title: A UPLC-MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside, isopsoralenoside, psoralen and isopsoralen from Psoralea corylifolia extract.
abstract: BACKGROUND
  The dried fruit of Psoralea corylifolia L. has been used to prevent and treat vitiligo, osteoporosis, arthralgia and asthma in Traditional Chinese Medicine for some 1600 years. Psoralen (P), isopsoralen (IP), psoralenoside (PO) and isopsoralenoside (IPO) are the major coumarins and coumarin-related benzofuran glycosides in Psoraleae Fructus, which have been reported to show estrogen-like activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. The first aim of this study is to develop a rapid, sensitive and selective ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) approach for simultaneous determination of PO, IPO, P and IP in rat plasma and samples collected from in vitro incubation experiments. The second aim is to investigate the pharmacokinetic properties of PO, IPO, P and IP after oral administration of Psoralea corylifolia extract (PCE) to rats. The third aim is to confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions.
  
  
  METHODS
  A UPLC-MS/MS method with a C18 column and a mobile phase of methanol-0.1% aqueous formic acid was validated according to the criteria in FDA guidelines about bioanalytical method, which was developed to investigate the pharmacokinetic behavior of PO, IPO, P and IP from PCE and the metabolic pathways of PO to P or IPO to IP.
  
  
  RESULTS
  The criteria for establishment of a new UPLC-MS/MS method including selectivity, linearity, accuracy, precision, extraction recovery, matrix effect and stability were validated. This method was successfully applied to the quantitative determination of PO, IPO, P and IP in biological samples collected from both in vitro incubations and in vivo rat experiments. After oral administration of PCE to rat, pharmacokinetic parameters of these four compounds indicated that in vivo biotransformation may occur between PO and P or IPO and IP. Purified benzofuran glycosides fraction (PBGF), containing only PO and IPO, was orally administered to rats to further confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions. An in vitro incubation study elucidated that PO and IPO were metabolized to P and IP by intestinal microflora through de-glucosylation.
  
  
  CONCLUSIONS
  This paper developed a rapid, sensitive and selective UPLC-MS/MS method for simultaneous determination of PO, IPO, P and IP from PCE in biological samples, and investigated on their comprehensive in vivo and in vitro pharmacokinetic studies. These obtained results showed that the metabolism by intestinal bacteria plays an important role in pharmacological effects of orally administered PCE.
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tjianjin 300457, China.
authors: Yue-Fei
authors: YF
authors: Wang
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tjianjin 300457, China.
authors: Ya-Nan
authors: YN
authors: Liu
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tjianjin 300457, China.
authors: Wen
authors: W
authors: Xiong
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China.
authors: Dong-Mei
authors: DM
authors: Yan
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tjianjin 300457, China.
authors: Yan
authors: Y
authors: Zhu
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China. Electronic address: gaoxiumei@tjutcm.edu.cn.
authors: Xiu-Mei
authors: XM
authors: Gao
authors: Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tjianjin 300457, China.
authors: Yan-Tong
authors: YT
authors: Xu
authors: College of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, 312 An Shan Xi Road, Tianjin 300193, China.
authors: Ai-Di
authors: AD
authors: Qi
mesh_terms: D000818:Animals; D001572:Benzofurans / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics*; D002853:Chromatography, Liquid; D005363:Ficusin / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D005638:Fruit / Q000737:chemistry; D011564:Furocoumarins / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D006027:Glycosides / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics*; D008297:Male; D015394:Molecular Structure; D010936:Plant Extracts / Q000737:chemistry*; D029907:Psoralea / Q000737:chemistry*; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry
chemical_list: D001572:Benzofurans; D011564:Furocoumarins; D006027:Glycosides; D010936:Plant Extracts; C511398:isopsoralenoside; C511397:psoralenoside; C011659:angelicin; D005363:Ficusin
keywords: AUC; Benzofuran glycosides; C(max); CE; CV; Coumarins; ESI; IP; IPO; IS; LLOQ; MRM; MRT; P; PBGF; PCE; PO; Pharmacokinetics; Psoralea corylifolia; Psoralea corylifolia extract; QC; RE; RSD; SD; SGF; SIF; T(max); TCM; UPLC–MS/MS; collision energies; cone voltages; electrospray ionization; elimination half-life; internal standard; isopsoralen; isopsoralenoside; lower limit of quantification; maximum plasma drug concentration; mean retention time; multiple reaction monitoring; psoralen; psoralenoside; purified benzofuran glycosides fraction; quality control; relative error; relative standard deviation; simulated gastric fluid; simulated intestinal fluid; standard deviation; t(1/2); the area under the plasma concentration versus time; the time to reach the maximum plasma drug concentration; traditional Chinese medicine; ultra performance liquid chromatography tandem mass spectrometry