HPLC同时分析金银花中绿原酸和黄酮类成分的方法建立及其应用

Study on simultaneous analysis of chlorogenic acid and flavonoid constituents in Flos Lonicerae by HPLC

目的建立应用反相高效液相色谱法同时测定金银花中绿原酸和4种黄酮类成分——芦丁、木犀草素7-O-β-D-半乳糖苷、忍冬苷、槲皮素含量的方法。方法色谱柱为AglientZorbax80A(4.6mm×250mm,5μm);流动相为乙腈-0.5%磷酸,梯度洗脱;流速1.0mL·min-1,检测波长为355nm。结果绿原酸、芦丁、木犀草素7-O-β-D-半乳糖苷、忍冬苷、槲皮素的线性范围分别为0.275-22μg(r=0.9998),7.725-772.5ng(r=0.9997),8.025-802.5ng(r=0.9999),7.725-772.5ng(r=0.9999),3.338-333.8ng(r=0.9999),平均回收率(n=9)分别为99.42%,99.11%,101.46%,98.61%,100.65%。结论本法准确、快速、重现性好,适合于同时测定各种金银花中绿原酸和黄酮类成分的含量。

OBJECTIVE: To develop a RP-HPLC method with gradient elution for the separation and simultaneously determination of chlorogenic acid and four flavonoids, rutin, luteolin-7-0-β-D-galactoside, lonicerin and quercetin in Flos Lonicerae. METHODS: The separation was performed on a Aglient Zorbax 80A column (4.6 mm × 250 mm, 5 μm). Acetonitril-0.5% H3PO 4 was used as the mobile phase with gradient elution. The flow rate was 1.0 mL&middotmin-1. The detection wavelength was at 355 nm. RESULTS: Good linearities of chlorogenic acid, rutin, luteolin-7-0-β-D- galactoside, lonicerin and quercetin were obtained over the ranges of 0.275-22 μg (r = 0.999 8), 7.725-772.5 ng (r = 0.999 7), 8.025-802.5 ng (r = 0.999 9), 7.725-772.5 ng (r = 0.999 9) and 3.338-333.8 ng (r = 0.999 9). The average recoveries(n = 9) were 99.42%, 99.11%, 101.46%, 98.61% and 100.65%, respectively. CONCLUSION: This method is accurate, rapid and reproducible, will be used to determine chlorogenic acid and flavonoids in the raw material of Flos Lonicerae.