Determination of hydroxytyrosol in plasma by HPLC.
Hydroxytyrosol (2-(3,4-dihydroxyphenyl)ethanol), a phenolic compound present in extravirgin olive oil, has been reported to contribute to the prevention of cardiovascular disease. The present study describes an accurate and reproducible reversed-phase HPLC method to measure hydroxytyrosol in plasma. This compound was extracted from acidified plasma by solid-phase extraction using an Oasis HLB copolymer. The plasma sample was rinsed with water and methanol in water (5:95; v/v). Hydroxytyrosol was eluted with methanol, which was subsequently evaporated under a nitrogen stream. Analysis by HPLC with diode array-UV detection was carried out using a C18 column and a gradient elution with acidified water and methanol/acetonitrile (50:50; v/v). The method was validated by the analyses of plasma samples spiked with pure hydroxytyrosol, obtaining a linear correlation (0.9986) and precision with a coefficient of variation ranging from 0.79 to 6.66%. The recovery was approximately 100%, and the limit of detection was 37 ng/mL. The oral administration of hydroxytyrosol to rats and its subsequent detection in plasma showed that the method is suitable for pharmacokinetic studies.
Ruiz-Gutiérrez
V
V
Instituto de la Grasa y sus derivados, Consejo Superior de Investigaciones Científicas, Sevilla, Spain.
Juan
M E
ME
Cert
A
A
Planas
J M
JM
D000818:Animals; D000975:Antioxidants / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007202:Indicators and Reagents; D010626:Phenylethyl Alcohol / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D051381:Rats; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D000975:Antioxidants; D007202:Indicators and Reagents; C005975:3,4-dihydroxyphenylethanol; D010626:Phenylethyl Alcohol