Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 152, Issue -, Pages 179-187Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jpba.2018.01.049
Keywords
Trimethylamine N-oxide; Choline; Betaine; Stroke; Plasma concentration; UPLC-MS/MS
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Funding
- National Key Research and Development Program [2017YFC0909303]
- Guangdong Provincial Key Laboratory of Construction Foundation [2011A060901014, 2017B030314030]
- Fundamental Research Funds for the Central Universities [16ykpy14]
- Science and Technology Planning Project of Guangdong Province [2017A020215147]
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Trimethylamine-N-oxide (TMAO) is derived from the gut microbiome and tissues metabolism of dietary choline and betaine. These molecules are closely related to the development of cardiovascular and cerebrovascular diseases. A rapid, sensitive and accurate method has been developed and validated for the simultaneous determination of trimethylamine N-oxide (TMAO), choline and betaine in human plasma using d9-trimethylamine N-oxide (TMAO), d9-choline, d9-betaine as the internal standard (IS). After methanol precipitation with 10 mu L plasma samples, the analytes were extracted and then separated on Amide column (2.1 x 100 mm, 1.7 mu m, waters) with an isocratic elution program consisting of acetonitrile-water (containing 10 mM ammonium formate pH = 3.0) at a flow of 400 mu L/min. The detection was achieved under the selected reaction monitoring (SRM) scan using positive electrospray ionization (ESI+) in 3 min. The mass transitions monitored were as follows: m/z 76.3 -> 58.4 for TMAO, m/z 104.2 -> 60.3 for choline, m/z 118.1 -> 58.3 for betaine, m/z 85.1 -> 66.3 for d9-TMAO, m/z 113.2 -> 69.3 for d9-choline, and m/z 127.1 -> 67.2 for d9-betaine, respectively. The method has been fully validated for specificity, lower limit of quantification, linearity, stability, intra-and inter-day accuracy and precision. This assay combines simple sample processing with a short run time and small plasma volumes, making it well suited for high-throughput routine clinical or research purposes. The newly developed method was successfully applied to the patients (n = 220) suffered from acute stroke, and the concentration of choline was firstly found to be closely related with the prognosis of these patients. (C) 2018 Elsevier B.V. All rights reserved.
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