How To Test Cephradine L-Arginine Raw Material PURPOSE
This raw material specification & analysis prescribes the requirements for How To Test Cephradine L-Arginine Raw Material (Sterile) which is used in Injection.
SCOPE
How To Test Cephradine L-Arginine Raw Material is applicable in Material Department, Product Development and Quality Assurance Department to confirm its physical and chemical specifications.
REFERENCES
USP/BP/Inhouse/ACS DOBFAR s.p.a of How To Test Cephradine L-Arginine Raw Material.
DEFINITIONS
Follow How To Test Cephradine L-Arginine Raw Material SOPs
SPECIFICATIONS
USP/BP/Inhouse/ACS DOBFAR s.p.a of How To Test Cephradine L-Arginine Raw Material SAP
PHYSICAL ANALYSIS
- Physical: Form Crystalline Powder
- Colour: White to Creamy White
- Color after reconstitution: Light yellow
- Reconstitution Time: Not more than 3 minute
- pH: After Reconstitution 8 to 9.6 at 30°C
- Moisture Contents: Not more than 5%
- Reconstitution: 2.1 ml (±0.1 ml)
- Clarity: Clear Solution without any visible contamination
- Bulk Density: 0.400 – 0.500 g/ml
- Tapped Density: 0.550 – 0.700 g/ml
CHEMICAL ANALYSIS
- Identification of Cephradine examination by IR spectrum Should comply the standard spectrum
- Identification Test of Cephradine Solution should become green in colour
- Identification by HPLC Method Retention time of sample should match with the retention time of standard ±10%
- Content of Cephradine 62-69%
MICROBIOLOGICAL ANALYSIS
Potency of Cephradine 62-69%
SAMPLING
Refer to the Incoming SOPs for sampling of determination How To Test Cephradine L-Arginine Raw Material
REJECTIONS:
If the Cephradine L-Arginine (Sterile) does not satisfy the specifications mentioned in 5.0 then it will be rejected. In addition to the specifications, Cephradine L-Arginine (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.
PACKAGING
Cephradine L-Arginine (Sterile) is received in polyethylene bag/thick aluminium foil bag placed in study, metallic container to protect it from moisture and light.
STORAGE: USP
Store in container for sterile solids as How To Test Cephradine L-Arginine Raw Material
HANDLING
10.1 Use gloves & mask during weighing and dispensing operation of Cephradine L-Arginine (Sterile) in the sterile area.
10.2 Keep the container of Cephradine L-Arginine (Sterile) well closed immediately after use.
10.3 Place the container of Cephradine L-Arginine (Sterile) back to its specified area.
ANALYSIS
11.1(A) SUITABLE MACHINERY/EQUIPMENT/APPARATUS
- Moisture Balance
- pH Meter
- Test Tubes
- Beaker
- Glass Rod
- Test tubes
- 100 ml Volumetric Flask
- Pipette 50 ml, 25 ml, 20 ml (Bulb Type)
- Stoppered Titration Flask 250 ml
- Burette 50 ml
- Analytical balance
- Magnetic Stirrer
- Funnel
- FTIR, mortar & pestle
- HPLC system
11.1(B)LIST OF REAGENT/CHEMICAL
- Ethanol (96%) 10 ml
- Chloroform 10 ml
- Potassium bromide IR grade 200 mg
- Ether 10 ml
- 1 N Sodium Hydroxide 120 ml
- 1 N Hydrochloric Acid 150 ml
- 0.1 N Iodine 60 ml
- 0.1 N Sodium Thiosulphate 50 ml
- Starch Solution 5 ml
- Potassium Cupri Tartrate Solution 2 ml
11.2 PHYSICAL ANALYSIS USP/BP/Inhouse/ACS DOBFAR s.p.a
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 creamy white.
11.2.3 Test of Reconstitution:
Weigh accurately on analytical balance 770 mg of sample on weighing balance, 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).
11.2.4 Test of Clarity:
Weigh accurately on analytical balance 770 mg of sample, put it in the vial, reconstitute it with 2 ml of Water for Injection. After reconstitution check the clarity; it should be clear solution and without any visible contamination.
11.2.5 Test of Reconstitution Time:
Weigh accurately on analytical balance 770 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 3 minute.
11.2.6 Colour after Reconstitution:
Weigh accurately on analytical balance 770 mg of sample on weighing balance, put it in the vial, reconstitute it with 2 ml of Water for Injection; after reconstitution colour should be light yellow.
11.2.7 Test of pH After Reconstitution:
Weigh accurately 10 mg of sample and make the volume upto 100 ml with distilled water in the beaker. Take 50 ml from the resulting solution and measure the pH of solution at 30°C on pH meter; it should be 8.0 to 9.6 at 30°C.
11.2.8 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 5%.
1.1.2.9 Bulk Density:
Take sample in the measuring cylinder, the surface level of the sample in the cylinder should be even, note the volume without tapping, it should be between 0.400 – 0.500 g/ml.
11.2.10 Tapped Density:
Take sample in the measuring cylinder, the surface level of the sample in the cylinder should be even, note the volume with tapping, it should be between 0.550 – 0.700 gm/ml.
