Quantification of 38 dietary polyphenols in plasma by differential isotope labelling and liquid chromatography...

Polyphenols constitute one of the most complex classes of phytochemicals in the human diet and have been suggested to play a role in the prevention of chronic diseases such as cardiovascular diseases, diabetes and cancers. However, evidence from epidemiological studies is still needed to better understand their role in disease prevention. To do so, robust methods for the accurate measurement of these molecules in large series of samples are needed. We report here the development of a highly-sensitive method based on differential isotope labelling with ¹³C- and ¹²C-dansyl chloride for the analysis of 38 structurally diverse polyphenols in 50 microL plasma by tandem mass spectrometry with limits of quantification varying between 0.11 to 44 nmol/L. Full validation of the method was achieved for 37 compounds out of the 38 tested. The method showed intra- and inter-batch coefficients of variations of 2.3-9.0% and 2.8-20.3% respectively depending on polyphenols when applied to 1163 plasma samples from the European Prospective Investigation on Cancer and Nutrition (EPIC) study. For the first time this method allowed to quantify with high accuracy and reproducibility a large selection of compounds representative of the main classes of dietary polyphenols in low volumes of plasma.