HPLC Determination of Chlorpropamide in Human Serum by Fluorogenic Derivatization Based on the Suzuki Coupling Reaction
- PDF / 263,127 Bytes
- 4 Pages / 595.276 x 790.866 pts Page_size
- 81 Downloads / 192 Views
SHORT COMMUNICATION
HPLC Determination of Chlorpropamide in Human Serum by Fluorogenic Derivatization Based on the Suzuki Coupling Reaction with Phenylboronic Acid Naoya Kishikawa • Sherin Farouk Hammad • Kaname Ohyama • Kimiko Kubo • Mokhtar Mohamed Mabrouk Kenichiro Nakashima • Naotaka Kuroda
•
Received: 21 December 2012 / Revised: 26 February 2013 / Accepted: 7 March 2013 / Published online: 9 April 2013 Ó Springer-Verlag Berlin Heidelberg 2013
Abstract A fluorogenic derivatization method for the determination of chlorpropamide in human serum was developed by means of high-performance liquid chromatography (HPLC) with fluorescence detection. The Suzuki coupling reaction with a non-fluorescent reagent, phenylboronic acid (PBA), was employed to convert chlorpropamide into highly fluorescent biphenyl derivative. Chlorpropamide was extracted from human serum by liquid–liquid extraction with toluene after addition of hydrochloric acid, and subsequently reacted with PBA. Because the fluorogenic derivatization was highly selective for aryl halide, the proposed method allowed sensitive and selective detection of chlorpropamide with a detection limit (at a signal to noise ratio of 3) of 0.5 ng mL-1. The sensitivity of our method was from 4 to 100 times better than HPLC–UV, gas chromatography, and LC-mass spectrometry. Keywords High-performance liquid chromatography Fluorogenic derivatization Phenylboronic acid Chlorpropamide
N. Kishikawa S. F. Hammad K. Ohyama K. Kubo N. Kuroda (&) Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8521, Japan e-mail: [email protected] S. F. Hammad M. M. Mabrouk Department of Pharmaceutical Analytical Chemistry, Tanta University, Tanta 31527, Egypt K. Nakashima Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo 859-3298, Japan
Introduction Chlorpropamide is one of sulphonylurea derivatives that are widely used as an oral hypoglycemic drug for the treatment of non-insulin-dependent diabetes mellitus [1]. Chlorpropamide can stimulate pancreatic beta-cell insulin production, which results in the reduction of glucose levels in blood. However, an excess dose of chlorpropamide can cause side effects including hypoglycemic symptoms and coma [2], because it possibly stimulates excessive insulin production [3]. Therefore, monitoring therapeutic blood levels is required for the management of side effects of chlorpropamide. Several chromatographic methods have been reported for the analysis of chlorpropamide in biological samples, such as high-performance liquid chromatography (HPLC) and micellar electrokinetic chromatography with UV detection [4–8]. However, because of low selectivity of UV detection, some interfering peaks appeared on the chromatogram. Other chromatographic methods with mass spectrometry (MS) were also developed [6, 9]; however, MS equipment is not available in many laboratories. On the other hand, we previously developed selective fluorescence derivatization techniques
Data Loading...
