Journal of Chromatography & Separation Techniques
Open Access

Sensitive high-performance liquid chromatography method for quantification of gliclazide in human plasma using monolithic column

3^rd International Conference and Exhibition on Advances in Chromatography & HPLC Techniques

July 13-14, 2017 Berlin, Germany

Afshin Zarghi

Shahid Beheshti University of Medical Sciences, Iran

Posters & Accepted Abstracts: J Chromatogr Sep Tech

Abstract :

Gliclazide is a second-generation sulfonylurea oral hypoglycemic agent used in the treatment of non-insulin-dependent diabetes mellitus (NIDDM). Gliclazide is readily absorbed from the gastro-intestinal tract with peak concentrations in plasma occurring about 2 to 4 hours and it is highly protein bound. In present study, a rapid and sensitive isocratic reversed phase HPLC method with UV detection using a monolithic column has been developed and validated for the determination of gliclazide in human plasma. The assay enables the measurement of gliclazide for therapeutic drug monitoring with a minimum quantification limit of 10 ngml-1. The method involves simple, one-step extraction procedure and analytical recovery was complete. The separation was carried out in reversed-phase conditions using a Chromolith Performance RP-18e, 100x4.6 mm column with an isocratic mobile phase consisting of 0.01 M disodium hydrogen phosphate buffer-acetonitrile (75:25 v/v, flow rate: 4 ml/min) adjusted to pH 4.0. The wavelength was set at 230 nm. The calibration curve was linear over the concentration range 100-5000 ngml-1. The coefficients of variation for interday and intra-day assay were found to be less than 6.0%. This method is well suited for routine application in the clinical laboratory because of the speed of analysis and simple extraction procedure. Owing to use of the monolithic column, which has lower separation impedance compared to the particulate packings, much faster separations are possible; the productivity of chromatographic processes can be increased by at least one order of magnitude as compared to traditional chromatographic columns packed with porous particles. Accordingly, the chromatographic elution step is undertaken in a short time (<5 min) with high resolution. Over 500 plasma samples were analyzed by this method without any significant loss of resolution. No change in the column efficiency and backpressure was observed over the entire study time, thus proving its suitability.

Biography :

Email: zarghi@sbmu.ac.ir