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How Can Test Vitamin B6 (Pyridoxine Hydrochloride)

1.0 PURPOSE

This raw material specification & analysis prescribes the requirements for Vitamin-B6 (Pyridoxine) Hydrochloride, which is one of the components use for Syrup, Injection, Tablet and Capsule.

2.0 SCOPE

It is applicable in Material Department, Product Development and Quality Assurance Department to confirm its physical and chemical specifications.

3.0 REFERENCES

USP/BP/Assay of vitamins in Pharmaceutical Preparation

4.0 DEFINITIONS

Follow SOP

5.0 SPECIFICATIONS USP/BP/BP/Assay of vitamins in Pharmaceutical Preparation

PHYSICAL ANALYSIS

1 Physical: Form Crystalline Powder
2 Colour: White to almost white
3 Melting Point: At about 205° with decomposition
4 Solubility: Freely soluble in water. Sparingly soluble in ethanol 96%.
5 pH: of 10% w/v solution 2.4 to 3.0 at 20°C to 25°C
6 Loss on Drying: Not more than 0.5%

CHEMICAL ANALYSIS

  • Identification of Vitamin B6 by IR spectrum Should comply the standard spectrum
  • Identification Test of Pyridoxine Hydrochloride Blue color should produce which is suppressed by the addition of the 5% boric acid solution.
  • Identification Test of Chloride Curdy white presipitate should produce which are insoluble in dilute nitric acid but are soluble in ammonia solution
  • Contents of Pyridoxine Hydrochloride 98.0 to 102% OAB

6.0 SAMPLING

Refer to the Incoming SOPs for sampling.

7.0 REJECTIONS:

If the Vitamin-B6 (Pyridoxine) Hydrochloride does not satisfy the specifications mentioned in 5.0 then it will be rejected. In addition to the specifications, Vitamin-B6 (Pyridoxine) Hydrochloride is also rejected due to the following reasons:

7.1 If the receiving raw material is cross contaminated due to any reason.
7.2 The material does not bear 75% of its stated shelf life at the time of receipt.

8.0 PACKAGING

Vitamin-B6 (Pyridoxine) Hydrochloride is received in polyethylene bag placed in cardboard container to protect it from moisture and light.

9.0 STORAGE:

BP: Store protected from light.

10.0 HANDLING

10.1 Use gloves & mask during weighing and dispensing operation of Vitamin-B6 (Pyridoxine) Hydrochloride.
10.2 Keep the container of Vitamin-B6 (Pyridoxine) Hydrochloride well closed immediately after use.
10.3 Place the container of Vitamin-B6 (Pyridoxine) Hydrochloride back to its specified area.

11.0 ANALYSIS

11.1 (A) SUITABLE Machinery/Equipment/Apparatus

PHYSICAL

1. Test Tubes
2. Beaker
3. Glass Rod
4. Moisture Balance
5. pH Meter
6. Melting Point Apparatus

CHEMICAL

1 Volumetric Flask 100 ml, 200 ml
2 Graduated Pipette 1 ml, 2 ml, 5 ml, 10 ml
3 Titration Flask 100 ml
4 U.V. Spectrophotometer
5 Analytical Balance
6 Magnetic Stirrer
7 FTIR,
8 Spatula
9 Ethanol 96% 10 ml
10 Isopropyl Alcohol 60 ml
11 Potassium bromide IR grade 200 mg
12 Conc. Nitric Acid 1 ml

Reagents

1. Dilute Ammonia Solution 1 ml
2. 0.04% w/v 2,6 Dichloroquinone Chlorimide 5 ml
3. Ammonia Ammonium Chloride Buffer Solution pH 6 6 ml
4. 5% Boric Acid 2 ml
5. Silver Nitrate TS 1 ml
6. Dilute nitric acid 1 ml

11.2 PHYSICALANALYSIS USP/BP/Assay of vitamins in Pharmaceutical Preparation

11.2.1 Test of Physical Form:

Check the sample and confirm its physical form; it should be crystalline powder.

11.2.2 Test of Colour:

Confirm the colour of the material by comparing with standard from colour chart; it should be white to almost white

11.2.3 Test of Solubility:

check the solubility of the sample in water & in Ethanol 96%.
It should be soluble in water, sparingly soluble in ethanol 96%.

11.2.4 Test of pH:

Weigh accurately on analytical balance 10 g of sample and make the volume upto 100 ml with distilled water. Take 50 ml from the resulting solution and measure the pH of solution at 20°C to 25°C on pH meter; it should be 2.4 to 3.0 at 20°C to 25°C.

11.2.5 Test of Loss on drying:

Weigh accurately 10 g of sample on moisture balance. Adjust the temperature at 105°C, start drying the sample. At the end balance show percentage loss of moisture; moisture contents should not be more than 0.5%.

11.2.6 Test of Melting Point:

Take the powdered sample in the capillary tube meant for the purpose, put it in the Melting Point Apparatus. Check the temperature at which the powdered sample melts; melting point should be at about 205°C with decomposition.

