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Stability of anthocyanins in vitro

Anthocyanins (ACNs) are abundant in berry fruits and are associated with a variety of health benefits to humans, including cardiovascular disease prevention, improved glucose metabolism, body weight regulation and improvement of vision and memory. However, ACNs are known to be unstable in vitro and this needs to be considered in the design of cell studies. The aim of this experiment was to investigate the stability of cyanidin-3-glucoside (C3G) and cyanidin-3-rutinoside (C3R) in vitro over a 24 h period. The methodological design was adapted from a study by Woodward et al.. ACNs (C3G and C3R) at 10mM in either RPMI medium alone or in RPMI medium with added foetal calf serum (FCS) and Penicillin-Streptomycin (AB) were incubated at 37 C in a 5% CO2 atmosphere. After 0, 0.5, 2, 4, 6, 12 and 24 h, aliquots were removed, acidified with formic acid, filtered and analysed using high-pressure liquid chromatography (HPLC). To evaluate the influence of freezing, half of the acidified aliquots were frozen at 80 C for 2 to 3 weeks prior to HPLC analysis.