Repaglinide
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C27H36N2O4 452.59
(S)-2-Ethyoxy-4-[2-[[methyl-1-[2-[(1-piperidinyl)phenyl]butyl]amino]-2-oxoethyl]-benzoic acid.
(+)-2-Ethoxy-a-[[(S)-a-isobutyl-o-piperidinobenzyl]carbamoyl]-p-toluic acid [135062-02-1].
»Repaglinide contains not less than 98.0percent and not more than 101.0percent of C27H36N2O4,calculated on the dried basis.
Packaging and storage— Preserve in tight containers.
Identification—
A: Infrared Absorption á197Kñ.
B: Ultraviolet Absorption á197Uñ—
Solution: 25µg per mL.
Medium: methanol.
Specific rotation á781Sñ :between +6.3and +7.3,at 20.
Test solution: 50mg per mL,in methanol.
Loss on drying (see Thermal Analysis á891ñ)— Determine the percentage of volatile substances by thermogravimetric analysis on an appropriately calibrated instrument,using about 30mg of Repaglinide,accurately weighed.Heat the specimen at the rate of 10per minute between 30and 210in an atmosphere of nitrogen at a flow rate of 200mLper minute.From the thermogram,determine the accumulated loss in weight between 30and 200:it loses not more than 0.7%of its weight.
Residue on ignition á281ñ: not more than 0.1%,an ignition temperature of 600±25being used.
Chromatographic purity—
Solution A— Prepare a filtered and degassed monobasic potassium phosphate solution (3in 1000).Adjust with 1Nsodium hydroxide to a pHof 7.0.
Solution B— Use filtered and degassed methanol.
Mobile phase— Use variable mixtures of Solution Aand Solution Bas directed for Chromatographic system.Make adjustments if necessary (see System Suitabilityunder Chromatography á621ñ).
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 RSin methanol to obtain a solution having known concentrations of about 10mg of USP Repaglinide RSper mLand 100µg each of the related compound Reference Standards per mL.
Test solution— Transfer about 100mg of Repaglinide,accurately weighed,to a 10-mLvolumetric flask,dissolve in and dilute with methanol to volume,and mix.
Standard solution— Transfer 0.1mLof the Test solutionto a 10-mLvolumetric flask,dilute with methanol to volume,and mix.
Chromatographic system (see Chromatography á621ñ)— 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
Chromatograph the System suitability solution,and record the peak areas as directed for Procedure:the relative retention times are about 0.3for repaglinide related compound B,0.9for repaglinide related compound C,1.0for repaglinide,and 1.6for 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%.
Procedure— Separately inject equal volumes (about 3µL)of the Test solutionand the Standard solutioninto 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 riis the peak response for each impurity obtained from the Test solution;and rSis 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 (1in 1000),and adjust with phosphoric acid to a pHof 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 RSand USP Repaglinide Related Compound B RSin methanol to obtain a solution having known concentrations of about 500µg per mLand 40µg per mL,respectively.
Standard preparation— Dissolve an accurately weighed quantity of USP Repaglinide RSin methanol,and dilute quantitatively,and stepwise if necessary,with methanol to obtain a solution having a known concentration of about 500µg per mL.
Assay preparation— Transfer about 25mg of Repaglinide,accurately weighed,to a 50-mLvolumetric flask,dissolve in and dilute with methanol to volume,and mix.
Chromatographic system(see Chromatography á621ñ)— 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 1.0mLper 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 1.0for repaglinide and 0.4for 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 10µL)of the Standard preparationand the Assay preparationinto the chromatograph,record the chromatograms,and measure the areas for the major peaks.Calculate the quantity,in mg,of C27H36N2O4in the portion of Repaglinide taken by the formula:
50C(rU/rS),
in which Cis the concentration,in mg per mL,of USP Repaglinide RSin the Standard preparation;and rUand rSare the peak responses obtained from the Assay preparationand the Standard preparation,respectively.
Auxiliary Information— Staff Liaison:Elena Gonikberg,Ph.D.,Scientist
Expert Committee:(PA4)Pharmaceutical Analysis 4
USP28–NF23Page 1710
Pharmacopeial Forum:Volume No.27(6)Page 3325
Phone Number:1-301-816-8251