高效液相色谱-电喷雾飞行时间质谱分析蟾酥有效成分

Rapid Quantification and Identification of Bioactive Components in Venenum Bufonis by HPLC-DAD/ESI-TOF-MS

目的采用高效液相色谱-高分辨电喷雾飞行时间质谱(HPLC-DAD/ESI-TOF-MS)联用技术,对蟾酥中的部分有效成分进行初步分析,探讨HPLC-DAD/ESI-TOF-MS联用技术作为蟾酥质量控制方法的可行性。方法采用Agilent C18色谱柱,以乙腈和水(含体积比0.1%醋酸)为流动相,采用梯度洗脱,流速为0.8mL·min-1,室温,DAD检测器检测波长为291nm,ESI-TOF-MS采用正离子模式。通过HPLC-ESI-TOF-MS在线测定部分化合物的精确分子量,结合文献报道,对蟾酥毒素进行鉴别。结果结合文献报道,在色谱峰当中对12个主要色谱峰进行鉴定。结论高效液相色谱-电喷雾飞行时间质谱联用技术是蟾酥质量控制的有效方法。

OBJECTIVE To identify the active components in Venenum Bufonis by high performance liquid chromatography （HPLC）- electrospray-time of flight mass spectrometry （ESI-TOF-MS）. METHODS The separation was performed on an Agilent Eclipse XDB C Is （4.6 mm×150mm, 5 μm） column with gradient elution of acetonitrile and water （with 0.1% acetic acid） as mobile phase, flow-rate was 0.8 mL·min^-1, with gradient elution at room temperature （about 25℃）. The detection wavelength was 291 nm and the UV spectra of peaks were obtained with a photodiode array detector （DAD）. On-line ESI-TOF-MS in positive mode was used to get the mass spectrum of some of the analytes. RESULTS 12 Effective components in Venenum Bufonis were primarily identified by comparison of the UV spectra and mass spectrum with literature and l0 were confirmed according to others＇ work results. CONCLUSION HPLC-ESI-TOF-MS is a very powerful tool for the identification of the effective components in Venenum Bufonis.