Quantification of artemisinin in Artemisia annua extracts by 1H-NMR
Quantification of artemisinin in Artemisia annua extracts by 1H-NMR
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DOI：
10.1002/pca.1053
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发表时间：
2008-07-01
影响因子：
3.3
通讯作者：
Rodrigues, Ana I.
中科院分区：
文献类型：
--
作者：
Castilho, Paula C.;Gouveia, Sandra C.;Rodrigues, Ana I.
青蒿素是一种多环倍半萜内酯，对恶性疟原虫的多药耐药株非常有效，恶性疟原虫是最严重形式的疟疾的病原体。青蒿素在源植物青蒿中的测定是一个具有挑战性的问题，因为该化合物以非常低的浓度存在，是热不稳定和不稳定的，并且缺乏发色团或荧光团。本研究的主要目的是建立一种简单的方法，用1H-1-NMR方法定量测定植物提取物中的青蒿素。通过用丙酮提取叶材料，用活性炭处理以除去叶绿素和除去溶剂来制备样品。以叔丁醇为内标，对溶解在氘代氯仿中的样品进行1H-1-NMR光谱测量。使用青蒿素的δ 5.864信号和叔丁醇的δ 1.276信号进行定量。该方法进行了优化，并充分验证了对青蒿素的参考标准。将结果与使用HPLC-折射率（RI）方法定量的相同样品中获得的结果进行比较。1H-NMR方法给出青蒿素在9.85-97.99 mM范围内的线性响应（r（2）= 0.9968）。用该方法从青蒿叶中提取青蒿素，产率为0.77-1.06%。annua杂种CPQBA x POP，这些值与使用HPLC-RI获得的值一致。版权所有（C）2008约翰威利父子有限公司
Artemisinin is a polycyclic sesquiterpene lactone that is highly effective against multidrug-resistant strains of Plasmodium falciparum, the etiological agent of the most severe form of malaria. Determination of artemisinin in the source plant, Artemisia annua, is a challenging problem since the compound is present in very low concentrations, is thermolabile and unstable, and lacks chromophoric or fluorophoric groups. The ain of this study was to develop a simple protocol for the quantification of artemisinin in a plant extract using an H-1-NMR method. Samples were prepared by extraction of leaf material with acetone, treatment with activated charcoal to remove chlorophylls and removal of solvent. H-1-NMR spectra were measured on samples dissolved in deuterochloroform with tert-butanol as internal standard. Quantification was carried out using the delta 5.864 signal of artemisinin and the delta 1.276 signal of tert-butanol. The method was optimised and fully validated against a reference standard of artemisinin. The results were compared with those obtained from the same samples quantified using an HPLC-refractive index (RI) method. The 1H-NMR method gave a linear response for artemisinin within the range 9.85-97.99 mM (r(2) = 0.9968). Using the described method, yields of artemisinin in the range 0.77-1.06% were obtained from leaves of the A. annua hybrid CPQBA x POP, and these values were in agreement with those obtained using an HPLC-RI. Copyright (C) 2008 John Wiley & Sons, Ltd.