Determination of glimepiride in human plasma using semi-microbore high performance liquid chromatography with column-switching

Abstract

A fully automated semi-microbore high performance liquid chromatographic (HPLC) method with column-switching using UV detection was developed for the determination of glimepiride from human plasma samples. Plasma sample (900 microl) was deproteinated and extracted with ethanol and acetonitrile. The extract (70 microl) was directly injected into a Capcell Pak MF Ph-1 pre-column where the primary separation occurred to remove proteins and retain drugs using a mixture of acetonitrile and 10mM phosphate buffer (pH 2.18) (20:80, v/v). The analytes were transferred from the pre-column to an intermediate column using a switching valve and then subsequently separated on an analytical column and monitored with UV detection at 228 nm. Glimepiride was eluted with retention time 34.9 min without interference of endogenous substance from plasma. The limit of quantification (LOQ) was 10 ng/ml for glimepiride. The calibration curves were linear over the concentration range of 10-400 ng/ml (r(2) = 0.9997). Moreover, inter- and intra-day precisions of the method were less than 15% and accuracies were higher than 99%. The developed method was successfully applied for the quantification of glimepiride in human plasma and was used to support a human pharmacokinetic study following a single oral administration of 2 mg glimepiride.

Bioequivalence assessment of the two brands of glimepiride tablets

“…The lack of interference with other peaks at the retention time of glimepiride, when blank serum samples were tested, supported the specificity of the method. According to the literature 18,19 the sensitivity of the HPLC method applied in this study (LLOQ, 0.01 µg/ml) was comparable to that of HPLC methodologies used by other investigators to determine concentrations of glimepiride. As expected, however, the HPLC method we used was less sensitive compared with HPLC coupled to tandem mass spectrometry 20,21 .…”

Bioanalytical Method by Column-Switching With Direct Injection of Human Plasma for Determination of Sulphonylureas

“…One study was found describing the determination of glimepiride (GM) in plasma using column-switching with RAM column. The total time spent to determine only one sulfonylurea was 40 minutes (24). In this context, the present study describes the optimization of a rapid and simple method, using a column-switching HPLC with RAM column (sample preparation) and fused core column (analytical separation), for the simultaneous determination of three sulfonylureas: glibenclamide (GB), GM and gliclazide (GZ), in human plasma.…”

Multi‐objective optimization strategy based on desirability functions used for electrophoratic separation and quantification of rosiglitazone and glimepiride in plasma and formulations

“…Because of the therapeutic important and clinical success of RSG and GLM, several methods have been developed for its quantitative determination in plasma samples and in pharmaceutical preparations. These methods for RSG alone include high performance liquid chromatography (HPLC),15–19 liquid chromatography‐tandem mass spectrometry (LC‐MS/MS),20 capillary electrophoresis,21 high performance tin layer chromatography (HPTLC),22, 23 or in combination with other anti‐diabetic drugs,24–29 and for GLM alone include derivative spectrophotometric technique,30 HPLC,31–33 LC‐MS/MS,34–37 or in combination with other anti‐diabetic drugs 38–41. A literature survey revealed that only one spectrophotometric estimation of RSG and GLM in tablet dosage form has been reported 42.…”