| GENERAL MATERIAL INFORMATION | |
| General Information | Vildagliptin and Metformin film coated tablets is consisting of Valsartan and Excipients. |
| Categories | Antidiabetic |
| Composition | Each film coated tablet contains:
Vildagliptin and Metformin ………50 mg &500 mg Vildagliptin and Metformin ………50 mg &1000 mg |
| Reference | Indian Pharmacopoeia 2022 |
| Sample Quantity | Chemical Analysis = 60 Tablets
Microbiology Analysis = 20 Tablets |
| Shelf Life | 24 Months
|
| Hazards and Precaution, If any | Use nose mask and hand gloves while sampling at bulk stage
|
| Storage | Store protected from moisture at a temperature not exceeding 30°.
|
| S.NO. | TESTS | SPECIFICATIONS | REFERENCE | ||
| 1. | Description | Orange color elongated shape film coated tablets having one side mid break line other side plain | IH | ||
| 2. | Average weight | 860 mg ± 3% | IH | ||
| 3. | Uniformity of weight | Within ± 5% of the Average weight | IH | ||
| 4. | Identification
|
In the assay, the principal peaks in the chromatogram obtained with the test solution (a) and test solution (b) correspond to the principal peaks in the chromatogram obtained with the reference solution (a) | IP | ||
| 5. | Hardness | NLT 4.0kg/cm2 | IH | ||
| 6. | Dimension | IH | |||
| 7. | Disintegration Time | NMT 30 min | IP | ||
| 8. | Leak Test | Should be Pass | IP | ||
| 9. | Dissolution | Q. Not less than 80% of the standard amount of Metformin and vildagliptin | IP | ||
| 10. | Related Substance | Determine by liquid chromatography | IP | ||
| 11. | Assay: | IP | |||
| Each filmcoated tablet contains: | Limit | ||||
| Claim | In mg | In % | |||
| Vildagliptin IP | 50 mg | 45 mg to 55 mg | 90% to 110% | ||
| Metformin IP | 500 mg | 450 mg to 550 mg | 90% to 110% | ||
| Pack size: | |||||
| Shelf life | 24 months | ||||
| Storage: Store in a cool, dark & dry place. | |||||
| Reference: As per I.P. | |||||
SPECIFICATION OF STABILITY ANALYSIS
| S.NO. | TESTS | SPECIFICATIONS | REFERENCE | ||
| 1. | Description | Orange color elongated shape film coated tablets having one side mid break line other side plain | IH | ||
| 2. | Average weight | 860mg ± 3% | IH | ||
| 3. | Uniformity of weight | Within ± 5% of the Average weight | IP | ||
| 4. | Identification
|
In the assay, the principal peaks in the chromatogram obtained with the test solution (a) and test solution (b) correspond to the principal peaks in the chromatogram obtained with the reference solution (a) | IP | ||
| 5. | Hardness | NLT 4.0 Kg/cm² | IH | ||
| 6. | Disintegration Time | NMT 30 min | IP | ||
| 7. | Dissolution | Q. Not less than 80% of the standard amount of Metformin and vildagliptin | IP | ||
| 8. | Related Substance | Determine by liquid chromatolgraphy | IP | ||
| 9. | Assay: | IP | |||
| Each filmcoated tablet contains: | Limit | ||||
| Claim | In mg | In % | |||
| Vildagliptin IP | 50 mg | 45 mg to 55 mg | 90% to 110% | ||
| Metformin IP | 500 mg | 450 mg to 550 mg | 90% to 110% | ||
*Related Substance: once in a year.
*Microbial Limit Test: once in a year.
