UPLC-PDA法同时测定连翘中4种成分的含量

Simultaneous Determination of Four Components in Forsythiae Fructus by UPLC-PDA

目的建立同时测定连翘中连翘酯苷A、连翘酯苷B、连翘苷、槲皮素4种化学成分的UPLC方法。方法采用Acquity UPLC BEH-C18(2.1 mm×100 mm,1.7μm)色谱柱;以0.2%甲酸溶液为流动相A,乙腈为流动相B,梯度洗脱(0~15 min,90%~70%A;15~25 min,70%~30%A);柱温35℃,流速0.3 ml/min,检测波长235 nm,进样量2μl。结果使用此种方法能够在25 min内实现了对连翘中4种化学成分的分离,连翘酯苷B、连翘酯苷A、连翘苷和槲皮素分别在0.42~2.10μg(r=0.9999),0.92~4.60μg(r=0.9999),1.02~3.06μg(r=0.9997),0.098~2.45μg(r=1.0000)范围内与色谱峰峰面积线性关系良好,加样回收率分别为93.48%(RSD=0.81%),97.79%(RSD=1.10%),97.53%(RSD=1.23%),96.29%(RSD=1.88%)。结论该方法操作简单,结果准确,专属性强,可用于连翘药材的质量控制。

Objective To establish a UPLC method for simultaneous determination of forsythia A,forsythia B,forsythia and quercetin in Forsythiae fructus.Methods Acquity UPLC BEH-C18(2.1 mm×100 mm,1.7μm)column was adopted.Gradient elution was performed,using 0.2%formic acid-acetonitrile solution as mobile phase(0-15 min,90%-70%A;15-25 min,70%-30%A).The column temperature was 35℃,the flow rate was 0.3 ml/min,the detection wavelength was 235 nm and the injection volume was 2μl.Results The four chemical constituents of Forsythiae fructus could be separated within 25 min by using this method.The linear range of forsythin B,forsythin A,phillyrin and quercetin was 0.42-2.10μg(r=0.9999),0.92-4.60μg(r=0.9999),1.02-3.06μg(r=0.9997)and 0.098-2.45μg(r=1.0000),respectively.The average recoveries were 93.48%(RSD=0.81%),97.79%(RSD=1.10%),97.53%(RSD=1.23%)and 96.29%(RSD=1.88%),respectively.Conclusion This method is simple,accurate and highly specific.It can be used for quality control of Forsythiae fructus.