engleski

Validation of HPLC-PDA method for determination of polyphenol profile of Croatian traditional apple cultivars

Polyphenols are molecules with strong antioxidant activity that plants synthesize in response to stress conditions. Apple cultivars containing higher levels of polyphenols, procyanidins, dihydrochalcones, flavan-3-ols, flavonols, and phenolic acids, are more resistant to blue mould caused by P. expansum. Furthermore, apple polyphenols are involved in the response to patulin contamination, as they neutralize the free radicals induced by patulin. For this reason, a HPLC method was established, optimized, and validated for the separation and quantitation of 19 polyphenols extracted from the Croatian traditional apple cultivars. A reversed-phase C18 column was used as stationary phase, and an acidified water and methanol were used as mobile phase. The polyphenols were detected using a photo-diode array detector (PDA) at 280, 320, 360 and 520 nm. For the evaluation of fitness for purpose, linearity, trueness and precision were determined and all validation parameters were acceptable for all determined polyphenols. Identification of the separated components was performed based on the retention time and comparison of the absorption spectra of the components in the apple extract with the spectra of the standards, while the quantification of the components was performed based on the external calibration method. The method was successfully applied for simultaneous analysis of procyanidins, dihydrochalcones, flavan-3-ols, flavonols, and phenolic acids from Croatian traditional apple cultivars.

polyphenols ; HPLC ; validation ; Croatian traditional apple cultivars

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