UPLC-MS/MS同时测定雷公藤悬浮细胞中8种萜类成分

Simultaneous detemination of 8 terpenoids compounds in tripterygium wilfordii suspension cells by UPLC-MS/MS

目的建立超高效液相色谱-三重四级杆串联质谱(ultra-high performance liquid chromatography triple quadrupole tandem mass spectrometry, UPLC-QQQ-MS/MS)同时测定雷公藤悬浮细胞中8种萜类成分的定量分析方法。方法采用Waters HSS T3 C18色谱柱,以0.1%甲酸水-乙腈为流动相,梯度洗脱,流速0.3 ml/min,柱温30 ℃。采用电离子喷雾源,选择正负离子交替扫描多反应离子检测模式。结果雷公藤甲素、雷公藤内酯酮、去氢松香酸、雷酚内酯、去甲泽拉木醛、雷公藤褔啶、扁塑藤素和雷公藤红素8种萜类成分的检测限和定量限范围分别为0.009 6~0.248 0 ng和0.019 2~0.496 0 ng,线性关系良好,线性相关系数r在0.996 6~0.999 3之间,平均加样回收率为95.04%~105.20%,RSD为2.14%~6.31%。结论该定量分析方法简便、快捷、灵敏度强、准确性高,可同时定量测定雷公藤悬浮细胞中8种萜类成分的含量,为雷公藤及其植物组织培养物的研究和质量控制提供方法学依据。

Objective To establish a UPLC-QQQ-MS/MS (ultra-high performance liquid chromatography triple quadrupole tandem mass Spectrometry) method for simultaneous detemination of 8 terpenoids compounds in Tripterygium wilfordii suspension cells. Methods The separation and analysis were performed on Water HSS T3 C18, with 0. 1% acetic acid solution-acetonitrile as the mobile phase for gradient elution. The column temperature was set at 30 ℃ with flow rate was 0.3 ml/min. The electrospray ionization (ESI) source was applied and operated in alternating positive and negative mode. Multi-reaction monitor (MRM) mode was used to quantify the 8 compounds. Results The limits of detection and limits of quantitation in 8 compounds (triptolide, triptonide, dehydroabietlc acid, triptophenolide, demethylzeylasteral, tripteridine, pristimerin, celastrol) were 0.009 6-0.248 0 ng and 0.019 2-0.496 0 ng, respectively. The correlation coefficents (r) of calibration curves were 0.996 6-0.999 3. The average recovery rates were 95.04%-105.20%, and the relative standard deviation was 2.14%-6.31%. Conclusions The established quantitative method is simple, rapid, sensitive and accuracy. It can be used for simultaneous analysis of 8 terpenoids compounds in Tripterygium wilfordii suspension cells by using UPLC-QQQ-MS/MS, which affords methodology evidence for research and quality control of Triptervgium wilfordii Hook. f. and its plant tissue cultures.