高效液相色谱-离子阱质谱法测定海洋微藻藻粉中的8种脂溶性毒素

Determination of eight typical lipophilic algae toxins in marine microalgae powder using high-performance liquid chromatography-ion trap mass spectrometry

建立了利用高效液相色谱-电喷雾离子阱质谱(HPLC-ESI-IT-MS)测定海洋微藻藻粉中8种典型脂溶性毒素的分析方法。藻粉样品经超声细胞破碎后,采用超声波辅助提取法对藻毒素进行提取,用HPLC-ESI-IT-MS多反应离子监测(MRM)模式对各种毒素(包括大田软海绵酸(OA)、鳍藻毒素1(DTX-1)、扇贝毒素2(PTX-2)、虾夷扇贝毒素(YTX)、原多甲藻酸1(AZA1)原多甲藻酸2(AZA2)、罗环内酯毒素(SPX),米氏裸甲藻毒素(GYM))进行测定。8种脂溶性藻毒素均在线性范围内线性关系良好(R2均在0.991以上),检出限均介于0.085～1.315 pg之间,加标回收率在88.5%～111.4%之间,方法重复性相对标准偏差(RSD)在4.82%～10.17%范围。应用该方法对利玛原甲藻干藻粉中的毒素进行了测定,分析结果良好,说明本方法是海洋微藻藻粉中脂溶性藻毒素测定的有效方法。

A method involving high-performance liquid chromatography coupled with electrospray ionization–ion trap mass spectrometry（HPLC–ESI–IT–MS） was developed to determine eight typical lipophilic algae toxins（okadaic acid（OA）, dinophysistoxin-1（DTX-1）, pectenotoxin-2（PTX-2）, yessotoxin（YTX）, azaspiracid1, 2（AZA1, AZA2）, spirolidel1（SPX1）, gymnodimine（GYM）） in marine microalgae powder. The powder was extracted using ultrasound-assisted extraction after the algae cell-breaking process was performed using ultrasonication. The crude extracts were then analyzed using HPLC–ESI–IT–MS in the multiple reaction monitoring mode. The results showed that the calibration curve for each lipophilic algae toxin was obtained with a good correlation coefficient（R^20.991） in its linear concentration. The detection limits of the eight target toxins ranged from 0.85 to 1.315 pg. The average recoveries of these toxins ranged from 88.5% to 111.4% with relative standard deviations in the range of 4.82%–10.17%. The proposed method was applied to determine typical lipophilic algae toxins in marine microalgae powder, and good results were obtained. The results also indicated that this method is a useful tool for determining lipophilic algae toxins in marine microalgae powder.