SIMULTANEOUS ESTIMATION OF GLIBENCLAMIDE, GLICLAZIDE, AND METFORMIN HYDROCHLORIDE FROM BULK AND COMMERCIAL PRODUCTS USING A VALIDATED ULTRA FAST LIQUID CHROMATOGRAPHY TECHNIQUE

Abstract

A rapid, precise, sensitive, economical, and validated analytical method is reported for simultaneous separation and quantification of three anti-diabetic drugs, viz., glibenclamide (GLB), gliclazide (GLC), and metformin hydrochloride (MHC) using ultra fast liquid chromatography (UFLC). The separation of the three drugs was achieved using a XR-ODS C18 column (30°C) with a mobile phase comprised of acetonitrile-water-trifluoroacetic acid-triethylamine (54:46:0.1:0.1v/v) in isocratic elution mode at a flow rate of 0.38 mL/min and detected at 230 nm. System suitability tests essential for the assurance of quality performance of the method were performed. The method was validated for accuracy, precision, reproducibility, robustness, detection (LOD), and quantification (LOQ) limits according to FDA and ICH guidelines. MHC (Rt = 0.98 min), GLC (Rt = 4.10 min), and GLB (Rt = 6.40 min) separated with good resolution in a single chromatographic run of 7.5 min. Linear relationship (r² > 0.999) was observed between the peak area and concentration for all the three compounds within the range of 5–50 µg/mL. Accuracy ranged from 98 to 103% and the coefficient of variation for precision was found to be less than 3%; in all cases. LOD and LOQ values were 10 ng/mL and 20 ng/mL, respectively, for GLC and GLB; whereas 25 ng/mL and 35 ng/mL, respectively, for MHC. The method was found to be robust with minor changes in injection volume and column temperature. Validation results indicated that the method shows satisfactory linearity, precision, accuracy, and ruggedness. The extremely low flow rate, short run time, and simple mobile phase composition makes the method cost effective, rapid, non-tedious, and can also be successfully employed for simultaneous analysis of the three anti-diabetic drugs from commercial products.

Keywords:

• glibenclamide
• gliclazide
• metformin hydrochloride
• simultaneous analysis
• UFLC
• validation

ACKNOWLEDGMENTS

The authors would like to thank Mrs. Elizabeth Abraham and Mrs. Doha Nabeel for their technical assistance.

Notes

†RSD of peak areas of five consecutive injections at a concentration of 10 µg/mL of each drug.

‡Resolution between (MHC & GLC) & (GLC & GLB).

*SD = Standard deviation (n = 3).

*RSD% = SD/Mean × 100.

†SEM = Standard error of mean.

*n = 3.

†RE (%) = % Relative error = (Mean assayed concentration – added concentration)/Added concentration × 100.

*Theoretical content of each drug = 40 µg/mL.

**n = 3.