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Assay of vitamin B 6 in human plasma with graphitic carbon column
Author(s) -
Kurioka Susumu,
Ishioka Noriaki,
Sato Junko,
Nakamura Junta,
Ohkubo Tatuo,
Matsuda Makoto
Publication year - 1993
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130070313
Subject(s) - chemistry , chromatography , human plasma , column (typography) , plasma , vitamin , carbon fibers , biochemistry , composite number , physics , quantum mechanics , materials science , structural engineering , connection (principal bundle) , engineering , composite material
A high performance liquid chromatographic (HPLC) procedure for measuring pyridoxal‐5′‐phosphate (PLP) and certain forms of B 6 vitamers in plasma is presented here. This HPLC procedure consisted of a single graphitic carbon column with a fluorescence detector employing an isocratic eluent (15% acetonitrile:1% perchloric acid: 0.05% sodium bisulfite). The graphitic carbon column is useful in acidic eluent without deteriorization. The relatively low fluorescent intensity of PLP under acidic conditions is improved by its derivatization with bisulfite in the eluent during chromatographic separation. Using this procedure, the detection limit of PLP is 50 fmol, and an aliquot of 5–50 μL of human plasma is required giving satisfactorily precise results within 5 min. We applied this method to the determination of PLP and certain B 6 vitamers in human plasma after oral supplementation of pyridoxine.