European Journal of Biomedical and Pharmaceutical Sciences

DIRECT CHIRAL SEPARATION AND INVESTIGATION OF CHIRAL DRUGS IN BIOLOGICAL MATRICES WITH PROTEIN PRECIPITATION TECHNIQUES BY RP-HPLC

R. Sathiyasundar*, J. Karthikeyan, K. Prabhu, K. Venkatesan and M. Swamivelmanickam

ABSTRACT

The current research described about direct chiral separation method on chiral drugs in plasma sample by RP-HPLC method. The combination of Ezetimibe and Tramadol both the drug prescribed in the obesity related rheumatoid arthritis. The separation was done with Chiralpak-ASH analytical column (150 x 4.6 i.d mm, 3 μm) by using acetonitrile: methanol: diethyl amine: formic acid (98.5/1.5/0.1/0.1% v/v/v/v) as mobile phase were delivered at a flow rate of 0.8 ml/min, the detection was measured in PDA detector at the isobestic point of 225 nm. The recovery of the spiked plasma was done with simple protein precipitation techniques. The mean, % recoveries achieved when analyzed plasma samples were, 97, 98% of (+) Ezetimibe and (-)Ezetimibe and 101, 102 % of (+)tramadol and (–)tramadol. The proposed method was validated as per the ICH guidelines and found to be specific, accuracy, precise, linear, selective, etc., The proposed method helpful for both pharmacokinetic and pharmacodynamic study at single stationary and mobile phase.

Keywords: Ezetimibe, Tramadol, Chiralpak-ASH, protein precipitation techniques.