Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry

Lamia Alhammadi; Maitha Aal Abdulla; Naila Ahli; Nahid Al Dhahouri; Anas Al Aidaros; Fatma Al-Jasmi; Osama Y. Al-Dirbashi

doi:10.24911/JBCGenetics/183-1563341940

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Original Article
Online Published: 23 Jan 2020

JBCGenetics. 2019; 2(2): 93-98

doi: 10.24911/JBCGenetics/183-1563341940

Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi.

Abstract
Background: We developed a novel method for measuring the concentrations of tricarboxylic acid (TCA) cycle intermediates in dried blood spots (DBS) using liquid chromatography-tandem mass spectrometry (LC-MS/ MS). Analytes were derivatized before analysis using 4-[2-(N,N-dimethylamino) ethylaminosulfonyl]-7-(2-aminoethylamino)-2,1,3-benzoxa-diazole (DAABD-AE), a reagent that imparts powerful chromatographic and mass spectrometric properties onto carboxyl group-containing analytes. Methodology: Extraction and derivatization of TCA cycle intermediates were achieved in a single step by incubating a 3.2 mm circle of the DBS samples with DAABD-AE for 1 hour at 65 C. From the resultant mixture, 1.0 µl was injected into the LC-MS/MS. Results: Total analytical run time to separate target analytes from other interfering components in the sample was 8 minutes. The peaks corresponding to malic, fumaric, citric, succinic, and 2-ketoglutaric acid appeared at 3.55, 3.62, 3.64, 3.67, and 3.68 minutes, respectively. The method was adequately reproducible with a coefficient of variation for intraday (n = 15) and inter-day (n = 13) studies of 5.2%-18.4%. Reference intervals in DBS from controls (n = 125) were as follow (µmol/l): citric (36.6-126.4), 2-ketoglutaric (9.1-42.1), succinic (1.2-2.4), fumaric (2.4-9.0), and malic acid (15.9-39.3). Compared to controls, the levels of citric, succinic, and malic acids were statistically different in patients (n = 7) with a p-value of< 0.05. No statistically significant difference was detected in concentrations of 2-ketoglutaric and fumaric acids. Conclusion: We describe a simple, quick, and sensitive method to measure TCA cycle intermediates in DBS samples. That TCA cycle plays a central role in cellular metabolism; this method should be useful in studying these metabolites in health and disease.

Key words: Tricarboxylic acid cycle, citric acid, 2-ketoglutaric acid, succinic acid, fumaric acid, malic acid, DBS, LC-MS/MS, DAABD-AE.

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How to Cite this Article

Pubmed Style

Alhammadi L, Abdulla MA, Ahli N, NAD, Aidaros AA, Al-Jasmi F, Al-Dirbashi OY. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBCGenetics. 2019; 2(2): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Web Style

Alhammadi L, Abdulla MA, Ahli N, NAD, Aidaros AA, Al-Jasmi F, Al-Dirbashi OY. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. https://www.jbcgenetics.com/?mno=57331 [Access: November 14, 2024]. doi:10.24911/JBCGenetics/183-1563341940

AMA (American Medical Association) Style

Alhammadi L, Abdulla MA, Ahli N, NAD, Aidaros AA, Al-Jasmi F, Al-Dirbashi OY. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBCGenetics. 2019; 2(2): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Vancouver/ICMJE Style

Alhammadi L, Abdulla MA, Ahli N, NAD, Aidaros AA, Al-Jasmi F, Al-Dirbashi OY. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBCGenetics. (2019), [cited November 14, 2024]; 2(2): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Harvard Style

Alhammadi, L., Abdulla, . M. A., Ahli, . N., , . N. A. D., Aidaros, . A. A., Al-Jasmi, . F. & Al-Dirbashi, . O. Y. (2019) Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBCGenetics, 2 (2), 93-98. doi:10.24911/JBCGenetics/183-1563341940

Turabian Style

Alhammadi, Lamia, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, and Osama Y. Al-Dirbashi. 2019. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. Journal of Biochemical and Clinical Genetics, 2 (2), 93-98. doi:10.24911/JBCGenetics/183-1563341940

Chicago Style

Alhammadi, Lamia, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, and Osama Y. Al-Dirbashi. "Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry." Journal of Biochemical and Clinical Genetics 2 (2019), 93-98. doi:10.24911/JBCGenetics/183-1563341940

MLA (The Modern Language Association) Style

Alhammadi, Lamia, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, and Osama Y. Al-Dirbashi. "Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry." Journal of Biochemical and Clinical Genetics 2.2 (2019), 93-98. Print. doi:10.24911/JBCGenetics/183-1563341940

APA (American Psychological Association) Style

Alhammadi, L., Abdulla, . M. A., Ahli, . N., , . N. A. D., Aidaros, . A. A., Al-Jasmi, . F. & Al-Dirbashi, . O. Y. (2019) Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. Journal of Biochemical and Clinical Genetics, 2 (2), 93-98. doi:10.24911/JBCGenetics/183-1563341940

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