| S.NO. | TESTS | SPECIFICATIONS | REFERENCE | ||
| 1. | Description | Orange color elongated shape film coated tablets having one side mid break line other side plain | IH | ||
| 2. | Average weight | 860mg ± 3% | IH | ||
| 3. | Uniformity of weight | Within ± 5% of the Average weight | IP | ||
| 4. | Identification
|
In the assay, the principal peaks in the chromatogram obtained with the test solution (a) and test solution (b) correspond to the principal peaks in the chromatogram obtained with the reference solution (a) | IP | ||
| 5. | Hardness | NLT 4.0 Kg/cm2 | IH | ||
| 6. | Dimension | ||||
| 7. | Disintegration Time | NMT 30 min | IP | ||
| 8. | Leak Test | Should be Pass | IP | ||
| 9. | Dissolution | Q.Not less than 80% of the standard amount of Metformin and vildagliptin | IP | ||
| 10. | Related Substance | Determine by liquid chromatography | IP | ||
| 11. | Assay: | IP | |||
| Each Filmcoated tablet contains: | Limit | ||||
| Claim | In mg | In % | |||
| Vildagliptin IP | 50 mg | 45 mg to 55 mg | 90% to 110% | ||
| Metformin IP | 1000 mg | 900 mg to 1100 mg | 90% to 110% | ||
SPECIFICATION OF STABILITY ANALYSIS
| S.NO. | TESTS | SPECIFICATIONS | REFERENCE | ||
| 1. | Description | Orange color elongated shape film coated tablets having one side mid break line other side plain | IH | ||
| 2. | Average weight | 860mg ± 3% | IH | ||
| 3. | Uniformity of weight | Within ± 5% of the Average weight | IP | ||
| 4. | Identification
|
In the assay, the principal peaks in the chromatogram obtained with the test solution (a) and test solution (b) correspond to the principal peaks in the chromatogram obtained with the reference solution (a) | IP | ||
| 5. | Hardness | NLT 4.0 Kg/Cm2 | IH | ||
| 6. | Disintegration Time | NMT 30 min | IP | ||
| 7. | Dissolution | Q.Not less than 80% of the standard amount of Metformin and vildagliptin | IP | ||
| 8. | Related Substance | Determine by liquid chromatography | IP | ||
| 9. | Assay: | IP | |||
| Each uncoated tablet contains: | Limit | ||||
| Claim | In mg | In % | |||
| Vildagliptin IP | 50 mg | 45 mg to 55 mg | 90% to 110% | ||
| Metformin IP | 1000 mg | 900 mg to 1100 mg | 90% to 110% | ||
*Related Substance: once in a year.
*Microbial Limit Test: once in a year .
Testing method
- Description: Visual
- Average Weight: Check weight of 20 tablets at randomly 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.
Tablets were weighed individually and the percentage of deviation of its weight from the average weight was determined for each tablet. Formula to calculate the percentage of deviation.
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 assay, the principal peaks in the chromatogram obtained with the test solution correspond to the principal 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) Dimension of Tablet:
Length, breadth and thickness are determined by vernier calliper in mm.
7.0) 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
8.0) 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.
9.0) Dissolion for Vildagliptin:
Medium : 900 ml, 0.01 M Hydrochloric acid
Apparatus : Paddle
Speed : 50rpm
Temperature: 37°C ±0.5°C
Time : 45 Min.
Dissolution medium: Transfer 0.85ml of Hydrochloric acid in 1000ml of volumetric flask and make up to volume with water and mix.
Chromatographic parameter
Column : C18, 250 mm x 4.6 mm ( 5µm)
Flow Rate : 1.0 ml/min.
Injection : 20 µl
Detection : 210nm
Column temperature: 25°C
Preparation of buffer: Dissolve 3.15gm of Ammonium dihydrogen orthophosphate and 0.68gm of Diammonium hydrogen orthophosphate in 1000ml of water. Adjust the pH 5.0 with orthophosphoric acid.
Preparation of Mobile Phase: Prepare a degassed mixture of 85 volume of buffer and 15 volume of Acetonitrile.
Preparation of Diluent: Water and Acetonitrile (90 +10)
Standard Solution: weigh accurately about 55.0 mg of vildagliptin WS into a 50ml volumetric flask, makeup the volume with diluents. Further dilute 5ml of the solution to 100ml with dissolution medium.
Sample Preparation: Place one tablet into each of the six dissolution vessels and start the dissolution test .At the specified sampling time, withdraw 20 ml from each dissolution vessel and filter.
Procedure: Separately inject 20 µl of black (Single injection). Standard solution (Five replicate injections), Sample solution (Single injection of each) and Bracketing Standard Solution (Single injection) into liquid chromatography and record the chromatograms. Measure the responses for Vildagliptin peaks. Calculate the quantity of Vildagliptin in percentage from the Vildagliptin peak areas of standard, sample and percentage potency of working standard used.
Dissolution for Metformin HCl :
Medium : 900 ml, phosphate buffer pH 6.8
Apparatus : Basket
Speed : 100rpm
Temperature: 37°C ±0.5°C
Time : 60 Min.
Dissolution medium: Dissolve 40.8 g of potassium dihydrogen phosphate and 4.8 g sodium hydroxide in 6000 ml of water; adjust the pH to 6.8 ± 0.05 with sodium hydroxide or orthophosphoric acid, if necessary.
Preparation of standard solution : E1% =806
Sample Solution for 500 mg : Set dissolution parameters and place one tablets into each vessel taking care to exclude air bubbles from the surface of the tablets and immediately start the apparatus. After 60 minutes, withdraw the sample of medium and filter through 0.45µ Nylon syringe filter, discard first few ml filtrate. Dilute 2 ml of the filtrates t0 200 ml with dissolution medium and mix well.
