1.0) Description: Visual
2.0) Average Weight: Check weight of 20 tablets at random and calculate the average weight by formula.
Average weight (mg) = wt of 20 tablets (gm) x 1000
20
3.0) Uniformity of weight:
Weigh 20 tablets selected at random and calculate the average weight. Not more than two of the individual weights deviate from the average weight by more than the percentage shown in table.
Weigh the tablets individually and calculate the percentage of deviation for each tablet by using formula:
Deviation (%) = Weight of each tablet – Average weight x 100
Average weight
| Average weight of tablets | Percentage deviation |
| 250mg or More | ±5% |
4.0) Identification Test:
In the assay, the principal peaks in the chromatogram obtained with the test solution corresponds to the peaks in the chromatogram obtained with the reference solution.
5.0) Hardness:
The standard method used for tablet hardness testing is compression testing. The tablet is placed between two jaws that crush the tablet. The machine measures the force applied to the tablet and detects when it fractures.
6.0) Disintegration Time:
Unless otherwise stated in the individual monograph, introduce one tablet into each tube and add a disc to each tube. The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1-litre beaker. The volume of liquid is such that the wire mesh at its highest point is at least 15 mm below the surface of the liquid, and at its lower point is at least 25 mm above the bottom of the beaker. At no time should the top of the basket-rack assembly become submerged. There is a thermostatic arrangement for heating the liquid and maintaining the temperature at 37±2°.
If 1 or 2 tablets fail to disintegrate, repeat the test on 12 additional tablets; not less than 16 of the total of 18 tablets tested disintegrate.
If the tablets adhere to the disc and the preparation under examination fails to comply, repeat the test omitting the disc. The preparation complies with the test if all the tablets in the repeat test disintegrate.
7.0) Dimension of Tablet:
Length, breadth and thickness are determined by vernier calliper in mm.
8.0) Friability:
The test is applicable to compressed tablets and is intended to determine the physical strength of tablets. Tablets with a unit weight equal to or less than 650 mg, take a sample of whole tablets corresponding as near as possible to 6.5 g. For tablets with a unit weight of more than 650mg, 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.
The test is run only once unless the results are difficult to interpret or if the weight loss is greater than the target value, in which case, the test is repeated twice and the mean of the three tests is determined.
% Friability = W1 – W2 X 100
W1
W1= Initial weight W2= Final weight
A maximum loss of weight (From a single test or from the mean of three tests) not greater than 1.0 percent is acceptable for most tablets. If obviously cracked, chipped or broken tablets are present in the sample after tumbling, the sample fails the test.
9.0) Leak test:
The apparatus is used to test for the integrity of packed strips, blisters and Alu-Alu Blister pack containing tablets. Ensure apparatus bath is filled with purified water upto mark indicated and add 0.5% crystal violet solution in water. Samples are placed into the desiccators and the lid is placed in position. The pump starts to produce a vacuum 15inHg inside the desiccators and the vacuum is held for 1 minute. The sample remains at the required vacuum level for given time interval buzzer will sound after time is over and will cut off the vacuum pump. As the package is immersed in a colored dye solution the venting of the desiccators will allow any holes to be penetrated by the dye and the contents of the flexible packaging will also be stained with the same coloring material.
Examine all the strips for any leakage by opening the pockets manually. If anyone pocket shows evidence of leakage, reject the sample, stop the Blister / Strip machine and immediately take corrective action.
10.0) Assay: By HPLC Method ( For Paracetamol + Nimesulide)
Determined by liquid chromatography.
Test solution: Weigh and powder 20 tablets. Disperse a quantity of powder containing 50 mg of Nimesulide with 50 ml of methanol with the aid of ultrasound and dilute to 100 ml with methanol. Filter the sample and dilute 2 ml of this solution to 25 ml with Mobile phase.
Reference solution: Weigh 50 mg of Nimesulide reference standard and 162.5 mg of Paracetamol reference standard in 100 ml volumetric flask. Add 50 ml methanol and dissolve the standard and make up to mark with methanol. Dilute this solution 2 ml to 25ml with mobile phase.
