采用HPLC同时进行连花清瘟颗粒多成分含量测定及其指纹图谱研究

摘要

Objective:To establish a RP-HPLC method for the content determination of 4 active compounds in Lianhua Qingwen granules,including Phillyrin,Forsythoside A,Chlorogenic Acid,Crytochlorogenic Acid and fingerprint analysis.Methods:The separation was performed on Swell chromplus TM-C18 column with a temperature at 30 ℃,mobile phase consisted of methanol -acetonitrile-0.3％ phosphoric acid(gradient elution)at flow rate of 1.0 mL· min-1,and detection wavelength at 278 nm.Resuhs:The linear ranges were 0.01632～0.1632 mL· min-1 for phillyrin with the correlation coefficient of 0.9996,0.05512～0.5512 mL·min-1 for forsythoside A with the correlation coefficient of 0.9999,0.0188～0.188 mL·min-1 for chlorogenic acid with the correlation coefficient of 0.9998,0.011556～0.11556 mL·min-1 for crytochlorogenic acid with the correlation coefficient of 0.9998.The recovery were 110.57％ (RSD =2.76％,n =6),107.02％ (RSD =1.45％,n =6),108.59％ (RSD =1.78％,n =6),109.77％ (RSD =3.49％,n =6),respectively.And fingerprint spectrum was established.10 batches of samples were detected and 14 peaks in the chromatogram were common.There was a high similarity and each chromatographic peak was obtained with good separation.Conclusion:The method had a fine conclusion and can be used in quality control of Lianhua Qingwen granules.%目的:建立高效液相色谱法同时进行连花清瘟颗粒中连翘苷、连翘酯苷A、绿原酸、新绿原酸等多种有效成分的含量测定及其指纹图谱研究.方法:采用Swell Chromplus TM-C18色谱柱(4.6 mm×250 mm,5μm),以0.3％的磷酸与乙腈的混合溶液为流动相,采用梯度洗脱方式,流速为1.0 mL· min-,检测波长278 nm,柱温30℃.结果:连翘苷(0.01632～0.1632 mL· min-1,r=0.9996)、连翘酯苷A(0.05512 ～0.5512 mL·min-,r=0.9999)、绿原酸(0.01880～0.1880 mL· min-,r =0.9998)、新绿原酸(0.011556～0.11556 mL· min-,r=0.9998)检测质量浓度在相应范围内与峰面积呈良好的线性关系(n=6),加样回收率分别为110.57％(RSD为2.76％)、107.02％(RSD为1.45％)、108.59％(RSD为1.78％)、109.77％(RSD为3.49％).10批连花清瘟颗粒样品指纹图谱测定结果确定了14个共有峰,各色谱峰分离度较好,相似度较高,符合中药色谱指纹图谱研究的技术要求.结论:本文所建立的方法可准确测定连花清瘟颗粒中四种有效成分的含量及其指纹图谱,方法可靠、重复性好,有助于完善其质量控制标准.