11.3 Record the result of Physical Analysis on quality report.
11.4 CHEMICAL ANALYSIS: USP/BP/Inhouse/ACS DOBFAR s.p.a
11.4.1 Examination of Cephradine L-Arginine by IR spectrum:
Take dried sample to 5mg of Cephradine L-Arginine 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 be complies the standard spectrum.
11.4.2 Identification Test of Cephradine:
Weigh accurately on analytical balance 20 mg of the sample, add 10 ml distilled water, 0.5 ml of Potassium Cupri Tartrate Solution; the solution becomes green in colour immediately.
11.4.3 Contents of Cephradine
11.4.3.1 Sample Preparation:
11.4.3.1.1 Weigh accurately on analytical balance 100 mg of powdered sample of Cephradine L-Arginine and make the volume upto 100-ml with distilled water in volumetric flask.
11.4.3.1.2 Take 25 ml from the step 11.4.3.1.1 and add 40 ml of 1 N Sodium Hydroxide Solution. Cover and allow to stand in the dark for 15 minutes. After 15 minutes, add 50 ml of 1N HCl and 20 ml of 0.1 N Iodine Solution. Again cover and allow to stand in the dark for 15 minutes.
11.4.3.1.3 Titrate the step 11.4.3.1.2 with the help of 50 ml burette with 0.1 N Sodium Thiosulphate Solution using Starch Solution as an indicator.
11.4.3.2 Standard Preparation:
11.4.3.2.1 Weigh accurately on analytical balance 100 mg of reference standard of Cephradine L-Arginine and make the volume upto 100 ml with distilled water in volumetric flask.
11.4.3.2.2 Take 25 ml from the step 11.4.3.2.1 and add 40 ml of 1 N Sodium Hydroxide Solution. Cover and allow to stand in the dark for 15 minutes. After 15 minutes, add 50 ml of 1N HCl and 20 ml of 0.1 N Iodine Solution. Again cover and allow to stand in the dark for 15 minutes.
11.4.3.2.3 Titrate the step11.4.3.2.2 with the help of 50 ml burette with 0.1 N Sodium Thiosulphate Solution using Starch Solution as an indicator.
11.4.3.3 Blank Preparation:
11.4.3.3.1 Take 40 ml of 1 N Sodium Hydroxide Solution and add 50 ml of 1 N Hydrochloric Acid and 20 ml of 0.1 N Iodine Solution.
11.4.3.3.2 Titrate the step 11.4.3.3.1 with the help of 50 ml burette with 0.1 N Sodium Thiosulphate Solution using Starch Solution as an indicator.
11.4.3.4 Observations:
a) Volume of 0.1 N Sodium Thiosulphate used for blank = P
b) Volume of 0.1 N Sodium Thiosulphate used for standard = Q
c) Volume of 0.1 N Sodium Thiosulphate used for sample = R
d) Volume of 0.1 N Iodine absorbed by standard = P – Q = S
e.) Volume of 0.1 N Iodine absorbed by sample = P – R = T
f) Record the reading.
11.4.3.5Calculations:
11.4.3.5.1 Calculate the contents of Cephradine as follows:
T
———————– x 100 = U
S
Limit: The contents of Cephradine should be 62% to 69%.
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.
11.6 MICROBIOLOGICAL ANALYSIS:
Follow SOPs
11.7 Alternate Method of Analysis for Cephradine on HPLC System:
11.8.1 Mobile Phase:
The mobile phase is constitutes of 600 ml of 0.01M sodium Acetate and 400 ml methanol, mix and maintain the pH 5.2 with glacial acetic acid.
After maintaining pH, degas and fitter through 0.45μm fitter paper the mobile phase before use.
11.8.2 Operation Parameter:
- Flow rate: 1.5 ml / min
- Detection: UV at 254nm
- Injection volume: 20µl
11.8.3 Standard preparation:
Weigh accurately equivalent to 40 mg of cephradine standard in a 50 ml of volume metric flask add 20 ml of mobile phase, sonicate it till dissolved and make up the volume up to 50ml with mobile phase.
Pippet out 5 ml of the above dilution in 50 ml of volume metric flask and make up the volume to 50 ml with mobile phase, filter through 0.22 µm when inject in to the HPLC and inject 20µl of standard preparation.
11.8.4 Sample Preparation:
Weight accurately equivalent to 40 mg of cephradine and transfer it into 50 ml volumetric flask flask, add 20 ml of mobile phase, sonicate it till dissolved and make up the volume to 50 ml.
Pippet out 5ml of the above dilution in 50 ml of volumetric flask and make up the volume to 50 ml with mobile phase.
Filter through 0.22 micron filter at time of injection in to the HPLC system and inject 20µl of sample preparation.
11.8.5 Identification of Cephradine by HPLC Method
Inject the standard and sample preparation respectively.
The retention time of sample should match with the retention time of standard ±10%.
11.8.6 Calculation:
Percentage of Cephradine contents:
Peak area of Sample x weight of Std. x Potency of standard
———————————————————————————– x 100
Peak area of std. X weight of sample
11.8.7 Limit: The contents of Cephradine should be between 95-100%.