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
1Departament of Analytical Chemistry and Drug Analysis
2Departament of Pharmaceutical Chemistry
*corresponding author: valentin.ion@umftgm.ro
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Abstract:
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.

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