Analysis on Metabolites and Metabolic Pathways of Limonin in Rats by UHPLC-Q-Orbitrap HRMS
10.13422/j.cnki.syfjx.20230563
- VernacularTitle:柠檬苦素在大鼠体内代谢产物与代谢途径的UHPLC-Q-Orbitrap HRMS分析
- Author:
Xuemin LI
1
;
Qianqian GONG
1
;
Zhenxing ZHENG
1
;
Li ZENG
1
;
Mengjiao ZHANG
1
;
Tingyin DUAN
1
;
Xin LIU
1
;
Fang DENG
1
Author Information
1. School of Pharmacy，State Key Laboratory of Southwestern Chinese Medicine Resources， Key Laboratory of Standardization of Chinese Herbal Medicine，Ministry of Education， Chengdu University of Traditional Chinese Medicine，Chengdu 611137，China
- Publication Type:Journal Article
- Keywords:
limonin;
ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS）;
metabolites;
metabolic pathways;
gender differences;
plasma;
animal tissues;
feces
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2023;29(20):126-133
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveUltra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used to identify the metabolites of limonin in rats， and to explore the gender differences in the distribution of prototype components and metabolites in rats after single dose intragastric administration of limonin， as well as to speculate the metabolic pathways. MethodThe separation was performed on a Thermo Scientific Accucore™ C18 column（3 mm×100 mm， 2.6 μm） with 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） as mobile phase in a gradient elution mode（0-1 min， 5%B； 1-6 min， 5%-20%B； 6-18 min， 20%-50%B； 18-23 min， 50%-80%B； 23-25 min， 80%-95%B； 25-30 min， 95%B） at a flow rate of 0.3 mL·min-1 and a column temperature of 30 ℃. MS data of biological samples were collected under the positive ion mode of electrospray ionization（ESI） and in the scanning range of m/z 100-1 500. Plasma， tissues（heart， liver， spleen， lung， kidney， stomach and small intestine）， urine and fecal samples were collected and prepared after intragastric administration， and the prototype component and metabolites of limonin were identified. ResultThe prototype component of limonin were detected in the feces， stomach， small intestine of female and male rats， and in the heart， liver， spleen， lung and kidney tissues of female rats. A total of 23 metabolites related to limonin were detected in rats， of which 2， 1， 5， 4， 23 metabolites were detected in liver， stomach， small intestine， urine and feces， respectively， and the main metabolic pathways were hydrolysis， reduction， hydroxylation and methylation， etc. The distribution of some metabolites differed between male and female rats. ConclusionThe prototype component of limonin are mainly distributed in the stomach and small intestine in rats， and the distribution of prototype component and some metabolites are different by gender. Limonin is mainly excreted through feces with phase Ⅰ metabolites as the main ones. The results of this study can provide a reference for further elucidation of the effect of gender differences on the metabolism of limonin in vivo and its mechanism of action.
