高效液相色谱法测定金银花中绿原酸和木犀草苷的含量

Determination of Chlorogenic Acid and Luteoloside in Lonicerae Japonicae Flos by HPLC

目的：建立高效液相色谱（HPLC）法测定金银花中绿原酸和木犀草苷的含量。方法：流动相：0．5％冰乙酸-乙腈，线性梯度洗脱：0—20min（90：10—78：22），20～35min（78：22～76：24），35—37min（76：24～65：35），37—40min（65：35—90：10）；流速：1．0ml／min；检测波长：绿原酸为326nm，木犀草苷为350nm；柱温：30℃；色谱柱：依利特HypersilODS2（4．6mm×250mm，5μm）；进样量：10μl。结果：绿原酸的质量浓度在0．0216～0．1080mg／ml内与峰面积具有良好的线性关系，回归方程为Y=26347606．4815X-76704．9000（R^2=0．9990）。木犀草苷的质量浓度在0．000416—0．002080mg／ml内与峰面积具有良好的线性关系，回归方程为Y=28566947．1154X＋22．1500（R^2=0．9991）。70％甲醇超声提取方法较好，金银花提取物中绿原酸和木犀草苷的含量分别为3．416％和0．124％，这2种成分在24h内稳定性良好。结论：该方法简便、准确、快速易行，且该色谱条件可测定金银花中2种成分的含量。

OBJECTIVE：To establish HPLC method for determination of chlorogenic acid and luteoloside in Lonicerae japonicae Flos. METHODS： The mobile phase consisting of 0. 5% glacial acetic acid and acetonitrile startde was given a linear gradient elution from 90：10 to 78：22 （0-20 min）, 78：22 to 76：24 （20-35 min）, 76：24 to 65：35 （35-37 min） and 65：35 to 90：10 （37-40 min） at a flow rate of 1.0 ml/min, detection wavelength of 326 nm for chlorogenic acid and 350 nm for luteoloside, column temperature of 30℃ and injection volume of 10 ul. The chromatographic column was Hypersil ODS2 （4.6 mm × 250mam, 5 μm） of Elite. RESULTS ： The linear range of chlorogenic acid stood at 0. 021 6-0. 108 0 mg/ml with regression equation Y = 26 347 606. 481 5 X - 76 704. 900 0 （R2 =0. 999 0） and the linear range of luteoloside stood at 0. 000 416-0. 002 080 mg/mlwith regression equation Y = 28 566 947.115 4X ＋ 22. 150 0 （ R^2 = 0. 999 1 ）. The contents of chlorogenic acid and luteoloside in Lonicerae japonicae Flos extracted ultrasonically using 70% methanol stood at 3. 416% and 0. 124%, respectively and the two components were stable in quality within 24 hrs. CONCLUSION： This method is simple, accurate, rapid and feasible and applicable for determination of chlorogenic acid and luteoloside in Lonicerae japonicae Flos under the above- mentioned chromatographic condition.