Action and use
Stimulates insulin release; treatment of diabetes mellitus.
DEFINITION
Repaglinide Tablets contain Repaglinide.
The tablets comply with the requirements stated under Tablets and with the following requirements.
Content of repaglinide, C27H36N2O4
95.0 to 105.0% of the stated amount.
IDENTIFICATION
A. Carry out the method for thin-layer chromatography, Appendix III A, using the following solutions prepared in methanol.
(1) Mix with the aid of ultrasound a quantity of powdered tablets containing 5 mg of Repaglinide with 15 mL of methanol. Dilute to 50 mL, filter (a 0.45-μm nylon filter is suitable) and use the filtrate.
(2) 0.01% w/v of repaglinide BPCRS.
CHROMATOGRAPHIC CONDITIONS
(a) Use as the coating silica gel F254 (Merck silica gel 60 F254 plates are suitable).
(b) Use the mobile phase as described below.
(c) Apply 25 μL of each solution.
(d) Develop the plate to 5 cm.
(e) After removal of the plate, dry in air and examine under ultraviolet light (254 nm).
MOBILE PHASE
25 volumes of methanol, 30 volumes of toluene and 45 volumes of dichloromethane.
CONFIRMATION
The principal spot in the chromatogram obtained with solution (1) is similar in position and size to that in the chromatogram obtained with solution (2).
B. In the Assay, the retention time of the principal peak in the chromatogram obtained with solution (1) is similar to that of the peak in the chromatogram obtained with solution (2).
TESTS
Dissolution
Comply with the dissolution test for tablets and capsules, Appendix XII B1.
TEST CONDITIONS
(a) Use Apparatus 2, rotating the paddle at 50 revolutions per minute.
(b) Use 900 mL of 0.1M hydrochloric acid, at a temperature of 37°, as the medium.
PROCEDURE
(1) After 30 minutes withdraw a sample of the medium and filter (a 0.45-μm Millex LCR PTFE filter is suitable). Use the filtered medium, diluted with the dissolution medium if necessary to produce a solution expected to contain 0.00006% w/v
of Repaglinide.
(2) 0.006% w/v of repaglinide BPCRS in methanol. Dilute 1 volume to 100 volumes with the medium.
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (12.5 cm × 4.6 mm) packed with end-capped octadecylsilyl silica gel for chromatography (10 μm) (Nucleosil C18 is suitable).
(b) Use isocratic elution and the mobile phase described below.
(c) Use a flow rate of 1 mL per minute.
(d) Use a column temperature of 40°.
(e) Use fluorimetric detection with an excitation wavelength of 244 nm and an emission wavelength of 348 nm.
(f) Inject 20 μL of each solution.
MOBILE PHASE
11 volumes of methanol, 40 volumes of 0.01M potassium dihydrogen orthophosphate adjusted to pH 2.3 with orthophosphoric acid and 49 volumes of acetonitrile.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (2), the symmetry factor of the peak due to repaglinide is between 0.8 and 1.8.
DETERMINATION OF CONTENT
Calculate the total content of repaglinide, C27H36N2O4, in the medium from the chromatograms obtained and using the declared content of C27H36N2O4, in repaglinide BPCRS.
LIMITS
The amount of repaglinide released is not less than 75% (Q) of the stated amount.
Related substances
Carry out the method for liquid chromatography, Appendix III D, using the following solutions, protected from light and prepared in a mixture of equal volumes of acetonitrile R1 and mobile phase A (solution A).
(1) Shake a quantity of the powdered tablets containing 5 mg of Repaglinide in 2 mL of water. Add 10 mL of solution A, mix with the aid of ultrasound and dilute to 20 mL. Centrifuge and use the supernatant liquid.
(2) Dilute 1 volume of solution (1) to 100 volumes. Further dilute 1 volume of this solution to 5 volumes.
(3) Dilute 1 volume of repaglinide impurity standard solution BPCRS to 10 volumes with solution A.
(4) 0.1% w/v of repaglinide for system suitability EPCRS.
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (15 cm × 4.6 mm) packed with end-capped extra-dense bonded octylsilyl silica gel for chromatography (5 μm) (Zorbax Eclipse XDB C8 is suitable).
(b) Use gradient elution and the mobile phase described below.
(c) Use a flow rate of 1.5 mL per minute.
(d) Use a column temperature of 40°.
(e) Use detection wavelengths of 210 nm and 240 nm.
(f) Inject 20 μL of each solution.
MOBILE PHASE
Mobile phase A: 0.02M potassium dihydrogen orthophosphate adjusted to pH 2.9 with orthophosphoric acid.
Mobile phase B: acetonitrile R1.
| Time (Minutes) | Mobile phase A (% v/v) | Mobile phase B (% v/v) | Comment |
| 0-4 | 75 | 25 | isocratic |
| 4-30 | 75→50 | 25→50 | linear gradient |
| 30-40 | 50 | 50 | isocratic |
| 40-41 | 50→75 | 50→25 | linear gradient |
| 41-50 | 75 | 25 | re-equilibration |
When the chromatograms are recorded under the prescribed conditions, the relative retentions with reference to repaglinide (retention time about 24 minutes) are: impurity A, about 0.1; impurity B, about 0.4; impurity C, about 0.6; impurity 1, about 1.05; and impurity D, about 1.6.
