无梗五加茎中3种酚酸类成分的HPLC-DAD-MS分析

Analysis of three phenolic acids in stems of Acanthopanax sessiliflorus by HPLC-DAD-MS

目的利用高效液相色谱-二极管阵列检测器-质谱法,对无梗五加茎中3种酚酸类成分进行定性定量分析。方法 RP-HPLC,色谱柱为Agilent Zorbax SB-C8(250 mm×4.6 mm,5μm);流动相为乙腈-甲醇-酸水溶液梯度洗脱;体积流量为1 mL/min;柱温为50℃;进样量为10μL;定性检测波长为270 nm,定量检测波长为260 nm、324 nm;离子源ESI,负离子扫描模式。结果原儿茶酸、绿原酸和咖啡酸在25 min内均被很好地分离,经光谱及质谱的分析得到确认。3种成分分别在0.102～2.04,0.256～5.12,0.295～5.90 mg/mL质量浓度范围内与色谱峰面积呈良好的线性关系(r≥0.999 8)。3种成分的精密度、重复性、稳定性的RSD均小于2.0%,平均回收率范围为99.2%～100.4%,RSD为1.0%～2.2%。结论 不同产地无梗五加茎中3种成分的含有量有较大的差别。方法准确、重现性好,可用于无梗五加茎中原儿茶酸、绿原酸、咖啡酸成分的分析测定。

AIM To develop a method for qualitatively and quantitatively analysing three constituents of phenolic acid in the stems of Acanthopanax sessifilorus by HPLC with diode array detection and mass spectrometer（HPLC-DAD-MS）.METHODS RP-HPLC analysis was achieved on an Agilent Zorbax SB-C8 column（250 mm× 4.6 mm,5 μm） with gradient elution using acetonitrile-methanol-acid aqueous solution as the mobile phase.The flow rate was 1.0 mL/min and the column temperature of 50 ℃,the detection wavelength of 260 nm,270 nm and 324 nm.An ESI source was used with a negative ionization scan mode.RESULTS Protocatechuic acid,chlorogenic acid and caffeic acid were separated and identified in 25 min by HPLC-DAD-MS.The calibration curves of three compounds showed good linearity（r ≥0.999 8） in the ranges of concentration,the linear ranges were 0.102-2.04,0.256-5.12,0.295-5.90 mg/mL,respectively.The RSDs of precision reproducibility and stability tests were less than 2.0%,the average recoveries were in the range of 99.2%-100.4% and the range of RSDs was within 1.0%-2.2%.CONCLUSION The contents of three compounds have difference in Acanthopanax sessiliflorus from various habitats.The established method is accurate and reproducible,which can be used to analyse the phenolic acids.