SOP for Diclofenac Sodium SR 100mg Tablet Tested UV and HPLC Methods to determine its purity and quality. Both UV and HPLC methods are widely used in pharmaceutical laboratories to test the quality and purity of diclofenac sodium tablets. These methods are highly accurate and reliable and are used to ensure that the tablets meet the required quality standards.
PURPOSE:
To describe the procedure for analysis at in-process and finished stage of Diclofenac Sodium SR 100mg Tablet.
SCOPE:
This SAP gives a detailed outline for the finished product analysis of Diclofenac Sodium SR 100mg Tablet and will cover In process testing activities on a physical, chemical & instrumental basis.
RESPONSIBILITY:
QC Analyst is responsible for physical / chemical testing and preparing standard analytical procedure.
It is the responsibility of QC Manager to assist and ensure Testing Procedure as per SAP and to make certain that this SAP is followed in its entirety, reviewed regularly and revised as necessary.
REFERENCE:
USP 43, NF 38
MATERIAL AND EQIUPMENT:
- HPLC
- Dissolution apparatus
- Friability apparatus
- Hardness apparatus
- Disintegration Tester
- Vernier Caliper
- Analytical Balance
- pH meter
- Moisture analyzer
- Mortar and pestle
- Spatula
- Filter Paper
- Magnetic Stirrer & Hot Plate
- Sonicator
- Vacuum Pump
- Glassware Ammonium Acetate
- Acetonitrile HPLC Grade
- Methanol HPLC Grade
- Distilled water
- Phosphoric acid
- Monobasic Potassium phosphate
- Ammonium Acetate
PROCEDURE:
FINAL MIX
Description:
white powder.
Procedure: Take 2.0 g of the test sample in a watch glass and observe visually with black background. Check the appearance of color, nature and any visible foreign particles.
Identification: (By HPLC)
A- The retention time of the major peak in the chromatogram of the assay sample preparation corresponds to that in the chromatogram of the standard preparation as obtained in the assay.
B- The UV (190-400 nm) spectrum of the diclofenac peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay.
Loss on Drying: (By Moisture Analyzer)
Procedure:
Turn on instrument 20 minute before the test. Take 2 g powder spread uniformly on Moisture analyzer plate. Close the lid and press start button.
NMT 5.0% when determined by LOD Apparatus, Use 2.0 g of powder.
Assay: (Limit: 90%-110%)Compression Weight of Powder/Tablet: 250 mg /tablet
HPLC testing involves separating the different components of diclofenac sodium using high-performance liquid chromatography (HPLC) and measuring the amount of diclofenac sodium present in the tablet. This method is more accurate and sensitive than the UV method.
In the HPLC method, a sample of the diclofenac sodium tablet is dissolved in a suitable solvent and injected into an HPLC system. The HPLC system consists of a mobile phase (a solvent or a mixture of solvents), a stationary phase (a column), and a detector. The mobile phase is pumped through the column, and the different components of the diclofenac sodium tablet are separated based on their chemical properties. The detector measures the amount of diclofenac sodium in the sample, and the concentration is calculated based on the peak area or peak height obtained.
Procedure:
Solution A: 0.7708 g/L of ammonium acetate in water. Adjust with acetic acid to a pH of 5.3. Pass through a suitable filter of 0.2-um pore size.
Solution B: Acetonitrile
Mobile phase: See Table 1.
Table 1
| Time (Minute） | Solution A (%) | Solution B (%) |
| 0 | 70 | 30 |
| 0.5 | 70 | 30 |
| 8.5 | 5 | 95 |
| 10 | 5 | 95 |
| 10.01 | 70 | 30 |
| 14 | 70 | 30 |
Diluent: Acetonitrile and water (50:50)
Standard solution: 0.2 mg/mL of USP Diclofenac Sodium RS in Diluent
Sample solution: Nominally 0.2 mg/mL of diclofenac sodium in Diluent prepared as follows. Transfer a suitable portion of diclofenac sodium to a suitable volumetric flask from NLT 20 finely powdered Tablets. Add Diluent equivalent to 50% of the flask volume. Dissolve with sonication for 25 min at 45° and fill with Diluentto volume. Pass the solution through a suitable filter of 0.22-um pore size.
[NOTE-Protect the Standards solution and Sample solution from light.]
Chromatographic system
Mode: LC
Detector: UV 280 nm. For Identification test B, use a photo-diode array detector.
Column:10-cm x 2.0-mm; 1.9-um packing L1
Column Temperatures: 35°C
Flow rate: 0.3 mL/min
Injection volume: 1 µL
System Suitability:
Samples: Standard solution
Tailing Factor: NMT 1.2
Relative standard deviation: NMT 2.8%.
