Quantitative analysis of Z-2-[4-(4-chloro-1,2-diphenyl-but-1-enyl)phenoxy]ethanol in human plasma using high-performance liquid chromatography

A highly sensitive and precise high-performance Liquid chromatography (HPLC ) assay was developed and validated for the quantitation of Z-2-[4-(4-chlor o-1,2-diphenyl-but-1-enyl) phenoxy]ethanol (FC-1271a) in human plasma. Plas ma samples (1.0 ml) containing FC-1271a and internal standard (toremifene c itrate; Fareston(R)) were extracted using a 2% 1-butanol, 98% hexane soluti on with an extraction efficiency of >97%. Samples were reconstituted in met hanol, irradiated with high intensity ultraviolet light (254 nm) for 1 min, and injected onto a C-18 reverse phase column. Samples were eluted isocrat ically at a flow-rate of 0.5 ml/min with a mobile phase consisting of 6.5% water and 0.5% triethylamine in methanol. The fluorescence of photochemical ly activated compounds was detected using a fluorometer set at an excitatio n wavelength of 266 nm and emission wavelength of 370 nm. Under these assay conditions, standard calibration curves were linear through a concentratio n range of 10-400 ng/ml. In summary, we have developed and validated an HPL C assay to quantitate FC-1271a in human plasma. (C) 1999 Elsevier Science B .V. All rights reserved.