Sample Solution For 850 mg: Set dissolution parameters and place one tablet into each vessel taking care to exclude air bubbles from the surface of the tablets and immediately start the apparatus. After 60 minutes, withdraw the sample of medium and filter through 0.45µ Nylon syringe filter, discard first few ml filtrate. Dilute 1 ml of the filtrates t0 200 ml with dissolution medium and mix well.
Sample Solution For 100 mg : Set dissolution parameters and place one tablet into each vessel taking care to exclude air bubbles from the surface of the tablets and immediately start the apparatus. After 60 minutes, withdraw the sample of medium and filter through 0.45µ Nylon syringe filter, discard first few ml of filtrate. Dilute 1 ml of the filtrates to 200 ml with dissolution medium and mix well.
Procedure: Measure the absorbance of the blank (dissolution medium), Standard solution and the test solution at about 233 nm and record the observations.
Calculation:
For 500 mg:
Area of Test 1000 900 200
—————–X————X————X———X100
Even Value 100 500 2
For 1000 mg:
Area of Test 1000 900 200
—————–X————X————X———X100
Even Value 100 1000 1
10.0) Related Substance:
Determine by liquid chromatography
Buffer solution: Disslove 1.7 g of potassium dihydrogen phosphate in 1000 ml of water and adjust to pH 3.0 with orthophosphoric acid, add 8 gm of ammonium hexafluorophosphate and mix.
Solvent mixture: 90 volumes of water, 10 volumes of acetonitrile and 0.1 volume of orthophosphoric acid .
Solution A. : 40 volumes of the buffer solution, 60 volumes of water and 2.5 volumes of acetonitrile.
Test solution (a). Disperse 10 intact tablets in the solvent mixture, with the aid of magnetic stirrer for 45 minutes, dilute to 500.0ml with the solvent mixture. Centrifuge to get a clear supernatant,filter. Dilute 10.0ml of the filtrate to 20.0ml with the solvent mixture.
Test solution(b). Dilute a suitable volume of test solution (a) with solution A to obtain a solution having 0.02 per cent w/v of Metformin Hydrochloride.
Reference solution(a). A solution containing 0.05per cent w/v of vildagliptin IPRS and 0.02 per cent w/v of metformin hydrochloride IPRS in the solvent mixture.
Reference solution (b). Dilute 1.0ml of reference solution(a) to 100.0ml with the solvent mixture.
Reference solution (c).A.0.005 per cent w/v solution of dicyandiamide IPRS in the solvent mixture.
Reference solution (d). Dilute 5.0ml of reference solution (b) and 0.2ml of reference solution (c) to 50.0ml with the solvent mixture.
Reference solution (e).A solution containing 0.0125 per cent w/v each of, vildagliptin impurities A,B,C IPRS and dicyandiamide IPRS in the solvent mixture. Dilute 2.0ml of the solution to 50.0ml with the reference solution (a).
Chromatographic system
-a stainless steel column 15cm X 4.6mm, packed with octadecylsilane bonded to spherical silica (3.0µm)(Such as waters, Atlantis d C18 100 Å),
-column temperature:35°,
-mobile phase A:a mixture of 40 volumes of the buffer solution and 60 volumes of water,
B:a mixture of 40 volumes of 40 volumes of the buffer solution and 60 volumes of acetonitrile,
-a gradient programme using the conditions given below,
-flow rate: 1ml per minute,
-spectrophotometer set at 210nm (for vildagliptin)and 218nm (for Metformin hydrochloride),
-injection volume: 10µl.
| Time (in min.) | Mobile Phase A (percent v/v) | Mobile Phase B (percent v/v) |
| 0 | 97 | 3 |
| 1 | 97 | 3 |
| 2 | 85 | 15 |
| 5 | 75 | 25 |
| 12 | 25 | 75 |
| 12.1 | 97 | 3 |
| 18 | 97 | 3 |
| Name | Relative Retention Time | Correction Factor |
| Dicyandiamide Cyanoguanidine | 0.21 | 0.36 |
| Metformin Hydrochloride | 0.41 | —- |
| Vildagliptin Impurity Cyclic amidine | 0.87 | 0.58 |
| Vildagliptin Impurity Amide | 0.83 | 0.77 |
| Vildagliptin Impurity Diketopiperazine | 0.95 | 0.68 |
| Vildagliptin | 1.0 | —- |
Inject reference solution (b) , (d) and (e) at 210 nm the test is not valid unless the resolution between the peaks due to vildagliptin impurity B and vildagliptin impurity A is not less than 1.8 in the chromatogram obtained with reference solution (e), the tailing factor is not more than 2.0 and the relative standard deviation for replicate injections is not more than 5.0 percent for both the peaks in the chromatogram obtained less than 10 in the chromatogram obtained with reference solution (d) for all the three peaks.
