title: Profiling and identification of the metabolites of calycosin in rat hepatic 9000×g supernatant incubation system and the metabolites of calycosin-7-O-β-D-glucoside in rat urine by HPLC-DAD-ESI-IT-TOF-MS(n) technique.
abstract: Calycosin and calycosin-7-O-β-D-glucoside are two main bioactive isoflavonoids in Astragali Radix. To profile the metabolites of calycosin in rat hepatic 9000×g supernatant incubation system and the metabolites of calycosin-7-O-β-D-glucoside in rat urine, high performance liquid chromatography with diode array detector and combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry (HPLC-DAD-ESI-IT-TOF-MS(n)) technique was used. Totally, 24 new in vitro metabolites of calycosin and 33 new in vivo metabolites of calycosin-7-O-β-D-glucoside were identified. Monoglucosylation, monopentosylation, demethylation, dehydroxylation, dimerization, and trimerization were found to be new in vitro metabolic reactions of calycosin; hydroxylation and hydrogenation were new metabolic reactions of calycosin-7-O-β-D-glucoside in vivo. The major metabolic reactions of calycosin in rat hepatic 9000×g supernatant incubation system were monohydroxylation on A-ring, dimerization (CO coupling), dimerization (CC coupling) and dehydroxylation; the major phase I metabolic reactions of calycosin-7-O-β-D-glucoside in rats were deglycosylation, hydroxylation, demethylation and dehydroxylation. Hydroxylation, dehydroxylation, and demethylation were common metabolic pathways to calycosin and calycosin-7-O-β-D-glucoside, and some of their metabolites formed through these reactions, such as 8-hydroxycalycosin (S10, M10), pratensein (5-hydroxycalycosin, S19, M27) and formononetin (S22, M28), daidzein (M22), 7,3',4'-trihydroxyisoflavone (S13, aglycon of M3 and M8), equol (aglycon of M19 and M20) had been reported to have many bioactivities related to the pharmacological effects of calycosin and calycosin-7-O-β-D-glucoside. These findings would enhance understanding of the metabolism and real active forms of calycosin and calycosin-7-O-β-D-glucoside.
authors: State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, No.38 Xueyuan Road, Beijing 100191, China.
authors: Ya-Zhou
authors: YZ
authors: Zhang
authors: Feng
authors: F
authors: Xu
authors: Jing
authors: J
authors: Dong
authors: Jing
authors: J
authors: Liang
authors: Yuki
authors: Y
authors: Hashi
authors: Ming-Ying
authors: MY
authors: Shang
authors: Dong-Hui
authors: DH
authors: Yang
authors: Xuan
authors: X
authors: Wang
authors: Shao-Qing
authors: SQ
authors: Cai
mesh_terms: D000284:Administration, Oral; D000818:Animals; D001711:Biotransformation; D002851:Chromatography, High Pressure Liquid; D003640:Dealkylation; D019281:Dimerization; D004365:Drugs, Chinese Herbal / Q000008:administration & dosage / Q000493:pharmacokinetics*; D005960:Glucosides / Q000008:administration & dosage / Q000737:chemistry / Q000493:pharmacokinetics* / Q000652:urine*; D006031:Glycosylation; D006900:Hydroxylation; D007529:Isoflavones / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics* / Q000652:urine*; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D015394:Molecular Structure; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D016482:Urinalysis
chemical_list: D004365:Drugs, Chinese Herbal; D005960:Glucosides; D007529:Isoflavones; C526426:calycosin-7-O-beta-D-glucoside; C121707:7,3'-dihydroxy-4'-methoxyisoflavone