Sensitive and Rapid UHPLC-MS/MS for the Analysis of Tomato Phenolics in Human Biological Samples
2015
Martinez-Huelamo, Miriam | Tulipani, Sara | Jauregui, Olga | Valderas-Martinez, Palmira | Vallverdu Queralt, Anna | Estruch, Ramón | Torrado, Xavier | Lamuela-Raventos, Rosa M. | Department of Nutrition and Food Science-XARTA-INSA, School of Pharmacy ; Universitat de Barcelona (UB) | Carlos III Health Institute ; Biomedical Research Center in Red-Physiopathology of Obesity and Nutrition (CIBERobn) | Biomedical Research Institute (IBIMA), Service of Endocrinology and Nutrition, Hospital Virgen de la Victoria, Teatinos Campus ; Universidad de Málaga [Málaga] = University of Málaga [Málaga] | Universitat Autònoma de Barcelona = Autonomous University of Barcelona = Universidad Autónoma de Barcelona (UAB) | Department of Internal Medicine, Hospital Clinic, Institute of Biomedical Investigation August Pi i Sunyer (IDIBAPS) ; Universitat de Barcelona (UB) | Sciences Pour l'Oenologie (SPO) ; Université Montpellier 1 (UM1)-Institut National de la Recherche Agronomique (INRA)-Université de Montpellier (UM)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro) | CICYT [AGL2010-22319-C03 ], [AGL2013-49083-C3-1-R ]
An UHPLC-MS/MS method for the quantification of tomato phenolic metabolites in human fluids was optimized and validated, and then applied in a pilot dietary intervention study with healthy volunteers. A 5-fold gain in speed (3.5 min of total run); 7-fold increase in MS sensitivity and 2-fold greater efficiency (50% peak width reduction) were observed when comparing the proposed method with the reference-quality HPLC-MS/MS system, whose assay performance has been previously documented. The UHPLC-MS/MS method led to an overall improvement in the limits of detection (LOD) and quantification (LOQ) for all the phenolic compounds studied. The recoveries ranged between 68% and 100% in urine and 61% and 100% in plasma. The accuracy; intra- and interday precision; and stability met with the acceptance criteria of the AOAC International norms. Due to the improvements in the analytical method; the total phenolic metabolites detected in plasma and urine in the pilot intervention study were 3 times higher than those detected by HPLC-MS/MS. Comparing with traditional methods; which require longer time of analysis; the methodology described is suitable for the analysis of phenolic compounds in a large number of plasma and urine samples in a reduced time frame.
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