摘要
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 A ultra - sensitive, high - performance and precise method for simultaneous determination of azaspiracid -2 （AZA2） and peetenotoxins （PTX） in bivalves was established by high performance liquid chromatography tandem mass spectrometry （HPLC/MS/MS） based on solid phase extraction （SPE）. Methods After three - time extraction by methanol, the sample extract was loaded on an Oasis HLB SPE column. Separation of the target compound was achieved on a reversed - phase CIs column with aeetonitrile - water containing formic acid and ammonium acetate as mobile phase for gradient elution. The sample loading, washing and elution conditions of SPE process were systematically optimized. Matrix effects of post - SPE extract and crude extract were seriously evaluated. Results Increased suppressive matrix effects in post - SPE extract counter- vailed the concentration purpose to some extent, but the suppression degree is not proportional to the concentration multiple. The contrast results indicated that a concentration multiple higher than 2 was helpful to improve the method sensitivity. The linear ranges of the target compounds were 0. 86 ng/ml - 55.2 ng/ml and 0. 128 ng/ml - 8.2 ng/ml. The LODs of the SPE - HPLC/ MS/MS method were in the range of 0.013 μg/kg ~0.085 μg/kg. The recoveries of fortified samples at three different concen- trations were 99% ~ 101%, and the preeisions were lower than 7%. Conclusion The SPE - HPLC/MS/MS method can be applied to determine PTX and AZA2 in bivalve molluscs simultaneously.
出处
《中国卫生检验杂志》
北大核心
2014年第16期2316-2320,共5页
Chinese Journal of Health Laboratory Technology
关键词
固相萃取
高效液相色谱串联质谱
扇贝毒素
原多甲藻酸
双壳类
Solid - phase extraction
High performance liquid chromatography tandem mass spectrometry
Pectenotoxins
Aza-spiracid - 2
Bivalves
