剧毒卡罗藻GM5株中标志性毒素4,5-dihydro-KmTx2的色谱制备

建立了剧毒卡罗藻GM5株中标志性毒素4,5-dihydro-KmTx2的分离纯化体系, 通过高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q-Orbitrap HRMS)对其纯化效果进行评价. 过滤后的培养液经大型固相萃取柱(YMC-GEL ODS-A-HG, 80 cm×10 cm, 12 nm, S-50 μm)进行富集, 依次用体积分数为10%、40%和60%的甲醇溶液进行淋洗, 体积分数为80%的甲醇溶液进行洗脱, 得到目标化合物粗提液(纯度为51.3％). 采用半制备型高效液相色谱技术进一步分离纯化, 使得目标化合物纯度高达93.2％. 该方法提取及纯化效率高, 可操作性强, 可以为4,5-dihydro- KmTx2的规模化生产及功能活性研究提供技术支持.

Chromatographic preparation of 4,5-dihydro-KmTx2 from Karlodinium veneficum GM5 strain

A separation and purification system for the toxin 4,5-dihydro-KmTx2 in Karlodinium veneficum GM5 strain at the end of the platform was established. The purification effect of the system was evaluated by high-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectro- metry (UHPLC-Q-Orbitrap HRMS). First, the collected culture solution following the removal of the algae was enriched on a large solid-phase extraction column (YMC-GEL ODS-A-HG, 10 cm×80 cm, 12 nm, S-50 μm). The results showed that leaching was accomplished in sequence by using 10%, 40％ and 60％ methanol solution. The purity of the crude extract obtained by elution with 80％ methanol was 51.3％. The crude extract was separated and purified by semi-preparative HPLC. The purity of the 4,5-dihydro-KmTx2 monomer was 93.2％ with gradient elution using a methanol-water mixture as the mobile phase. The method has high extraction efficiency and operability, which can provide technical support for large-scale production and functional activity research of 4,5-dihydro-KmTx2.