ENANTIOSEPARATION OF INDAPAMIDE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY USING OVOMUCOID GLYCOPROTEIN AS CHIRAL SELECTOR
ANCA GABRIELA CÂRJE1, VALENTIN ION1*, DANIELA-LUCIA MUNTEAN1, GABRIEL HANCU2, ALINA BALINT1, SILVIA IMRE1
University of Medicine and Pharmacy, Faculty of Pharmacy, Tîrgu Mureş, Romania
1.Departament of Analytical Chemistry and Drug Analysis
2.Departament of Pharmaceutical Chemistry
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The purpose of this study was to separate the enantiomers of indapamide with the help of high performance liquid chromatography on an ovomucoid protein based chiral selector as stationary phase. Enantioseparation of indapamide was performed on an Agilent 1100 Series HPLC system (Agilent Technologies, USA) equipped with UV-VIS detector. An Ultron ES Ovomucoid, 150 x 4.6 mm, 5 μm (Shinwa Chemical Industries LTD, Agilent Technologies) column has been used for the separation of the two optical isomers. The detection was set at a value of 242 nm, chosen as a specific wavelength of indapamide. Under an isocratic elution with a mobile phase composed of phosphate buffer (Na2HPO4 10 mM) and acetonitrile as organic modifier, different experimental conditions (proportion of the organic modifier in the mobile phase, buffer pH, and column temperature) were approached in a univariate manner. The effect of the organic modifier under its structure properties, the buffer pHs and the column temperature, over the selectivity (α) and the resolution (Rs) were investigated. It was found that a pH = 3.1 of the mobile phase, under an isocratic elution (flow = 1 mL/min) with a mobile phase containing 10% organic modifier (acetonitrile) and 90% sodium diphosphate buffer and a column compartment temperature of 20°C were the optimum chromatographic conditions for the enantioseparation of this racemic mixture. The proposed HPLC method, due to its simplicity and high efficiency turned to be an ideal method for the chiral separation of racemic indapamide.