Development and validation of a high throughput micro solid phase extraction method coupled with ultra high performance liquid chromatography quadrupole time of flight mass spectrometry for rapid identification and quantification of phenolic metabolites in human plasma and urine.
A rapid and high throughput micro solid phase extraction (m SPE) method coupled with ultra high performance liquid chromatography quadrupole time of flight mass spectrometry (UHPLC Q TOF MS) analysis was optimized and validated for the quantification of 67 (poly)phenol metabolites in human plasma and urine using authentic standards. The method was fully validated in terms of specificity, linearity, method detection limit (MDL) method quantification limit (MQL) repeatability, intra and inter day precision, accuracy and matrix effects. The method proved to be specific and results showed linearity of responses for all compounds, with MDL ranging between 0.04nM and 86nM in plasma and between 0.01nM and 136nM in urine. MQL ranged between 0.14nM and 286nM in plasma and between 0.03nM and 465nM in urine. Repeatability varied between 1.7 and 9.2% in plasma and between 2.2% and 10.4% in urine. Median precision values of 8.7 and 11.5% (intra day) and 10.8% and 10.0% (inter day) were obtained in plasma and urine, respectively. The median recovery was 89% in both biological matrices. Matrix effects were determined and median values of 1.2% and 6.8% in plasma and urine were obtained. After method validation, 49 and 57 compounds, including phase II and gut microbial metabolites, were quantified in plasma and urine, respectively, following cranberry juice consumption. This methodology can be applied to large scale human dietary intervention trials allowing for high sample throughput.
Journal of chromatography. A