Inject reference solution (b) and test solution (a) at 210 nm. In the chromatogram obtained with test solution (a), the area of any peak corresponding to vildagliptin impurity A and vildagliptin impurity C, each of, is not more than the area of vildagliptin peak in the chromatogram obtained with reference (b) (0.2 per cent), the area of any other secondary peak is not more than 0.2 times the area of the vildagliptin peak in the chromatogram obtained with the reference solution (b) (0.2 per cent) and the sum of the areas of all the secondary peak is not more than 3.5 times the area of vildagliptin peak in the chromatogram obtained with reference solution (b) (3.5 per cent). Ignore the peaks due to Metformin Hydrochloride and any peak with an area less than 0.05 times the area of the vildagliptin peak in the chromatogram obtained with reference solution (b) (0.05 per cent).
Inject reference solution (b) and test solution (b) at 218 nm. In the chromatogram obtained with the test solution (b), the area of any peak corresponding to dicyandiamide is not more than 0.02 times the area of principal peak in the chromatogram obtained with reference solution (b) (0.02 per cent), the area of any other secondary peak is not more than 0.1 the area of the Metformin Hydrochloride peak in the chromatogram obtained with reference solution (b) (0.1 per cent), and the sum of the areas of all the secondary peaks is not more than 0.6 times the area of Metformin Hydrochloride peak in the chromatogram obtained with reference solution (b) (0.6 percent). Ignore the peak due to vildagliptin.
11.0) Assay: Determine by liquid chromatography
- For Metformin HCl:
By UV spectroscopy
Weight and powder 20 tablets. Weigh a quantity of the powder containing about 0.1 g
of Metformin Hydrochloride, shake with 70 ml of water for 15 minutes, dilute to
100.0 ml with water and filter. Dilute 10.0 ml of the filtrate to 100.0 ml with water.
Further dilute 10.0 ml to 100.0 ml with water and measure the absorbance of the
resulting solution at the maximum at about 232 nm. Calculate the content of
Metformin HCl taking 798 as specific absorbance at 232 nm.
Calculation:
Test area 1000 100 100 100
Metformine HCl = ——————x ———— x ———x——-x——-x Avg wt of tablet
Specific absorbance 100 test wt. 10 10
Acceptance criteria: 90.0%-110.0%
- For Vildagliptin: Determine by liquid chromatography.
Chromatographic Parameters
Column : C18, 250mm X4.6mm (5µm)
Flow rate : 1.0 ml/minute
Injection volumes : 20 µl
Detection : 210 nm
Column Temp. : 35° C
Preparation of buffer: Dissolve 3.15 gm of Ammonium dihydrogen orthophosphate and 0.68 gm of diammonium hydrogen orthophosphate in 1000 ml of water. Adjust the pH to 5.0 with orthophosphoric acid.
Preparation of mobile phase: Prepare a degassed mixture of 85 volumes of buffer and 15 volumes of acetonitrile.
Preparation of Diluent: Water Acetonitrile (90:10)
Standard solution: Weigh accurately about 50.0 mg of vildagliptin working standard into a 50 ml volumetric flask, and 30 ml diluents, sonicate to 10 minutes and make up the volume with diluents. Further diluents 5 ml of the solution to 50 ml with diluents.
Sample preparation: Weigh accurately equivalent to 50.0 mg of sample into a 50 ml volumetric flask add 30 ml diluents, socinate to 10 minutes and makeup the volume with diluents. Further diluents 5 ml of the solution to 50 ml with diluents.
Procedure: Inject five replicate injections of Standard Solution and duplicate injections of sample solution into the chromatograph and record the chromatogram.
Sequence of injection:
| Sequence Table | ||
| Sr. No. | Sample | No. of Injection |
| 1 | Blank | 1 |
| 2 | Standard Solution | 5 |
| 3 | Sample Solution | 2 |
| 4 | Bracketing Standard | 1 |
Calculation for Vildagliptin:
Test area Std. wt. 5 50 5 Potency
———– x ———— x——–x ——– x———–x——– x Avg. wt of tablet
Std.area 50 50 Test wt. 50 100
Acceptance criteria: 90.0%-110.0%
10.0) 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
11.0) Abbreviations:
Wt.: Weight HCl: Hydrochloride
Mg: Milligram O.P.A.: Orthophosphoric acid
Ml: Milliliter Abs.: Absorbance
STD: Standard CFU: Colony forming unit
InHg: Inch of Mercury RPM: Rotations per minute
Temp. : Tempecature
