丹参水提取物成分的定性与半定量研究

A qualitative and semi-quantitative study of Salvia miltiorrhiza aqueous extracts

目的利用Q-TOF-MS技术对丹参水提取物中的成分进行定性研究,并且利用Q-Trap-MS技术对部分未知成分进行半定量研究。方法采用Waters Acquity UPLC HSS T3(100 mm×2.1 mm,1.8μm)色谱柱,以乙腈-0.1%甲酸水体系梯度洗脱,体积流量0.3 mL/min,柱温30℃。Q-TOF-MS检测采用负离子ESI模式,电压2.5 kV,离子源温度110℃,雾化气温度350℃,雾化气流量800 L/h。Q-Trap-MS检测采用负离子ESI模式,离子喷射电压:-4 500 V;温度450℃;源内气体1(GS1,N2)压力310 kPa;源内气体2(GS2,N2)压力310 kPa;气帘气体(N2)压力138 kPa;应用多重反应监测(MRM)模式进行半定量分析。结果鉴定出丹参水提取物中成分26个,其中有15个为常见的酚酸类物质,另有11个含量较低且研究较少的已知物质;在Q-Trap-MS中进行了方法学的考察,并测定出了11种未知成分的含量。结论实现了对丹参水提取物高效快速的物质定性,同时也为无对照品且含量较低的成分的含量测定提供了新的思路。

Objective To qualitative research the composition of Salvia miltiorrhiza aqueous extracts by using Q-TOF-MS and to establish a semi-quantitative analysis method of some active components in S.miltiorrhiza aqueous extracts based on Q-Trap-MS.Methods The chromatographic separation was performed on a Waters Acquity UPLC HSS T3(100 mm×2.1 mm,1.8μm)column with the mobile phase consisting of acetonitrile and 0.1%formic acid for gradient elution.The flow rate was 0.3 mL/min,and the column temperature was 30℃.In Q-TOF-MS,electrospray ionization(ESI)source was applied and operated in negative ion mode.The capillary voltage was set at 2.5 kV.The nebulization gas was set to 800 L/h at 400℃,and the source temperature was 100℃.In Q-Trap-MS,electrospray ionization(ESI)source was applied and operated in negative ion mode.The ionspray voltage was set to-4 500 V at 450℃.The pressure of source gas 1(GS1,N2)and source gas 2(GS2,N2)were both 45 psi,and the pressure of curtain gas(N2)was 20 psi.Semi-quantification analysis was performed with multiple reaction monitoring(MRM)mode.Results A total of 26 components of S.miltiorrhiza aqueous extracts were identified,of which 15 components were common phenolic acids.The two reference substances had good linearity ranges(r≥0.997 6),and the values of precision,repeatability,stability and recovery met the demand of Chinese Pharmacopoeia.The content of 11 unknown compositions was determined.Conclusion The composition identification of S.miltiorrhiza aqueous extracts was realized with a high effective and rapid method,and a new idea for the content determination of unknown components was provided.