Highly sensitive analytical method for the accurate determination of 5-methyltetrahydrofolic acid monoglutamate in various volumes of human plasma using isotope dilution ultra-high performance liquid chromatography-mass spectrometry.
J Chromatogr B Analyt Technol Biomed Life Sci
; 1179: 122725, 2021 Aug 01.
Article
em En
| MEDLINE
| ID: mdl-34311437
ABSTRACT
One predominant and bioactive folate vitamer circulating in the blood is 5-methyltetrahydrofolate (5-Me-THF). In this study, a method for the accurate determination of 5-Me-THF in human plasma samples of various volumes was established using isotope dilution ultra-high performance liquid chromatography-mass spectrometry (ID-UPLC-MS). For this purpose, 500⯵L of homogeneous human plasma was initially employed, and the 5-Me-THF and the 13C5-5-Me-THF standard solutions prepared using 1% ascorbic acid in water gave the calibration solution and spiking sample. The desired amount of 13C5-5-Me-THF standard solution was spiked into the sample followed by sample pretreatment. The method was validated for its repeatability, reproducibility, recovery, and limits of detection and quantification. Subsequently, it was applied to smaller volumes of human plasma samples (i.e., 50 and 10⯵L), the results of which corresponded well with those obtained using 500⯵L. The feasibility of the method was further confirmed using 10⯵L of a standard reference material, SRM 3949, which is a human serum sample containing three different levels of 5-Me-THF. The established ID-UPLC-MS method was successfully applied to various volumes of human plasma or serum ranging from 500 to 10⯵L, which exhibited particularly good sensitivity in addition to reliable results for the quantification of 5-Me-THF. Our method therefore expands on the ability to obtain accurate quantitative results for 5-Me-THF using small volumes of blood.
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Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Massas
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Tetra-Hidrofolatos
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Cromatografia Líquida de Alta Pressão
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Técnicas de Diluição do Indicador
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article