title: An analytical strategy to investigate Semen Strychni nephrotoxicity based on simultaneous HILIC-ESI-MS/MS detection of Semen Strychni alkaloids, tyrosine and tyramine in HEK 293t cell lysates.
abstract: A Previous metabolomics study has demonstrated that tyrosine metabolism might be disrupted by treating with Semen Strychni on the cell nephrotoxicity model. To investigate the relationship between Semen Strychni alkaloids (SAs) and endogenous tyrosine, tyramine under the nephrotoxicity condition, an HILIC-ESI-MS/MS based analytical strategy was applied in this study. Based on the established Semen Strychni nephrotoxicity cell model, strychnine and brucine were identified and screened as the main SAs by an HPLC-Q Exactive hybrid quadrupole Orbitrap mass system. Then, a sensitive HILIC-ESI-MS/MS method was developed to simultaneously monitor strychnine, brucine, tyrosine and tyramine in cell lysate. The analytes were separated by a Shiseido CAPCELL CORE PC (150mm×2.1mm, 2.7μm) HILIC column in an acetonitrile/0.1% formic acid gradient system. All the calibration curves were linear with regression coefficients above 0.9924. The absolute recoveries were more than 80.5% and the matrix effects were between 91.6%-107.0%. With the developed method, analytes were successfully determined in cell lysates. Decreased levels of tyrosine and tyramine were observed only in combination with increased levels of SAs, indicating that the disturbance of tyrosine metabolism might be induced by the accumulation of SAs in kidney cell after exposure of Semen Strychni. The HILIC-ESI-MS/MS based analytical strategy is a useful tool to reveal the relationships between the toxic herb components and the endogenous metabolite profiling in the toxicity investigation of herb medicines.
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors: Liqiang
authors: L
authors: Gu
authors: Thermo Fisher Scientific, Shanghai, China.
authors: Pengyi
authors: P
authors: Hou
authors: Stem Cell Institute, Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, 777 Shelby Interdisciplinary Biomedical Building, 1825 University Boulevard, Birmingham, AL, 35294-2182, USA.
authors: Ruowen
authors: R
authors: Zhang
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors: Ziying
authors: Z
authors: Liu
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
authors: Kaishun
authors: K
authors: Bi
authors: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China. Electronic address: cxh_syphu@hotmail.com.
authors: Xiaohui
authors: X
authors: Chen
mesh_terms: D000470:Alkaloids / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000633:toxicity*; D057809:HEK293 Cells; D006801:Humans; D016014:Linear Models; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014439:Tyramine / Q000633:toxicity*; D014443:Tyrosine / Q000633:toxicity*
chemical_list: D000470:Alkaloids; D004365:Drugs, Chinese Herbal; D014443:Tyrosine; D014439:Tyramine
keywords: HILIC-ESI-MS/MS; Nephrotoxicity; Semen Strychni alkaloids; Tyrosine