Hashemi Mousavi Syed Husain, Kobarfard Farzad, Husain Syed Waqif, Tehrani Mohammad Saber, Abroomand Azar Parviz, Ahmadkhaniha Reza, Mehdizadeh Amir
Department of Chemistry, Science and Research Branch, Islamic Azad University, Tehran, Iran .
Department of Medicinal Chemistry, School of Pharmacy, Phytochemistry Research Center, SBMU, Tehran, Iran.
Iran J Pharm Res. 2012 Winter;11(1):59-67.
A fast, accurate, sensitive, selective and reliable method using reversed-phase high performance liquid chromatography coupled to electrospray ionization ion trap mass spectrometry was developed and validated for the determination of finasteride in human plasma. After protein precipitation with perchloric acid, satisfactory separation was achieved on a Zorbax Eclipse(®) C8 analytical column using a mobile phase consisted of acetonitrile, 2 mM ammonium formate buffer (58:42, pH adjusted at 2.5 using formic acid); the flow rate was 0.25 mLmin(-1) and the column oven was set to 50°C. Tamoxifen citrate was used as internal standard. This method involved the use of M +H ions of finasteride and IS at m/z 373 and 372 respectively with the selected ion monitoring (SIM) mode. The calibration curve was linear over the range of 0.1-60 ng mL(-1). The limit of quantification for finasteride in plasma was 0.1 ng mL(-1). The intra-day and inter-day repeatability (precision) were 2.68-13.87% and 2.14-14.69% respectively. Intra-day and inter-day accuracy were 98-101.57% and 99.7-110%. The assay method has been successfully used to estimate the pharmacokinetics of finasteride after oral administration of a 5 mg tablet of finasteride in 12 healthy volunteers.
建立了一种快速、准确、灵敏、选择性好且可靠的反相高效液相色谱-电喷雾电离离子阱质谱联用法，并对其进行了验证，用于测定人血浆中非那雄胺的含量。用高氯酸进行蛋白沉淀后，在Zorbax Eclipse(®) C8分析柱上实现了良好的分离，流动相由乙腈、2 mM甲酸铵缓冲液（58:42，用甲酸将pH调至2.5）组成；流速为0.25 mLmin(-1)，柱温箱设定为50°C。柠檬酸他莫昔芬用作内标。该方法采用非那雄胺和内标的M +H离子，分别在m/z 373和372处进行选择离子监测（SIM）模式。校准曲线在0.1 - 60 ng mL(-1)范围内呈线性。血浆中非那雄胺的定量限为0.1 ng mL(-1)。日内和日间重复性（精密度）分别为2.68 - 13.87%和2.14 - 14.69%。日内和日间准确度分别为98 - 101.57%和99.7 - 110%。该测定方法已成功用于评估在12名健康志愿者口服5 mg非那雄胺片后非那雄胺的药代动力学。
