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LC with Coulometric Detection for Analysis of 5-Methyltetrahydrofolate in Human Plasma

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Abstract

An isocratic high-performance liquid chromatographic method with coulometric electrochemical detection has been used for analysis of 5-methyltetrahydrofolate (5-MTHF) in human plasma. A 250 mm × 4.6 mm i.d., 5-μm particle, C18 column was used with 12:88 (v/v) acetonitrile −35 mM sodium phosphate buffer pH 3.8 as mobile phase at a flow rate of 1.0 mL min−1. The method was validated for 5-MTHF plasma concentrations in the range 2.5–100.0 nM. The method was characterized by a good linearity (regression coefficient r ≥ 0.9989) and limits of detection and quantification of 0.72 and 2.16 nM, respectively. Mean recovery at low and high concentrations ranged from 89.1 to 96.3%, respectively, with a relative standard deviation <4.6%. Between-run imprecision (4.2%) was higher than within-run imprecision (3.4%). The proposed separation and detection procedures were successfully applied to analysis of 5-MTHF in human plasma.

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Authors and Affiliations

  1. Department of Toxicology, Medical University of Gdańsk, J. Hallera 107, 80-416, Gdańsk, Poland

    Marcin Leszek Marszałł, Sylwia Hinc & Wojciech Czarnowski

  2. Department of Psychology, University of Gdańsk, Pomorska 68, 40-343, Gdańsk, Poland

    Ryszard Makarowski

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  1. Marcin Leszek Marszałł
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  2. Ryszard Makarowski
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  3. Sylwia Hinc
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  4. Wojciech Czarnowski
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Correspondence to Marcin Leszek Marszałł.

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Marszałł, M.L., Makarowski, R., Hinc, S. et al. LC with Coulometric Detection for Analysis of 5-Methyltetrahydrofolate in Human Plasma. Chroma 69, 829–835 (2009). https://doi.org/10.1365/s10337-009-0952-z

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