Preconcentration and Determination of Clomiphene Citrate Using Hollow- Fiber Liquid-phase Mixroextraction Coupled with HPLC-UV Detection

Abstract

Background: Currently, ovulation stimulation with clomiphene citrate is the most commonly used treatment in infertility. It stimulates the secretion of gonadotropins necessary for the release of an egg. Objective: In this study, a microextraction method of liquid phase was utilized using hollow fiber with high-performance liquid chromatography and ultraviolet (HPLC-UV) detector for pre-concentration and measuring clomiphene citrate in plasma. Method: Clomiphene citrate was extracted from a 15 ml sample solution with pH=11 in an organic solvent (Octanol), which was located in the vent wall of the fiber. Subsequently, the drug was introduced into an internal organic solvent into an acidic pH=2 receptor that was located inside the fiber. In the following, effective factors in microextraction including pH of the donor phases and acceptor phases, type of organic solvent, ion phase strength, extraction time and stirring speed were investigated and optimized. Results: After extracting the drug with an optimal pre-concentration factor of 212, extraction recovery was 91%, the detection limit was 1.50 ng/ml, limiting the linearity of 5-1000 ng/mL and %RSD=6.98. Conclusion: The proposed method was successfully implemented and optimized for trace detection of clomiphene citrate in plasma, therefore the clinical and investigational application of this method is facilitated by this study.