Fluorometric method based on molecular recognition solid-phase extraction for determination of riboflavin in milk and infant formula

Riboflavin (vitamin B₂) is involved in several biological processes, particularly in energy production, and it is acquired from food ingestion, principally from supplemented food during the first years of life. Therefore, a simple, fast and cost-effective high-throughput method for determination of riboflavin in milk and infant formula is proposed, based on selective extraction using commercially available molecularly imprinted polymers targeted to riboflavin, followed by direct fluorometric determination. Several aspects were studied, namely microplate assay conditions, the composition of eluting solution and the stability of riboflavin in the eluate. Hence, elution using 1% (v/v) acetic acid in methanol or in acetonitrile is recommended, followed by immediate analysis or solvent evaporation, with reconstitution and analysis within 24 h. The proposed method provided a LOD of 0.03 mg L^?1, with working range for undiluted samples between 0.125 and 2 mg L^?1, and sample throughput of 24 h^?1. It was successfully applied to certified reference material NIST-1846 and also to commercial milk and infant formula samples.