JBCGenetics |

Original Article

Volume: 2 | Issue: 2 | Published: Jan 23, 2020 | Pages: 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

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

Article Info

Authors

Lamia Alhammadi

UAE University, Al-Ain, United Arab Emirates

Maitha Aal Abdulla

UAE University, Al-Ain, United Arab Emirates

Naila Ahli

UAE University, Al-Ain, United Arab Emirates

Nahid Al Dhahouri

UAE University, Al-Ain, United Arab Emirates

Anas Al Aidaros

UAE University, Al-Ain, United Arab Emirates

Fatma Al-Jasmi

UAE University, Al-Ain, United Arab Emirates, Tawam Hospital, Al-Ain, United Arab Emirates

Osama Y. Al-Dirbashi

UAE University, Al-Ain, United Arab Emirates, University of Ottawa, Ottawa, ON, Canada, Children&apos;s Hospital of Eastern Ontario, Ottawa, ON, Canada

ORCID

Publication History

Received: July 17, 2019

Revised: November 22, 2019

Accepted: December 12, 2019

Published: January 23, 2020

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.

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

Pubmed Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBC Genetics. 2020; 23 (January 2020): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Web Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. https://www.jbcgenetics.com/articles/2179 [Access: April 27, 2025]. doi:10.24911/JBCGenetics/183-1563341940

AMA (American Medical Association) Style

Vancouver/ICMJE Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi. Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBC Genetics. (2020), [cited April 27, 2025]; 23 (January 2020): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Harvard Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi (2020) Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. JBC Genetics, 23 (January 2020): 93-98. doi:10.24911/JBCGenetics/183-1563341940

Chicago Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi. "Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry." 23 (2020), 93-98. doi:10.24911/JBCGenetics/183-1563341940

MLA (The Modern Language Association) Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi. "Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry." 23.January 2020 (2020), 93-98. Print. doi:10.24911/JBCGenetics/183-1563341940

APA (American Psychological Association) Style

Lamia Alhammadi, Maitha Aal Abdulla, Naila Ahli, Nahid Al Dhahouri, Anas Al Aidaros, Fatma Al-Jasmi, Osama Y. Al-Dirbashi (2020) Analysis of tricarboxylic acid cycle intermediates in dried blood spots by ultraperformance liquid chromatography-tandem mass spectrometry. , 23 (January 2020), 93-98. doi:10.24911/JBCGenetics/183-1563341940