波长切换高效液相色谱法同时测定青翘抗毒颗粒中7个成分的含量

Simultaneous determination of seven constituents in Qingqiaokangdu Granules by HPLC with switching wavelengths

目的：建立高效液相色谱法同时测定青翘抗毒颗粒中绿原酸、马钱苷、连翘苷、葛根素、（R,S）-告依春、射干苷、咖啡酸的含量。方法：采用岛津InertSustain C18柱（250 mm×4.6 mm,5μm）,以甲醇-0.3%磷酸水溶液为流动相进行梯度洗脱;流速为1.0mL·min-1;进样量为5μL;检测波长为240 nm（0-25 min,检测（R,S）-告依春）、327 nm（25-39 min,检测绿原酸和咖啡酸）、240 nm（39-45 min,检测葛根素和马钱苷）、265 nm（45-53 nm,检测射干苷）、228 nm（53-65 min,检测连翘苷）;柱温为30℃。结果：绿原酸、马钱苷、连翘苷、葛根素、射干苷、咖啡酸、（R,S）-告依春7个成分的进样质量浓度分别在4.56-228μg·mL-1（R2=0.999 9）、4.04-202μg·mL-1（R2=0.999 8）、8.10-405μg·mL-1（R2=0.999 9）、8.06-161μg·mL-1（R2=0.999 9）、2.10-105μg·mL-1（R2=0.999 9）、3.46-173μg·mL-1（R2=0.999 9）、4.58-229μg·mL-1（R2=0.999 9）与峰面积呈良好的线性关系;各组分分离度良好;加样回收率（n=6）分别为99.35%（RSD为2.6%）、100.76%（RSD为1.5%）、100.34%（RSD为0.9%）、100.63%（RSD为1.5%）、100.31%（RSD为1.7%）、100.70%（RSD为1.6%）、99.03%（RSD为1.7%）。结论：本研究建立的含量测定方法符合方法学验证要求,可用于青翘抗毒颗粒7个指标性成分的同时测定。

OBJECTIVE To establish an HPLC method for simultaneous determination of seven constituents（chlorogenic acid,loganin,phillyrin,puerarin,belamcandin,caffeic acid,epigoitrin）in Qingqiaokangdu Granules.METHODS Separation was performed on a Shimadzu Insertsustain-C18（250 mm×4.6 mm,5μm）column with gradient elution of methanol-0.3%phosphoric acid solution at flow rate of 1.0ml·min-1.Sample volume was 5μl,UV detection wavelength was set at 240 nm for epigoitrin in 0-25 min,327 nm for chlorogenic acid and caffeic acid in 25-39 min,240 nm for puerarin and loganin in 39-45 min,265 nm for belamcandin in45-53 min,228 nm for phillyrin in53-65 min.Column temperature was set at 30℃.RESULTS Ranges for linear correlation of chlorogenic acid,loganin,phillyrin,puerarin,belamcandin,caffeic acid and epigoitrin were4.56-228μg·mL-1（R2=0.999 9）,4.04-202μg·mL-1（R2=0.999 8）,8.10-405μg·mL-1（R2=0.999 9）,8.06-161μg·mL-1（R2=0.999 9）,2.10-105μg·mL-1（R2=0.999 9）,3.46-173μg·mL-1（R2=0.999 9）,4.58-229μg·mL-1（R2=0.999 9）,respectively.Excellent chromatographic separation was achieved.Average recoveries（n=6）of seven components were 99.35%（RSD=2.6%）,100.76%（RSD=1.5%）,100.34%（RSD=0.9%）,100.63%（RSD=1.5%）,100.31%（RSD=1.7%）,100.70%（RSD=1.6%）and 99.03%（RSD=1.7%）.CONCLUSION This method is consistent with method validation requirements,and can simultaneously determine the above 7 index components of Qingqiaokangdu Granules.