雪卡毒素的提取纯化方法初步研究

Preliminary Study on Extraction and Purification of Ciguatoxin

雪卡毒素(ciguatoxin)是一类具有神经毒性的海洋藻类毒素,高纯度雪卡毒素是开展其相关研究的物质基础。以石斑鱼肝脏为原料,对其脂溶性粗提物经Florisil柱和Sephadex LH-20柱纯化后,用高效液相色谱/质谱(HPLC/MS)和细胞毒性方法对其进行鉴定,证明为太平洋雪卡毒素(P-CTX-1)。HPLC鉴定分析提取物与太平洋雪卡毒素(P-CTX-1)具有相同的保留时间。HPLC/MS检测表明纯化样品在m/z=1 111.3和m/z=1 133.0处出现雪卡毒素[M+H]+峰和[M+Na]+峰,与已有的P-CTX-1分子离子峰完全吻合。四甲基偶氮唑蓝法(MTT)细胞毒性试验证明了雪卡毒素样品对人神经母细胞瘤(SK-N-SH)具有毒性作用,毒素剂量与细胞生长抑制率成线性关系。细胞毒性试验进一步确认该提取物为雪卡毒素,且纯度较高。

Ciguatoxin is a kind of neurotoxic marine algal toxins. However, ciguatoxin in high purity is the material foundation for related research. In this paper, ciguatoxin was extracted from Epinephelussp liver and purified by Florisil and Sephadex LH-20 chromatography. Furthermore, this ciguatoxin was identified as Pacific ciguatoxin （P-CTX-1） by high-performance liquid chromatography/mass spectrometry （HPLC/ MS） and cytotoxicity assay. This ciguatoxin has the same retention time comparing to the reference com- pound P-CTX-1 in HPLC. The mass spectra of this ciguatoxin showed the mass peaks of 1 111.3 （I-M＋ HI＋） and 1 133.0 （[-M＋Na]＋）, which was fully consistent with the reported mass peaks of P-CTX-1. The cytotoxicity of ciguatoxin on the human neuroblastoma cell（SK-N-SH ）was determined by MTT as- say, and the results indicated a linear dose-dependent inhibition effect of ciguatoxin on proliferation of the cells. The cytotoxicity assay further confirmed it as ciguatoxin in high purity.