Ingkaninan K, de Best C M, van der Heijden R, Hofte A J, Karabatak B, Irth H, Tjaden U R, van der Greef J, Verpoorte R
Division of Pharmacognosy, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, Leiden University, The Netherlands.
J Chromatogr A. 2000 Mar 3;872(1-2):61-73. doi: 10.1016/s0021-9673(99)01292-3.
A high-performance liquid chromatography (HPLC) method with on-line coupled ultraviolet (UV), mass spectrometry (MS) and biochemical detection for acetylcholinesterase (AChE) inhibitory activity has been developed. By combining the separation power of HPLC, the high selectivity of biochemical detection, and the ability to provide molecular mass and structural information of MS, AChE inhibitors can be rapidly identified. The biochemical detection was based on a colorimetric method using Ellman's reagent. The detection limit of galanthamine, an AChE inhibitor, in the HPLC-biochemical detection is 0.3 nmol. The three detector lines used, i.e., UV, MS and Vis for the biochemical detection were recorded simultaneously and the delay times of the peaks obtained were found to be consistent. This on-line post-column detection technique can be used for the identification of AChE inhibitors in plant extracts and other complex mixtures such as combinatorial libraries.
已开发出一种用于乙酰胆碱酯酶（AChE）抑制活性的高效液相色谱（HPLC）方法，该方法在线联用紫外（UV）、质谱（MS）和生化检测。通过结合HPLC的分离能力、生化检测的高选择性以及提供MS分子量和结构信息的能力，可以快速鉴定AChE抑制剂。生化检测基于使用埃尔曼试剂的比色法。在HPLC - 生化检测中，AChE抑制剂加兰他敏的检测限为0.3 nmol。同时记录使用的三条检测线，即用于生化检测的UV、MS和可见光谱，发现所得峰的延迟时间一致。这种在线柱后检测技术可用于鉴定植物提取物和其他复杂混合物（如组合文库）中的AChE抑制剂。
