Determination of medium- and short-chain fatty acids in feces by UHPLC-QTOF-MS/MS with pre-column derivatization
10.16438/j.0513-4870.2019-0631
- VernacularTitle:粪便内中短链脂肪酸柱前衍生化UHPLC-QTOF-MS/MS分析方法研究
- Author:
Long-Jie CONG
1
;
Yu-jian SHAO
2
;
Hong SHEN
3
;
Dong-zhou KANG
4
;
Jin-di XU
3
;
Shan-shan ZHOU
1
;
Song-lin LI
5
Author Information
1. Department of Pharmaceutical Analysis, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine to Nanjing University of Chinese Medicine, Nanjing 210028, China
2. Department of Pharmaceutical Analysis, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine to Nanjing University of Chinese Medicine, Nanjing 210028, China; Yanbian University, Yanji 133000, China
3. Department of Metabolomics, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Traditional Chinese Medicine, Nanjing 210028, China
4. Yanbian University, Yanji 133000, China
5. Department of Pharmaceutical Analysis, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine to Nanjing University of Chinese Medicine, Nanjing 210028, China; Department of Metabolomics, Jiangsu Province Academy of Traditional Chinese Medicine and Jiangsu Branch of China Academy of Traditional Chinese Medicine, Nanjing 210028, China
- Publication Type:Research Article
- Keywords:
medium- and short-chain fatty acid;
UPLC-QTOF-MS/MS;
pre-column derivatization;
feces;
etermination
- From:
Acta Pharmaceutica Sinica
2020;55(1):131-138
- CountryChina
- Language:Chinese
-
Abstract:
A pre-column derivatization and ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS) method was developed for qualitative and quantitative determination of medium- and short-chain fatty acids in mice feces, and was further applied to evaluate variations in the feces of mice before and after antibiotic treatment. This animal experiment had been approved by Animal Experimental Ethics Committee of Jiangsu Province Academy of Traditional Chinese Medicine. By optimizing the derivatization conditions and UHPLC-QTOF-MS/MS parameters a new UHPLC-QTOF-MS/MS method with 3-nitrophenylhydrazine as the derivatization reagent was developed for simultaneous determination of 16 medium- and short-chain fatty acids. Validation studies showed that the linearity of the calibration curves was good (R2>0.99), the RSD of intra-day and inter-day precision was less than 10%, the repeatability RSD was less than 6%, the recovery rate was between 80% - 120% at three spiked levels, and the stability RSD was less than 7% within 36 h. The types and amounts of the detected medium- and short-chain fatty acids in feces significantly changed after the mice were treated with antibiotics. The content of formic acid, acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and lactic acid decreased, whereas that of heptanoic acid and succinic acid increased significantly. All these results suggest that the newly established method is accurate and reliable, and can be used for determination of medium- and short-chain fatty acids in feces.
