论文部分内容阅读
目的建立一种超灵敏、高效、准确并能同时测定双壳贝类中扇贝毒素和原多甲藻酸的固相萃取-高效液相色谱-串联质谱(SPE-HPLC/MS/MS)分析方法。方法样品经甲醇三次提取,提取液过Oasis HLB固相萃取小柱,所得滤液采用反相C18柱作为色谱分离柱,含甲酸和乙酸铵的乙腈水作为流动相,梯度洗脱方式分离待测化合物。优化了固相萃取的上样、淋洗及洗脱条件,考察了粗提取液及过固相萃取小柱提取液的基质效应。结果过固相萃取小柱提取液的基质抑制效应抵消了一部分因浓缩而提高的灵敏度。基质抑制效应不与SPE浓缩倍数成正比,浓缩倍数>2即可提高方法灵敏度。由此得到方法线性范围为0.86 ng/ml~55.2 ng/ml及0.128 ng/ml~8.2 ng/ml,检出限为0.013μg/kg~0.085μg/kg,精密度低于7%,回收率为99%~101%。结论本方法适用于双壳贝类中PTX2和AZA2的同时测定。
OBJECTIVE To establish a SPE-HPLC / MS / MS method for the simultaneous determination of scallop toxin and protocatechuic acid in bivalve molluscs by ultra-sensitive, efficient and accurate method. . Methods The sample was extracted three times with methanol and the extract was passed through the Oasis HLB cartridge. The filtrate was separated on a C18 reversed-phase column using a mobile phase of acetonitrile and water containing formic acid and ammonium acetate as the mobile phase. The analytes were separated by gradient elution . The sample loading, elution and elution conditions of solid phase extraction (SPE) were optimized. The matrix effects of crude extract and SPE solid phase extraction cartridges were investigated. Results The matrix inhibition effect of SPE-based cartridge offset some of the increased sensitivity due to concentration. The matrix inhibition effect is not proportional to the SPE concentration multiples, concentration ratio> 2 can increase the sensitivity of the method. The linear range of the method was 0.86 ng / ml ~ 55.2 ng / ml and 0.128 ng / ml ~ 8.2 ng / ml with the detection limit of 0.013 μg / kg ~ 0.085 μg / kg and the precision of less than 7% 99% ~ 101%. Conclusion This method is suitable for the simultaneous determination of PTX2 and AZA2 in bivalve molluscs.