Simultaneous quantification of 21 water soluble vitamin circulating forms in human plasma by liquid chromatography-mass spectrometry.

Journal of chromatography. A (2015-11-03)
Karine Meisser Redeuil, Karin Longet, Sylvie Bénet, Caroline Munari, Esther Campos-Giménez
ABSTRAKT

This manuscript reports a validated analytical approach for the quantification of 21 water soluble vitamins and their main circulating forms in human plasma. Isotope dilution-based sample preparation consisted of protein precipitation using acidic methanol enriched with stable isotope labelled internal standards. Separation was achieved by reversed-phase liquid chromatography and detection performed by tandem mass spectrometry in positive electrospray ionization mode. Instrumental lower limits of detection and quantification reached <0.1-10nM and 0.2-25nM, respectively. Commercially available pooled human plasma was used to build matrix-matched calibration curves ranging 2-500, 5-1250, 20-5000 or 150-37500nM depending on the analyte. The overall performance of the method was considered adequate, with 2.8-20.9% and 5.2-20.0% intra and inter-day precision, respectively and averaged accuracy reaching 91-108%. Recovery experiments were also performed and reached in average 82%. This analytical approach was then applied for the quantification of circulating water soluble vitamins in human plasma single donor samples. The present report provides a sensitive and reliable approach for the quantification of water soluble vitamins and main circulating forms in human plasma. In the future, the application of this analytical approach will give more confidence to provide a comprehensive assessment of water soluble vitamins nutritional status and bioavailability studies in humans.

MATERIAŁY
Numer produktu
Marka
Opis produktu

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Methanol, anhydrous, 99.8%
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Ethyl alcohol, Pure, 190 proof, ACS spectrophotometric grade, 95.0%
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Hydrochloric acid, ACS reagent, 37%
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2-Propanol, BioReagent, for molecular biology, ≥99.5%
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Acetonitrile, anhydrous, 99.8%
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2-Propanol, ACS reagent, ≥99.5%
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Hydrochloric acid, ACS reagent, 37%
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Methanol, ACS reagent, ≥99.8%
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2-Propanol, anhydrous, 99.5%
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Trichloroacetic acid, ACS reagent, ≥99.0%
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Hydrogen chloride solution, 4.0 M in dioxane
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Hydrochloric acid solution, 1.0 N, BioReagent, suitable for cell culture
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DL-Dithiothreitol solution, BioUltra, for molecular biology, ~1 M in H2O
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Hydrochloric acid, 36.5-38.0%, BioReagent, for molecular biology
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Isopropyl alcohol, ≥99.7%, FCC, FG
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DL-Dithiothreitol solution, 1 M in H2O
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Trichloroacetic acid solution, 6.1 N
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Nicotinic acid, ≥98%
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(−)-Riboflavin, from Eremothecium ashbyii, ≥98%
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Pyridoxal 5′-phosphate hydrate, ≥98%
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