Hunan Engineering Technology Research Center of Veterinary Drugs, Hunan Agricultural University, Furong District, Changsha, 410128, Hunan, China.
College of Veterinary Medicine, Hunan Agricultural University, Furong District, Changsha, 410128, Hunan, China.
Anal Bioanal Chem. 2019 Jul;411(17):3857-3870. doi: 10.1007/s00216-019-01859-2. Epub 2019 May 9.
The present paper describes a novel two-dimensional liquid chromatography (2D-LC) system, which is comprised of a first-dimensional ion exchange chromatography (IEX1) column, trap column, and second-dimensional reversed-phase chromatography (RP2) column system. The biological sample is separated by the first-dimensional LC using an IEX column to remove interferences. The analytes are transferred to the trap column after heart-cutting. Then, the analytes are transferred to the second-dimensional LC using an RP2 column for further separation and ultraviolet detection. This 2D-LC system can offer a large injection volume to provide sufficient sensitivity and exhibits a strong capacity for removing interferences. Here, the determination of three monoterpene indole alkaloids (MIAs; gelsemine, koumine, and humantenmine) from Gelsemium in biological matrices (plasma, tissue, and urine) was used this 2D-LC system. After a rapid and easy sample preparation method based on protein precipitation, the sample was injected into the 2D-LC. The method was developed and validated in terms of the selectivity, LOD, LOQ, linearity, precision, accuracy, and stability. The sample preparation time for the three MIAs was 15 min. The LOD for these compounds was 10 ng/mL, which was lower than the developed HPLC methods. The results showed that this method had good quantitation performance and allowed the determination of gelsemine, koumine, and humantenmine in biological matrices. The method is rapid, exhibits high selectivity, has good sensitivity, and is low-cost, thus making it well-suited for application in the pharmaceutical and toxicological analysis of Gelsemium. Graphical abstract.
本文描述了一种新型二维液相色谱（2D-LC）系统，该系统由第一维离子交换色谱（IEX1）柱、捕集柱和第二维反相色谱（RP2）柱系统组成。生物样品通过第一维 LC 采用 IEX 柱分离以去除干扰。心切后将分析物转移到捕集柱上。然后，采用 RP2 柱将分析物转移到二维 LC 中进行进一步分离和紫外检测。该 2D-LC 系统可以提供大的进样体积以提供足够的灵敏度，并具有很强的去除干扰的能力。在此，使用该 2D-LC 系统从钩藤中生物基质（血浆、组织和尿液）中测定三种单萜吲哚生物碱（MIAs；钩藤碱、钩藤宁和胡蔓藤碱）。在基于蛋白质沉淀的快速简便的样品制备方法之后，将样品注入 2D-LC。该方法在选择性、LOD、LOQ、线性、精密度、准确度和稳定性方面进行了开发和验证。三种 MIAs 的样品制备时间为 15 分钟。这些化合物的 LOD 为 10ng/mL，低于已开发的 HPLC 方法。结果表明，该方法具有良好的定量性能，可用于生物基质中钩藤碱、钩藤宁和胡蔓藤碱的测定。该方法快速、选择性好、灵敏度高、成本低，因此非常适合在钩藤的药物和毒理学分析中应用。
