[78] Fluorometric determination of pyridoxal and pyridoxal 5′-phosphate in biological materials by the reaction with cyanide

Tamura, Z.; Takanashi, Seiji

doi:10.1016/0076-6879(71)18346-2

Cited by 11 publications

(5 citation statements)

References 2 publications

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“…In the cyanide derivatization method [2,3], pyridoxal (PL) is oxidized to 4-pyridoxolactone; unfortunately, the lactone's fluorescence is maximal at pH 9-10 and markedly decreases at lower pH [7]. This might explain why acidic, isocratic cyanide methods fail to detect PL [2,3].…”

Section: Resultsmentioning

confidence: 95%

Use of chlorite to improve HPLC detection of pyridoxal 5′-phosphate

Ericson

Mahuren

Zubovic

et al. 2005

Journal of Chromatography B

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Section: Resultsmentioning

confidence: 95%

Use of chlorite to improve HPLC detection of pyridoxal 5′-phosphate

Ericson

Mahuren

Zubovic

et al. 2005

Journal of Chromatography B

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“…The use of cyanide to produce a highly¯uorescent derivative of PLP was ®rst used in 1959 (Ohishi & Fukui, 1968, Adams, 1969, Tamura & Takanaski, 1970 and enabled more sensitivity than other derivatives. Cyanide derivatisation of PLP was used in combination with HPLC in methods by Naoi et al (1988), who determined PLP concentrations in puri®ed enzymes, human brain homogenates and tissue cultured cells, but not human plasma.…”

Section: Introductionmentioning

confidence: 99%

High performance liquid chromatography method for the determination of pyridoxal-5-phosphate in human plasma: How appropriate are cut-off values for vitamin B6 deficiency?

1999

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Objectives: Application of a HPLC (high performance liquid chromatography) method, using cyanide derivatisation, to the determination of plasma pyridoxal-5-phosphate (PLP) concentrations as an indicator of vitamin B 6 adequacy. Setting: The study was performed at the Institute of Food Research, Norwich, UK. Blood samples were taken at the Institute, at Health Centres, or in the volunteer's home. Subjects: 51 adolescent, 131 adult, 68 non-institutionalized elderly and 44 aged ( b 73 y) volunteers were recruited from local authority schools, local Health Centres and General Practitioners. Results: The mean PLP recovery was 92.8%. The intra-and inter-assay coef®cients of variation were 2.8% and 5.2% respectively. Mean PLP concentrations for males and females, respectively, were: adolescents (13 ± 14 y), 36.4 and 43.5 nM; adults (20 ± 64 y), 39.2 and 40.0 nM; elderly (68 ± 73 y), 34.8 and 35.3 nM; aged ( b 73 y), 57.8 and 49.0 nM. Percentages of subjects with PLP concentrations`34.4 nM were over 26% in all population groups. Mean vitamin B 6 intakes (mgag protein intake), as assessed by weighed dietary records, were all above reference nutrient intakes (15 mgag protein). Conclusions: An HPLC method, using cyanide derivitisation, has been applied to the determination of plasma PLP. Comparisons of results for local population groups with current cut-off values for plasma PLP, show large numbers of volunteers at risk of vitamin B 6 de®ciency although this is not re¯ected by vitamin B 6 intakes calculated from food tables. The 34.4 nM cut-off value for value for plasma PLP, indicating de®ciency, is questioned.

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“…A sensitive assay for pyridoxal is a chemical method, in which it is oxidized to a highly ‰uorescent product, 4pyridoxolactone, by the catalytic action of cyanide. 8) However, the reaction is not speciˆc for pyridoxal. Thus, pyridoxal 5?-phosphate in the samples has to be removed by ion-exchange chromatography before the reaction.…”

Section: Poundsmentioning

confidence: 99%

An Enzymatic Fluorometric Assay for Pyridoxal with High Specificity and Sensitivity

Trongpanich

Mito

Yagi

2002

Bioscience, Biotechnology, and Biochemistry

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An enzymatic fluorometric assay for pyridoxal with pyridoxal dehydrogenase was developed. The detection limit was about 10 pmol: the calibration curve of pyridoxal showed high linearity (r=0.993). The values obtained by this method correlated well with those by the HPLC method. The enzyme had a high specificity for pyridoxal, and thus animal samples could be directly analyzed without separation of pyridoxal 5'-phosphate by column chromatography.

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[78] Fluorometric determination of pyridoxal and pyridoxal 5′-phosphate in biological materials by the reaction with cyanide

Cited by 11 publications

References 2 publications

Use of chlorite to improve HPLC detection of pyridoxal 5′-phosphate

Use of chlorite to improve HPLC detection of pyridoxal 5′-phosphate

High performance liquid chromatography method for the determination of pyridoxal-5-phosphate in human plasma: How appropriate are cut-off values for vitamin B6 deficiency?

An Enzymatic Fluorometric Assay for Pyridoxal with High Specificity and Sensitivity

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