Simultaneous determination of L-tryptophan and L-kynurenine in rat plasma by LC-MS/MS
10.11665/j.issn.1000-5048.20160614
- VernacularTitle:LC-MS/MS法同时测定大鼠血浆中L-色氨酸和L-犬尿氨酸浓度的研究
- Author:
Kaicheng XU
1
;
Ruilian YU
;
Yunfei YANG
;
Zhihui BIAN
;
Jiawei SHAO
Author Information
1. 南京中医药大学药学院
- Publication Type:Journal Article
- Keywords:
LC-MS/MS;
plasma;
L-tryptophan;
L-kynurenine;
endogenous substances;
drug metabolism analysis;
mechanism of Chinese medicine;
antagonistic adjustment
- From:
Journal of China Pharmaceutical University
2016;47(6):714-718
- CountryChina
- Language:Chinese
-
Abstract:
To establish a simple and sensitive LC-MS/MS method for simultaneous determination of the concentration for L-tryptophan(L-Try)and L-kynurenine(L-Kyn)in rat plasma. The changesin the process of the liver tumors formation may provide a basis for the diagnosis of liver cancer. 3-Nitro-L-tyrosine(3-NT)were added as the internal standard for the determination of two active substances and the chromatographic analysis was performed on a RRHD Eclipse Plus C18 column(3. 0 mm×100 mm, 1. 8 μm). The mobile phase was composed of water and acetonitrile(containing 0. 1%formic acid)(90 ∶10)at a flow rate of 0. 25 mL/min, and the injection volume is 5 μL. Detection and quantification were performed by mass spectrometry in multiple reaction monitoring mode with m/z 205. 12→146. 10(L-Try), m/z 209. 09 →146. 10(L-Kyn), m/z 227. 09→181. 10(3-NT), respectively. The results show that thelinear ranges were 9. 670-9 670 ng/mL for L-Try, and 9. 973-9973 ng/mL for L-Kyn(r2≥0. 9990). The limit of quantitation were 9. 670 ng/mL for L-Try, and 9. 973 ng/mL for L-Kyn, respectively. The intra- and inter-day precisions were all less than15%; the recoveries ofthe two analytes were more than 81. 17% and severe matrix effect was not observed. The ratio of L-Try/L-Kyn determined by LC-MS/MS in rat plasma showed an overall downward trend, which could used effectively for the drug metabolism studies and researches on the action mechanism of medicine on liver cancer. A rapid, simple, sensitive and specific LC-MS/MS method has been successfully developed and could also be used effectively for the drug metabolism studies and researches on the action mechanism of medicine on liver cancer.
