Metabolic Profiling of Carboxylic Acids and Amino Acids in the Biological Fluids of Patients Diagnosed with Endometriosis Using Liquid (HPLC-UV) and Gas (GC–MS) Chromatography

Е. А. Бессонова, А. Т. Арасланова, А. И. Лазаретова, Igor Govorov, С. И. Ситкин, Л. А. Карцова
In: Journal of Analytical Chemistry · 2023 · vol. 78(11) , pp. 1469–1479 · doi:10.1134/s1061934823100040 · W4389060073
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⚙ AI-generated summary by claude@2026-06, 2026-06-08 ⓘ

This study developed and optimized HPLC and GC-MS methods to analyze carboxylic acids and amino acids in patient blood serum, revealing characteristic metabolic profiles for endometriosis.

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⚙ AI-generated deep summary by claude@2026-06, 2026-06-08 ⓘ

The paper developed and optimized chromatographic methods to quantify low–molecular-weight fatty acids and amino acids in blood serum from patients diagnosed with endometriosis, using HPLC with UV detection for 23 dansyl-chloride–derivatized amino acids and GC–MS for organic acids. Key findings include that GC-based analysis of silyl derivatives lacked the sensitivity needed due to volatility-driven losses during sample preparation, whereas GC–MS conditions without derivatization on a polar stationary phase (with an optimized temperature gradient) enabled organic-acid profiling after protein precipitation and lipid removal. The developed workflow produced characteristic organic-acid profiles in serum samples from endometriosis patients compared with a uterine myoma reference group, with the main caveat being that the GC silyl-derivative approach was irreproducible and insufficiently sensitive under the tested conditions. This paper is centrally about endometriosis — it reports metabolite-profiling methods and serum organic-acid profiles in patients diagnosed with endometriosis.

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Abstract

Approaches to the highly sensitive determination of low-molecular-weight fatty acids and amino acids in blood serum samples of patients diagnosed with endometriosis by gas chromatography with mass spectrometric detection (GC–MS) and HPLC with a diode array detector were proposed. Conditions for the selective determination of 23 amino acids in the form of dansyl chloride derivatives by reversed-phase HPLC with spectrophotometric detection were found, and the main factors influencing the separation parameters (pH of the mobile phase, the solvent and the buffer solution, and the gradient profile) were identified. It was shown that the traditional GC determination of metabolites in the form of silyl derivatives did not provide the required sensitivity: the high volatility of derivatives already at the stage of sample preparation led to significant losses and, as a consequence, to irreproducible results. Conditions for the determination of organic acids without derivatization using GC–MS on a polar stationary phase were optimized. A procedure for preparing blood serum for analysis (precipitation of proteins and removal of lipids) and conditions for the selective separation of analytes (temperature gradient, 70−230°C) were proposed. The developed approaches made it possible to obtain characteristic profiles of organic acids in the blood serum samples of patients with endometriosis and uterine myoma (as a comparison reference group).

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endometriosis

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