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Extraction, purification, methylation and GC–MS analysis of short-chain carboxylic acids for metabolic flux analysis.
- Source :
-
Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences . Aug2016, Vol. 1028, p165-174. 10p. - Publication Year :
- 2016
-
Abstract
- Dynamic metabolic flux analysis requires efficient and effective methods for extraction, purification and analysis of a plethora of naturally-occurring compounds. One area of metabolism that would be highly informative to study using metabolic flux analysis is the tricarboxylic acid (TCA) cycle, which consists of short-chain carboxylic acids. Here, we describe a newly-developed method for extraction, purification, derivatization and analysis of short-chain carboxylic acids involved in the TCA cycle. The method consists of snap-freezing the plant material, followed by maceration and a 12–15 h extraction at −80 °C. The extracts are then subject to reduction (to stabilize β-keto acids), purified by strong anion exchange solid phase extraction and methylated with methanolic HCl. This method could also be readily adapted to quantify many other short-chain carboxylic acids. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 15700232
- Volume :
- 1028
- Database :
- Academic Search Index
- Journal :
- Journal of Chromatography B: Analytical Technologies in the Biomedical & Life Sciences
- Publication Type :
- Academic Journal
- Accession number :
- 116864989
- Full Text :
- https://doi.org/10.1016/j.jchromb.2016.05.042