title: Development and validation of a rapid and sensitive liquid chromatography-tandem mass spectrometry method for benvitimod quantification in human plasma.
abstract: Benvitimod is a newly synthesized non-steroid small molecule being developed as a candidate drug for the treatment of inflammatory skin diseases. Here a rapid, sensitive and specific high performance liquid chromatography-tandem mass spectrometry (LC/ESI/MS/MS) method was developed for the determination of benvitimod in human plasma. The samples were alkalified with disodium tetraborate firstly, and then extracted by methyl tert-butyl ether. Fluorophenyl-benvitimod was used as internal standard (I.S.). Chromatographic separation was performed on an Ultra C(18) column (150mm×2.1mm, 5.0μm). The mixed mobile phase delivered at 300μl/min was CH3CN/H2O, 76.65:23.35 (v/v), containing 0.2mmol/L NH(4)COOH. Detection and quantitation was performed by electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the negative ion mode. The most intense [M-H](-) MRM transition of benvitimod at m/z 253.1→211.0 was used for benvitimod quantitation and the transition at m/z 270.9→229.2 was used to monitor I.S. The calibration curve was linear within the concentration range of 0.1-10.0ng/mL (r>0.99). The lower limit of quantification (LLOQ) was 0.1ng/mL. The extraction recovery was above 80%. The accuracy expressed as relative error (RE) was less than 1.03%. The intra- and inter-day precisions were less than 11.81%. The freeze-thaw stability was also investigated and it was found that both benvitimod and the I.S. were quite stable. This method is especially useful for the pharmacokinetic study of benvitimod.
authors.affiliation: Department of Pharmacy, Peking University People's Hospital, Beijing 100044, China.
authors.forename: Libo
authors.initials: L
authors.lastname: Zhao
authors.forename: Baoying
authors.initials: B
authors.lastname: Zhu
authors.forename: Xin
authors.initials: X
authors.lastname: Chen
authors.forename: Genghui
authors.initials: G
authors.lastname: Chen
authors.forename: Haibo
authors.initials: H
authors.lastname: Chen
authors.forename: Yuzhen
authors.initials: Y
authors.lastname: Li
authors.forename: Shan
authors.initials: S
authors.lastname: Jing
authors.forename: Yi
authors.initials: Y
authors.lastname: Fang
mesh_terms: D000287:Administration, Topical; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D011565:Psoriasis / Q000188:drug therapy; D016032:Randomized Controlled Trials as Topic; D015203:Reproducibility of Results; D012118:Resorcinols / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D012680:Sensitivity and Specificity; D013267:Stilbenes / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D012118:Resorcinols; D013267:Stilbenes; C571829:tapinarof