1.0) Description: Yellow color elongated shape, biconvex, uncoated tablet having one side mid break line and other side plain.
2.0) Average Weight: Check weight of 20 tablets at randomly. And calculate the average weight by formula :
wt of 20 tablets (gm) x 1000 / 20. found avg. wt in mg
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 = (experimental weight – theoretical weight) x 100%
Theoretical weight
| Average weight of tablets | Percentage deviation |
| More than 80mg but Less than 250mg | 7.5% |
| 250mg or More | 5% |
- Identification Test:
- Determined by thin-layer chromatography, using silica gel R6 as the coating substance and a mixture of 40 volumes of light petroleum R1, 10 volumes of ethyl acetate R and 5 volumes of glacial acetic acid R as the mobile phase. Apply separately to the plate 10μl of each of the following two solutions in acetone R. For solution (A) shake a quantity of the powdered tablets containing about 10 mg of Artemether (about 60 mg of Lumefantrine) for 5 minutes with 10 ml, filter, and use the clear filtrate. For solution (B) use 1 mg of artemether reference standard and 6 mg of lumefantrine reference standard per ml. After removing the plate from the chromatographic chamber, allow it to dry exhaustively in air or in a current of cool air. Examine the chromatogram in ultraviolet light (254 nm).
The principal spot obtained with solution A corresponds in position, appearance, and intensity to that obtained with solution B (identifying lumefantrine).
Spray the plate with sulfuric acid/methanol TS. Heat the plate for 10 minutes at 140°C. Examine the chromatogram in daylight.
The principal spot obtained with solution A corresponds in position, appearance, and intensity to that obtained with solution B (identifying artemether; a faint spot due to lumefantrine may also be visible).
- Carry out the test Thin-layer chromatography, using the conditions described above under test A but using silica gel R5 as the coating substance After removing the plate from the chromatographic chamber, allow it to dry exhaustively in air or in a current of cool air. Spray with sulfuric acid/methanol TS. Heat the plate for 10 minutes at 140°C, allow it to cool and expose to iodine vapours for 20 minutes. Examine the chromatogram immediately in daylight. The principal spots obtained with solution A correspond in position, appearance, and intensity to those obtained with solution B.
- 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. Although compressive force is applied to the tablet, along the diameter of the tablet at right angles to the applied force.
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 15mm below the surface of the liquid, and at its lower point is at least 25mm 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 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 6.5g. 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.
Formula: initial wt. – after friability wt. x100 / initial wt.
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.
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) Related substances:
Protect samples from light, also during chromatography.
Carry out the test as described under identification thin-layer chromatography, using silica gel R5 as the coating substance and a mixture of 40 volumes of light petroleum R1, 10 volumes of ethyl acetate R and 5 volumes of glacial acetic acid R as the mobile phase.
Prepare the following solutions in the solvent consisting of 1 volume of water R and 1 volume of acetonitrile R. For solution (1), weigh and powder 20 tablets. To a quantity of the powder containing 100 mg of Artemether add 20 ml of the solvent sonicate for 15 minutes and centrifuge.
Filter a portion of the supernatant through a 0.45-μm filter, discarding the first few ml of the filtrate. For solution (2) dissolve 5 mg of each of artemether reference standard, artenimol reference standard and α-artemether reference standard in 50ml of the solvent. For solution (3) dilute 2.0 ml of solution (2) to 20 ml with the solvent. For solution (4) dilute 3.0 ml of solution (2) to 20 ml with the solvent. For solution (5) dilute 5.0 ml of solution (2) to 20 ml with the solvent. For solution (6) dilute 1.0 ml of solution (2) to 2 ml with the solvent. For solution (7) dilute 3.0 ml of solution (2) to 4 ml with the solvent.
Apply separately to the plate 20μl of each of solutions (1), (3), (4), (5), (6) and (7). After application allow the spots to dry for 15 minutes in a current of cool air. Develop over a path of 12 cm. After removing the plate from the chromatographic chamber, allow it to dry exhaustively in air or in a current of cool air. Dip the plate in vanillin/sulfuric acid TS2. Heat the plate for 10minutes at 140°C. Examine the chromatogram in daylight. Artemether and related substances have the following Retention factor values: impurity A about 0.25; impurity B (artenimol) about 0.3; impurity C about 0.35; impurity D (α-artemether) about 0.4; artemether about 0.55. The test is not valid unless the chromatogram obtained with solution (3) shows three clearly separated spots.
In the chromatogram obtained with solution (1):
- Any spot corresponding to impurity A is not more intense than the principal spot in the chromatogram obtained with solution (7) (1.5%),
- any spot corresponding to impurity B is not more intense than the spot due to artenimol in the chromatogram obtained with solution (6) (1.0%),
- any spot corresponding to impurity C is not more intense than the principal spot in the
Chromatogram obtained with solution (5) (0.5%),
- any spot corresponding to impurity D is not more intense than the spot due to α-artemether
in the chromatogram obtained with solution (4) (0.3%),
- any other spot is not more intense than the principal spot in the chromatogram obtained
with solution (3) (0.2%). Disregard any spot remaining at the point of application.
