不同产地高良姜的质量分析及其抗肿瘤活性部位筛选

Quality analysis of Alpinia officinarum Hance from different areas and screening of active antitumor fractions

目的对来自不同产地的高良姜饮片进行质量分析,确定优质高良姜药材来源,在此基础上,筛选其对6种肿瘤细胞的抗肿瘤活性部位。方法以高良姜素含量为指标,利用HPLC法对不同产地的高良姜进行质量分析,色谱柱为Agilent C_(18)(4.6 mm×250 mm,5μm)柱,检测波长266 nm,柱温25℃,流动相甲醇-0.2%磷酸(55∶45),流速1 mL·min^(-1)。采用石油醚、二氯甲烷、乙酸乙酯、正丁醇依次对高良姜85%乙醇提取物进行萃取,将得到的各提取部位用MTT法评价其体外细胞毒活性,进行抗肿瘤活性筛选。结果广东产高良姜中高良姜素含量高于其他2个产地,为14.17 mg·g^(-1)。高良姜石油醚萃取部位和二氯甲烷萃取部位对6种肿瘤细胞均有明显抑制作用,活性强于乙酸乙酯萃取部位、正丁醇萃取部位与水部位,且呈现时间和浓度依赖关系。对人宫颈癌细胞株Hela抑制效果最好,石油醚萃取部位24 h与48 h的IC_(50)分别为7.88、6.55μg·mL^(-1);二氯甲烷萃取部位24 h与48 h的IC_(50)分别为6.13、4.78μg·mL^(-1)。结论 HPLC法优选方便、高效,高良姜抗肿瘤活性的有效部位为石油醚萃取部位和二氯甲烷萃取部位。

Objective To analyze the quality of Alpinia officinarum Hance from different areas, to determine the source of high quality Alpinia officinarum Hance, and to screen its active antitumor fractions against 6 types of tumor cells. Methods We detemined the galangin in content of Alpinia officinarum Hance from different areas by HPLC method. The chromatographic column was Agilent C（18） column（4.6 mm×250 mm, 5 μm） with detection wavelength at 266 nm. The column temperature was 25 ℃; the mobile phase： methanol and 0.2% phosphoric acid, the ratio was 55 ∶ 45 and the flow rate was 1 mL·min^（-1）. The 85% ethanol extract of galangal was extracted by petroleum ether, dichloromethane, ethyl acetate and n-butanol in turn. The in vitro cytotoxic activity of the extract was evaluated by MTT method. Results The content of galangin（14.17 mg·g^（-1）） in Alpinia officinarum Hance from Guangdong was higher than that from the other two areas. The extracts of dichloromethane and petroleum ether had obvious inhibition on the tested 6 types of tumor cells in a dose and time dependent manner, whose activity was stronger than that of the ethyl acetate fractions, the n-butanol fractions and the water fractions. The best inhibition was on Hela cells. The IC（50） of petroleum ether extracts at 24 h and 48 h was 7.88 μg·mL^（-1） and 6.55 μg·mL^（-1）, while the IC（50） of dichloromethane extracts at 24 h and 48 h was 6.13 μg·mL^（-1） and 4.78 μg·mL^（-1）. Conclusion HPLC optimization is convenient and efficient. The active antitumor fractions of Alpinia officinarum Hance include petroleum ether extract fraction and dichloromethane extract fraction.