evaporating dish with dried residue on analytical balance. Note down the weight into test data sheet (W2)

Calculate the residue on evaporation as per formula given below:

 

( W2 – W1) X100

% Residue on Evaporation = —————————

Sample taken in ml

 

Specification: Maximum 0.003 %

 

 

16. Particulate contamination

Sample shall be send to Public Testing Laboratory.

1.6000 per container equal to or greater than 10 µm

2.600 per container equal to or greater than 25 µm.

17.Sterility

Procedure : Follows the Respective Microbiology SOP

Specification: Should be sterile.

18. Bacterial endotoxins

Procedure : Follows the respective Microbiology SOP

Specification: Less than 0.25 IU/ml.