AI Article Synopsis
- Liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) is highlighted as a reliable method for detecting metabolites, but quantifying polar compounds like poly-carboxylic acids in urine remains challenging.
- Chemical derivatization is suggested as a solution to enhance the sensitivity and performance of these analyses, with a focus on two agents: o-benzylhydroxyl amine (oBHA) and 2-picolyl amine (2-PA).
- The study finds that oBHA outperforms 2-PA, offering better sensitivity, more stable compounds, and successful detection in stored urine samples, confirming its preference for quantifying (poly)-carboxylic acids.
Article Abstract
Due to its high sensitivity and specificity, liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) could be considered as the gold-standard in targeted metabolomics. Although LC-MS/MS allows for the direct detection of a large number of molecules, the proper quantification of highly polar compounds such as poly-carboxylic acids in complex matrices like urine is still a challenge. Chemical derivatization offers a suitable way to improve chromatographic behavior and sensitivity for these compounds. Several derivatizing agents have been proposed for the LC-MS/MS determination of carboxylic acids but studies dealing with their comparison in challenging scenarios are scarce. Here we present the evaluation of two different derivatization agents; o-benzylhydroxyl amine (oBHA) and 2-picolyl amine (2-PA); for the quantification of the (poly)-carboxylic acids belonging to the tricarboxylic acid cycle in urine. The suitability of both derivatizating agents was compared by validation of the two approaches. Derivatization with oBHA showed important advantages against 2-PA derivatization such as (i) providing better sensitivity, (ii) more stable derivatives and (iii) allowing for the proper validation of a larger number of analytes. Moreover, while 2-PA derivatization failed in the determination of the target analytes in some stored urine samples, oBHA derivatization successfully allowed for their appropriate determination in the same samples. A comparison between the concentrations obtained using oBHA derivatization and those provided by an external laboratory using UV and GC-MS detection revealed a satisfactory agreement between both results. Additionally, the concentrations obtained by the oBHA method for a set of 38 urines are in agreement with those previously reported in the literature. As a conclusion, our results show that the use of oBHA is preferred against 2-PA for the detection and quantification of (poly)-carboxylic acids in urine.
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| http://dx.doi.org/10.1016/j.jpba.2018.10.055 | DOI Listing |
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