title: Simultaneous quantification of Akebia saponin D and its five metabolites in human intestinal bacteria using ultra-performance liquid chromatography triple quadrupole mass spectrometry.
abstract: A rapid and sensitive ultra-performance liquid chromatography triple quadrupole mass spectrometry (UPLC-TQ/MS) method was developed for simultaneous quantification of Akebia saponin D (ASD) and its five metabolites in intestinal mixtures of bacteria from human feces. After protein precipitation, the analytes and internal standard (IS), glycyrrhetinic acid, were determined in selected ion recording (SIR) mode with negative ion ESI source. Chromatographic separation was carried out on an ACQUITY UPLC™ BEH C18 column (100mm×2.1mm, 1.7μm) using gradient elution. The mobile phase consisted of solvents A (acetonitrile) and B (0.1% aqueous formic acid) at the flow rate of 0.4mL/min. Each sample was chromatographed within 10.5min including equilibration time. The linearity ranged from 0.1 to 100μg/mL for ASD, and 2-1000ng/mL for five metabolites, Dipsacus saponin A (M1), HN-saponin F (M2), hederagenin-28-O-β-d-glucopyranoside (M3), Akebia saponin PA (M4), hederagenin (M5). The limits of detection (LOD) were 0.41, 0.59, 0.61, 0.55, 0.52 and 0.31ng/mL for ASD, M1, M2, M3, M4 and M5, respectively. The intra- and inter-day precision was all within 11.1% and accuracy ranged from -8.33% to 12.47%. The conversion rate of five metabolites was 41.21% in 24h. The method was validated and successfully applied to quantification of ASD and its five metabolites in human intestinal bacteria.
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Yan
authors.affiliation: Jiangsu Provincial Center for Research and Development of Marine Drugs, Nanjing University of Traditional Chinese Medicine, Nanjing 210029, PR China.
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China; Jiangsu Kanion Pharmaceutical Co. Ltd., Lianyungang 222001, PR China.
authors.forename: Zhaoqing
authors.initials: Z
authors.lastname: Meng
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China.
authors.forename: Yongliang
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: Jiangsu Kanion Pharmaceutical Co. Ltd., Lianyungang 222001, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiao
authors.affiliation: Jiangsu Kanion Pharmaceutical Co. Ltd., Lianyungang 222001, PR China.
authors.forename: Zhenzhong
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Jiangsu Kanion Pharmaceutical Co. Ltd., Lianyungang 222001, PR China.
authors.forename: Wenze
authors.initials: W
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: yzl1950@126.com.
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, PR China. Electronic address: zhangchunfeng67@126.com.
authors.forename: Chunfeng
authors.initials: C
authors.lastname: Zhang
mesh_terms: D001419:Bacteria / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005243:Feces / Q000382:microbiology; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D013058:Mass Spectrometry / Q000379:methods*; D009828:Oleanolic Acid / Q000031:analogs & derivatives / Q000032:analysis; D015203:Reproducibility of Results; D012503:Saponins / Q000032:analysis* / Q000378:metabolism*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D012503:Saponins; C534004:akebia saponin D; D009828:Oleanolic Acid; C025763:hederagenin
keywords: Akebia saponin D; Human intestinal bacteria; Metabolites; UPLC-TQ/MS