HPLC法测定桑叶和金银花中绿原酸及四种黄酮类化合物  

Determination of Chlorogenic Acid and Four Flavonoids from Flos Lonicerae and Mulberry Leaf by HPLC

作　　者：何聪杰 李舸远 HE Cong-jie;LI Ge-yuan(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,南京210023

出　　处：《海峡药学》2023年第9期25-32,共8页Strait Pharmaceutical Journal

摘　　要：目的建立一种同时测定绿原酸和黄酮类成分的含量测定方法,并应用于桑叶和金银花试样,为中药材和天然产物的提取工艺研究、质量分析探索适应性强的新分析方法,并探索更为环保的中药材水分测定方法。方法运用HPLC法,色谱柱为Agilent extend C_(18)柱(4.6 mm×250 mm,5μm),流动相为甲醇-0.2%磷酸,采用梯度洗脱,325 nm和350 nm双波长检测,柱温30℃,流速1 mL·min^(-1);以色谱级甲醇为提取溶剂,采用超声辅助提取方法制备供试液,随后分别采用常压干燥法和减压干燥法测定了桑叶和金银花试样的水分含量,用于计算试样干品中绿原酸及四种黄酮类化合物的含量。结果绿原酸、木犀草苷、芦丁、槲皮素、木犀草素浓度分别在0.11~53.70μg·mL^(-1)、0.12~58.20μg·mL^(-1)、0.12~58.50μg·mL^(-1)、0.13~63.30μg·mL^(-1)、0.10~50.90μg·mL^(-1)范围内线性良好,相关系数r分别为0.9993、0.9999、0.9999、1.0000、0.9995;供试液和混合对照品溶液在24 h内稳定性良好。结论建立的HPLC梯度洗脱方法准确度高、重现性好、精密度优,可作为桑叶、金银花中绿原酸和黄酮类成分的含量测定方法;此外,桑叶、金银花样品提取方法操作简便、溶液稳定性好。OBJECTIVE To establish a method for the simultaneous determination of chlorogenic acid and flavonoids,and apply it to mulberry leaves and honeysuckle flower samples,to explore a new analytical method with strong adaptability for the extraction technology and quality analysis of Chinese medicinal materials and natural products,and to explore a more environmentally friendly method for the determination of water content of Chinese medicinal materials.METHODS The determination was performed on Agilent extend C_(18)column(4.6 mm×250 mm,5μm)with mobile phase consisted of methanol-0.2%phosphoric acid by gradient elution and dual-wavelength detection at 325 nm and 350 nm.The column temperature was 30℃and the flow rate was 1 mL·min^(-1).Using chromatography-grade methanol as extraction solvent,the test solution was prepared by ultrasonic-assisted extraction method.Then,the water content of mulberry leaves and honeysuckle samples was determined by atmospheric drying method and vacuum drying method respectively,which was used to calculate the content of chlorogenic acid and four kinds of flavonoids in the samples.RESULTS The concentrations of chlorogenic acid,luteolin,rutin,quercetin and luteolin had good linearity respectively in the range of 0.11-53.70μg·mL^(-1),0.12-58.20μg·mL^(-1),0.12-58.50μg·mL^(-1),0.13-63.30μg·mL^(-1)and 0.10-50.90μg·mL^(-1),the correlation coefficients r were 0.9993,0.9999,0.9999,1.0000 and 0.9995 in turn.The test solution and the mixed control solution had good stability within 24 h.CONCLUSION The established HPLC gradient elution method has high accuracy,good reproducibility and excellent precision,and can be used for the determination of chlorogenic acid and flavonoids in mulberry leaves and honeysuckle.In addition,the extraction method of mulberry leaf and honeysuckle was simple and stable.

关 键 词：HPLC 金银花 桑叶 绿原酸 木犀草苷 木犀草素 芦丁 槲皮素 稳定性 

分 类 号：R927[医药卫生—药学]