title: On-cartridge derivatization coupled with solid-phase extraction for the ultra-sensitive determination of minodronic acid in human plasma by LC-MS/MS method.
abstract: Minodronic acid (MA) is a third-generation bisphosphonate (BP). Its high potency allows lower doses to be administered in clinical settings compared with other BPs, which results in extremely low systemic exposure. Therefore, it is essential to develop an ultra-sensitive bioassay for pharmacokinetics studies of MA. In this work, we used on-cartridge derivatization of MA with trimethylsilyldiazomethane to extract MA from plasma samples and improve its LC-MS/MS behavior. The reaction produced a known derivative, tetramethylated MA, and a novel derivative, pentamethylated MA (PMMA). PMMA exhibited a better signal-to-noise ratio, and was monitored for the quantification of MA. However, the derivatization yield of d4-PMMA was much lower and more variable than that of PMMA, which decreased the effectiveness of its correction function as an internal standard. Therefore, a two-cycle derivatization approach was introduced to increase its yield and improve the reproducibility. The calibration curves of MA showed good linearity over the range of 10.0-1000 pg/mL. A lower limit of quantification of 10.0 pg/mL was achieved with acceptable precision (<10.5%) and accuracy (5.0%). The intra- and inter-batch precision of quality control samples was <9.5%, and the accuracy ranged from -2.8% to 0.6%. The stability results showed that MA was stable in human plasma for 6h at room temperature (25°C), for 115 days at -20°C, during three freeze/thaw cycles (from -20°C to 25°C), and in post-preparative samples for 24h at 4°C. The method was successfully used to characterize the pharmacokinetic profile of MA following an oral dose of 1.0mg MA hydrate to healthy volunteers (n=12). The proposed derivatization procedure was also extended to measure other BPs (risedronic acid and zoledronic acid) in human plasma at low pg/mL.
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Cai
authors.initials: C
authors.lastname: Liu
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China.
authors.forename: Dafang
authors.initials: D
authors.lastname: Zhong
authors.affiliation: Center for Drug Metabolism and Pharmacokinetics, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 501 Haike Road, Shanghai 201203, PR China. Electronic address: xychen@mail.shcnc.ac.cn.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Chen
mesh_terms: D000925:Anticoagulants / Q000737:chemistry; D019540:Area Under Curve; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D003978:Diazomethane / Q000031:analogs & derivatives / Q000737:chemistry; D004164:Diphosphonates / Q000032:analysis / Q000097:blood*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000032:analysis / Q000097:blood*; D057230:Limit of Detection; D013058:Mass Spectrometry; D008745:Methylation; D009584:Nitrogen / Q000737:chemistry; D015203:Reproducibility of Results; D000068296:Risedronic Acid / Q000032:analysis; D052616:Solid Phase Extraction / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014297:Trimethylsilyl Compounds / Q000737:chemistry; D000077211:Zoledronic Acid
chemical_list: D000925:Anticoagulants; D004164:Diphosphonates; D007093:Imidazoles; D014297:Trimethylsilyl Compounds; C087958:YM 529; D003978:Diazomethane; D000077211:Zoledronic Acid; D000068296:Risedronic Acid; D009584:Nitrogen; C060070:trimethylsilyldiazomethane
keywords: Bisphosphonate; Derivatization; LC–MS/MS; Minodronic acid; Trimethylsilyldiazomethane