11.3 Record the result of Physical Analysis on quality report.

11.4 CHEMICALANALYSIS: USP/BP/Assay of vitamins in Pharmaceutical Preparation 

11.4.1 Examination of Vitamin B6 by IR spectrum:

Take dried sample to 5mg of Vitamin B6 and thoroughly mix with 95mg of dried standard of potassium bromide in pastel mortar.

Fill Macro cups with provided funnel like instrument and draw the spectrum it should comply with the standard spectrum.

11.4.2 Identification Vitamin B6:

Weigh accurately on analytical balance 200-mg g of the sample, put it in the test tube, and add 10 ml of distilled water. Take 1 ml from the resulting solution, add 1 ml of 0.04% w/v Dichloroquinone Chlorimide Solution and 0.5 ml of dilute ammonia solution; blue colour should produce which is suppressed by the addition of 5% boric acid solution.

11.4.3 Identification Test of Chloride:

Weigh accurately on analytical balance 50 mg of the sample, put it in test tube, add 5 ml of distilled water, 1 ml of Silver Nitrate Solution and 0.5 ml of nitric acid; curdy white precipitate should produced which are insoluble in dilue nitric acid, but soluble in ammonia solution.

11.4.4 Contents of Vitamin B6 (Pyridoxine) Hydrochloride:

11.4.4.1 Standard Preparation:

11.4.4.1.1 Weigh accurately on analytical balance 100 mg reference standard of Vitamin B6 (Pyridoxine) Hydrochloride equivalent to 100 mg of Vitamin B6 (Pyridoxine) Hydrochloride, make the volume upto 100 ml with distilled water in volumetric flask.

11.4.4.1.2 Take 1 ml from the step 11.4.4.1.1 and make the volume upto 100 ml with distilled water in the volumetric flask and filter it in 100 ml titration flask.

11.4.4.1.3 Take 5 ml from the step 11.4.4.1.2 and make the volume upto 30 ml with isopropyl alcohol in 100 ml volumetric cylinder.

11.4.4.1.4 Take 6 ml from the step 11.4.4.1.3 in the test tube and add 2 ml of ammonia ammonium chloride buffer pH 6 solution, 1 ml of distilled water and 1 ml of 0.04% 2,6 dichloroquinone chlorimide solution.

11.4.4.1.5 Measure the absorption of step 11.4.4.1.4 at 650 nm on U.V Spectrophotometer within 80-90 seconds.

11.4.4.2 Sample Preparation:

11.4.4.2.1 Weigh accurately on analytical balance 100 mg of powdered sample of Vitamin B6 (Pyridoxine) Hydrochloride, make the volume upto 100 ml with distilled water in volumetric flask.

11.4.4.2.2 Take 1 ml from the step 11.4.4.2.1 and make the volume upto 100 ml with distilled water in the volumetric flask and filter it in 100 ml titration flask.

11.4.4.2.3 Take 5 ml from the step 11.4.4.2.2 and make the volume upto 30 ml with isopropyl alcohol in 100 ml volumetric cylinder.

11.4.4.2.4 Take 6 ml from the step 11.4.4.2.3 in the test tube and add 2 ml of ammonia ammonium chloride buffer pH 6 solution, 1 ml of distilled water and 1 ml of 0.04% 2,6 dichloroquinone chlorimide solution.

11.4.4.2.5 Measure the absorption of step 11.4.4.2.4 at 650 nm on U.V Spectrophotometer within 80-90 seconds.

11.4.4.3 Blank Preparation:

11.4.4.3.1 Weigh accurately 100 mg of powdered sample of Vitamin B6 (Pyridoxine) Hydrochloride, make the volume upto 100-ml with distilled water in volumetric flask.

11.4.4.3.2 Take 1 ml from the step 11.4.4.1.1 and make the volume upto 100 ml with distilled water in the volumetric flask and filter it in 100 ml titration flask.

11.4.4.3.3 Take 5 ml from the step 11.4.4.3.2 and make the volume upto 30 ml with isopropyl alcohol in 100 ml volumetric cylinder.

11.4.4.3.4 Take 6 ml from the step 11.4.4.3.3 and add 2 ml of ammonia ammonium chloride buffer pH 6 solution, 1 ml of 5% boric acid and 1 ml of 0.04% 2,6 dichloroquinone chlorimide solution.

11.4.4.3.5 Measure the absorption of step 11.4.4.3.4 at 650 nm on U.V Spectrophotometer within 80-90 seconds.

11.4.4.4 Observations:

a) Absorption of Sample Preparation = A
b) Absorption of Standard Preparation = B
c) Record the reading.

11.4.4.4.1 Calculations:

Calculate the contents of Vitamin B6 (Pyridoxine) Hydrochloride as follows:
                     A
——————————— x 100 = C
                    B
Limit: The contents of Vitamin B6 (Pyridoxine) Hydrochloride should be between 98-102%.OAB

11.5 Record the result of Chemical Analysis on quality report.

Note. Incase of any non- conformance observed during Inspection and testing , Please Follow Quality System Procedure for Control of Non-Conforming Product.

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