血浆中河豚毒素的衍生气相色谱质谱分析  

Determination of tetrodotoxin in plasma using gas chromatography /mass spectrometry after derivatization

作　　者：柳洁[1] 丁文婕[2] 何碧英[1] 黄蔷[1] 罗兰[1] 

机构地区：[1]深圳市南山区疾病预防控制中心,广东深圳518054 [2]深圳职业技术学院,广东深圳518055

出　　处：《中国卫生检验杂志》2013年第14期2880-2882,共3页Chinese Journal of Health Laboratory Technology

基　　金：深圳市南山区卫生科技计划项目(2009039)

摘　　要：目的建立血浆样品中河豚毒素(TTX)的气相色谱-质谱分析方法。方法采用3%乙酸甲醇溶液提取,Oasis MCX固相柱净化,样品液中加入3 mol/L KOH溶液和甲醇、DMF将河豚毒素转化成C9碱,经Oasis HLB固相萃取柱净化、再用硅烷化衍生剂(BSTFA+TMCS)衍生,用气相色谱-质谱联用法进行定性定量分析鉴定。结果方法定性的特征离子为m/z318、m/z376、m/z392、m/z407,定量离子为m/z392;定量测定的线性范围在5 ng/ml^200 ng/ml,相关系数在0.9998,检出限为1.0 ng/ml,定量限为5.0 ng/ml,血浆样品加标回收率为82.2%~94.7%,相对标准偏差为7.6%~8.8%。结论建立的分析方法快速、准确、选择性好,可适用于公共卫生突发事件的应急检测和临床中毒病人的诊断确证。Objective To develop a highly selective and sensitive method for determination of tetrodotoxin （ TTX ） in human plasma by gas chromatography/mass spectrometry. Methods TTX was extracted from plasma samples with methanol containing 3% acetic acid and cleaned up through Oasis MCX cartridges. Then the compound was converted to 2 -amino -6 -hydroxymethyl -8 -hydroxyquinazoline（ C9 -base） under alkaline conditions and extracted with Oasis HLB extraction cartridge. The C9 - base was analyzed after BSTFA ＋ TMCS derivatization by gas ehromatography/mass spectrometry with SIM mode. Results The characteristic mass spectral peaks of TTX- C9 -TMS were observed at m/z318, m/z376, m/z392, m/z407 and quantitative ion was m/z392. The calibration curve for TTX - C9 - TMS was linear from 5 ng/ml to 200 ng/ml with the correlation coefficient of 0. 9998. The detection limit was 1.0 ng/ml and the limit of quantification was 5.0 ng/ml. The recovery rates and RSD of the TFX - C9 - TMS for spiked plasma samples were 82.2% - 94.7% and 7.6% - 8.8% , respectively. Conclusion The method was selective, reliable and accurate, so it is suitable for emergency detection of public health events and clinical diagnostic confirmation of poisoned cases.

关 键 词：河豚毒素 气相色谱-质谱 血浆 固相萃取 

分 类 号：O657.63[理学—分析化学]