Anthocyanin determination in blueberry extracts from various cultivars and their antiproliferative and apoptotic properties in B16-F10 metastatic murine melanoma cells

• Published in: Phytochemistry (Oxford) Vol. 95; pp. 436 - 444
• Main Authors: Bunea, Andrea, Rugină, Dumitriţa, Sconţa, Zoriţa, Pop, Raluca M., Pintea, Adela, Socaciu, Carmen, Tăbăran, Flaviu, Grootaert, Charlotte, Struijs, Karin, VanCamp, John
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.11.2013
• Subjects: Animals; Anthocyanins; Anthocyanins - isolation & purification; Anthocyanins - pharmacology; Anthocyanins - therapeutic use; Antineoplastic Agents, Phytogenic - isolation & purification; Antineoplastic Agents, Phytogenic - pharmacology; Antineoplastic Agents, Phytogenic - therapeutic use; antioxidant activity; Antioxidants - isolation & purification; Antioxidants - pharmacology; Antioxidants - therapeutic use; Apoptosis; Apoptosis - drug effects; blueberries; Blueberry extracts; Blueberry Plants - chemistry; Cell culture; Cell Proliferation - drug effects; chemoprevention; cultivars; electrospray ionization mass spectrometry; Glucosides - isolation & purification; Glucosides - pharmacology; Glucosides - therapeutic use; high performance liquid chromatography; L-Lactate Dehydrogenase - metabolism; melanoma; Melanoma, Experimental - drug therapy; Melanoma, Experimental - metabolism; metastasis; Mice; Phytotherapy; Plant Extracts - pharmacology; Plant Extracts - therapeutic use; Species Specificity; Romania; Cell culture; Anthocyanins; Blueberry extracts; Apoptosis
• ISSN: 0031-9422; 1873-3700
• DOI: 10.1016/j.phytochem.2013.06.018

A crude methanolic extract was obtained from fresh blueberries of each seven cultivars tested. The crude extract was then purified over an Amberlite XAD-7 resin and a Sephadex LH-20 column, in order to obtain the anthocyanin rich fraction (ARF). High performance liquid chromatography followed by electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to identify and quantify individual anthocyanins. The richest anthocyanin fraction obtained from cultivar Torro (ARF-T) was used to evaluate the antiproliferative and apoptotic properties in B16-F10 metastatic murine melanoma cells. ARF-T showed an inhibition effect on B16-F10 cell proliferation and stimulated apoptosis (TUNEL). •Twelve anthocyanins were identified and quantified from seven blueberry cultivars.•Cultivar Toro had the highest anthocyanins concentration and antioxidant scores.•Anthocyanins could inhibit proliferation and induce apoptosis in B16-F10 cells. Blueberry consumption is associated with health benefits contributing to a reduced risk for cardiovascular disease, diabetes and cancer. The aim of this study was to determine the anthocyanin profile of blueberry extracts and to evaluate their effects on B16-F10 metastatic melanoma murine cells. Seven blueberry cultivars cultivated in Romania were used. The blueberry extracts were purified over an Amberlite XAD-7 resin and a Sephadex LH-20 column, in order to obtain the anthocyanin rich fractions (ARF). The antioxidant activity of the ARF of all cultivars was evaluated by ABTS, CUPRAC and ORAC assays. High performance liquid chromatography followed by electrospray ionization mass spectrometry (HPLC–ESI-MS) was used to identify and quantify individual anthocyanins. The anthocyanin content of tested cultivars ranged from 101.88 to 195.01mg malvidin-3-glucoside/100g fresh weight. The anthocyanin rich-fraction obtained from cultivar Torro (ARF-T) was shown to have the highest anthocyanin content and antioxidant activity, and inhibited B16-F10 melanoma murine cells proliferation at concentrations higher than 500μg/ml. In addition, ARF-T stimulated apoptosis and increased total LDH activity in metastatic B16-F10 melanoma murine cells. These results indicate that the anthocyanins from blueberry cultivar could be used as a chemopreventive or adjuvant treatment for metastasis control.