商陆中9种核苷类物质的快速检测方法

Determination of Nine Nucleosides in Phytolacca acinosa Roxb. and the Effect of Extraction Condition on the Nucleosides

建立高效液相色谱法测定核苷类物质,探究含量并比较提取工艺对核苷类物质含量的影响。采用Waters Symmetry C18色谱柱(4.6 mm×150 mm),流动相A相为甲醇;流动相B相为纯水,梯度洗脱,流速1.0 mL/min, VWD检测波长260 nm;柱温37℃。检测方法显示, 9种核苷类物质的检测线性范围为0.05~100 mg/L;检出限(S/N=3)为0.002~0.018 mg/kg;在加标浓度分别为1.0, 5.0, 10.0和20.0 mg/g条件下,加标回收率(N=6)为87.12%~104.22%,检测方法灵敏有效。检测结果表明,不同提取工艺得到的商陆中叶尿嘧啶、胞苷、次黄嘌呤、尿苷、腺嘌呤、次黄嘌呤核苷、鸟苷、胸苷和腺苷9种商陆核苷类物质的总含量以超临界萃取最高,超临界萃取过程没有加热处理,最大程度地保留了商陆核苷类物质。结果表明,从商陆核苷类物质的含量及物质特性保留的角度考虑,选择商陆的提取工艺以超临界萃取为宜。

The influence of different extraction condition on contents of nine kinds of the nucleosides(uracil,cytidine,hypoxanthine,uridine,adenine,inosine,guanosine,thymidine and adenosine)in Phytolacca acinosa Roxb.was compared by using HPLC.The experiment was based on Waters Symmetry C18 column(4.6 mm×150 mm)with mobile phase A of methanol and mobile phase B of water,and gradient eluted with the flow rate of 1.0 mL/min,VWD detection wavelength of 260 nm and column temperature of 37℃.Detection method showed sensitively and effectively,with nine kinds of nucleosides linear range of 0.05-100 mg/L,the detection limit(S/N=3)between 0.002 and 0.018 mg/kg,the recoveries(N=6)between 87.12%and 104.22%,under the condition of spiked concentrations of 1.0,5.0,10.0 and 20.0 mg/g.The results showed that the total content of nine nucleosides(uracil,cytidine,hypoxanthine,uridine,adenine,inosine,guanosine,thymidine and adenosine)in Phytolacca acinosa Roxb.was the highest when using supercritical extraction among all the extraction methods,due to no heating treatment during supercritical extraction,which could keep the nucleosides in Phytolacca acinosa Roxb.by maximum.It was shown as well that from the aspects of increasing nucleosides content and retaining material properties,it was the best to choose supercritical extraction to extract the Phytolacca acinosa Roxb.