沙棘籽粕原花青素制备、体外抗氧化及细胞活力评价

Technological conditions,antioxidant activities in vitro and cell viability of the proanthocyanidins extracted from seabuckthorn( Hippophae rhamnoides L.) seed residues

本实验优化了沙棘籽粕原花青素的提取方法,条件为80%乙醇,提取温度35℃,液料比8∶1 m L/g,时间1.5 h,p H3.0,在该条件下的原花青素提取率为(97.31±0.48)mg/g沙棘籽粕;对提取得到的原花青素提取物进行了DPPH自由基和羟自由基清除能力实验,并分析得到IC_50,发现沙棘籽粕原花青素提取物的DPPH自由基清除能力较维生素C强;羟自由基清除能力较弱;MTT法测定了沙棘籽粕原花青素提取物对小鼠B16黑色素瘤细胞活力的抑制作用,分析得到24、48、72 h处理时间下提取物IC_50分别为542.78、199.25、82.58μg/m L。结论:沙棘籽粕提取物对小鼠B16黑色素瘤细胞具有一定的抑制作用,随着剂量和时间的增加,呈逐渐增强的趋势。

The optimal conditions of proanthocyanidins extracted from seabuckthorn （Hippophae rhamnoides L.） seed residues were obtained by orthogonal test,on the basis of one factor tests.The optimal condition of ethanol extraction was ethanol concentration 80% ,temperature 35 ℃, liquid to solid ratio 8：1 mL/g, extraction time 1.5 h, pH3.0.Under this condition, proanthocyanidins was 97.31 ± 0.48 mg/g seabuckthorn （Hippophae rhamnoides L.） seed residues. Antioxidant activities were tested, using crude extract lyophilized power. Antioxidant activities of proanthocyanidins extracted from seabuckthorn （Hippophae rhamnoides L.）seed residues were investigated with the method of scavenging hydroxyl and DPPH. IC50 was calculated. Results indicated that the proanthocyanidin extracts had higher antioxidant activity than vitamin C on the scavenging ability of DPPH.The ability on scavenging hydroxyl was low.Methods MTT assay was employed to determine the cell viability of proanthocyanidin extracts in B16 cells.The IC50 was 542.78,199.25 and 82.58 μg/mL when B16 cells was exposed for 24,48,and 72 h, respectively. It was suggested that the B16 cell viability followed the dose-and time-dependent manner.