Raloksifen'in Biyolojik Materyalden ve Farmasötik Preparattan Analizi İçin Gaz Kromatografisi-Kütle Spektrometrisi Yönteminin Geliştirilmesi ve Valide Edilmesi

Abstract

Raloxifene is selective estrogen modulator (SERM) drug which is used in the treatment of postmenopausal osteoporosis in women and used for doping in men. A new gas chromatography-mass spectrometry (GC-MS) method was developed for the determination of Raloxifene in human urine and pharmaceutical preparation. Raloxifene was silylated prior to GC-MS analysis. Derivatization reaction was optimized to investigate parameters such as different derivatization reagents, reaction temperatures and times. It was efficiently derivatized using MSTFA/ß-mercaptoethanol/NH4I at 80 °C for 30 min. Methyltestosterone was selected as internal Standard (IS). Analysis was performed in selected ion monitoring (SIM) mode. The ions m/z 578 for raloxifene and m/z 446 for IS were selected for quantitation. Chromatographic variables (initial and final oven temperature, flow rate of gas) were investigated to optimize the chromatographic method. Raloxifene was extracted from urine with carbonate buffer (pH 9.0) and methyl ter-butyl ether. The developed method was validated in terms of system suitability tests, specificity, sensitivity, linearity, accuracy, precision, recovery, stability, robustness and ruggedness for analysis of Raloxifene in each urine and pharmaceutical formulation. Linearity was established in the range of 10-200 ng mL-1, limit of quantification (LOQ) was 10.0 ng mL-1 and limit of detection (LOD) was 8.0 ng mL-1 for urine. Linearity was found 10-400 ng, LOQ was 10.0 ng and LOD was 5.0 ng for pharmaceutical analysis. Developed method was sensitive, precise, accurate, specific, sensitive, rugged and robust due to the validation study results. The developed and validated method was successfully applied to analysis of the pharmaceutical preparation containing raloxifene HCl. The results were compared to the method in literature and no significant difference was found between them.