Simultaneous quantification of five major biologically active ingredients of saffron by high-performance liquid chromatography

A simple, sensitive and specific high-performance liquid chromatography-UV (HPLC-UV) method has been developed for the first time to simultaneously qu antify the five major biologically active ingredients of saffron, namely cr ocin 1, crocin 2, crocin 3, crocin 4 and crocetin. Calibration curves were derived by spiking authentic compounds and internal standard, 13-cis-retino ic acid, into herbal samples prior to extraction. Extraction was conducted simply by stirring dried herb (20 mg) with 80% aqueous methanol (5 ml) at a mbient temperature in the dark for 2 h. The HPLC assay was performed on a r eversed-phase C-18 column with linear gradient elution using methanol and 1 % aqueous acetic acid. Calibrations were linear (r(2) = 0.999) for all five analytes, with overall intra- and inter-day RSDs of less than 11%. The ass ay was successfully applied to the determination of four crocins and crocet in in three saffron samples and two Zhizi, another crocin-containing herb. Results indicate that the developed HPLC assay can be readily utilized as a quality control method for crocin-containing medicinal herbs. (C) 1999 Els evier Science B.V. All rights reserved.