狭叶柴胡愈伤组织和悬浮细胞成分的UPLC/Q-TOF-MS分析

Analysis on compounds of callus and suspension cell of Bupleurum scorzonerifolium Willd.by UPLC/Q-TOF-MS

以柴胡皂苷d(SSd)为检测指标,基于超高效液相色谱联用四级杆串联飞行时间质谱仪(UPLC/Q-TOF-MS),建立狭叶柴胡愈伤组织及悬浮细胞成分定性分析的方法。采用C18色谱柱(50mm×2.1mm,1.7μm),以0.1%甲酸-乙腈为流动相梯度洗脱,使用ESI离子源,正离子模式下采集数据,结合对照品SSd谱图,分析相关文献数据对照。结果表明:在UPLC/Q-TOF-MS检测下,狭叶柴胡悬浮细胞、细胞发酵液及愈伤组织在洗脱时间为11.22、11.19及11.17min时均与标准品SSd(11.12min)有相同的裂解碎片,愈伤组织及悬浮细胞中均含有活性成分SSd。相比传统的高效液相检测方法,该方法快速、准确且灵敏度好,适用于柴胡组织培养物中活性成分的检测和分析。

To establish the qualitative analysis method using UPLC/Q-TOF-MS for bioactive compound of saikosaponin d （SSd） in callus and suspension cell of Bupleurum scorzonerifolium Willd.. C18（50 mm×2.1 mm, 1.7 μm） column was adopted under the ESI positive mode eluting gradiently with 0.1% formic acid -methyl cyanide. The data were collected and comparatively analyzed with references saikosaponin d and relevant literatures. The results showed that suspension cell, cell supernatant and callus had the same fragments as SSd （11.12 min） at 11.22, 11.19 and 11.17 min, respectively. This indicated that callus and suspension cell can produce SSd. Compared with traditional HPLC method, UPLC/Q-TOF-MS method was quick, precise and highly sensitive, which could be suitable for detection and analysis of bioactive compounds of tissue cultures of B. scorzonerifolium Willd.