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Development and Validation of Stereo Selective Method for the Separation of Razoxane Enantiomers in Hydrophilic
Ch Thirupathi1, K Nagesh Kumar1, G Srinivasu1
1Daicel Chiral Technologies (India) Pvt Ltd, IKP Knowledge Park, Hyderabad 500078, India.
Abstract:
A novel sensitive and high throughput chiral hydrophilic interaction chromatographic (HILIC) method was developed to separate razoxane enantiomers namely levrazoxane (R-isomer) and dexrazoxane (DEX) in pharmaceutical active ingredient samples. A systematic chiral chromatographic screening system was employed in using multiple HPLC chromatographic modes on various polysaccharide based chiral columns to obtain a potential separation between enantiomers. HPLC separation was achieved using a mobile phase of aqueous 10 mM ammonium bicarbonate and mixture of organic modifiers (70/30, v/v) in the ratio of (5/95, v/v) on an immobilized polysaccharide based chiral stationary phase namely CHIRALPAK IE-3. The chromatographic resolution between the enantiomers was found to be not <8 in the developed method. The values of the limit of detection and limit of quantification of DEX and levrazoxane were found to be 0.0037, 0.011 and 0.0043, 0.013 μgmL-1, respectively. The validated method yielded good results regarding precision, linearity, selectivity and found to be superior in sensitivity when compared to reported method for the accurate quantification of undesired enantiomer.
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