基于UPLC/Q-TOF-MS/MS分析鳖血柴胡皂苷类化学成分

Analysis of the Chemical Constituents of Bupleurum Saponins in Bupleuri radix with Turtle Blood by UPLC/Q-TOF-MS/MS

以鳖血柴胡(Bupleuri radix with turtle blood)炮制品为原料,基于超高效液相色谱-飞行时间质谱(UPLC/Q-TOF-MS/MS)分析手段,建立皂苷类成分快速分析方法。采用Waters ACQUITY UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7μm),以0.01%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,体积流量0.3 mL/min,进样量2μL,柱温40℃,ESI采用负离子扫描模式采集数据。结果表明:利用已建立的筛查数据库将满足质量误差小于5×10-^6、同位素分布正确且含有二级质谱裂解碎片的离子作为分析目标化合物,结合软件Formula Finder,Mass Calculators等功能、在线数据库(Human Metabolome Database、Pub Chem、Mass Bank、ChemSpider等)及二级碎片裂解规律,从鳖血柴胡中共分离和鉴定出44个皂苷类成分,主要为环氧醚型、异环双烯型、12-烯型、12-烯-28-羧酸型柴胡皂苷。综上分析,UPLC/Q-TOF-MS/MS方法能快捷、准确、较全面地鉴定鳖血柴胡中皂苷类成分,为鳖血柴胡的质量评价指标选择和药效物质基础的研究提供可行方法。

A rapid method for the analysis of saponins based on UPLC/Q-TOF-MS/MS was established by using Bupleuri radix with Turtle Blood processed products as raw materials.The analysis was performed on an Acquity UPLC BEH C18 column(100 mm×2.1 mm,1.7μm)by gradient elution.The mobile phase consisted of 0.01%formic acid solution(A)-acetonitrile(B)at a flow rate of 0.3 mL/min,and the injection volume was 2μL,the column temperature was at 40℃.The mass spectrometer was operated in negative mode.Results showed that the compounds with a mass error of less than 5×10^-6,a correct isotope distribution and fragment ion were used as the target compounds,using the established screening database.Combined with software Formula Finder,Mass Calculators and other functions,online databases(Human Metabolome Database,PubChem,MassBank,ChemSpider,etc.),and two stage fragmentation rule,a total of 44 saponins were separated and identified in Bupleuri radix with Turtle Blood,including epoxy ether type,isophthalene type,12-ene-type and 12-ene-28-carboxylic acid type saikosaponins.In conclusion,the results demonstrate that UPLC/Q-TOF-MS/MS method is quick,accurate,and efficient for the identification of saponins in Bupleuri radix with Turtle Blood,and provides a feasible method for the selection of quality control indicators and the efficacy of active compounds in Bupleuri radix with Turtle Blood.