Calculations:
Average Sample AUC x Standard Concentration x Ave. Weight of Tablet x Potency
Average Standard AUC x Sample Concentration x Label Claim
(Limit for Diclofenac Sodium: 90%-110%of the labeled amount)
UV Testing:
UV testing involves measuring the absorbance of diclofenac sodium at a particular wavelength using a UV-visible spectrophotometer. The absorbance of diclofenac sodium is measured at a wavelength of 276 nm, which is the maximum absorption wavelength of diclofenac sodium. The absorbance value obtained is used to calculate the concentration of diclofenac sodium in the tablet.
ALTERNATE METHOD
SOP for Diclofenac Sodium SR 100mg Tablet Tested UV and HPLC Methods
Standard Solution:
Take 50 mg of Diclofenac Sodium WS dissolved in methanol and make up the volume to 50mL with methanol. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 2 mL from standard stock solution and make up the volume to 100 mL with water.
Sample solution:
Take weight of finally grinded granules equivalent to 100mg of Diclofenac Sodium and dissolved in methanol and make up the volume to 100mL with methanol. Shake by mechanical mean for 10 minutes to disperse and allow any insoluble matter to settle.
Pass a portion of the sample stock solution through a suitable filter of 0.45um pore size.
Transfer the 2 mL of filtrate to a 100mL volumetric flask make up volume with water. Take reading at UV spectrophotometer at 276 nm using water as blank.
Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim
(Limit for Diclofenac Sodium: 90%-110%of the labeled amount)
Uniformity of Dosage Units: (By Weight Variation)
Weigh 20 tablets individually and calculate the average weight as:
Average weight = (Weight Of 20 Tablets)/20
Weigh all these tablets individually and mark among these weights maximum (W Max) & minimum (W Min) weight tablets. Calculate the maximum and minimum variation in percent as:
Maximum% age variation =((WMax) – (WAvg.))/WAvg×100
Minimum % age variation ((WMin) – (WAvg.))/WAvg×100
Hardness Test & Dimensions:
Perform the hardness test on 10 tablets and take the average. Power on the instrument and regulate zero adjustment and preset the ‘’HARDNESS’’ and “THICKNESS” mode. Place the tablet on the test plate, begin testing and read the hardness thickness and diameter. Clean the shattered tablet and print out test results, perform the test on 10 tablets and take the average.
Hardness Standard Limit: NMT 10 Kg
Friability Test:
For tablets with a unit mass equal to or less than 650 mg, take a sample of whole tablets corresponding to 6.5 g. For tablets with a unit mass of more than 650 mg, take a sample of 10 whole tablets. The tablets should be carefully dedusted prior to testing. Accurately weigh the tablet sample, and place the tablets in the drum. Rotate the drum 100 times, and remove the tablets. Remove any loose dust from the tablets as before, and accurately weigh. Generally, the test is run once. If obviously cracked, cleaved, or broken tablets are present in the tablet sample after tumbling, the sample fails the test. If the results are doubtful or if the weight loss is greater than the targeted value, the test should be repeated twice and the mean of the three tests determined. A maximum mean weight loss from the three samples of not more than 1.0% is considered acceptable for most products.
% FRIABILITY= W1-W2 /W1 x 100
Dissolution Test:
Dissolution Medium:
USP Apparatus: USP Apparatus II Paddle use wire sinkers.
Medium: 0.05 M phosphate buffer, pH 7.5;
Speed: 50rpm
Volume: 900mL
Recommended Sampling Time: 1, 5, 10, 16, and 24 h
Diluent: Dissolution medium
Standard solution: Take 27.7 mg of Diclofenac Sodium WS dissolved in medium and make up the volume to 50mL with medium. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 2 mL from standard stock solution and make up the volume to 50 mL with medium.
Analysis: Pass portions of the solution under test through a suitable filter.
Dilute 5mL to 25 mL with Medium.
Instrumental Conditions
Mode: UV
Analytical wavelength: UV 276 nm
Cell length: 10 mm
Blank: Medium
Calculations:
Calculate the amount of Diclofenac Sodium dissolved by the formulae
Result = (Au/As) x Cs x V x D x (1/L) X 100.
Where,
Au= Absorbance of sample
As = Absorbance of WS
Cs= concentration in mg/ml of the working standard solution.
V = volume in ml of medium (900ml),
D = dilution factor of the sample solution
L = label claim (mg/Tablet).
Tolerance:
| Time in hours | Amount Dissolved |
| 1 | 15%-35% |
| 5 | 45%-65% |
| 10 | 65%-85% |
| 16 | 75%-95% |
| 24 | NLT 80% |