Bazoti Fotini N, Gikas Evangelos, Tsarbopoulos Anthony
Department of Pharmacy, University of Patras, Panepistimiopolis, Rio, Greece.
Biomed Chromatogr. 2010 May;24(5):506-15. doi: 10.1002/bmc.1319.
Oleuropein (OE) is the cardinal bioactive compound derived from Olea europaea and possesses numerous beneficial properties for human health. However, despite the plethora of analytical methods that have studied the biological fate of olive oil-derived bioactive compounds, no validated methodology has been published to date for the simultaneous determination of OE, along with all its major metabolites. In this study, a liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI MS/MS) method has been developed and validated for the quantification of OE, simultaneously with its main metabolites hydroxytyrosol, 2-(3,4-dihydroxyphenyl)acetic acid, 4-(2-hydroxyethyl)-2-methoxy-phenol or homovanillyl alcohol, 2-(4-hydroxy-3-methoxyphenyl)acetic acid or homovanillic acid, and elenolic acid in rat plasma matrix. Samples were analyzed by LC-ESI MS/MS prior to and after enzymatic treatment. A solid-phase extraction step with high mean recovery for all compounds was performed as sample pretreatment. Calibration curves were linear for all bioactive compounds over the range studied, while the method exhibited good accuracy, intra- and inter-day precision. The limit of detection was in the picogram range (per milliliterof plasma) for HT and OE and in the nanogram range (per milliliter of plasma) for the other analytes, and the method was simple and rapid. The developed methodology was successfully applied for the simultaneous quantification of OE and its aforementioned metabolites in rat plasma samples, thus demonstrating its suitability for pharmacokinetics, as well as bioavailability and metabolism studies.
橄榄苦苷（OE）是从油橄榄中提取的主要生物活性化合物，对人体健康具有多种有益特性。然而，尽管有大量分析方法研究了橄榄油衍生生物活性化合物的生物学命运，但迄今为止，尚未发表经过验证的方法来同时测定OE及其所有主要代谢产物。在本研究中，开发并验证了一种液相色谱-电喷雾电离-串联质谱（LC-ESI MS/MS）方法，用于在大鼠血浆基质中同时定量测定OE及其主要代谢产物羟基酪醇、2-(3,4-二羟基苯基)乙酸、4-(2-羟乙基)-2-甲氧基苯酚或高香草醇、2-(4-羟基-3-甲氧基苯基)乙酸或高香草酸以及橄榄酸。在酶处理前后，通过LC-ESI MS/MS对样品进行分析。作为样品预处理，进行了所有化合物平均回收率高的固相萃取步骤。在所研究的范围内，所有生物活性化合物的校准曲线均呈线性，该方法具有良好的准确性、日内和日间精密度。羟基酪醇和橄榄苦苷的检测限在皮克范围内（每毫升血浆），其他分析物的检测限在纳克范围内（每毫升血浆），该方法简单快速。所开发的方法成功应用于大鼠血浆样品中OE及其上述代谢产物的同时定量测定，从而证明了其适用于药代动力学以及生物利用度和代谢研究。
