摘要
软骨藻酸(Domoic acid, DA)是由海洋硅藻产生的一种可致人记忆缺失的亲水性毒素,对海产养殖业以及人类健康造成严重威胁。本研究建立了一种高效富集海水中微量DA的新方法,结合高效液相色谱-紫外检测/质谱分析实现了养殖海水中DA的检测。利用磺化苯乙烯-乙烯基苯共聚物材质的固相萃取膜盘,对经甲酸酸化处理的海水样品中的DA进行富集、净化;采用反相C_(18)色谱柱分离DA,以含2.0 mmol/L乙酸铵和0.1%(V/V)甲酸的90.0%(V/V)乙腈为流动相,紫外检测波长为242 nm。在最佳实验条件下,对海水中DA的富集倍数可达2000倍,平均回收率89.3%,具有良好的精密度(相对标准偏差(RSD)≤4.0%);海水中DA的检出限(S/N=3)为2.5 ng/L,使高效液相色谱-紫外检测能满足实际海水中DA测定的灵敏度要求。采用本方法对中国东海部分海域海水中的DA进行了检测,在两个站位海水样品中检出DA,含量最高可达2.7μg/L。本方法操作简单、灵敏度高,是近海养殖海水环境中DA测定的有效方法。
Domoic acid(DA), a type of hydrophilic toxin produced by marine diatoms, can cause memory loss, which poses a serious threat to marine culture and even human health. In this study, a new method was developed for the highly efficient accumulation and detection of trace DA in seawater by high performance liquid chromatography-ultraviolet detection/mass spectrometry(HPLC-UV/MS). In this method, a solid phase extraction disk made of sulfonated styrene-vinyl benzene copolymer was used for DA enrichment and purification from acidified seawater samples. A C_(18) chromatographic column was employed for DA separation using acetonitrile-water solution(90%, V/V) containing 2 mmol/L ammonium acetate and 0.1%(V/V) formic acid as mobile phase. Detection wavelength was set at 242 nm. Under optimal conditions, the average recovery could still maintain at 89.3% with good precision(relative standard deviation(RSD) ≤4.0%) while enrichment factor could reach up to 2000. The limit of detection(S/N=3) was 2.5 ng/L. The results showed that the HPLC-UV detection developed here could meet the sensitivity requirements of DA determination in real seawater samples. This method was used to determine the seawater samples obtained from East China Sea, and it was found that the DA content in two station seawater samples was up to 2.7 μg/L. This method was efficient in determination of the concentration of DA in coastal aquaculture seawater, with easy-operation and high sensitivity.
引文
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