engleski

Validation of videodensitometric method for determination of oleuropein on TLC plates

Oleuropein is the primary phenolic compound found in olive products. There are several chromatographic methods for determination of oleuropein, HPLC being the most used. However, its use is somewhat limited by relatively high cost and long time of analysis. Videodensitometry combined with TLC is gaining popularity due to very low cost of analysis and low toxicity to environment as compared to HPLC.1 Hereby we report validation of videodensitometric method for determinig oleuropein olive leaf extracts using trial version of Sorbfil program for chromatogram analysis. The whole method was validated by the assay tests of detection (LOD) and quantification limit (LOQ), calibration curve, repeatability and recovery. The extracts were prepared using varying amount of ethanol and water (0%-100% v/v) at different temperatures (20-60 C). For TLC, Pre-Coated Silica gel 60 F-254 plates (Merck, DE) were used. As mobile phase ethyl-acetate:dioxan:formic acid (30:10:0.3 v/v/v) was employed. LOD and LOQ were 4.5910-4 g and 1.5310-3 g, respectively. The calibration curve equation was y = 6106x - 3000, 2 where x is weight (g) and y area under curve (arbitrary units). Repeatability and recovery were 3.3% and 103.9%, respectively. The analysis of 18 extracts was done in approximately 2 hours using 50 ml of solvent mixture which represents considerable savings in time and cost of the analysis compared to HPLC. The results showed that the tested chromatographic conditions, combined with videodensitometric analysis, were applicable for the quantification of oleuropein in the olive leaf extracts.

validation; TLC; oleuropein

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