SYSTEM SUITABILITY
The test is not valid unless:
At 210 nm
in the chromatogram obtained with solution (2), the signal to noise ratio of the peak due to repaglinide is at least 40;
At 240 nm
in the chromatogram obtained with solution (3), the resolution between the peaks due to repaglinide and impurity 1 is at least 1.8.
LIMITS
At 210 nm
Identify any peak corresponding to impurity C in the chromatogram obtained with solution (1), using the chromatogram obtained with solution (4), and multiply the area of this peak by a correction factor of 2.2.
In the chromatogram obtained with solution (1):
the area of any peak corresponding to impurity C is not greater than 2.5 times the area of the peak in the chromatogram obtained with solution (2) (0.5%).
At 240 nm
Identify any peak corresponding to impurities A and B in the chromatogram obtained with solution (1), using the chromatogram obtained with solution (4), and multiply the areas of these peak by a correction factor of: 0.6 and 0.7 respectively.
In the chromatogram obtained with solution (1):
the area of any peak corresponding to impurity A is not greater than 2.5 times the area of the peak in the chromatogram obtained with solution (2) (0.5%);
the area of any other secondary peak is not greater than the area of the principal peak in the chromatogram obtained with solution (2) (0.2%).
Disregard any peak with an area less than half the area of the principal peak in the chromatogram obtained with solution (2) (0.1%).
The sum of all impurities, including impurity C, is not greater than 1.0%.
Uniformity of content
Repaglinide Tablets containing less than 2 mg and/or less than 2% w/w of Repaglinide comply with the requirements stated under Tablets using the following method of analysis. Carry out the method for liquid chromatography, Appendix III
D, using the following solutions.
(1) To one tablet add 3 mL of the mobile phase, mix with the aid of ultrasound and shake for 15 minutes. Dilute to 5 mL with mobile phase and filter (a 0.45-μm Millex LCR PTFE filter is suitable). Dilute the filtrate, if necessary, with mobile phase to produce a solution containing the equivalent of 0.01% w/v of repaglinide.
(2) 0.1% w/v of repaglinide BPCRS in methanol. Dilute 1 volume to 10 volumes with the mobile phase.
(3) Dilute 1 volume of repaglinide impurity standard solution BPCRS to 10 volumes with the mobile phase.
CHROMATOGRAPHIC CONDITIONS
(a) Use a stainless steel column (15 cm × 4.6 mm) packed with end-capped extra-dense bonded octylsilyl silica gel for chromatography (5 μm) (Zorbax Eclipse XDB C8 is suitable).
(b) Use isocratic elution and the mobile phase described below.
(c) Use a flow rate of 1 mL per minute.
(d) Use a column temperature of 45°.
(e) Use a detection wavelength of 242 nm.
(f) Inject 10 μL of each solution.
MOBILE PHASE
25 volumes of 0.007M potassium dihydrogen orthophosphate adjusted to pH 2.5 with orthophosphoric acid and 75 volumes of methanol.
When the chromatograms are recorded under the prescribed conditions, the retention time of repaglinide is about 6 minutes.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3), the resolution between impurity 1 and repaglinide is at least 2.0.
DETERMINATION OF CONTENT
Calculate the content of repaglinide, C27H36N2O4, in the tablets from the chromatograms obtained and using the declared content of C27H36N2O4 in repaglinide BPCRS.
ASSAY
For tablets containing the equivalent of less than 2 mg and/or less than 2% w/w of Repaglinide
Use the average of the individual results determined in the test for Uniformity of content.
For tablets containing the equivalent of 2 mg or more and 2% w/w or more of Repaglinide
Carry out the method for liquid chromatography, Appendix III D, using the following solutions.
(1) Add 75 mL of the mobile phase to 10 whole tablets, mix with the aid of ultrasound and shake for 15 minutes. Dilute to 100 mL with mobile phase and filter (a 0.45-μm Millex LCR PTFE filter is suitable). Further dilute this solution, if necessary, with the mobile phase to produce a solution containing 0.01% w/v of Repaglinide.
(2) 0.1% w/v of repaglinide BPCRS in methanol. Dilute 1 volume to 10 volumes with the mobile phase.
(3) Dilute 1 volume of repaglinide impurity standard solution BPCRS to 10 volumes with solution A.
CHROMATOGRAPHIC CONDITIONS
The chromatographic conditions described under Uniformity of content may be used.
SYSTEM SUITABILITY
The test is not valid unless, in the chromatogram obtained with solution (3), the resolution between impurity 1 and repaglinide is at least 2.0.
DETERMINATION OF CONTENT
Calculate the content of repaglinide, C27H36N2O4, in the tablets from the chromatograms obtained and using the declared content of C27H36N2O4 in repaglinide BPCRS.
IMPURITIES
The impurities limited by the requirements of this monograph include those listed under Repaglinide and:
1. 2-ethoxy-4-(2-{(1S)-1-[2-(pyrolidin-1-yl)phenyl]-2-phenylethylamino}-2-oxoethyl)benzoic acid.
