High-performance liquid chromatographic determination of yohimbine in serum by chemiluminescence detection

An ion-pair high-performance liquid chromatography with chemiluminescence detection using tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)(3)(2+)] was developed for the determination of yohimbine (YOH) in human serum. A sample solution, containing narceine as an internal standard, was treated with a Bond-Elut Certify disposable column, and the purified fraction was separated on a TSKgel ODS-80Ts column. The recovery of YOH at 50 ng/ml was 97.2+/-4.4% (mean+/-SD, n=5). The detection limit was calculated to be 30 ng/ml of YOH in serum (S/N=3). The method was successfully applied to drug monitoring of the YOH level in the serum of healthy volunteers.