免疫亲和柱净化-超高效亲水色谱-三重四极杆质谱联用法测定人尿液和血浆中河豚毒素
摘要
An ultra-performance hydrophilic interaction liquid chromatography-triple quadrupole mass spectrometric (UPLC-MS/MS) method was developed for the determination of tetrodotoxin (TTX) in human urine and plasma. After the sample was cleaned-up and concentrated by immunoaffinity column, the separation of the TTX was carried out on an Acquity UPLC BEH amide column (100 mm×2.1 mm, 1.7 μm) with gradient elution using mobile phases of 0.1% ( V/V ) formic acid in water and acetonitrile. The analyte was detected by positive electrospray ionization mass spectrometry in the multiple reaction monitoring (MRM) mode, and quantified by external solvent standard calibration. The measuring ranges of TTX in urine and plasma were 0.05-400 μg/L. The average recoveries were 92%-95% and 91%-96% for TTX respectively spiked in urine and plasma with relative standard deviations of 3.3%-7.2% and 3.9%-7.8% ( n =5). The limits of detection (LOD, S/N =3) and limits of quantitation (LOQ, S/N =10) of TTX were 0.02 μg/L and 0.05μg/L for urine and plasma, respectively. This method is suitable for the detection of TTX in urine and plasma for both forensic and clinical purposes.