Document Detail


Fluorometric determination of pantothenic acid in human urine by isocratic reversed-phase ion-pair high-performance liquid chromatography with post-column derivatization.
MedLine Citation:
PMID:  19539544     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe here a method for the determination of pantothenic acid, vitamin B(5), in human urine by isocratic reversed-phase ion-pair HPLC with post-column derivatization. Pantothenic acid in urine was separated using a Tosoh ODS-80Ts (4.6 i.d. x 250 mm) column with phosphate-sodium hydroxide buffer (pH 7.0) containing dodecyltrimethylammonium chloride. Following the isolation of pantothenic acid it was decomposed to pantoic acid and beta-alanine by alkali treatment. The product beta-alanine was post-derivatized to the fluorescent 1-alkylthio-2-alkylisoindole with orthophthaldialdehyde in the presence of 3-mercaptopropionic acid. In the proposed method, a urine sample can be directly injected into a HPLC system without any pre-clean up treatment. The limit of detection was 3 pmol (ca. 650 pg) per 20 microL of urine at a signal-to-noise ratio of 5:1 and the limit of quantification was 5 pmol (ca. 1000 pg) per 20 microL of urine, which was sufficiently sensitive for the determination of pantothenic acid in human urine. The total time required for the analysis was ca. 25 min. The proposed method can be used to assess the pantothenic acid content of human urine as an alternative to the standard microbioassay for pantothenic acid.
Authors:
Kei Takahashi; Tsutomu Fukuwatari; Katsumi Shibata
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-06-06
Journal Detail:
Title:  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences     Volume:  877     ISSN:  1873-376X     ISO Abbreviation:  J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.     Publication Date:  2009 Jul 
Date Detail:
Created Date:  2009-06-29     Completed Date:  2009-10-08     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101139554     Medline TA:  J Chromatogr B Analyt Technol Biomed Life Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  2168-72     Citation Subset:  IM    
Affiliation:
Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture, 2500 Hassaka, Hikone, Shiga 522-8533, Japan.
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MeSH Terms
Descriptor/Qualifier:
Chromatography, High Pressure Liquid / instrumentation,  methods*
Fluorometry / methods*
Humans
Pantothenic Acid / urine*
Sensitivity and Specificity
Chemical
Reg. No./Substance:
79-83-4/Pantothenic Acid

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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