Chromatographic system:
- A stainless steel column 250 mm X 4.6 mm, packed with octadecylsilane bonded to porous silica (5µm),
- Mobile Phase: Methanol: water (80:20) pH 4.5 with G.A.A.
- Flow rate: 1.5 ml per minute,
- Spectrophotometer set at 276 nm,
- Injection volume: 20 µl
Inject the reference solution and test solution.
Calculate the contents of Paracetamol and Nimesulide in the tablets.
Formula:
Area of Test Std Wt.(mg) 2 100 25 Potency
—————–X—————-X——–X—————X———-X————-X Average wt.
Area of STD 100 25 Test Wt.(mg) 2 100
Alternative Method: By UV Spectrophotometer
For Paracetamol
Test Solution: Weigh and powder 20 tablets. Weight sample equivalent to 50 mg of Paracetamol 100 ml volumetric flask add 20 ml 0.1 M HCl into it and sonicate to dissolve the test sample. Make up to mark with 0.1 M HCl. Filter and dilute 1 ml of this solution to 50 ml in 0.1 M HCl.
Reference Solution: Weigh 50 mg of reference standard of Paracetamol 100 ml of volumetric flask. Add 20 ml 0.1 M HCl into it and sonicate to dissolve. Make up to mark with 0.1 M HCl. Dilute 1 ml of this solution to 50 ml with 0.1 M HCl.
Check Absorbance of test as well as reference standard at about 243 nm in UV Spectrophotometer. Calculate the content of Paracetamol in the tablets.
Formula:
Abs. of Test STD Wt.(mg) 1 100 50 Potency
—————–X————–X——-X—————X——–X———–X Average wt.
Abs. of STD 100 50 Test Wt.(mg) 1 100
Acceptance criteria: 90.0%-110.0%
For Nimesulide
Test Solution: Weigh and powder 20 tablets. Weight sample equivalent to 25 mg of Nimesulide 100 ml volumetric flask add 20 ml 0.1 M NaOH into it and sonicate to dissolve the test sample. Make up to mark with 0.1 M NaOH. Filter and dilute 1 ml of this solution to 25 ml in 0.1 M NaOH.
Reference Solution: Weigh 25 mg of reference standard of Nimesulide 100 ml of volumetric flask. Add 20 ml 0.1 M NaOH into it and sonicate to dissolve. Make up to mark with 0.1 M NaOH. Dilute 1 ml of this solution to 25 ml with 0.1 M NaOH.
Check Absorbance of test as well as reference standard at about 391 nm in UV Spectrophotometer. Calculate the content of Nimesulide in the tablets.
Formula:
Abs. of Test STD Wt.(mg) 1 100 25 Potency
—————–X————–X——-X—————X——–X———–X Average wt.
Abs. of STD 100 25 Test Wt.(mg) 1 100
Acceptance criteria: 90.0%-110.0%
B) Assay of Serratiopeptidase( by UV)
The tyrosine units are dissolved in sodium carbonate solution, which is alkaline in nature. The folin s ciocalteau reagent helps in color development where the tyrosine units bind to the copper molecule in the reagent and causes the reduction of phosphomolybdate which is present in the reagent .There is formation of tyrosine-copper molybdate colored sample .The intensity of color depends upon the tyrosine units present which is read at 660nm.
Disodium tetraborate buffer .Dissolve 19.0 gm of disodium tetraborate in 900 ml of water adjusted to pH 9.0 with 1 M hydrochloric acid and dilute to 1000 ml with water
Casein Hammerstein substrate. Dissolve 1.2 gm of casein Hammerstein to 100ml of the sodium tetraborate buffer.Allow the casein to dissolve and form homogeneous solution.Boil the solution in boling water bath for 2 minutes.After removing from boling water wath,cool the casein substrate immediately in ice cold water .filter this solution and dilute to 200ml with water
Trichoroacetic acid reagent: dissolve 18gm of trichloroacetic acid, 30gm of anhydrous sodium acetate and 20ml of glacial acetic acid in 1000 ml of water,
Diluted folin’s ciocalteau reagent: Dilute 1 ml of Folin’S Ciocalteau reagent with 2 ml of water to get 3 ml of the reagent, Note: “Folin’s Ciocalteau reagent should be of the protein stimation grade and
Not the indicator grade. Folin;s reagent should be diluted to just before addition to the tube since it is sensitive to light.do not prepare this solution at the beginning of the assay”
Tyrosine reference solution .Dissolve 160 mg of tyrosine in 100ml of 0.2 M hydrochloric acid Dilute 1ml of this solution to 100 ml with 0.2 M hydrochloric acid. Use 2 ml of this solution for color development.
