title: Simultaneous quantification of three active alkaloids from a traditional Chinese medicine Ramulus Mori (Sangzhi) in rat plasma using liquid chromatography-tandem mass spectrometry.
abstract: Fagomine, 1-deoxynojirimycin (DNJ) and 1,4-dideoxy-1,4-imino-d-arabinitol (DAB) are the major bioactive constituents in the active fraction of alkaloids from the traditional Chinese medicine mulberry twig (Ramulus Mori, Chinese name Sang Zhi), which has a strong activity on α-glucosidase in vitro and in vivo. A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of DNJ, fagomine and DAB in rat plasma. Plasma samples were prepared using a simple protein precipitation by the addition of 1% volume of Tris and two volumes of methanol-acetonitrile. The analytes and internal standard (IS, miglitol) were chromatographed in an XBridge™ amide column with a gradient mobile phase of acetonitrile-water (0.1% ammonium hydroxide) at a flow rate of 0.7mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer equipped with electrospray ionization (ESI) source in positive ion mode by multiple reaction monitoring (MRM) mode. Linear detection responses were obtained for DNJ ranging from 5.00 to 5000.00ng/mL, 10.00 to 2500.00ng/mL for fagomine and DAB. The lower limits of quantification (LLOQs) were 5.00, 10.00, 10.00ng/mL for DNJ, fagomine and DAB, respectively. Intra-day and inter-day precisions (R.S.D.%) were within 10% for three analytes with accuracies (R.E.%) less than 12%. The mean recoveries of analytes were greater than 85%. All analytes were proved to be stable during the sample storage, preparation and analytic procedures. The method was successfully applied to the pharmacokinetic study of the three alkaloids in rats after oral administration of the active fraction of alkaloids from mulberry twig.
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China. Electronic address: yangsh@imm.ac.cn.
authors.forename: Shuang
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Baolian
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutics, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Xia
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Xue
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Pharmaceutics, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Renyun
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Li
authors.initials: L
authors.lastname: Sheng
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Pharmaceutics, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Drug Metabolism, Beijing Key Laboratory of Non-clinical Drug Metabolism and PK/PD Study, State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, PR China. Electronic address: yanli@imm.ac.cn.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
mesh_terms: D000470:Alkaloids / Q000097:blood*; D000818:Animals; D001089:Arabinose / Q000032:analysis; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004365:Drugs, Chinese Herbal / Q000032:analysis*; D050113:Imino Furanoses / Q000032:analysis; D050112:Imino Pyranoses / Q000032:analysis; D007202:Indicators and Reagents; D008297:Male; D031625:Morus / Q000737:chemistry*; D011786:Quality Control; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015203:Reproducibility of Results; D013048:Specimen Handling; D013402:Sugar Alcohols / Q000032:analysis; D053719:Tandem Mass Spectrometry
chemical_list: D000470:Alkaloids; D004365:Drugs, Chinese Herbal; D050113:Imino Furanoses; D050112:Imino Pyranoses; D007202:Indicators and Reagents; D013402:Sugar Alcohols; C058137:1,4-dideoxy-1,4-iminoarabinitol; C105643:fagomine; D001089:Arabinose
keywords: 1,4-Dideoxy-1,4-imino-d-arabinitol; 1-Deoxynojirimycin; Fagomine; LC–MS/MS; Ramulus Mori