method validation of clonidine hydrochloride in human plasma by lc-ms technique

A simple and sensitive high performance liquid chromatography-electrospray ionization mass spectrometry method has been evaluated for the assignment of clonidine hydrochloride in human plasma. the mobile phase composed of acetonitrile–water 60:40 (v/v), and 0.2% formic acid 20 μl of sample was chromatographically analyzed using a repacked zorbax-xdb-ods c18 column (2.1 mm×30 mm, 3.5 micron). detection of analytes was achieved by tandem mass spectrometry with electrospray ionization (esi) interface in positive ion mode was operated under the multiple-reaction monitoring mode (m/z 230.0 →213). sample pretreatment involved in a one-step protein precipitation (ppt) with methanol and percholoric acid (hclo4) of 0.15 ml plasma. standard curve was linear (r = 0.998) over the concentration range of 0.01-10.0 ng/ml and showed suitable accuracy and precision. the limit of quantification (loq) was 0.01ng/ml. the method is rapidly, simple, very steady and precise for the separation, assignment, evaluation of clonidine healthy in human plasma.