Improved liquid chromatographic tandem mass spectrometric determination and pharmacokinetic study of glimepiride in human plasma

Independent Research and Laboratory Solutions (ILS), 240 Klisthenous Str., 153 44, Gerakas, Athens, Greece.
Biomedical Chromatography (Impact Factor: 1.72). 06/2005; 19(5):394-401. DOI: 10.1002/bmc.465
Source: PubMed


An improved liquid chromatographic tandem mass spectrometric method for the determination of glimepiride in human plasma has been developed and fully validated. The article describes in detail the bioanalytical procedure and summarizes the validation results obtained. The samples were extracted using liquid--liquid extraction with a mixture of 1-chlorobutane-isopropanol-ethyl acetate (88:2:10, v/v/v). The chromatographic separation was performed on a reversed-phase Hypersil ODScolumn (250 x 4.6 mm i.d.; 5 microm particle size) using a mobile phase consisting of formic acid 0.05 M-acetonitrile (28:72, v/v), pumped at a flow rate of 0.3 ml min(-1) heated to 25 degrees C. The analytes were detected using an API 3000 triple quadrupole mass spectrometer with positive electrospray ionization in multiple reaction monitoring mode. Tandem mass spectrometric detection enabled the quantitation of glimepiride down to 0.50 ng mL(-1). Calibration graphs were linear (r better than 0.998, n=1), in concentration range 0.50--1000 ng mL(-1), and the intra- and inter- day RSD values were less than 10.37 and 11.55% for glimepiride. The method was successfully applied to a kinetic study in order to assess the main pharmacokinetic parameters of glimepiride.

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    ABSTRACT: A simple and sensitive high performance liquid chromatographic (HPLC) method for glimepiride determination in human serum is described. The assay involves one‐step liquid‐liquid extraction with dichloromethane in acidified serum. Glibenclamide is used as the internal standard. Detection is done at 228 nm and limit of quantification is less than 10 ng/mL for glimepiride. The calibration curves are linear over the concentration range tested (10–1000 ng/mL). Accuracy, precision, and stability studies are performed.This method is applied to the analysis of glimepiride serum samples of 41 Lebanese male volunteers after oral administration of a single glimepiride 3 mg tablet. Pharmacokinetic analysis of the data is done using a noncompartmental approach with WinNonlin software (WinNonlin Professional, version 4.1).
    No preview · Article · Dec 2005 · Journal of Liquid Chromatography & Related Technologies
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    ABSTRACT: A sensitive and specific liquid chromatography-positive electrospray ionization-tandem mass spectrometry method has been developed and validated for the determination of glimepiride (GPD) in human plasma. GPD and the internal standard (IS, glibenclamide) were extracted from a small aliquot of human plasma (200 microL) by a simple liquid-liquid extraction technique using ethyl acetate as extraction solvent. The compounds were separated on a YMC Propack, C18, 4.6x50 mm column using a mixture of ammonium acetate buffer, acetonitrile and methanol (30:60:10, v/v) as mobile phase at 0.5 mL/min on an API 4000 Sciex mass spectrometer connected to an Agilent HPLC system. Method validation and pre-clinical sample analysis was performed as per FDA guidelines and the results met the acceptance criteria. GPD and IS were detected without any interference from human plasma matrix. The method was proved to be accurate and precise at linearity range of 0.02-100.00 ng/mL with a correlation coefficient of 0.999. The method was robust with a lower limit of quantitation of 0.02 ng/mL. Intra- and inter-day accuracies for GPD were 88.60-113.50 and 96.82-103.93%, respectively. The inter-day precision was better than 12.21%. This method enabled faster and reliable determination of GPD in a pre-clinical study.
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    No preview · Article · Feb 2009 · Journal of Analytical Chemistry
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