HPLC法同时测定维吾尔药材百里香中6种成分的含量

目的:建立HPLC法同时测定维吾尔药材百里香中绿原酸、咖啡酸、百里香酚、芦丁、迷迭香酸和蒙花苷的含量。方法:采用Agilent ZORBX SB-C₁₈色谱柱(250 mm×4.60 mm,5 μm),以乙腈(A) -0.2%磷酸水溶液(B)为流动相,梯度洗脱(0~10 min,90%B→85%B;10~16 min,85%B→80%B;16~25 min,80%B→50%B;25~35 min,50%B→30%B;35~40 min,30%B→20%B;40~45 min,20%B→10%B;45~60 min,10%B→90%B),流速1.0 mL·min^-1,检测波长300 nm,柱温30℃,进样量10 μL。结果:绿原酸、咖啡酸、百里香酚、芦丁、迷迭香酸和蒙花苷线性范围分别为0.033~0.198 mg·mL^-1(r=0.9999)、0.034~0.204 mg·mL^-1(r=0.9999)、0.008~0.048 mg·mL^-1(r=0.9997)、0.0155~0.093 mg·mL^-1(r=0.9996)、0.0302~0.1812 mg·mL^-1(r=0.9991)和0.0901~0.5406 mg·mL^-1(r=0.9994),加样回收率在99.25%~101.4%之间,RSD为0.8%~1.9%。3批百里绿原酸5.009、5.106、5.185 mg·g^-1,咖啡酸4.006、4.152、4.220 mg·g^-1,香药材中百里香酚含量分别为1.024、1.036、1.106 mg·g^-1,芦丁1.822、1.863、1.879 mg·g^-1,迷迭香酸7.004、7.054、7.164 mg·g^-1,蒙花苷20.001、20.16、20.926 mg·g^-1。结论:该方法专属性强,能够快速、准确地测定百里香药材中6种指标性成分含量,为更好地控制维吾尔药材百里香的质量并评价其药用价值提供科学数据。

Objective:To establish an HPLC method for simultaneous determination of chlorogenic acid, coffee acid,thymol,rutin,rosmarinic acid and linarin in Thymus marschallianus Willd.Methods:An Agilent ZORBX SB-C₁₈ column(250 mm×4.60 mm,5 μm)was adopted in the study.The mobile phase consisted of acetonitrile(A)and 0.2% phosphoric acid water solution(B)with gradient elution(0-10 min,90%B→85%B; 10-16 min,85%B→80%B;16-25 min,80%B→50%B,25-35 min;50%B→30%B;35-40 min,30%B→20%B;40-45 min,20%B→10%B;45-60 min,10%B→90%B)at a flow rate of 1.0 mL·min^-1,the detection wavelength was 300 nm,the column temperature was controlled at 30℃,and the injection volume was 10 μL.Results:The linear ranges of six compositions in Thymus marschallianus were 0.033-0.198 mg·mL^-1 (r=0.999 9)for chlorogenic acid,0.034-0.204 mg·mL^-1(r=0.999 9)for coffee acid,0.008-0.048 mg·mL^-1 (r=0.999 7)for thymol,0.015 5-0.093 mg·mL^-1(r=0.9996)for rutin,0.030 2-0.181 2 mg·mL^-1(r=0.9991) for rosmarinic acid,and 0.090 1-0.540 6 mg·mL^-1(r=0.9994)for linarin.The average recovery range of six components was 99.25%-101.4% with RSD of 0.8%-1.9%.The contents of 3 batches of thyme herb were 5.009, 5.106,and 5.185 mg·g^-1 for chlorogenic acid,4.006,4.152,and 4.220 mg·g^-1 for coffee acid,1.024,1.036, and 1.106 mg·g^-1 for thymol,1.822,1.863,and 1.879 mg·g^-1 for rutin,7.004,7.054,and 7.164 mg·g^-1 for rosmarinic acid,and 20.001,20.16,20.926 mg·g^-1 for linarin.Conclusion:The established method has high specificity and be used to quickly and accurately determine the content of 6 kinds of index components of thyme herbs,and provide the scientific data evaluation for quality control of Uygur medicine thyme.