Determination of Folate Patterns in Mouse Plasma, Erythrocytes, and Embryos by HPLC Coupled with a Microbiological Assay
- 1 December 1998
- journal article
- Published by Elsevier BV in Analytical Biochemistry
- Vol. 265 (1), 157-166
- https://doi.org/10.1006/abio.1998.2865
Abstract
Folates are important cofactors in one-carbon metabolism. Disturbances in folate homeostasis and metabolism may be related to an increased risk of cardiovascular disease and carcinogenesis and may lead to congenital malformations, namely neural tube defects. Determination of these compounds in biological samples is often a problem due to the existence of numerous folate metabolites, their relative instability, and the low contents in serum and most tissues. As existing methods have distinct limitations, we developed a method, which facilitates the separation as well as the sensitive detection of eight folates by coupling HPLC with a microbiological assay. After a simple sample preparation, including deproteinization and enzymatic hydrolysis of folate polyglutamates, extracts were chromatographed, fractions were collected on microtiter plates, and folates were quantitated using the Lactobacillus casei assay. The raw data were processed using a computing system after reconstructing the HPLC chromatogram with the bacterial growth data. Using the described method, the eight physiologically occurring folate monoglutamates could be simultaneously determined. The detection limits were 2-20 fmol per injection. The application of the method was demonstrated with the analysis of the folate pattern in milligram or sub-milligram quantities of plasma, erythrocyte, and embryos of pregnant mice during organogenesis.Keywords
This publication has 38 references indexed in Scilit:
- Blood folic acid and vitamin B12 in relation to neural tube defectsBJOG: An International Journal of Obstetrics and Gynaecology, 1996
- Folate Levels and Neural Tube DefectsJAMA, 1995
- A Quantitative Assessment of Plasma Homocysteine as a Risk Factor for Vascular DiseaseJAMA, 1995
- Prevention of the First Occurrence of Neural-Tube Defects by Periconceptional Vitamin SupplementationNew England Journal of Medicine, 1992
- Maternal vitamin levels during pregnancies producing infants with neural tube defectsThe Journal of Pediatrics, 1992
- Valproic Acid‐Induced Neural Tube Defects in Mouse and Human: Aspects of Chirality, Alternative Drug Development, Pharmacokinetics and Possible MechanismsBasic & Clinical Pharmacology & Toxicology, 1991
- Prevention of neural tube defects: Results of the Medical Research Council Vitamin StudyThe Lancet, 1991
- Is disordered folate metabolism the basis for the genetic predisposition to neural tube defects?Clinical Genetics, 1987
- Maternal serum folate and vitamin B12 concentrations in pregnancies associated with neural tube defects.Archives of Disease in Childhood, 1985
- Vitamin dificiencies and neural tube defects.Archives of Disease in Childhood, 1976