A simple, rapid, cost-effective and green analytical method is developed based on ultrasound-assisted dispersive liquid-liquid microextraction (US-DLLME) coupled to thin-layer chromatography (TLC)-image analysis for the simultaneous determination of two major alkaloids of Strychnos nux-vomica L i.e., strychnine and brucine. The method is composed of three steps, namely (i) US-DLLME by injecting a mixture of 100-μL chloroform (extraction solvent) and 1-mL methanol (disperser solvent) in 5 mL of aqueous sample, followed by ultrasonication and centrifugation, (ii) TLC of 20 μL of sedimented phase with methanol: ammonia (100:1.5, v/v) as the mobile phase and visualization under ultraviolet radiation (254 nm) and (iii) photography of TLC plate and quantification of spots by image analysis using freely available imageJ software (National Institute of Health, Bethesda, MD, USA). The limit of detection and limit of quantification for both alkaloids were found to be in the range of 0.12-0.15 and 0.36-0.48 μg/spot, respectively. The method was found to be linear in the range of 0.5-5 μg/spot with correlation coefficient (R2) of 0.995 and 0.997 for strychnine and brucine, respectively. The developed method was successfully applied for the determination of strychnine and brucine in Ayurvedic formulations and blood samples. The method does not require any sophisticated instrument and handling skills and can be adopted for rapid analysis of strychnine and brucine in forensic toxicological laboratories.

中文翻译：

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一种基于超声辅助分散液-液微萃取-薄层色谱-图像分析的同时测定血液和阿育吠陀药品中马兜铃生物碱的简单，经济高效的方法。

基于超声辅助分散液-液微萃取（US-DLLME）结合薄层色谱（TLC）图像分析开发了一种简单，快速，经济高效的绿色分析方法，用于同时测定两种主要生物碱。马兜铃属马钱子碱和马钱子碱。该方法包括三个步骤，即（i）US-DLLME：在5 mL水性样品中注入100μL氯仿（萃取溶剂）和1-mL甲醇（分散剂）的混合物，然后进行超声和离心分离， （ii）用甲醇：氨水（100：1.5，v / v）作为流动相，并在紫外辐射（254 nm）下可视化，以及（iii）TLC板照相和使用免费提供的imageJ软件（美国马里兰州贝塞斯达卫生研究院，美国国立卫生研究院）通过图像分析对斑点进行定量。两种生物碱的检出限和定量限分别在0.12-0.15和0.36-0.48μg/点的范围内。发现该方法在0.5-5μg/ spot范围内是线性的，对于士的宁和布鲁辛的相关系数（R2）分别为0.995和0.997。所开发的方法已成功地用于阿育吠陀制剂和血液样品中甲苯丙氨酸和马钱子碱的测定。