Determination of vitamin B₅ in human urine by high‐performance liquid chromatography coupled with mass spectrometry

Abstract

In the present work, we have developed a simple and rapid liquid chromatography/mass spectrometry (LC/MS) method for the identification and quantification of vitamin B5 in human urine. Urine was spiked with vitamin B5 internal standard, hopantenic acid (HOPA), and then diluted with the LC mobile phase prior to its analysis by LC/MS. The quantification was performed in single ion monitoring mode. The calibration curve was linear (r2 = 0.999) between 0.25 to 10 microg/mL. With a limit of detection of 0.1 microg/mL the method was sensitive enough to determine low levels of vitamin B5 in urine. The overall quantitative efficiency of the method was evaluated by spiking urine samples with four different concentrations of vitamin B5; the intra-assay coefficient of variation was below 5% and the recoveries were between 96 to 108%. The results of the present study show that the proposed method is selective and sensitive enough for the quantification of vitamin B5 in urine.

Fluorometric determination of pantothenic acid in human urine by isocratic reversed-phase ion-pair high-performance liquid chromatography with post-column derivatization

“…[12][13][14]; and liquid chromatography-tandem mass spectrometry [15] have also been reported. Whilst these methods may be associated with high precision, the analytical systems required are expensive and difficult to maintain.…”

Comprehensive characterization of amino acids and water‐soluble vitamins in a pentylenetetrazole‐induced seizures rat model

“…Dan Bylund and colleagues developed an HPLC-MS/MS method for the analysis of amino acids in a mixed mode [22]. Olivier Heudi et al determined vitamin B5 in human urine by LC-MS [23]. However, to the best of our knowledge, up to now there are few studies comprehensively exploring the roles of amino acids and WSVs in epilepsy, which will be helpful in finding effective treatments especially for intractable epilepsy.…”

Method development and validation of an HPLC assay for the detection of hopantenic acid in human plasma and its application to a pharmacokinetic study on volunteers

“…There are only a few publications demonstrating the application of gas chromatography-mass spectrometry (MS) methods with multiple ion detection for the determination of hopantenic acid in natural products and biological samples, including plasma and urine [3][4][5][6][7]. Reversedphase liquid chromatography (RP-HPLC)-MS with electrospray ionization [8], RP-HPLC with UV detection [9], and MS [10] detection have been used for the analysis of pantothenic acid, which is similar in structure to hopantenic acid. In some of these works, hopantenic acid was used as an internal standard (IS) for pantothenic acid analysis in plasma [8,10].…”

“…Reversedphase liquid chromatography (RP-HPLC)-MS with electrospray ionization [8], RP-HPLC with UV detection [9], and MS [10] detection have been used for the analysis of pantothenic acid, which is similar in structure to hopantenic acid. In some of these works, hopantenic acid was used as an internal standard (IS) for pantothenic acid analysis in plasma [8,10]. No validated methods for the determination of hopantenic acid in human plasma have been reported.…”