Simultaneous determination of metformin and gliclazide in human plasma by liquid chromatography–tandem mass spectrometry: application to a bioequivalence study of two formulations in healthy volunteers

Abstract

A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated to simultaneously determine gliclazide and metformin in human plasma using huperzine A as the internal standard (IS). After acetonitrile-induced protein precipitation of the plasma samples, gliclazide, metformin and the IS were subjected to LC/MS/MS analysis using electro-spray ionization (ESI). Chromatographic separation was performed on a Hypersil BDS C18 column (50 mm x 2.1 mm, i.d., 3 microm). The method had a chromatographic running time of 2.0 min and linear calibration curves over the concentration ranges of 10-10,000 ng ml(-1) for gliclazide and 7.8-4678.9 ng ml(-1) for metformin. The recoveries of the method were found to be 71-104%. The lower limits of quantification (LOQ) of the method were 10.0 and 7.8 ng ml(-1) for gliclazide and metformin, respectively. The intra- and interday precision was less than 15% for all quality control samples at concentrations of 100, 500, and 2000 ng ml(-1). The validated LC/MS/MS method has been used to study bioequivalence in healthy volunteers. These results indicate that the method was efficient with a very short running time (2.0 min) for metformin and gliclazide compared to the methods reported in the literature. The presented method had acceptable accuracy, precision and sensitivity and was used in clinical bioequivalence study.

Multi-component plasma quantitation of anti-hyperglycemic pharmaceutical compounds using liquid chromatography–tandem mass spectrometry

“…Methods include LC procedures for metformin and glipizide with a lower limit of quantitation (LLQ) of 5.0 and 22.5 ng/mL [23], or metformin and glimepride with a LLQ of 22 and 39 ng/mL, respectively [24]. Methods based on LC/MS/MS have been reported for metformin and gliclazide (LLQ range of 7.8-10.0 ng/mL) [25]. Simultaneous quantitation of eight sulfonylureas based on LC-MS/MS was also reported utilizing LLE and obtaining LLQ in the range of 7.8-78 ng/mL [26].…”

“…The reported methods are applicable to analysis of human plasma, equine plasma and urine, and tablet formulations [23][24][25][26][27][28]. Methods include LC procedures for metformin and glipizide with a lower limit of quantitation (LLQ) of 5.0 and 22.5 ng/mL [23], or metformin and glimepride with a LLQ of 22 and 39 ng/mL, respectively [24].…”

Simultaneous Determination of Metformin and Three Gliptins in Pharmaceutical Formulations Using RP HPLC: Application to Stability Studies on Linagliptin Tablet Formulation

“…Type 2 diabetes is progressive in nature and glycemic control become difficult in most patients, requiring either increased Metformin dosage or supplementary (add-on) therapy. 6 A survey of the literature revealed some HPLC and LC-MS/MS methods for analysis of MET alone [7][8][9][10][11][12][13] and in combination with other drugs. However, quite a few HPLC methods have been reported for simultaneous determination of MET with SIT [14][15][16][17][18][19][20][21][22] or VLD [23][24][25][26][27][28] or LIN 29,30 in formulations, but, no simple and rapid method is yet established for simultaneous determination of Metformin, Sitagliptin, Vildagliptin and Linagliptin using single analytical procedure.…”

Simultaneous determination of metformin and rosiglitazone in human plasma by liquid chromatography/tandem mass spectrometry with electrospray ionization: Application to a pharmacokinetic study

“…Most of these methods are tedious and time-consuming involving complex sample preparation, such as equilibrium dialysis [6,7], ultrafiltration [1], solid phase extraction [3] and liquid-liquid extraction [5]. Recently, some new methods for analysis of metformin and rosiglitazone using liquid chromatography/mass spectrometry or tandem mass spectrometry with improved sensitivity and efficiency have been published [10][11][12][13][14][15]. However, up to now, there is no report on the use of LC/MS/MS method for simultaneous determination of both metformin and rosiglitazone in human plasma.…”