1H-qNMR for direct quantification of stachydrine in Leonurus japonicus and L. cardiaca
1H-qNMR for direct quantification of stachydrine in Leonurus japonicus and L. cardiaca
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DOI：
10.1016/j.fitote.2014.03.023
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发表时间：
2014-07-01
期刊：
影响因子：
3.4
通讯作者：
Wilhelm-Rauwald, Hans
中科院分区：
文献类型：
--
作者：
Kuchta, Kenny;Ortwein, Jutta;Wilhelm-Rauwald, Hans
利用H-1-qNMR谱对益母草(益母草，中国Ph.2010，DAB2012)地上部分中具有药理活性的水苏碱((2S)-1，1-dimethylpyrrolidinium-2-carboxylic-acid)进行了定量研究，发现益母草可治疗多种妇科和心血管疾病。关于甜菜碱的药理学出版物描述了心血管、降压和组织保护作用。然而，它的药典分析造成了严重的困难，因为它不包含任何适合高效液相色谱紫外检测的生色团。来自三个国家的9个样品被制成煎剂并冷冻干燥。在D2O中记录了H-1-核磁共振波谱。直接定量的H-1-q核磁共振程序是使用增量为3.03 ppm的N-CH3-单线态进行的，与顺丁烯二酸的两个乙烯基质子的增量为6.18 ppm的单线态相比较，后者被确定为最有利的内标。水苏碱的定量限为0.44 mg/g药材。既不需要校准曲线的参比化合物，也不需要样品的预纯化。这一方案显示，所测试的益母草样品的水苏碱含量在0.09%到1.01%(w/w)之间。此外，在首次检测到水苏碱成分的L果实药材(益母草，崇味子；中国Ph.2010)中发现了0.18%至0.21%的水苏碱。在四个共同调查的密切相关和类似使用的欧洲草本植物益母草中，证实了更高的含量，高达1.55%。所提出的定量H-1-q核磁共振方法被证明是关于浓度的精确的，并且在一系列日间重复中产生高度重复性的数据。从方法上讲，H-1-q核磁共振可能是水苏碱植物和中草药质量保证的有力工具，特别是对于工业常规方案。(C)2014爱思唯尔B.V.保留所有权利。
H-1-qNMR-spectroscopy was successfully applied to quantify the pharmacologically active alkaloid stachydrine ((2S)-1,1-dimethylpyrrolidinium-2-carboxylic-acid) in aerial parts of Leonurus japonicus (Leonuri herba, yimucao; Chin.Ph.2010, DAB2012) which are used in TCM and Kampo for the treatment of various gynaecological and cardiovascular disorders. Pharmacological publications on this betaine describe cardiovascular, hypotensive, and tissue-protective effects. However, its pharmacopeial analytics poses severe difficulties as it does not contain any chromophore suitable for HPLC UV-detection. Nine samples from three countries were prepared as decoctions and freeze-dried. H-1-NMR-spectra were recorded in D2O. The direct-quantitative H-1-qNMR-procedure was carried out using the N-CH3-singlet at delta 3.03 ppm in comparison to the delta 6.18 ppm singlet of the two vinylic protons of maleic-acid, which was identified as a most favourable internal standard. The quantification limit of stachydrine was 0.44 mg/g drug material. Neither reference-compounds for calibration-curves nor sample-pre-purification was necessary. This protocol revealed stachydrine contents in the range from 0.09 up to 1.01% (w/w) for the tested yimucao samples. Furthermore, between 0.18 and 0.21% of stachydrine was found in the L japonicus fruit-drug (Leonuri fructus, chongweizi; Chin.Ph.2010) which was examined for this constituent for the first time. In four co-investigated samples of the closely related and similarly used European herb Leonurus cardiaca Ph.Eur., even higher contents up to 1.55% were attested. The presented quantitative H-1-qNMR-method was shown to be precise with respect to concentration, and yielded highly reproducible data in a series of inter-day repetitions. Methodically, H-1-qNMR may be a powerful tool for quality assurance of stachydrine containing plants and herbal drugs, especially for industrial routine protocols. (C) 2014 Elsevier B.V. All rights reserved.