微小亚历山大藻中麻痹性贝类毒素的冻融法提取优化

Optimization of extraction of paralytic shellfish poisoning from Alexandrium minimum by melting-freezing method

为增加微小亚历山大藻的细胞破壁率,降低提取过程中毒素组成的改变,提高麻痹性贝类毒素(PSP)提取液的毒力,选用冻融法进行PSP提取并进行方法优化。首先对提取液p H、冷冻温度、解冻温度、冷冻时间和冻融次数等进行单因素试验,考察冻融过程中藻细胞破壁率和相对荧光值以确定优化范围;再以相对荧光值为响应值进行Box-Behnken中心组合响应面优化,得到最佳工艺条件为:冻融次数3.45,解冻温度36.99℃,p H7.66,该条件下预测相对荧光值为200.347,毒力为7 034 MU。验证试验设定藻破壁毒素提取条件为:冻融3次,解冻温度37℃,p H 7.8,该条件下相对荧光值为188.923(±10.96),提取液的毒力为6 616(±101)MU,与预测值基本相符。

To enhance the broken rate of Alexandrium minimum, to keep the composition unchanged and the toxicity of Paralytic Shellfish Poisoning （PSP）in the extraction process, the freezing-melting extraction method for PSP was used in this paper. Firstly, five single factor experiments were conducted to study the effects of pH, freezing temperature, melting temperature, freezing time and melting times on broken rate and relative fluorescence value, respectively. Then Box-Behnken response surface optimization was used to obtain the optimum conditions, with the relative fluorescence value as response value. The results showed that, the optimum conditions for extraction of PSP by melting-freezing method were as follows： pH 7.66, melting times of 3.45, at 36.99℃. Under these conditions, the re｜ative fluorescence value and virulence were 200.347, 7 034 MU, respectively. In the verified experiment,the conditions for PSP extraction from A. minimum were： pH 7.8, melting times of 3, at 37℃, under these conditions, the relative fluorescence value and virulence were 188.923 （ ± 10.96）, 6 616 （ ±101 ） MU, respectively.