High Performance Liquid Chromatographic Analysis of Riboflavin (Vitamin B2) with Visible Absorbance Detection in Italian Cheeses

A high-performance liquid chromatographic method for riboflavin determination in cheese samples was developed. Reverse phase conditions were used, i.e., a C-18 column isocratically eluted with an acetonitrile water mixture (20:80). A shift of the detection wavelength from 267 to 446 nm was proposed as a simple and very effective means for circumventing the problem of incomplete resolution of riboflavin from interfacing peaks. The detection limit of riboflavin was 2.5 ng, a value well above the level required for determining the riboflavin content in cheese samples. From both high- and low-fat cheese, the riboflavin recovery was the same (90.2%). The riboflavin content of several different Italian cheeses was determined.