Trisodium phosphate buffer ph 6.8: weigh 19g of trisodium phosphate and 6.4 ml of hydrochloric acid in 1000 ml of water adjusted to PH 6.8.
Test Solution: weigh and powder 20 tablets. Disperse a quantity of powder containing about 120mg of serratiopeptidase with 100ml of trisodium phosphate buffer pH 6.8.Stir on magnetic stirrer for 1 hour. Further dilute 5.0ml to 25 ml with 5.0 per cent ammonium sulphate solution.stir for 5 minutes and filter.Dilute 2 ml of the filtrate to 100ml with sodium tetraborate buffer. 1 ml of resulting solutions is used for analysis .set the water wath to 37ᵒ
For each sample, keep four test tubes. mark two test tubes as test and two as blank add 5 ml of casein substrate to the tubes and two as blank Add 5 ml of casein substrate to the tubes marked as test and 5 ml. of TCA to the test tube marked as blank.Prewarm these tubes for five minutes along
With the tubes containing the sample dilutions. Add 1 ml of enzyme solution to all these tubes and vertex the tubes for exactly 10 seconds. Keep the test tubes in the water bath at 37ᵒ for 20 minutes for the reactions. After exactly 20 minutes add 5 ml of TCA to tubes marked as test and 5 ml of casein substrate to the tube marked as blank .Vertex all the tubes exactly for 30 seconds. Keep the tubes in the water bath at 37ᵒ for 30 minutes for precipitations; filter Mark tube as test as test, blank, tyrosine and tyrosine blank
Add the following respectively
Test 2 ml of the test filtrate.
Blank -2 ml of blank filtrate.
Tyrosine– 2 ml of tyrosine standard solution
Tyrosine blank –0.2M hydrochloric acid
Add 5 ml of 6 percent sodium carbonate solution to all tubes
Add 1 ml of dilute folin’s reagent to all the tube .keep the tubes at 37ᵒ for the 30 minutes for color development .Measure the absorbance at about 660 nm
Calculation: A1-A2 1 100 25 100
Serratiopeptidase Units/Tablets = ———-X176X——-X————x ——-x———x Avg. wt in gm
A3-A4 20 SPL.wt(g) 5 2
Serratippeptidase Units/ Tablets
% of L.A. = ————————————– x 100
L.A.
Where 176 = Conversion co-efficient of Tyrosine to Serratipeptidase
20 = Reaction time (Min.)
A1 = Absorbance of test solution
A2 = Absorbance of blank solution
A3= Absorbance of tyrosine reference solution
A4 = Absorbance of 0.2M Hydrochloric acid
L.A. = 30 000units/tablets
11.0) MICROBIOLOGICAL PURITY:
Perform the test according to requirements of IP,
Total aerobic Microbial count (TAMC): NMT 103 CFU/g
Total combined yeasts/Moulds count (TYMC): NMT 102 CFU/g
Pathogens: in 1gm drug.
Escherichia Coli – Should be absent
Pseudomonas aeroginosa – Should be absent
Salmonella – Should be absent
Staphylococcus aureus– Should be absent
Abbreviations:
Wt.: Weight
mg: Milligram
ml: Millilitre
STD: Standard
inHg: Inch of Mercury
rpm: Rotations per minute
CFU: Colony forming unit
O.P.A.: Orthophosphoric Acid
HCl: Hydrochloric Acid
NaOH: Sodium Hydroxide
G.A.A.: Glacial Acetic acids
