Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1994 Jul;47(7):639–641. doi: 10.1136/jcp.47.7.639

Automated microbiological assay of thiamin in serum and red cells.

G Icke 1, D Nicol 1
PMCID: PMC502097  PMID: 8089221

Abstract

AIMS--To develop a sensitive, direct, automated method for the measurement of serum and red cell thiamin. METHODS--A microbiological assay using a chloramphenicol resistant strain of Lactobacillus fermenti as the test organism was developed. Addition of chloramphenicol and cycloheximide to the assay medium suppressed bacterial and yeast contamination and enabled tests to be automated without recourse to aseptic procedures. Evaluation of the assay included precision analysis and estimation of thiamin recovery. Results obtained on red cell extracts were compared with an established colorimetric (thiochrome) method. RESULTS--Acceptable intrabatch and interbatch precision was obtained and good recovery of thiamin added to serum was obtained. Non-parametric reference ranges based on the results from 505 healthy people were: serum thiamin 11.3-35.0 nmol/l and red cell thiamin 190-400 nmol/l. Results were not age or gender related. The method gave results for red cell thiamin which were significantly higher than those obtained with an established thiochrome method. CONCLUSIONS--This automated microbiological assay is sensitive to 2.0 nmol/l of thiamin and allows tests to be set up at the rate of 100 per hour and after 20-22 hours allows incubation results to be read at 60 per hour. The method has proved reliable, suitable for the assay of large numbers of samples, and relatively inexpensive to perform.

Full text

PDF
639

Selected References

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

  1. ANNEAR D. I. Recoveries of bacteria after drying on cellulose fibres. A method for the routine preservation of bacteria. Aust J Exp Biol Med Sci. 1962 Feb;40:1–8. doi: 10.1038/icb.1962.1. [DOI] [PubMed] [Google Scholar]
  2. BAKER H., FRANK O., FENNELLY J. J., LEEVY C. M. A METHOD FOR ASSAYING THIAMINE STATUS IN MAN AND ANIMALS. Am J Clin Nutr. 1964 Apr;14:197–201. doi: 10.1093/ajcn/14.4.197. [DOI] [PubMed] [Google Scholar]
  3. BRIN M. Erythrocyte transketolase in early thiamine deficiency. Ann N Y Acad Sci. 1962 Apr 26;98:528–541. doi: 10.1111/j.1749-6632.1962.tb30574.x. [DOI] [PubMed] [Google Scholar]
  4. Bettendorff L., Grandfils C., De Rycker C., Schoffeniels E. Determination of thiamine and its phosphate esters in human blood serum at femtomole levels. J Chromatogr. 1986 Oct 31;382:297–302. doi: 10.1016/s0378-4347(00)83533-1. [DOI] [PubMed] [Google Scholar]
  5. DEIBEL R. H., EVANS J. B., NIVEN C. F., Jr Microbiological assay for thiamin using Lactobacillus viridescens. J Bacteriol. 1957 Dec;74(6):818–821. doi: 10.1128/jb.74.6.818-821.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DREYFUS P. M. Clinical application of blood transketolase determinations. N Engl J Med. 1962 Sep 20;267:596–598. doi: 10.1056/NEJM196209202671204. [DOI] [PubMed] [Google Scholar]
  7. Davis R. E., Nicol D. J., Kelly A. An automated method for the measurement of folate activity. J Clin Pathol. 1970 Feb;23(1):47–53. doi: 10.1136/jcp.23.1.47. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kimura M., Itokawa Y. Determination of thiamine and its phosphate esters in human and rat blood by high-performance liquid chromatography with post-column derivatization. J Chromatogr. 1985 Sep 20;332:181–188. doi: 10.1016/s0021-9673(01)83295-7. [DOI] [PubMed] [Google Scholar]
  9. Leevy C. M., Cardi L., Frank O., Gellene R., Baker H. Incidence and significance of hypovitaminemia in a randomly selected municipal hospital population. Am J Clin Nutr. 1965 Oct;17(4):259–271. doi: 10.1093/ajcn/17.4.259. [DOI] [PubMed] [Google Scholar]
  10. Leveille G. A. Modified thiochrome procedure for the determination of urinary thiamin. Am J Clin Nutr. 1972 Mar;25(3):273–274. doi: 10.1093/ajcn/25.3.273. [DOI] [PubMed] [Google Scholar]
  11. O'Broin S., Kelleher B. Microbiological assay on microtitre plates of folate in serum and red cells. J Clin Pathol. 1992 Apr;45(4):344–347. doi: 10.1136/jcp.45.4.344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Pelletier O., Madère R. New automated method for measuring thiamine (vitamin B 1 ) in urine. Clin Chem. 1972 Sep;18(9):937–942. [PubMed] [Google Scholar]
  13. Roser R. L., Andrist A. H., Harrington W. H., Naito H. K., Lonsdale D. Determination of urinary thiamine by high-pressure liquid chromatography utilizing the thiochrome fluorscent method. J Chromatogr. 1978 Jul 1;146(1):43–53. doi: 10.1016/s0378-4347(00)81288-8. [DOI] [PubMed] [Google Scholar]
  14. Tallaksen C. M., Bøhmer T., Bell H., Karlsen J. Concomitant determination of thiamin and its phosphate esters in human blood and serum by high-performance liquid chromatography. J Chromatogr. 1991 Mar 8;564(1):127–136. doi: 10.1016/0378-4347(91)80075-n. [DOI] [PubMed] [Google Scholar]
  15. Weber W., Kewitz H. Determination of thiamine in human plasma and its pharmacokinetics. Eur J Clin Pharmacol. 1985;28(2):213–219. doi: 10.1007/BF00609694. [DOI] [PubMed] [Google Scholar]

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

RESOURCES