固相萃取-气质联用结合同位素峰形校正检索技术分析钩吻中的钩吻碱和钩吻碱子

目的:建立固相萃取-气质联用并结合同位素峰形检索技术(SPE-GC/MS-CLIPS)分析钩吻中的钩吻碱和钩吻碱子的方法。方法:样品采用1%盐酸超声提取,经PCX固相萃取小柱净化,再用氨化甲醇进行洗脱,洗脱液浓缩后用丙酮定容。供试液通过气相色谱质谱联用仪结合毛细管柱(HP-5MS0.25 mm×30 m×0.25μm),用全扫描模式(m/z 40~500)采集数据后,利用同位素-C峰形检索技术对目标峰进行校正分析。结果:根据钩吻植物的生物碱提取物所得到的质谱谱图,通过与NIST标准库的匹配与同位素峰形检索技术推算的分子式,确定了钩吻中的钩吻碱和钩吻碱子。该方法成功应用于钩吻中毒司法鉴定案例2例,共10批样品。结论:固相萃取-气质联用结合同位素峰形校正检索技术可以准确分析钩吻中的钩吻碱和钩吻碱子,能应用于钩吻的植物鉴定和钩吻中毒司法鉴定工作。

Objective:To establish a method for the analysis of gelsemine and koumine in Gelsemium elegans by solid phase extraction(SPE)-gas chromatography-mass spectrometry(GC/MS)-calibrated lineshape isotope profile search(CLIPS). Methods:The crude sample was extracted by 1% hydrochloric acid with ultrasound-assisted extraction,then purified by PCX-SPE column,and eluted with ammoria-methanol(5:95). After concentration, the residue was dissolved by acetone solution and analyzed by GC/MS. With the conditions of capillary chromatographic column(HP-5MS 0.25 mm×30 m×0.25μm), SCAN acquired mode from m/z 40-500. The target peaks were calibrated by CLIPS. Results:By combining analysis of the mass spectrogram matching with NIST mass spectrogram database and molecular structure prediction with CLIPS, the gelsemine and koumine contained in Gelsemium elegans were identified. The method has been applied successfully for 2 suspected Gelsemium elegans poisoning cases, with 10 samples involved. Conclusion:This method can be used to determine the gelsemine and koumine in Gelsemium elegans, which can be applied in plant identification and poisoning judicial authentication of Gelsemium elegans.