Comparison of HPLC Methods for Determination of Anthocyanins and Anthocyanidins in Bilberry Extracts

An HPLC method and an acid hydrolysis HPLC method for the analysis of anthocyanins and
anthocyanidins in bilberry extracts have been developed. The HPLC method coupled with a mass
detector has identified 11 anthocyanins in bilberry extracts. The method provides anthocyanin profiles
that are very useful in verifying the identity of botanical raw materials, monitoring the consistency of
the raw material source, and quantitating the total anthocyanins. The acid hydrolysis HPLC method
greatly simplifies the anthocyanin profile in bilberry samples and converts anthocyanins to five major
anthocyanidin aglycones:  delphinidin, cyanidin, petunidin, peonidin, and malvidin. Each of these
aglycones can be separated completely and quantitated accurately with external standards. Various
extraction and hydrolysis conditions were investigated, and the advantages and disadvantages of
the HPLC and acid hydrolysis methods are discussed.
Keywords: HPLC; HPLC-MS; anthocyanin; anthocyanidin; bilberry extract