title: A novel liquid chromatography method using diode-array detector for the determination of oleuropein in dietary supplements.
abstract: A simple and fast chromatographic method using ultraviolet diode-array detector (UV-DAD) was developed for the automatic high performance liquid chromatography (HPLC) determination of the title of oleuropein in a new dietary supplements in form of effervescent granules. The chromatographic separations were performed on a C18 core-shell column with detection at λ=232nm. The mobile phase consisted of deionized water with 0.1% TFA and acetonitrile under gradient conditions at a flow-rate of 0.8mL/min. Oleuropein and oleuroside present in the raw material were characterized by high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). The validation of the analytical procedure has been performed determining the following parameters: specificity, linearity, repeatability, reproducibility, accuracy, limit of quantification (LOQ), stability of the standard and sample solutions. Linear response was observed in fortified placebo solutions (determination coefficient: 0.9998). Intra-day precision (relative standard deviation, RSD) was ≤5.0% for peak area and for retention times (tR) without significant differences between intra- and inter-day data. The limits of quantitation (LOQ) was about 5μg/mL and 9pmol/inject. Oleuropein recovery studies gave good results (99.9%) with a R.S.D. of 0.5%. The speed of analysis and the stability of the solutions with a fluctuation Δ (%) ≤2.0 at room temperature means an undoubted advantage of the method allowing the simultaneous preparation of many samples and consecutive chromatographic analyses by using an autosampler. The developed method is suitable for the quality control of oleuropein in raw material and industrial products. The method can be applied in any analytical laboratory not requiring a sophisticated instrumentation.
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123, Trento, TN, Italy.
authors.forename: Tiziana
authors.initials: T
authors.lastname: Bertolini
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123, Trento, TN, Italy.
authors.forename: Lorenza
authors.initials: L
authors.lastname: Vicentini
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123, Trento, TN, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Boschetti
authors.affiliation: E-Pharma Trento S.p.A., Research and Development, via Provina 2, 38123, Trento, TN, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Andreatta
authors.affiliation: Department of Pharmacy & Biotechnology, Alma Mater Studiorum - Università di Bologna, Via Belmeloro 6, 40126 Bologna, Italy. Electronic address: rita.gatti2@unibo.it.
authors.forename: Rita
authors.initials: R
authors.lastname: Gatti
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019587:Dietary Supplements; D057888:Iridoid Glucosides; D039823:Iridoids / Q000737:chemistry*; D057230:Limit of Detection; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D057888:Iridoid Glucosides; D039823:Iridoids; C002769:oleuropein
keywords: Dietary supplements; HPLC UV-DAD; HPLC-ESI–MS/MS; Kinetex column; Olea europea dry extract; Oleuropein