Repaglinide
452.59(S)-2-Ethyoxy-4-[2-[[methyl-1-[2-[(1-piperidinyl)phenyl]butyl]amino]-2-oxoethyl]-benzoic acid.
(+)-2-Ethoxy-
-[[(S)--isobutyl-o-piperidinobenzyl]carbamoyl]-p-toluic acid
[135062-02-1].» Repaglinide contains not less than 98.0 percent and not more than 101.0 percent of C27H36N2O4, calculated on the dried basis.
Packaging and storage—
Preserve in tight containers.
USP Reference standards 
11
—
USP Repaglinide RS.
USP Repaglinide Related Compound A RS.
USP Repaglinide Related Compound B RS.
USP Repaglinide Related Compound C RS.
11—
USP Repaglinide RS.
USP Repaglinide Related Compound A RS.
USP Repaglinide Related Compound B RS.
USP Repaglinide Related Compound C RS.
Identification—
Loss on drying (see Thermal Analysis 891)—
Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument, using about 30 mg of Repaglinide, accurately weighed. Heat the specimen at the rate of 10
per minute between 30 and 210 in an atmosphere of nitrogen at a flow rate of 200 mL per minute. From the thermogram, determine the accumulated loss in weight between 30 and 200: it loses not more than 0.7% of its weight.
891)—
Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument, using about 30 mg of Repaglinide, accurately weighed. Heat the specimen at the rate of 10 per minute between 30 and 210 in an atmosphere of nitrogen at a flow rate of 200 mL per minute. From the thermogram, determine the accumulated loss in weight between 30 and 200: it loses not more than 0.7% of its weight.
Residue on ignition 281:
not more than 0.1%, an ignition temperature of 600 ± 25 being used.
281:
not more than 0.1%, an ignition temperature of 600 ± 25 being used.
Heavy metals, Method II 231:
0.001%.
231:
0.001%.
Chromatographic purity—
Solution A—
Prepare a filtered and degassed monobasic potassium phosphate solution (3 in 1000). Adjust with 1 N sodium hydroxide to a pH of 7.0.
Solution B—
Use filtered and degassed methanol.
Mobile phase—
Use variable mixtures of Solution A and Solution B as directed for Chromatographic system. Make adjustments if necessary (see System Suitability under Chromatography 621).
621).
System suitability solution—
Dissolve accurately weighed quantities of USP Repaglinide RS,USP Repaglinide Related Compound A RS, USP Repaglinide Related Compound B RS, and USP Repaglinide Related Compound C RS in methanol to obtain a solution having known concentrations of about 10 mg of USP Repaglinide RS per mL and 100 µg each of the related compound Reference Standards per mL.
Test solution—
Transfer about 100 mg of Repaglinide, accurately weighed, to a 10-mL volumetric flask, dissolve in and dilute with methanol to volume, and mix.
Standard solution—
Transfer 0.1 mL of the Test solution to a 10-mL volumetric flask, dilute with methanol to volume, and mix.
Chromatographic system (see Chromatography 621)—
Prepare as directed in the Assay, except to program the chromatograph as follows.
Chromatograph the System suitability solution, and record the peak areas as directed for Procedure: the relative retention times are about 0.3 for repaglinide related compound B, 0.9 for repaglinide related compound C, 1.0 for repaglinide, and 1.6 for repaglinide related compound A. Chromatograph the Standard solution, and record the peak areas as directed for Procedure: the relative standard deviation for replicate injections is not more than 10%.
621)—
Prepare as directed in the Assay, except to program the chromatograph as follows.
| Time (minutes) |
Solution A
(%) |
Solution B
(%) |
Elution |
| 0 | 50 | 50 | equilibration |
| 0–2 | 50®30 | 50®70 | linear gradient |
| 2–8 | 30 | 70 | isocratic |
| 8–12 | 30®5 | 70®95 | linear gradient |
| 12–15 | 5 | 95 | isocratic |
Procedure—
Separately inject equal volumes (about 3 µL) of the Test solution and the Standard solution into the chromatograph, record the chromatograms, and measure the peak areas. Calculate the percentage of each impurity, other than repaglinide related compound A, in the portion of Repaglinide taken by the formula:
ri / rS
in which ri is the peak response for each impurity obtained from the Test solution; and rS is the peak response of repaglinide obtained from the Standard solution. For repaglinide related compound A, use the same formula, but multiply the result by a response factor equal to 2.0: not more than 0.1% of any individual impurity is found, and not more than 0.5% of total impurities is found.
Assay—
Buffer solution—
Prepare a monobasic potassium phosphate solution (1 in 1000), and adjust with phosphoric acid to a pH of 2.5.
Mobile phase—
Prepare a filtered and degassed mixture of methanol and Buffer solution (80:20).
System suitability preparation—
Dissolve accurately weighed quantities of USP Repaglinide RS and USP Repaglinide Related Compound B RS in methanol to obtain a solution having known concentrations of about 500 µg per mL and 40 µg per mL, respectively.
Standard preparation—
Dissolve an accurately weighed quantity of USP Repaglinide RS in methanol, and dilute quantitatively, and stepwise if necessary, with methanol to obtain a solution having a known concentration of about 500 µg per mL.
Assay preparation—
Transfer about 25 mg of Repaglinide, accurately weighed, to a 50-mL volumetric flask, dissolve in and dilute with methanol to volume, and mix.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 240-nm detector and a 4.6-mm × 12.5-cm column that contains 5-µm packing L1. The flow rate is about 1.0 mL per minute. The column temperature is maintained at 45. Chromatograph the System suitability preparation, and record the peak areas as directed for Procedure: the relative retention times are about 1.0 for repaglinide and 0.4 for repaglinide related compound B. Chromatograph the Standard preparation, and record the peak areas as directed for Procedure: the relative standard deviation for replicate injections is not more than 2.0%.
621)—
The liquid chromatograph is equipped with a 240-nm detector and a 4.6-mm × 12.5-cm column that contains 5-µm packing L1. The flow rate is about 1.0 mL per minute. The column temperature is maintained at 45. Chromatograph the System suitability preparation, and record the peak areas as directed for Procedure: the relative retention times are about 1.0 for repaglinide and 0.4 for repaglinide related compound B. Chromatograph the Standard preparation, and record the peak areas as directed for Procedure: the relative standard deviation for replicate injections is not more than 2.0%.
Procedure—
Separately inject equal volumes (about 10 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the areas for the major peaks. Calculate the quantity, in mg, of C27H36N2O4 in the portion of Repaglinide taken by the formula:
50C(rU / rS)
in which C is the concentration, in mg per mL, of USP Repaglinide RS in the Standard preparation; and rU and rS are the peak responses obtained from the Assay preparation and the Standard preparation, respectively.
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
Chromatographic Column—
| Topic/Question | Contact | Expert Committee |
| Monograph | Elena Gonikberg, Ph.D.
Senior Scientist 1-301-816-8251 |
(MDGRE05) Monograph Development-Gastrointestinal Renal and Endocrine |
| Reference Standards | Lili Wang, Technical Services Scientist 1-301-816-8129 RSTech@usp.org |
USP32–NF27 Page 3484
Pharmacopeial Forum: Volume No. 27(6) Page 3325
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.