11.0) Assay:
11.1) Determine by High-performance liquid chromatography, using a stainless steel column (15 cm x 3.9 mm) packed with particles of silica gel, the surface of which has been modified with chemically bonded octadecylsilyl groups (5μm). Use the following conditions for gradient elution:
Mobile phase A: 700 volumes of ion pair reagent and 300 volumes of acetonitrile R.
Mobile phase B: 300 volumes of ion pair reagent and 700 volumes of acetonitrile R.
Prepare the ion pair reagent by dissolving 5.65 g of sodium hexanesulfonate R and 2.75 g of sodium dihydrogen phosphate R in about 900 ml of water R. Adjust the pH to 2.3 using phosphoric acid (~105 g/l) TS, dilute to 1000 ml and filter through a 0.45-μm filter.
| Time
(min)
|
Mobile phase A
(% v/v)
|
Mobile phase B
(% v/v)
|
Comments |
| 0-28 | 60 | 40 | Isocratic |
| 28-29 | 60-0 | 40-100 | Linear gradient |
| 29-45 | 0 | 100 | Isocratic |
| 45-46 | 60-60 | 100-40 | Return to initial
composition |
| 46-55 | 60 | 40 | Isocratic re-equilibration |
Prepare the following solutions in the solvent which is obtained by mixing 200 ml of ion pair
reagent, 60 ml of water R and 200 ml of 1-propanol R and diluting to 1000 ml with acetonitrile R. Prepare and keep both solutions at a temperature not below 20°C. For solution (1), weigh and powder 20 tablets. Transfer a quantity of the powder containing about 20 mg of Artemether (about 120 mg of Lumefantrine), accurately weighed, to a 100-ml volumetric flask. Add approximately 85 ml of the solvent, sonicate for 20 minutes, allow to cool to room temperature and dilute to volume with the solvent. Filter through a 0.45 μm-filter, discarding the first few ml of the filtrate. For solution (2), accurately weigh 20 mg of artemether RS and 120 mg of lumefantrine RS in a 100 ml volumetric flask. Add approximately 85 ml of solvent, sonicate until dissolved, allow to cool to room temperature and dilute to volume. Operate with a flow rate of 1.3 ml per minute. As a detector use an ultraviolet spectrophotometer set at a wavelength of about 210 nm for the first 28 minutes and then switch to about 380 nm.
Inject alternately 20μl each of solutions (1) and (2). (The peak for artemether is eluted at retention time of approximately 19 minutes, and that for lumefantrine at a retention time of approximately 34
minutes.) Measure the areas of the peak responses obtained in the chromatograms from solutions (1) and (2), and calculate the content of artemether (C16H26O5) and lumefantrine (C30H32Cl3NO) in the tablets.
11.2) Alternate method
For Artemether: (By HPLC)
Test solution: Weigh and powder 20 tablets. Weight sample containing 40mg Artemether in 100ml of volumetric flask. Add 10ml of mobile phase and sonicate the sample to dissolve and makeup to the mark with mobile phase. Filter the solution and inject.
Reference solution: Weigh 20mg of Artemether reference standard in 50ml volumetric flask. Add 10ml of mobile phase and dissolve. Sonicate the sample & shake by mechanical means for 5 min, add 40ml of mobile phase.
Chromatographic system:
- A stainless steel column 15cm X4.6mm, packed with octadecylsilane bonded to porous silica (5µm) (such as thermo quest hypersil), C18
- Mobile Phase: Acetonitrile + water (60:40) and filter.
- Flow rate: 1.2ml per minute,
- Spectrophotometer set at 216nm,
- Injection volume: 20µl
Inject reference solution. The test is not valid unless the relative standard deviation for replicate injections for each peak is not more than 2.0 percent.
Inject reference solution and the test solution.
Calculate the content of Artemether in the tablet.
Formula:
Area of Test STD Wt.(mg) 100 Potency
—————–X——————-X—————–X————–X Average weight
Area of STD 50 Test Wt.(mg) 100
Acceptance criteria: 90.0%-110.0%
For Lumefantrine: (By UVspectroscopy)
Reference solution: Weigh accurately about 50 mg of Lumefantrine reference standard, in 100 ml volumetric flask and dissolve in 10ml 0.1M methanolic HCl and make up the volume with 0.1M methanolic HCl. Take 1 ml of this solution in 25 ml volumetric flask and make up the volume with 0.1M methanolic HCl.
Test solution: Weigh and powder 20 tablets. Take sample equivalent to 50mg of Lumefantrine in 100 ml volumetric flask and dissolve in 10ml 0.1M methanolic HCl, sonicate the sample and make up volume with 0.1 M methanolc HCl. Filter, and take 1 ml of this solution in 25 ml volumetric flask and make up the volume with 0.1M methanolic HCl
Scan the dilutions from 200nm to 500nm in UV spectrophotometer. The solution gives absorbance maxima at about 303nm.
Formula:
Abs. of Test STD Wt.(mg) 1 100 25 Potency
—————–X————–X———X—————–X——–X———–X Average weight
Abs. of STD 100 25 Test Wt.(mg) 1 100
Acceptance criteria: 90.0%-110.0%
12.0) 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: Miligram
ml: Milileter
HCl: Hydrochloric acid
STD: Standard
inHg: Inch of Mercury
CFU: Colony forming unit
rpm: rounds per minute
UV: Ultraviolet
Abs.: Absorbance
Ion pair solution:
