East China University of Science and Technology, Shanghai 200237, China.
Phytochem Anal. 2010 Nov-Dec;21(6):524-30. doi: 10.1002/pca.1226.
Artemisia annua is a rich source of biologically active substances such as terpenoids, coumarins and polyacetylenes. These chemicals have been reported to show beneficial pharmacological properties such as antitumor and antibacterial activities. In genetically transformed root cultures of A. annua, three bioactive metabolites, namely, ponticaepoxide (an insecticidal polyacetylene, 1), drimartol A (an anticancer sesquiterpene coumarin, 2) and (Z)-7-acetoxy-methyl-11-methyl-3-methylene-dodeca-1,6,10-triene (a new anticancer sesquiterpene, 3) were isolated and identified in our recent work. However, no quantitative analysis methods for any of them are yet available, nor for their simultaneous analysis.
To develop an HPLC-PAD method for simultaneous determination of 1, 2 and 3 in hairy root cultures of A. annua.
HPLC operating conditions were optimised and the chromatographic separation was performed on a C(18) column with a gradient acetonitrile : water as mobile phase.
Linear relationships within the range of investigated concentrations were observed for the three metabolites with their correlation coefficients greater than 0.997. The method was validated for repeatability (RSD <3.59%) and intra- and inter-day precision (RSD <3.1%) with recovery between 94.8 and 107.6% and the RSD less than 3.40%. The method was successfully applied to the time-course of accumulation of the bioactive compounds in genetically transformed root cultures of A. annua.
The HPLC-PAD method developed for the simultaneous determination of three bioactive metabolites 1, 2 and 3 was simple, reproducible and sensitive.
青蒿是生物活性物质的丰富来源，如萜类、香豆素和多炔类。这些化学物质已被报道具有有益的药理特性，如抗肿瘤和抗菌活性。在青蒿遗传转化根培养物中，我们最近的工作分离并鉴定了三种生物活性代谢物，即蓬蒂亚烯氧化物（一种杀虫多炔，1）、二氢马桑毒素 A（一种抗癌倍半萜香豆素，2）和（Z）-7-乙酰氧基甲基-11-甲基-3-亚甲基-十二-1,6,10-三烯（一种新的抗癌倍半萜，3）。然而，目前还没有任何一种方法可以定量分析它们，也没有同时分析它们的方法。
建立青蒿毛状根培养物中 1、2 和 3 的同时 HPLC-PAD 测定方法。
优化了 HPLC 操作条件，采用 C（18）柱，以乙腈-水为流动相进行梯度洗脱，实现了色谱分离。
在所研究的浓度范围内，三种代谢物均呈良好的线性关系，相关系数均大于 0.997。该方法的重复性（RSD<3.59%）和日内、日间精密度（RSD<3.1%）良好，回收率在 94.8%至 107.6%之间，RSD 小于 3.40%。该方法成功应用于遗传转化青蒿根培养物中生物活性化合物的时间过程积累。
建立了一种用于同时测定三种生物活性代谢物 1、2 和 3 的 HPLC-PAD 方法，该方法简单、重现性好、灵敏度高。
