Pantoprazole Sodium 40mg Tablet Testing is performed to check the quality and potency of the tablets. The testing is done using various methods such as HPLC, UV, and dissolution testing.

HPLC method is used to determine the amount of Pantoprazole Sodium in the tablet. The SOP for Pantoprazole Sodium 40mg Tablet Testing HPLC and UV Method is used to identify the presence of Pantoprazole Sodium in the tablet. Dissolution testing is used to determine the rate at which the active ingredient is released from the tablet.

PURPOSE:

To describe the procedure for analysis at in-process and finished stage of Pantoprazole Sodium 40 mg Tablet.

SCOPE:

This SAP gives a detailed outline for the finished product analysis of Pantoprazole Sodium 40 mg 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 procedures.
It is the responsibility of the QC Manager to assist and ensure the 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:

• UV Spectrophotometer
• HPLC
• Dissolution apparatus
• Friability apparatus
• Hardness apparatus
• Disintegration Tester
• Vernier Caliper
• Analytical Balance
• Moisture analyzer
• Mortar and pestle
• Spatula
• Filter Paper
• Magnetic Stirrer & Hot Plate
• Sonicator
• Vacuum Pump
• Glassware
• Methanol AR Grade
• Distilled water

PROCEDURE:

FINAL MIX

Description:

A 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 UV SPECTROPHOTOMETER)

The spectrum of the sample preparation corresponds to spectrum of the standard preparation 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: 200 mg /tablet

Procedure:

Solution A: Dissolve 3.85 g of ammonium acetate and 1.1 g of tetrabutylammonium hydrogen sulfate in 1 L of water, and adjust with ammonium hydroxide solution diluted with 1:1 water to a pH of 7.9.

Mobile Phase: Prepare mixture of Acetonitrile and Solution A (35: 65)

Diluent: Mixture of Acetonitrile and 0.02 N sodium hydroxide (1:1)

Standard Solution: Transfer a weighed quantity of Pantoprazole Sodium WS to a suitable volumetric flask, add 0.02 N sodium hydroxide to about 60% of the final volume, sonicate for 5 min to dissolve, add about 2% of Acetonitrile, and dilute with 0.02 N sodium hydroxide to volume to obtain a solution having a known concentration of about 0.2 mg/mL of Pantoprazole sodium.

Sample solution: Transfer 5 Tablets into a 100mL volumetric flask. Add Diluent to about 60% of the final volume, shake mechanically for about 60 min, and dilute with Diluent to volume. Pass through a suitable filter, and dilute the filtrate with 0.02 N sodium hydroxide to obtain a solution having a known concentration of about 0.2 mg/mL of Pantoprazole, based on the label claim.

Chromatographic system

• Mode: LC
• Detector: UV 290 nm.
• Column:4.6-mm x 25-cm; 5-µm packing L1
• Temperatures: Column: 30°C
• Flow rate: 1.0 mL/min
• Injection volume:20 µL

System Suitability:

• Samples: Standard solution
• Tailing Factor: NMT 2.0
• Relative standard deviation: NMT 2.0%

Calculations:
Average Sample AUC x Standard Concentration x Ave. Weight of Tablet x Potency
Average Standard AUC x Sample Concentration x Label Claim x Factor

ALTERNATE METHOD By UV

SOP for Pantoprazole Sodium 40mg Tablet Testing HPLC and UV Method

Standard Solution:

Take 45.1 mg of Pantoprazole sodium equivalent 40 mg of Pantoprazole WS dissolved in water and make up the volume to 100mL with water. 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 water.

Sample solution:

Take compression weight of finally grinded granules and dissolved in water and make up the volume to 200mL with water. 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 25mL volumetric flask make up volume with water. Take reading at UV spectrophotometer at 271 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 x Factor

Dissolution Test:

Acid Stage:

USP Apparatus: USP Apparatus II Peddle
Medium: 1000mL 0.1N HCl
Speed: 75 rpm
Time: 120 Minute
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: Pantoprazole: 120 Minute
Diluent: Prepare a mixture of pH 6.8 phosphate buffer and 0.5 N sodium hydroxide (1:1).
Mobile phase: Acetonitrile, triethylamine, and water (40:1:60). Adjust with phosphoric acid to a Ph of 7.0±0.05.
Standard Stock Solution: Transfer about 20 mg of Pantoprazole Sodium WS to a 50-mL volumetric flask. Add about 30 mL of 0.02 N sodium hydroxide, and sonicate until dissolved. Add 2 mL of Acetonitrile, and dilute with 0.02 N sodium hydroxide to volume.
Standard Solution: Transfer 1.0 mL of the Standard stock solution to a 20-mL volumetric flask, and dilute with Diluent to volume.
Sample solution: After 120 min, withdraw an aliquot, pass through a suitable filter of 0.45µmpore size, and immediately dilute a portion of the filtrate by a factor of 2 with 0.5 N sodium hydroxide. Transfer the Tablets to the vessels containing the Buffer stage medium.
Tolerances: NMT 10% of the labeled amount of pantoprazole is dissolved.

