Rapid, Validated UPLC-MS/MS Method for Determination of Glibenclamide in Rat Plasma.

Mohd Aftab Alam,Abdullah Mohammed Al-Mohizea,Fahad Ibrahim Al-Jenoobi

doi:10.1155/2018/2569027

Mohd Aftab Alam, Abdullah Mohammed Al-Mohizea + Show 1 more

Open Access

https://doi.org/10.1155/2018/2569027

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Abstract

Quick and specific bioanalytical methods are required for analyzing drugs in biological samples. A simple, quick, sensitive, and specific UPLC-MS/MS method was developed and validated for glibenclamide determination in plasma samples. The plasma samples were processed by protein precipitation technique. Glimepiride was used as internal standard (IS). Glibenclamide and glimepiride were eluted on C18 column (Acquity UPLC®BEH). Mobile phase consisting of acetonitrile (0.1% formic acid) and water (0.1% formic acid) was pumped in binary gradient mode at flow rate of 150 μL/min. Glibenclamide and IS elution time was about 1.0 min, and total run time was 2.0 min. The mass spectrometer (triple-quadrupole) was operated in positive electrospray ionization mode. Sodium adducts [M + Na]+ of glibenclamide and IS were monitored in MRM mode. A linear calibration curve was obtained in the range of 10-1280 ng/mL, with regression equation Y = 0.0076 X – 0.0165 and linear regression coefficient r2 = 0.999. Lower limit of quantitation was 10 ng/mL. Accuracy of the method at LQC, MQC, and HQC was 109.7% (± 6.7), 93.6% (± 0.4), and 99.3% (± 1.9), respectively. The coefficient of variation for precision at all QC concentrations was less than 6%. Recovery at LLQC, MQC, and HQC was 104.2% (± 4.9), 100.6% (± 0.9), and 102.9% (± 5.8), respectively. The method was successfully implemented for pharmacokinetic investigations (in-house data).

Highlights

Glibenclamide is a hypoglycemic agent of class sulphonylureas
Glibenclamide and glimepiride were eluted on Acquity UPLC5BEH C18 (1.7 μm, 2.1 x 50 mm) column in gradient mode using acetonitrile (0.1% formic acid) and water (0.1% formic acid) as mobile phase
Mass detector (TQD) was tuned in positive electrospray ionization (ESI+) mode, and the sodium ion adducts of glibenclamide [M + Na]+ and glimepiride [M + Na]+ were monitored in multireaction monitoring (MRM) mode

Summary

Introduction

Glibenclamide (glyburide) is a hypoglycemic agent of class sulphonylureas. Glibenclamide (Glynase PresTab, micronized tablet) is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes mellitus (http://www.accessdata.fda.gov/drugsatfda docs/label/2015/020051s021lbl.pdf). The high pressure liquid chromatography in tandem with ultraviolet, or fluorescence, or diode array, or mass spectrometer detector has been the method of choice. Khatri et al developed a fluorescence based HPLC method for determination of glyburide in human plasma samples [1]. Hoizey et al published a LC-Ion-Trap tandem mass spectrometry method for determination of eight sulfonylureas in blood samples. The samples were prepared by liquid-liquid extraction method with a recovery of < 88%. The method showed good sensitivity, and the glibenclamide elution time was about 5.3 min [2]. The drugs from plasma samples were extracted using liquid-liquid extraction procedure. Retention time of glibenclamide was 20.7 min [3]

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