title: A HPLC-MS/MS method for the quantitation of free, conjugated, and total HDND-7, a novel hesperetin derivative, in rat plasma and tissues: Application to the pharmacokinetic and tissue distribution study.
abstract: A sensitive and reliable HPLC-MS/MS method was developed and validated for the determination of free (unconjugated), glucuronidated, sulfated, and total (free and conjugated) HDND-7 in rat plasma and tissues. Plasma and tissues samples were treated prior to and after the enzyme hydrolysis. Chromatographic separation was achieved on a Phenomenex Luna C18 column (150 × 4.6mm, 3 μm), using isocratic mobile phase consisting of 0.1% formic acid-acetonitrile (50:50, v/v) at a flow rate of 300 μl/min. The detection was performed on a triple quadruple tandem mass spectrometer using positive electrospray ionization (ESI) source with a chromatographic run time of 5.0 min. The detection was operated by multiple reaction monitoring (MRM) of the transitions of m/z 429.3 → 223.9 for HDND-7 and 272.9 → 152.9 for naringenin (IS), respectively. This method was validated in terms of specificity, linearity, precision, accuracy, and stability. The calibration curves for plasma and tissues were linear over a wide concentration range of 0.02-40 μg/ml with a lower limit of quantification (LLOQ) of 0.02 μg/ml. Mean extraction recoveries in plasma and tissues ranged from 87.4 to 97.1% and from 54.2 to 70.5%, respectively. The intra- and inter-day precision values were below 15% and the accuracy was within ± 15%. The samples were stable under all the tested conditions. This method has been successfully applied to the pharmacokinetic study following oral doses of 25, 50 and 100mg/kg and intravenous dose of 25mg/kg, and tissue distribution study following oral dose of 50mg/kg.
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Chenlin
authors.initials: C
authors.lastname: Shen
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Ruonan
authors.initials: R
authors.lastname: Chen
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Zhengyue
authors.initials: Z
authors.lastname: Qian
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Huang
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Meng
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Taotao
authors.initials: T
authors.lastname: Ma
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China; Anhui Provincial Hospital, Hefei, 230001 Anhui, China.
authors.forename: Zhaolin
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Basic and Clinical Pharmacology, School of Pharmacy, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Basic and Clinical Pharmacology, School of Pharmacy, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Li
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zang
authors.affiliation: School of Pharmacy, Institute for Liver Diseases of Anhui Medical University (AMU), Anhui Key Laboratory of Bioactivity of Natural Products, Anhui Institute of Innovative Drugs, Anhui Medical University, Hefei, 230032 Anhui, China. Electronic address: lj@ahmu.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006569:Hesperidin / Q000032:analysis* / Q000097:blood* / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014018:Tissue Distribution / Q000502:physiology
chemical_list: D006569:Hesperidin; C013015:hesperetin
keywords: Conjugated metabolites; HDND-7; HPLC–MS/MS; Hesperetin derivative; Pharmacokinetics; Tissue distribution