Buffer Stage:

USP Apparatus: USP Apparatus II Peddle
Medium: 1000mL;Phosphate Buffer pH 6.8.
Speed: 75 rpm
Time: 30 Minute
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: Pantoprazole: 30 Minute

Analysis: After 30 min, withdraw an aliquot, pass through a suitable filter of 0.45µ pore size, and immediately dilute a portion of the filtrate by a factor of 2 with 0.5 N sodium hydroxide. Determine the amount of Pantoprazole dissolved in the Buffer stage using the same procedure as for the Acid stage.

Instrumental Conditions
Mode: LC
Detector: UV 290 nm.
Column: 4.6-mm x 7.5-cm; 3-µm packing L1
Temperature: Column: 30°C
Flow Rate: 1.0 mL/min
Injection Volume: 10 µL

Calculations:
Sample Absorbance x Standard Concentration x Potency
Standard Absorbance x Sample Concentration x Factor
Tolerance: NLT 75% (Q) of the labeled amount of Pantoprazole is dissolved.

Test 2:

If the product complies with this test, the labeling indicates that the product meets USP Dissolution Test 2.Proceed as directed for Dissolution (711), Procedure, Apparatus7andApparatus 2, Delayed-Release Dosage Forms, Method B Procedure.

Acid stage

Acid stage medium: 0.1 N hydrochloric acid; 1000 mL
Apparatus 2: 100 rpm
Time: 2 h

Standard stock solution:
Transfer a quantity of USP Pantoprazole Sodium RS to a suitable volumetric flask. Dissolve first in 0.1 N sodium hydroxide, using 10% of the final volume, then dilute with pH 6.8 phosphate buffer to volume, to obtain a solution having a known concentration of about 0.46 mg of pantoprazole sodium per mL Mix well until a clear solution is obtained.

Calculate the concentration in mg of pantoprazole per mL, the molecular weights of pantoprazole and, pantoprazole sodium being 383.37 and 405.35,respectively.

Acid stage standard solution:
Dilute an appropriate volume of the Standard stock solution to 1 L with Acid stage medium in such a way as to obtain a final concentration of about 10% of the Tablet label claim per L.

Sample solution:
Pass a portion of the solution under test through a suitable filter of 10-um pore size.

Analysis:
Determine the amount of pantoprazole dissolved by using UV absorption at the wavelength of about 305 nm on portions of the Sample solution in comparison to the Acid stage working standard solution using a 4-cm path length cell and Acid stage medium as blank. Drain the Acid stage medium from each vessel and replace with Buffer stage medium.

Calculate the amount of pantoprazole dissolved:
Result= (Au/As) x Cs x Vx (100/L)
Au= absorbance of the Sample solution
As =absorbance of the Standard solution
Cs =concentration of pantoprazole in the Acid stage standard solution (mg/mL)
V =volume of Medium, 1000 mL
L =Tablet label claim of pantoprazole (mg)
Tolerances: NMT 10% of the labeled amount of pantoprazole is dissolved.

Buffer stage
Buffer stage medium: pH 6.8 phosphate buffer;1000 mL
Apparatus 2: 100 rpm
Time: 45 min

Buffer stage standard solution:
Dilute an appropriate volume of the Standard stock solution as described under Acid stage to 250 mL with Buffer stage medium in such away as to obtain a final concentration of about 100% of the Tablet label claim per L.

Sample solution:
Pass a portion of the solution under test through a suitable filter of 10-µm pore size.

Analysis:
Determine the amount of pantoprazole dissolved by using UV absorption at the wavelength of maximum absorbance at about 288 nm on portions of the Sample solution in comparison to Buffer stage Standard solution using a 0.5-cm path length cell and Buffer stage medium as blank.

Calculate the amount of pantoprazole dissolved:
Result= (Au/As) x Cs x V x (100/L)
Au=absorbance of the Sample solution
As=absorbance of the Buffer stage standard solution
Cs=concentration of pantoprazole in the Buffer stage standard solution (mg/mL)
V=volume of the Buffer stage medium, 1000 mL
L= Tablet label claim (mg)

Tolerances: NLT 75% (Q) of the labeled amount of pantoprazole is dissolved.

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 (WMax) & minimum (WMin) 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.

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

Leak Test:

Take 06 blisters of tablet 03 filled and 03 empty. Dip these blisters in the bowl of leak test apparatus containing colored water. Create vacuum 250 mmHg and hold for one minute. Release vacuum and check the penetration of water inside the blisters. All blisters should be free of water.

CONTENT UNIFORMITY TEST: By Weight variation
Calculation of Acceptance Value (AV):

Limits:
L1= ≤ 15 (For 10 Unit)
L2= ≤ 25 (For 30 Unit)
General Formula = M-x̄ ±ks
Acceptability Constant = (k)
Standard Deviation (SD) = s
Number of Units = (n)
Reference Value = M
Mean of Individual Content = x̄