HPLC-tandem mass spectrometric method to characterize resveratrol metabolism in humans.
BACKGROUND
Nutritional biomarkers are alternatives to traditional dietary assessment tools. We sought to develop a method for nutritional analysis of resveratrol, a phenolic compound with purported health-promoting properties, and to determine all resveratrol metabolites.


METHODS
We obtained LDL and urine samples from 11 healthy male volunteers who had consumed 250 mL of Merlot red wine. We measured resveratrol and its metabolites with 96-well solid-phase extraction plates coupled with HPLC-tandem mass spectrometry. Hexestrol was used as the internal standard. Gradient chromatography in multiple reaction monitoring mode was performed on a Luna C(18) column, maintained at 40 degrees C; m/z transitions were as follows: resveratrol, 227/185; resveratrol glucosides, 389/227; resveratrol glucuronides, 403/227; resveratrol sulfates, 307/227; taxifolin, 303/285; and hexestrol, 269/134.


RESULTS
Standard calibration curves were linear at 4.4-3289.5 nmol/L. Residual analyses were 100% (3.2) for trans-resveratrol and 100% (11.1) for trans-piceid. In both matrices, imprecision (CV) was <10.8% at all concentrations. Detection limits for resveratrol were 0.2 nmol/L (LDL), 0.3 nmol/L (synthetic urine), and 4.0 nmol/L (blank urine). Resveratrol and metabolites were checked for stability, and no degradation was observed.


CONCLUSIONS
The HPLC-tandem mass spectrometry method enabled us to identify resveratrol sulfates in human LDL and to characterize the complete profile of resveratrol metabolism in human LDL and urine. This method provides an accurate index of exposure to resveratrol and its metabolites, which can be used as nutritional biomarkers for evaluating the biological effects of moderate wine intake on human health.
Urpi-Sarda
Mireia
M
Nutrition and Food Science Department, CeRTA, Pharmacy School, University of Barcelona, Barcelona, Spain.
Zamora-Ros
Raul
R
Lamuela-Raventos
Rosa
R
Cherubini
Antonio
A
Jauregui
Olga
O
de la Torre
Rafael
R
Covas
Maria Isabel
MI
Estruch
Ramon
R
Jaeger
Walter
W
Andres-Lacueva
Cristina
C
D000293:Adolescent; D000328:Adult; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011786:Quality Control; D000077185:Resveratrol; D012680:Sensitivity and Specificity; D013237:Stereoisomerism; D013267:Stilbenes / Q000737:chemistry / Q000378:metabolism* / Q000652:urine; D053719:Tandem Mass Spectrometry; D014920:Wine
D008077:Lipoproteins, LDL; D013267:Stilbenes; D000077185:Resveratrol