For misoprostol, identification, assay, and related substances were considered essential, given the link to efficacy and the known instability of the product. SOP for Finished Product Analysis of Misoprostol 200mcg Tablet by HPLC Method due to the low ratio of API content to tablet mass and therefore higher risk of non-homogeneity within batches, content uniformity was included. Dissolution or disintegration was not included as misoprostol is a highly soluble substance (19), and anecdotally these parameters are rarely found out of specification (OOS) for misoprostol products. Misoprostol tablets were analyzed for the selected parameters using the methods and specifications described in the International Pharmacopoeia
PURPOSE:
To describe the procedure for analysis at the in-process and finished stage of Misoprostol 200 mcg Tablet.
SCOPE:
This SAP gives a detailed outline for the finished product analysis of Misoprostol 200 mcg 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:
INTERNATIONAL PHARMACOPOEIA
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
- Distilled water
- 0.1 N Hydrochloric acid
- Ammonium Acetate
- Glacial Acetic Acid
- Methanol
PROCEDURE:
FINAL MIX
SOP for Finished Product Analysis of Misoprostol 200mcg Tablet by HPLC Method
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:
The retention time of the misoprostol peak of the sample solution corresponds to that of the standard solution, as obtained in the Assay.
Assay By HPLC
(Limit: 90%-110%)
Procedure:
Mobile phase: (Acetonitrile 45: Water: 55)
Standard solution: 0.02mg/mL of Misoprostol RS in Mobile phase
Sample solution: weigh and powder 20 tablets weigh accurately a quantity of powder equivalent to 400mcg of misoprostol in 20mL volumetric flask. Add about 10mL and sonicate for 10 minutes. Ensure that the temperature of sonication bath is below the room temperature to avoid degradation of misoprostol and make up volume. Filter a portion of this solution, discarding first few mL of the filtrate.
Chromatographic system
- Mode: LC
- Detector: UV 200 nm
- Column: 4.6-mm x 15-cm; 5-um packing L1
- Column Temperature: 35oC
- Auto sampler: 4oC
- Flow rate: 1.5 mL/min
- Injection size: 100 uL
System suitability
- Sample: Standard solution
Suitability requirements
- Tailing factor: Not less than 0.8 NMT 1.5 for misoprostol
- Relative standard deviation: NMT 2.0% for misoprostol
Analysis
- Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of misoprostol(C22H38O5) in the portion of Tablets taken:
- Result= (Ru/Rs) x (Cs/Cu) x 100
- Ru= peak response from the Sample solution
- Rs= peak response from the Standard solution
- Cs= concentration of Misoprostol RS in the Standard solution(mg/mL)
- Cu= nominal concentration of misoprostol in the Sample solution(mg/mL)
Limit: Misoprostol: 90%-110%of the labeled amount
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:
Thickness Standard Limit:
Length/Diameter Standard Limit:
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 x 100/W1
Disintegration Test:
Place the beaker filled with water into the beaker stand inside the bath and. Switch on the instrument, set the temperature at 37ºC± 2º C and wait till the temperature of the beaker reaches to the 37ºC± 2º C. Engage the basket on basket hook put the tablets in 6 tubes individually and place the discs then start the oscillations and timer. Note the disintegration time of all the six tablets.
Note the Disintegration time of 1st tablet to the last tablet.
Disintegration Time: NMT 15 minutes at 370C ± 20C
Acceptance criteria of disintegration for tablets:
If 1 or 2 tablets fail to disintegrate completely repeat the test on 12 additional tablets, not less than 16 of the totals of 18 tablets tested disintegrate completely.
BLISTER STAGE:
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.
DISSOLUTION TEST:
Apparatus: Apparatus II Peddle
Medium: Water; 500 mL
Speed: 50 rpm
Time: 30 Minute
Medium Temperature: 37º ± 0.5º
Recommended Sampling Time: Misoprostol: 30 Minute
Diluent: Dissolution medium
Determine the amount of misoprostol (C22H38O5)dissolved by using one of the following methods.
Spectrometric method
Standard stock solution: 0.08mg/mL of Misoprostol RS in Medium prepared as follows.
Dissolve a suitable quantity in volumetric flask, then dilute with Medium to volume.
Standard solution: Dilute the Standard stock solution with Medium to obtain a final concentration of0.0004 mg/mL.
Sample solution: Pass a portion of the solution under test through a suitablefilterof0.45um pore size.
Mobile Phase: Acetonitrile : Water 45:55
Chromatographic system
- Mode: LC
- Detector: UV 200 nm
- Column: 4.6-mm x 15-cm; 5-um packing L1
- Column Temperature: 35oC
- Autosampler: 4oC
- Flow rate: 1.5 mL/min
- Injection size: 250 uL
System suitability
- Sample: Standard solution
Suitability requirements
- Tailing factor: Not less than 0.8 NMT 1.5 for misoprostol
- Relative standard deviation: NMT 2.0% for misoprostol
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of misoprostol in the portion of Tablets taken:
Analysis
Calculate the percentage of the labeled amount of Misoprostol (C22H38O5)dissolved:
Result = (Au/As) x (Cs/L) x Vx 100
Au = Area under curve of the Sample solution
As = Area under curve of the Standard solution
Cs = concentration of the Standard solution(mg/mL)
L = label claim (mg/Tablet)
V = volume of Medium, 500 mL
Tolerance:
The amount of drug dissolve in solution for each tablet is not less Than 80% Q of the amount stated on the label for Misoprostol at 30 minutes.
CONTENT UNIFORMITY TEST:
Proceed as directed in assay
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