Affiliation:
1. Qualisud Mixed Research Unit, Faculté de Sciences Pharmaceutiques Université de Montpellier Montpellier France
2. School of Pharmacy, Trinity College University of Dublin Dublin Ireland
Abstract
AbstractIntroductionOlive leaves, abundant by‐products of the olive oil industry, are a rich source of oleuropein, an important polyphenol in olive leaves. So far, no published methods have been validated using matrix standards for oleuropein quantification in olive leaves.ObjectivesThe study aimed to develop an HPLC method for oleuropein determination in olive leaves using spiked matrix standards prepared from a blank olive leaf matrix, to validate the method with respect to aqueous standards, and cross‐validate the HPLC method with UPLC‐MS and UPLC‐UV techniques.MethodologyOleuropein was extracted into methanol and analysed by HPLC with fluorescence detection (FLD; excitation and emission wavelengths 281 and 316 nm, respectively) and by UPLC‐MS‐UV. For validation, calibration curves of spiked matrix standards (0.4 to 4.8 mg/g) were analysed by the three methods over several days. Oleuropein was then analysed in French olive varieties.ResultsFor the HPLC‐FLD method, repeatability and intermediate precision were less than 5% RSD and linearity was demonstrated by the Fischer test. Differences in results of the spiked placebos by the three methods were non‐significant, as confirmed by ANOVA. Extraction recovery was >90%, and there was a strong linear relationship between authentic and spiked matrix standards. The determination of oleuropein in French olive varieties is reported, including analysis in “Olivière” cultivar for the first time, leaves of which contained twice the amount of oleuropein compared with “Picholine”.ConclusionAccurate quantification of oleuropein is possible using aqueous standards. Cross‐validation indicates that selective analysis can equally be carried out by HPLC or by UPLC‐MS techniques.
Subject
Complementary and alternative medicine,Drug Discovery,Plant Science,Molecular Medicine,General Medicine,Biochemistry,Food Science,Analytical Chemistry