摘要
目的：建立高效液相色谱法检测织纹螺中毒素的方法。方法：取织纹螺肉20g，用60ml含1％醋酸的甲醇溶液分3次抽提，离心分离合并上清液，在40℃真空旋转蒸发器中浓缩至近干，加蒸馏水溶解，三氯甲烷脱脂，水层经过活性炭柱吸附，用1％醋酸乙醇洗脱，再经过Bio-Gal P-2（2×98cm）柱中精制，精制液用Bio-Rex70（1×100cm）酸性凝胶柱控制流量在0．5ml／min下采用线性为0～0．03mol／L甲酸进行分割，收集的分割液经冷冻干燥，用1％醋酸定容，在高效液相色谱仪的反相色谱柱RP-18分离，荧光检测器定量检测。结果：本方法测定TTX的RSD为8．75％，平均样品加标回收率为82．5％，最低检出限2．1μg；anh-TTX的RSD为5．25％，平均样品加标回收率为90．1％，最低检出限12．5μg；GTX的RSD为7．23％，平均样品加标回收率为83．0％，最低检出限1．7μg；STX的RSD为5．93％，平均样品加标回收率为101．3％，最低检出限4．2μg。结论：（1）采用活性炭吸附，经Bio-Gal P-2（2×98cm）柱和Bio-Rex70（1×100cm）柱处理的样品，在液相色谱检测时，可以有效消除来自样品基体背景的严重干扰，使样品待测组分分析图谱与加标方法制备的标准分析图谱具有同样的抗干扰性。（2）分别采用两种流动相可以准确分析TTX、anh-TTX、GTX和STX毒素。（3）使用改制的柱后衍生系统能够方便、价廉的满足检测工作需要。解决了样品前处理中有效排除样品基体干扰物质的问题，使净化的待测定毒素成分能够采用高效液相色谱仪的反相色谱柱定量检测。
Objective：To establish a method to determine the toxins in nassarius Sp with high performance liquid chromatography. Methods：Twenty grams of nassarius Sp meat was extracted with 60 ml of acetic acid：methanol solution （1：99）. The mixture was centrifuged and supernatant was concentrated almost dry in a rotary vacuum evaporator at 40℃. The residual materials was solved in distilled water and degreased with trichloroethylene. The aqueous layer was adsorbed with activated charcoal and washed with acetic acid aqueous：ethanol solution（1：99） and refined in Bio -Gal P -2（2 - 98 cm） columns. The refined solution was partitioned in acid gel columns at flow speed of 0. 5 ml/min with formic acid at a linear of 0 - 0.03 mol/L. The collected partitioned solution was freeze -dried and solved in 1% acetic acid aqueous and separated with reversed phase columns RP - 18 in HPLC. The concentration was determined with fluorescence detector. Results： The RSD, average recovering rate of standard sample and detection limit for TTX are 8.75%, 82： 5% and 2. 1 μg, respectively. The results arze 5.25%, 90. 1% and 12.5 μg for anh -TTX, 4. 23%, 83.0% and 1.7 μg for GTX, and 5.93%, 101.3% and 4. 2 μg for STX, respectively. Conclusion： （1） The noise in matrix background is reduced in samples adsorbed with activated charcoal and treated with columns of Bio -Gal P- 2（2 - 98 cm） and Bio -Rex 70（ 1 - 100 cm）. Anti -interference can be achieved in spectra of both samples and standards. （2） Accurate determination results of TTX, anh - TTX, GTX and STX can be got with two kinds of mobile phase respectively. （ 3 ） Modified post - column derivation can facilitate the detection at low price and make it possible to determine toxins with reversed phase columns in HPLC.
出处
《中国卫生检验杂志》
CAS
2008年第7期1341-1343,共3页
Chinese Journal of Health Laboratory Technology
基金
宁波市医学科技项目(2004057)
关键词
织纹螺
毒素
测定
Nassarius SP
Toxins
Determination
