Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Granada, Fuentenueva s/n, E-18071, Granada, Spain.
Unidad de Resonancia Magnetica Nuclear, Centro de Instrumentacion Cientifica, Universidad de Granada, Fuentenueva s/n, E-18071, Granada, Spain.
Phytochem Anal. 2019 Jan;30(1):89-94. doi: 10.1002/pca.2793. Epub 2018 Sep 14.
L-Dopa, a key neurotransmitter used to treat neural disorders such as Parkinson's disease, is found in the seeds of the genus Mucuna at a sufficient concentration for possible commercial use.
To develop a simple and reliable method to extract L-Dopa from M. pruriens seeds in an aqueous medium and then quantitate this compound using a H qNMR method (internal standard); and also to evaluate the accuracy and reproducibility of this method with an NMR calibration curve.
The extraction method of L-Dopa from M. pruriens was optimized. The quantitation with single point quantitative NMR (qNMR) and NMR calibration curve was based on the resonance properties of the main functional groups of the L-Dopa molecule, in particular the signals of the three aromatic protons, which were compared with the signal of an internal standard such as syringic acid. The accuracy (precision and trueness) and reproducibility of both NMR techniques were evaluated.
The methods of single point qNMR and NMR calibration curve, applied to the seeds of two M. pruriens varieties, gave very similar L-Dopa contents: 3.0-3.2% and 3.0-3.1%, respectively.
The statistical analysis confirmed the accuracy and reproducibility of this single point qNMR method (internal standard) for determining L-Dopa, as well as other commercial preparations of this species, without performing an NMR calibration curve.
L-多巴是一种重要的神经递质,用于治疗帕金森病等神经疾病,在黎豆属的种子中以足够高的浓度存在,可用于商业用途。
开发一种从黎豆种子中在水性介质中提取 L-多巴的简单可靠的方法,并使用 1 H qNMR 方法(内标)定量该化合物;并使用 NMR 校准曲线评估该方法的准确性和重现性。
优化了从黎豆中提取 L-多巴的方法。单点定量 NMR(qNMR)和 NMR 校准曲线的定量基于 L-多巴分子主要官能团的共振特性,特别是三个芳香质子的信号,与内标物如丁香酸的信号进行比较。评估了两种 NMR 技术的准确性(精密度和准确度)和重现性。
单点 qNMR 法和 NMR 校准曲线法应用于两种黎豆品种的种子,得到非常相似的 L-多巴含量:分别为 3.0-3.2%和 3.0-3.1%。
统计分析证实了单点 qNMR 方法(内标)测定 L-多巴的准确性和重现性,以及其他该物种的商业制剂,无需进行 NMR 校准曲线。