无梗五加茎中3种酚酸类成分的HPLC-DAD-MS分析

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10 μL；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.

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

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.