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

Abstract

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. 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%. 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.