Skip to main content
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1983 Jun;36(6):701–706. doi: 10.1136/jcp.36.6.701

A fluorimetric method for the measurement of pyridoxal and pyridoxal phosphate in human plasma and leucocytes, and its application to patients with sideroblastic marrows.

G P Smith, D Samson, T J Peters
PMCID: PMC498354  PMID: 6853735

Abstract

A highly sensitive fluorimetric assay for the measurement of pyridoxal and pyridoxal phosphate in biological tissues is described. The method involves the enzymic hydrolysis of pyridoxal phosphate to pyridoxal. The pyridoxal (free or total) is separated on an anion-exchange column, concentrated by cation-exchange chromatography and reacted with potassium cyanide under slightly alkaline conditions to form 4-pyridoxolactone, a highly fluorescent compound. The method is applied to the measurement of pyridoxal, pyridoxal phosphate and total pyridoxal in plasma and neutrophils from control subjects and patients with sideroblastic marrow and identified the patient with pyridoxine-responsive sideroblastic anaemia.

Full text

PDF
702

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson B. B., Newmark P. A., Rawlins M., Green R. Plasma binding of vitamin B6 compounds. Nature. 1974 Aug 9;250(5466):502–504. doi: 10.1038/250502a0. [DOI] [PubMed] [Google Scholar]
  2. Anderson B. B., Peart M. B., Fulford-Jones C. E. The measurement of serum pyridoxal by a microbiological assay using Lactobacillus casei. J Clin Pathol. 1970 Apr;23(3):232–242. doi: 10.1136/jcp.23.3.232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhagavan H. N., Koogler J. M., Jr, Coursin D. B. Enzymatic microassay of pyridoxal-5'-phosphate using L-tyrosine apodecarboxylase and L-(1-14C) tyrosine. Int J Vitam Nutr Res. 1976;46(2):160–164. [PubMed] [Google Scholar]
  4. Brophy M. H., Siiteri P. K. Pyridoxal phosphate and hypertensive disorders of pregnancy. Am J Obstet Gynecol. 1975 Apr 15;121(8):1075–1079. doi: 10.1016/s0002-9378(16)33591-8. [DOI] [PubMed] [Google Scholar]
  5. French S. W., Castagna J. Some effects of chronic ethanol feeding on vitamin B 6 deficiency in the rat. Lab Invest. 1967 Apr;16(4):526–531. [PubMed] [Google Scholar]
  6. Hines J. D., Cowan D. H. Studies on the pathogenesis of alcohol-induced sideroblastic bone-marrow abnormalities. N Engl J Med. 1970 Aug 27;283(9):441–446. doi: 10.1056/NEJM197008272830901. [DOI] [PubMed] [Google Scholar]
  7. Hines J. D. Effect of pyridoxine plus chronic phlebotomy on the function and morphology of bone marrow and liver in pyridoxine-responsive sideroblastic anemia. Semin Hematol. 1976 Apr;13(2):133–140. [PubMed] [Google Scholar]
  8. LEEVY C. M., BAKER H., TENHOVE W., FRANK O., CHERRICK G. R. B-COMPLEX VITAMINS IN LIVER DISEASE OF THE ALCOHOLIC. Am J Clin Nutr. 1965 Apr;16:339–346. doi: 10.1093/ajcn/16.4.339. [DOI] [PubMed] [Google Scholar]
  9. Li T. K., Lumeng L., Veitch R. L. Regulation of pyridoxal 5'-phosphate metabolism in liver. Biochem Biophys Res Commun. 1974 Nov 27;61(2):677–684. doi: 10.1016/0006-291x(74)91010-9. [DOI] [PubMed] [Google Scholar]
  10. Lumeng L., Cleary R. E., Li T. K. Effect of oral contraceptives on the plasma concentration of pyridoxal phosphate. Am J Clin Nutr. 1974 Apr;27(4):326–333. doi: 10.1093/ajcn/27.4.326. [DOI] [PubMed] [Google Scholar]
  11. Lumeng L., Li T. K. Vitamin B6 metabolism in chronic alcohol abuse. Pyridoxal phosphate levels in plasma and the effects of acetaldehyde on pyridoxal phosphate synthesis and degradation in human erythrocytes. J Clin Invest. 1974 Mar;53(3):693–704. doi: 10.1172/JCI107607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Merrill A. H., Horiike K., McCormick D. B. Evidence for the regulation of pyridoxal 5-phosphate formation in liver by pyridoxamine (pyridoxine) 5 -phosphate oxidase. Biochem Biophys Res Commun. 1978 Aug 14;83(3):984–990. doi: 10.1016/0006-291x(78)91492-4. [DOI] [PubMed] [Google Scholar]
  13. RINEHART J. F., GREENBERG L. D. Arteriosclerotic lesions in pyridoxine-deficient monkeys. Am J Pathol. 1949 May;25(3):481–491. [PMC free article] [PubMed] [Google Scholar]
  14. Rustin G. J., Peters T. J. Studies on the subcellular organelles of neutrophils in chronic granulocytic leukaemia with special reference to alkaline phosphatase. Br J Haematol. 1979 Apr;41(4):533–543. doi: 10.1111/j.1365-2141.1979.tb05891.x. [DOI] [PubMed] [Google Scholar]
  15. Smith G. P., Peters T. J. Subcellular localization and properties of pyridoxal phosphate phosphatases of human polymorphonuclear leukocytes and their relationship to acid and alkaline phosphatase. Biochim Biophys Acta. 1981 Oct 13;661(2):287–294. doi: 10.1016/0005-2744(81)90017-6. [DOI] [PubMed] [Google Scholar]
  16. Solomon L. R., Hillman R. S. Vitamin B6 metabolism in idiopathic sideroblastic anaemia and related disorders. Br J Haematol. 1979 Jun;42(2):239–253. doi: 10.1111/j.1365-2141.1979.tb01129.x. [DOI] [PubMed] [Google Scholar]
  17. Spector R., Greenwald L. L. Transport and metabolism of vitamin B6 in rabbit brain and choroid plexus. J Biol Chem. 1978 Apr 10;253(7):2373–2379. [PubMed] [Google Scholar]
  18. TURNER J. M. Pyridoxal phosphate breakdown by an alkaline-phosphatase preparation. Biochem J. 1961 Sep;80:663–668. doi: 10.1042/bj0800663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Taylor F. B., Lee D. B., Vanheuvelen W., Grossman I., Hasson H. H., Mitchell H. C., Olive D. R., Pearl E. H., Price J. W., Shull I. F. Environmental Health in Community Growth: An Administrative Guide to Metropolitan Area Sanitation Practices Committee, Engineering and Sanitarian Section, APHA. Am J Public Health Nations Health. 1963 May;53(5):802–822. doi: 10.2105/ajph.53.5.802. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES