Application of RP‐HPLC and Spectrophotometry in Standardization of Bilberry Anthocyanin Extract

Abstract

V. myrriNus extracts were examined both qualitatively and quantitatively by UV-VIS spectrophotometry and a new RP-HPLC method. The results and Standardislerung der Anthocyanlne In Heldelbeerextrakten durch RP-HPLC und Spektrophotometrie the problems arising from application of these methods for bilbeny anthocyanins determination are discussed. The quality and purity of various extracts is compared.Die Heidelbeerextrakte werden mit Hilfe der Spektrophotometrie und einer neuen RP-HPLC Methode untersucht. Die Anwendung der beiden Methoden in der Bestimmung der Anthocyane wird diskutiert. Die Qualitiit und die Reinheit der Extrakte verschiedener Produzenten werden verglichen. Fruit of bilberry (Vacciniwn myrtillus L., Ericaceae) contain complex mixtures of anthocyanins, i.e. delphinidin, cyanidin, petunidin, and malvidin glycosides'). From the analytical point of view, the anthocyanins have provided a considerable challenge, mainly because of their relative lability. The methds for separation of these compounds require strong acidic media as anthocyanins are stable in such conditions in the form of mi flavylium ions2).The solutions of anthocyanins in acidified methanol or acidified ethanol absorb in the UV-region (maximum about 280 nm) and exhibit a characteristic maximum in the visible region (within the range of 510-560 m~)~) .The results of UV-VIS spectrophotometry of bilberry extracts are shown in Table 1. The extracts varied considerably in the anthocyanin content. The qualities of extracts R and F were much lower than those of extracts P and I. Absorbance values in the W depend also on the presence of other compounds (phenolic acids, tannins etc.). Relatively high values of A"lm of extracts R and F suggest the presence of large amounts of other polyphenols in these samples.Successful separation of phenolic acids and flavonoids by Rp-HPLC led to similar analytical methods for anthocyanidins and anth~cyanins~~). Chromatographic data of anthocyanin separation by our system are specified in Table 2.Phenolic acids, catechins and some proanthocyanidins were eluted during 40 min. The peaks of anthocyanins appeared within the next 50 min. However, when the sample abounded in tannins most of them were eluted together with anthocyanins and the base line drifted.The composition of the extracts, based on RP-HPLC determination, is presented in Table 3. The preponderance of delphinidin glycosides was observed in all the extracts. Extracts I and P had similar composition, moreover we did not find cyanidin-3-0-glucoside in extract P. The,content of

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Study of diversity of anthocyanin composition in bilberry (Vaccinium myrtillus L.) fruits

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