Determination of folic acid in milk, milk powder and energy drink by an indirect immunoassay

J Sci Food Agric. 2012 Aug 30;92(11):2297-304. doi: 10.1002/jsfa.5625. Epub 2012 Mar 1.

Abstract

Background: Folic acid (FA) is essential for healthy people (reference daily intake 400 µg day⁻¹) and pregnant women (600 µg day⁻¹). Insufficient intake of FA will increase the risk of neural tube defects in newborns. In this study an indirect enzyme-linked immunosorbent assay was developed for rapid and convenient detection of FA in vitamin-fortified foods.

Results: A carbodiimide-modified active ester method was used to synthesise the immunogen (FA-bovine serum albumin (BSA) conjugate) to raise polyclonal antibodies for FA. The coupling ratio of FA with BSA was determined to be 14:1 (molar ratio). The detection limit of the immunoassay was 3.0 ng mL⁻¹ in buffer, 3.52 ng mL⁻¹ in energy drink, 11.91 ng mL⁻¹ in milk and 16.50 ng mL⁻¹ in milk powder. Intra- and inter-assay variability ranged from 6.6 to 15.1%. Analytical recoveries of FA-spiked samples were 88.3-108.9%.

Conclusion: The immunoassay developed in this study can be used as a simple, rapid and accurate method for fast semi-quantitative and quantitative on-site analysis of FA in food products.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity
  • Beverages / analysis*
  • Cross Reactions
  • Folic Acid / analogs & derivatives
  • Folic Acid / analysis*
  • Folic Acid / chemistry
  • Food Inspection / methods*
  • Food, Fortified / analysis*
  • Food, Preserved / analysis*
  • Haptens / chemistry
  • Immunoassay
  • Limit of Detection
  • Milk / chemistry*
  • Reproducibility of Results

Substances

  • Haptens
  • Folic Acid