Application of UPLC-MS/MS for separation and quantification of 3α-Hydroxy Tibolone and comparative bioavailability of two Tibolone formulations in healthy volunteers

• Published in: Journal of pharmaceutical analysis Vol. 3; no. 4; pp. 270 - 277
• Main Authors: Shinde, Vijay P., Pudage, Ashutosh, Jangid, Arvind, Mistri, Hiren, Patel, P.K.
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.08.2013; Accutest Research Laboratories (I) Pvt. Ltd., Unit II, Opp. The Grand Bhagwati Hotel, S.G. Highway, Bodakdev,Ahmedabad 380059, India%Accutest Research Laboratories (I) Pvt. Ltd., Unit II, Opp. The Grand Bhagwati Hotel, S.G. Highway, Bodakdev,Ahmedabad 380059, India%Kadi Sarva Vishvavidyalaya, Sarva Vidyalaya Campus, Sector 15/23, Gandhinagar, India; Kadi Sarva Vishvavidyalaya, Sarva Vidyalaya Campus, Sector 15/23, Gandhinagar, India; Department of Chemistry, M.G. Science Institute, Navrangpura, Ahmedabad 380009, India; Xi'an Jiaotong University
• Subjects: p-toulenesulfonyl isocyanate; Pharmacokinetics; Tibolone; UPLC; 健康; 分离; 制剂; 应用; 志愿者; 生物利用度; 羟基; 超高效液相色谱; Pharmacokinetics; UPLC; Tibolone; p-toulenesulfonyl isocyanate
• ISSN: 2095-1779; 2214-0883
• DOI: 10.1016/j.jpha.2013.02.006

A novel, fast, sensitive and robust method based on ultra-performance liquid chromatography coupled to atmospheric pressure electrospray ionization tandem mass spectrometry （UPLC-ESI-MS/MS） has been developed to separate two Tibolone stereoisomers i.e., 3α-Hydroxy Tibolone and 3β-Hydroxy Tibolone and to quantify 3α-Hydroxy Tibolone using p-toulenesulfonyl isocyanate （PTSI） as a derivatizing reagent in human plasma. 3α-Hydroxy Tibolone-13CD3 was used as an internal standard （IS）. The analyte and IS were extracted from human plasma by liquid-liquid extraction using ethyl acetate. Extracted samples were analyzed by UPLC-ESI-MS/MS. Chromatography was performed using binary gradient on UPLC analytical column. A linear calibration curve over the range of 0.100-35.000 ng/ mL was obtained and lower limit of quantification （LLOQ） was 0.100 ng/mL demonstrating acceptable accuracy and precision. This method was successfully applied to a pharmacokinetic study in order to compare a test Tibolone 2.5 mg formulation vs. a reference 2.5 mg Tibolone tablet formulation in 50 post-menopausal/surgical menopause female human volunteers under fasting conditions. It is concluded that test formulation of Tibolone is bioequivalent to reference formulation of Tibolone.