title: Simultaneous Determination of Bosentan, Glimepiride, HYBOS and M1 in Rat Plasma by UPLC-MS-MS and its Application to Pharmacokinetic Study.
abstract: A rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method for the simultaneous determination of bosentan (BOS), glimepiride (GLP), hydroxyl bosentan (HYBOS) and hydroxyl glimepiride (M1) in rat plasma using one-step protein precipitation was developed and validated. After addition of ambrisentan as an internal standard (IS), protein precipitation by acetonitrile was used in sample preparation. Chromatographic separation was achieved on a Waters ACQUITY UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm particle size, Waters Corp., Milford, MA, USA) and inline 0.2 μm stainless steel frit filter (Waters Corp.) with acetonitrile-0.1% formic acid as the mobile phase at a flow rate of 0.4 mL/min with gradient elution. The column temperature was maintained at 40°C. Only 4 min was needed for an analytical run. The retention times were ∼3.29 min for BOS, 3.56 min for GLP, 1.42 min for HYBOS, 1.53 min for M1 and 3.22 min for IS. Electrospray ionization source was employed and operated in positive-ion mode; multiple reaction monitoring mode was applied to target fragment ions m/z 552 → 202, m/z 568 → 202, m/z 491 → 352, m/z 507 → 352 and m/z 379 → 347 for BOS, HYBOS, GLP, M1 and IS, respectively. The assay was validated over concentration ranges of 25-5,000 ng/mL (r(2) = 0.9984) for BOS, 1-200 ng/mL (r(2) = 0.9999) for GLP, 0.5-100 ng/mL (r(2) = 0.9999) for HYBOS and 0.1-20 ng/mL (r(2) = 0.9984) for M1. Intra- and interday precision values for replicate quality control samples were within 14.2% for all analytes during the assay validation. Mean quality control accuracy values were within -3.3 to 14.4% of nominal values for all analytes. The mean recoveries of BOS, GLP, HYBOS, M1 and ambrisentan from the plasma exceeded 90.4%. The analytes were stable in rat plasma for at least 2 h at room temperature, 30 days at -40°C and following at least three freeze-thaw cycles (-40°C to room temperature). This method was successfully applied to a pharmacokinetic study of coadministeration of BOS and GLP in rats.
authors.affiliation: The 2nd Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China School of Pharmacy of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Mengchun
authors.initials: M
authors.lastname: Chen
authors.affiliation: The 2nd Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Song
authors.affiliation: The Laboratory of Clinical Pharmacy, People's Hospital of Lishui City, Lishui, Zhejiang 323000, China.
authors.forename: Shuanghu
authors.initials: S
authors.lastname: Wang
authors.affiliation: School of Pharmacy of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Qiulei
authors.initials: Q
authors.lastname: Chen
authors.affiliation: School of Pharmacy of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Peipei
authors.initials: P
authors.lastname: Pan
authors.affiliation: School of Pharmacy of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
authors.affiliation: School of Pharmacy of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
authors.forename: Guoxin
authors.initials: G
authors.lastname: Hu
authors.affiliation: The 2nd Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China zhe_zhi2000@aliyun.com.
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Zheng
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000077300:Bosentan; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards*; D006900:Hydroxylation; D008297:Male; D015588:Observer Variation; D010666:Phenylpropionates / Q000097:blood; D011489:Protein Denaturation; D011724:Pyridazines / Q000097:blood; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000097:blood* / Q000493:pharmacokinetics; D013453:Sulfonylurea Compounds / Q000097:blood* / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards*
chemical_list: D010666:Phenylpropionates; D011724:Pyridazines; D013449:Sulfonamides; D013453:Sulfonylurea Compounds; C104273:hydroxyglimepiride; C057619:glimepiride; C467894:ambrisentan; D000077300:Bosentan