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Open AccessArticle

Development and Validation of a Rapid High-Performance Liquid Chromatography–Tandem Mass Spectrometric Method for Determination of Folic Acid in Human Plasma

1
Department of Medicinal Chemistry and Pharmacognosy, Faculty of Pharmacy, Jordan University of Science and Technology, Irbid 22110, Jordan
2
Pharmaceutical Research Unit, Amman 11910, Jordan
*
Author to whom correspondence should be addressed.
Pharmaceuticals 2018, 11(2), 52; https://doi.org/10.3390/ph11020052
Received: 14 May 2018 / Revised: 22 May 2018 / Accepted: 23 May 2018 / Published: 27 May 2018
There are health concerns associated with increased folic acid intake from fortified food and supplements. Existing analytical methods, however, which can be employed to carry out epidemiological and bioavailability studies for folic acid involve laborious sample preparation and/or lengthy chromatographic analysis. In this paper we describe a simple, rapid, and sensitive high-performance liquid chromatography–electrospray ionisation-tandem mass spectrometry (HPLC–ESI-MS/MS) method for determination of unmetabolised folic acid in human plasma using folic acid-d4 as an internal standard. The method required only a simple sample preparation step of protein precipitation and had a total run time of 3.5 min, which is the shortest run time reported to date for HPLC–MS/MS method employed for quantifying folic acid in plasma. The analytes were separated on a C18 column (3 µm; 50 × 3.00 mm) using an isocratic mobile phase consisting of ammonium acetate (1 mM)-acetic acid-acetonitrile (9.9:0.1:90, v/v/v). The method was fully validated in terms of accuracy, precision, linearity, selectivity, recovery, matrix effect, and stability. The short run time and the minimal sample preparation makes the method a valuable tool for performing high-throughput analyses. To demonstrate the applicability of the method in real conditions, it was applied successfully in a bioavailability study for the determination of unmetabolised folic acid levels in vivo in human plasma after oral administration of folic acid. View Full-Text
Keywords: high-performance liquid chromatography–tandem mass spectrometry; electrospray ionisation; folic acid; validation; human plasma high-performance liquid chromatography–tandem mass spectrometry; electrospray ionisation; folic acid; validation; human plasma
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MDPI and ACS Style

Zayed, A.; Bustami, R.; Alabsi, W.; El-Elimat, T. Development and Validation of a Rapid High-Performance Liquid Chromatography–Tandem Mass Spectrometric Method for Determination of Folic Acid in Human Plasma. Pharmaceuticals 2018, 11, 52.

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