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用 ERETIC ¹H NMR 法测定黄连根茎中的原小檗碱型生物碱。

Determination of protoberberine alkaloids in Rhizoma Coptidis by ERETIC ¹H NMR method.

机构信息

Department of Chemistry, Manufacturing, and Controls (CMC), Pharma Research & Early Development (pRED) China, F. Hoffmann-La Roche Ltd., Shanghai 201203, PR China.

出版信息

J Pharm Biomed Anal. 2012 Feb 23;60:44-50. doi: 10.1016/j.jpba.2011.10.030. Epub 2011 Nov 4.

Abstract

An alternative quantification approach called ERETIC (Electronic REference To access In vivo Concentrations) utilizing electronic reference-based proton nuclear magnetic resonance (¹H NMR) spectroscopy techniques has been successfully introduced in our present study to simultaneously determine the contents of five major active protoberberine alkaloids (berberine, coptisine, jatrorrhizine, palmatine and epiberberine) in Rhizoma Coptidis, one of the most commonly used traditional Chinese medicines (TCM). The NMR experimental conditions including deuterated solvent, relaxation delay time, and ERETIC transmitter power level were optimized, and the developed method was validated in terms of precision, reproducibility, stability, accuracy, recovery, and limit of quantification (LOQ). The recoveries of the five tested alkaloids ranged between 89.94 and 97.72% for berberine, 90.87 and 100.05% for coptisine, 98.35 and 107.57% for jatrorrhizine, 95.37 and 101.26% for palmatine, and 93.18 and 98.00% for epiberberine, respectively, and LOQ was 0.1 mM for berberine. The high universality, accuracy, reproducibility, and efficiency of the ERETIC method demonstrated in this work suggest that this method could serve as a highly potential quantification alternative for quality assessment of TCM.

摘要

本研究成功引入了一种称为 ERETIC(电子参考以获取体内浓度)的替代定量方法,该方法利用基于电子参考的质子核磁共振(¹H NMR)光谱技术,可同时测定黄连中五种主要活性原小檗碱类生物碱(小檗碱、黄连碱、药根碱、巴马汀和表小檗碱)的含量,黄连是最常用的中药之一。对 NMR 实验条件(氘代溶剂、弛豫延迟时间和 ERETIC 发射器功率水平)进行了优化,并从精密度、重现性、稳定性、准确度、回收率和定量限(LOQ)等方面对所建立的方法进行了验证。五种测试生物碱的回收率范围为小檗碱 89.94%至 97.72%,黄连碱 90.87%至 100.05%,药根碱 98.35%至 107.57%,巴马汀 95.37%至 101.26%,表小檗碱 93.18%至 98.00%,小檗碱的 LOQ 为 0.1mM。本工作中 ERETIC 方法表现出的高通用性、准确性、重现性和高效性表明,该方法可作为中药质量评估的一种有潜力的定量替代方法。

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