Hua Y, Lu W, Henry M S, Pierce R H, Cole R B
Department of Chemistry, University of New Orleans, Louisiana 70148, USA.
Anal Chem. 1995 Jun 1;67(11):1815-23. doi: 10.1021/ac00107a010.
On-line high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESMS) has been successfully applied to the separation and identification of brevetoxins associated with "red tide" algae. Brevetoxins are toxic polyethers produced by the marine dinoflagellate Gymnodinium breve. They are responsible for fish kills, and they pose certain health risks to humans. The LC-MS method employs reversed-phase microbore HPLC on a C18 column with a mobile phase consisting of 85:15 methanol/water, a flow rate of 8 microL/min, and a postcolumn split ratio of 3:1 (UV absorbance detector/mass spectrometer). A brevetoxin culture sample was found to contain at least six components, including two well-separated peaks corresponding to the brevetoxins PbTx-2 and PbTx-1, as well as several unknown compounds, including one with a molecular mass of 899 Da (possibly an isomer of PbTx-9). The brevetoxin molecules exhibited a high tendency to bind to alkali cations in positive ion ESMS. For standard PbTx-9, PbTx-2, and PbTx-1 brevetoxins analyzed on our LC-MS system, the detection limits (employing mass spectrometer scans of 100 m/z units) were determined to be less than 600 fmol, 1 pmol, and 50 fmol, respectively (S/N = 3); the total analysis time was about 35 min.
在线高效液相色谱 - 电喷雾电离质谱法（HPLC - ESMS）已成功应用于与“赤潮”藻类相关的短裸甲藻毒素的分离和鉴定。短裸甲藻毒素是由海洋双鞭毛藻短裸甲藻产生的有毒聚醚。它们导致鱼类死亡，并对人类构成一定的健康风险。该液相色谱 - 质谱方法采用反相微孔HPLC，使用C18柱，流动相为85:15甲醇/水，流速为8微升/分钟，柱后分流比为3:1（紫外吸收检测器/质谱仪）。发现一个短裸甲藻毒素培养样品至少含有六种成分，包括两个分离良好的峰，分别对应短裸甲藻毒素PbTx - 2和PbTx - 1，以及几种未知化合物，其中一种分子量为899 Da（可能是PbTx - 9的异构体）。在正离子ESMS中，短裸甲藻毒素分子表现出与碱金属阳离子结合的高度倾向。对于在我们的液相色谱 - 质谱系统上分析的标准PbTx - 9、PbTx - 2和PbTx - 1短裸甲藻毒素，检测限（采用100 m/z单位的质谱仪扫描）分别确定为小于600 fmol、1 pmol和50 fmol（信噪比 = 3）；总分析时间约为35分钟。
