Ferreira Jorge F S, Gonzalez Javier M
Appalachian Farming Systems Research Center, U.S. Department of Agriculture Agricultural Research Service, 1224 Airport Road, Beaver, WV 25813, USA.
Phytochem Anal. 2009 Mar-Apr;20(2):91-7. doi: 10.1002/pca.1101.
Although high-performance liquid chromatography with ultraviolet detection (HPLC-PAD) is widely available, it has not been used for artemisinin (1) analysis because of the lack of UV absorption reported for this lactone. Increased Artemisia annua cultivation for production of 1 requires an affordable and reliable method to analyse 1 and its precursors dihydroartemisinic acid (2) and artemisinic acid (3) simultaneously from underivatized plant extracts.
To validate HPLC-PAD for the quantification of underivatized artemisinin from A. annua and artemisinin-based drugs.
Dried A. annua leaves were extracted with petroleum ether, dried, reconstituted in acetonitrile, and analysed by HPLC-PAD at 192 nm using an isocratic mobile phase (60:40, acetonitrile:0.1% acetic acid). HPLC-PAD was evaluated through accuracy, precision, recovery and comparison with HPLC with evaporative light scattering detection (HPLC-ELSD).
HPLC-PAD proved accurate, precise and reproducible for the direct quantification of 1 and related compounds, and was more sensitive than ELSD for most of the compounds tested. The limit of quantification of 1-3 from plants was 0.048, 0.024 and 0.008 g/100 g dry weight, respectively. Recoveries were over 98%, with good intra- and inter-day repeatability. HPLC-PAD correlated significantly (r(2 )= 0.99, p < 0.001) with HPLC-ELSD for artemisinin analysis. HPLC-PAD was also reliable for the analysis of dihydroartemisinin, artesunate and artelinic acid.
HPLC with ultraviolet detection was validated for the quantification of underivatized 1, 2, and 3 from crude plant samples, and is readily applicable for the quality control of herbals and artemisinin-related pharmaceutical compounds.
尽管配备紫外检测的高效液相色谱法（HPLC-PAD）已广泛应用，但由于该内酯类青蒿素（1）缺乏紫外吸收，尚未用于其分析。为生产青蒿素而增加的青蒿种植需要一种经济且可靠的方法，以便从未经衍生化的植物提取物中同时分析青蒿素及其前体二氢青蒿酸（2）和青蒿酸（3）。
验证HPLC-PAD用于定量分析青蒿中未衍生化青蒿素及基于青蒿素药物的可行性。
将干燥的青蒿叶用石油醚萃取、干燥，再用乙腈复溶，然后采用等度流动相（60:40，乙腈:0.1%乙酸）在192 nm波长下通过HPLC-PAD进行分析。通过准确度、精密度、回收率以及与蒸发光散射检测高效液相色谱法（HPLC-ELSD）进行比较来评估HPLC-PAD。
HPLC-PAD被证明对于直接定量分析青蒿素及相关化合物准确、精密且可重现，并且对于大多数测试化合物而言，其比ELSD更灵敏。植物中青蒿素-3的定量限分别为0.048、0.024和0.008 g/100 g干重。回收率超过98%，日内和日间重复性良好。在青蒿素分析方面，HPLC-PAD与HPLC-ELSD具有显著相关性（r² = 0.99，p < 0.001）。HPLC-PAD对于二氢青蒿素、青蒿琥酯和青蒿酸的分析也可靠。
配备紫外检测的高效液相色谱法经验证可用于从未经衍生化的植物样品中定量分析青蒿素-3，并且易于应用于草药及青蒿素相关药物化合物的质量控制。
