Cefazolin Sodium (Sterile) Testing Procedure
1.0 PURPOSE
This raw material specification and analysis prescribes the requirements for Cefazolin Sodium (Sterile) which is used in Injection.
2.0 SCOPE
2.1 It is applicable in Material Department, Product Development and Quality Assurance Department to confirm its physical and chemical specifications.
2.2 Item code
3.0 REFERENCES
BP/USP/Inhouse
4.0 DEFINATIONS
Follow Sop
5.0 SPECIFICATIONS BP/USP/Inhouse
TESTS SPECIFICATIONS
1 Physical: Form Very Hygroscopic Powder
2 Colour: White or almost White
3. Solubility: Freely soluble in water, very slightly soluble in alcohol , practically insoluble in Ether.
4. Specific Optical Rotation: -15° to -24° on dried basis at 20.0°C
5 Reconstitution: 2.1 ml (±0.1 ml)
6 Reconstitution Time: Not more than 1 minute
7 pH: after reconstitution 4 to 6 (100 mg per ml) at (25±2°C)
8 Colour: after reconstitution Colourless
9 Clarity: Clear Solution without any visible contamination
10 Moisture Contents: Not more than 6%
11 Water Contents(by Karl Fischer): Not more than 6%.
CHEMICAL ANALYSIS
12 Identification of Cefazolin Sodium: examination by IR spectrum Should comply the standard spectrum
13 Identification Test of Sodium: Golden yellow colour flame should produce
14 Identification Test of Cefazolin Sodium: Yellow colour should develop
MICROBIOLOGICAL ANALYSIS
16 Potency of Cefazolin Sodium: 89.1% to 110.1% (On anhydrous basis)
6.0 SAMPLING
Refer to the Incoming SOPs for sampling.
7.0 REJECTIONS:
If the Cefazolin Sodium (Sterile) does not satisfy the specifications mentioned in 5.0 then it will be rejected. In addition to the specifications, Cefazolin Sodium (Sterile) 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
Cefazolin Sodium (Sterile) is received in polyethylene bag/thick aluminum foil bag placed in sturdy, metallic container to protect it from moisture and light.
9.0 STORAGE BP/USP/Inhouse
9.1 Store in an airtight container, protected from light.
9.2 Keep the container of the Cefazolin Sodium (Sterile) at a temperature not exceeding 30°C.
10.0 HANDLING
10.1 Use gloves & mask during weighing and dispensing operation of Cefazolin Sodium (Sterile) in the sterile area.
10.2 Keep the container of Cefazolin Sodium (Sterile) well closed immediately after use.
10.3 Place the container of Cefazolin Sodium (Sterile) back to its specified area.
11.0 ANALYSIS
11.1 (A) Suitable Machinery/Equipment/Apparatus
PHYSICAL
1. Moisture Balance
2. pH Meter 100 ml Volumetric Flasks
3. Test Tubes Pipette 1ml, 2ml, 10ml (Bulb Type)
4. Beaker UV-Visible Spectrophotometer
5. Glass Rod Analytical balance
6. Polarimeter Magnetic Stirrer
7. Funnel
8. FTIR, Mortar & Pastel
Chemicals
1. Ether 10 ml
2. Alcohol 10ml
3. KBr ( IR Grade ) 200mg
4. HCl (Concentrated) 0.5ml
Reagents
1. 0.1M Sodium Bicarbonate 500 ml
2. Sulphuric acid, Formaldehyde (Mix 2 ml of Formaldehyde solution with 100ml of H2SO4 (96% w/w) 2.0ml
11.2.1 Test of Physical Form:
Check the sample and confirm its physical form; it should be very Hygroscopic Powder.
11.2.2 Test of Colour:
Confirm the colour of the material by comparing with standard from colour chart; it should be white or almost white.
11.2.3 Test of Solubility:
Check the solubility of sample in water, Alcohol and Ether. It should be Freely soluble in water, very slightly soluble in alcohol , practically insoluble in Ether.
11.2.4 Test of Specific Optical Rotation:
Dissolve 1.25 g of sample to be examined in 25ml of water. The specific optical rotation is -15° to -24° on dried basis at 20°C.
11.2.5 Test of Reconstitution:
Weigh accurately on analytical balance 500 mg of Cefazolin Sodium put it in the vial, reconstitute it with 2 ml of Water for Injection; after reconstitution it should be 2.1 ml (± 0.1 ml).
11.2.6 Test of Reconstitution Time:
Weigh accurately on analytical balance 500 mg of sample, put it in the vial, reconstitute it with 2 ml of Water for Injection and note the time; it should not be more than 1 minute.
11.2.7 Test of pH after reconstitution:
Weigh accurately 5 g of sample and make the volume upto 50 ml with distilled water in the beaker and measure the pH of solution at 25±2°C; it should be 4 to 6.
11.2.8 Test of Colour after reconstitution:
Check the colour of the solution after reconstitution it should be Colourless.
11.2.9 Test of Clarity:
Weigh accurately on analytical balance 500 mg of sample, put it in the vial, and reconstitute it with 2 ml of Water for Injection. After reconstitution check the clarity; it should be clear solution and without any visible contamination.
11.2.10 Test of Moisture Contents:
Weigh accurately 10 g of sample on moisture balance. Adjust the temperature at 85°C, start drying the sample. At the end balance show percentage loss of moisture; moisture contents should not be more than 6%.
11.2.11 Water Contents:
Weigh accurately 50mg of sample and titrate for Karl Fischer water determination it should be not more than 6%.
11.2.12 Record the result of Physical Analysis on quality report.
11.3 CHEMICAL ANALYSIS: BP/USP/Inhouse
11.3.1 Examination of Cefazolin Sodium by IR spectrum:
Take dried sample to 5mg of Cefazolin Sodium 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 the standard spectrum.
11.3.2 Identification Test of Cefazolin Sodium:
Place about 2.0mg of sample in test tube add 0.05ml of water and add 2ml of Sulphuric acid Formaldehyde reagent. Mix the contents of the tube by swirling, the solution is pale yellow. Place the test-tube in a water-bath for 1min; a yellow colour develops.
11.3.3 Identification Test of Sodium:
Make the paste of 500 mg of Cefazolin sodium with 0.5 ml of HCl (Conc.). Moist the platinum wire with the paste and introduce it into the flame of Bunsen burner a golden yellow coloured flame should produce.
11.3.4 Contents of Cefazolin Sodium:
11.3.4.1 Standard Preparation:
Weigh accurately on analytical balance equivalent to 100 mg of Cefazolin Sodium and make the volume upto 100 ml with distilled water in volumetric flask.
Take 2 ml from the above step and make the volume upto 100 ml with 0.1M sodium bicarbonate solution in volumetric flask.
Measure the absorption of the standard at 272 nm using 0.1M sodium bicarbonate solution as a blank.
11.3.4.2 Sample Preparation:
Weigh accurately on analytical balance 100 mg of powdered sample of Cefazolin Sodium and make the volume upto 100 ml with distilled water in volumetric flask.
Take 2 ml from the above step and make the volume upto 100 ml with 0.1M sodium
bicarbonate solution in volumetric flask.
Measure the absorption of the sample at 272 nm using 0.1M sodium bicarbonate solution as a blank.
11.3.4.3 Observation:
a) Absorption of sample = X
b) Absorption of standard = Y
c) Record the readings
11.3.4.4 Calculations:
Calculate the contents of Cefazolin sodium as follows:
X
——————– x 100 = Z
Y
11.4 Limit: The contents of Cefazolin Sodium should be between 89.1% to 110.1% (On anhydrous basis)
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.