微小亚历山大藻麻痹性贝类毒素小鼠生物检测

Mouse Bioassay for the Paralytic Shellfish Poisoning(PSP)from Alexadrium minutum

应用小鼠生物检测法对培养的微小亚历山大藻(LJX02株系)麻痹性贝类毒素进行了检测。首先建立微小亚历山大藻培养技术,利用对数期藻细胞接种,可使藻细胞处于快速生长,并在7d内藻细胞密度最高达到30000ml-1。从培养株系中提取的藻毒素,经液相质谱分析主要为GTX1/4和GTX2/3,其含量分别为12.23μg/ml和9.742μg/ml,12L藻体培养液中可获得18.35μgGTX1/4和14.61μgGTX2/3毒素量。采用小鼠生物检测法建立了藻毒素剂量-小鼠死亡时间曲线,其曲线方程为Y=74.017X-1.5896,将死亡时间t变为时间倒数1/t,GTX毒性MU取对数为logMU,得直线回归方程为Y=3.7009X-0.0858(R2=0.9824),求得了昆明小鼠一个鼠单位所代表的GTX1/4和GTX2/3毒素剂量分别为0.026μg和0.021μg。

In this paper,the extract and detection of the paralytic shellfish poisoning （PSP） from Alexadrium minutum were studied. Firstly,the Alexadrium minutum culture method was established. The cell growth rate kept higher in initial 8-10 days,and during that time the highest density of algae cells was up to 30 000ml^-1 The crude toxin was isolated from Alexadrium minutum and identified to be GTX2/3 and GTX1/4 with hquid chromatography- mass spectrometry （LC-MS）technology. The concentrations of GTX1/4 and GTX2/3 were about 0.02968μmol/ml and 0.02472μmol/ml respectively. And the total content reached up to 18.35μg（GTX1/4）and 14.61μg（GTX2/3）from 12L cultures respectively. With the mouse bioassay of the GTX2/3 and GTX1/4 toxicity,the relation between dosage and death time was established,and its curve equation was Y=74.017X^-1.5896. The curve could be transferred into a straight line when the GTX toxin dosage （MU）was represented by logarithm （logMU）and the death time （t） by reciprocal of death time （1/t）, thus the straight line equation is Y=3.7009X-0.0858 （R^2=0.9824）. According to the experiment,0.026μg （GTX1/4）and 0.021μg（GTX2/3） was obtained that was presented by.