高效液相梯度洗脱法测定灵杞黄斑颗粒中毛蕊花糖苷和丹酚酸B含量  

Determination of verbascoside and salvianolic acid B in Lingqihuangban granules by HPLC gradient elution method

作　　者：姚慧娟[1] 胡道德[1] 金剑[1] 刘皋林[1] 

机构地区：[1]上海交通大学附属第一人民医院临床药学研究室,上海200080

出　　处：《中国新药与临床杂志》2010年第2期123-126,共4页Chinese Journal of New Drugs and Clinical Remedies

基　　金：上海中医药科研基金项目(2006Y004A)

摘　　要：目的建立同时测定灵杞黄斑颗粒中毛蕊花糖苷和丹酚酸B含量的反相高效液相梯度洗脱法。方法采用Agilent Eclipse XDB-C18(250mm×4.6mm,5μm)色谱柱,流动相:乙腈(A)-0.1%磷酸水溶液(B)进行梯度洗脱,梯度洗脱程序为进样0～8min,调节流动相A∶B(20∶80,V/V);8～20min,流动相A∶B(30∶70,V/V);20～23min,流动相A∶B(20∶80,V/V);流速:1.0mL·min-1,检测波长:334nm,柱温:30℃,进样量:10μL。结果毛蕊花糖苷和丹酚酸B分别在1.50～14.97mg·L-1和18.92～189.23mg·L-1范围内与峰面积呈良好的线性关系,相关系数分别为0.9998和0.9999,平均加样回收率毛蕊花糖苷为(99.7±s1.1)%,RSD=1.12%,丹酚酸B为(99.5±1.0)%,RSD=1.02%,3批样品中毛蕊花糖苷平均含量均不低于36.9μg·g-1,RSD均不大于1.54%,丹酚酸B平均含量均不低于344μg·g-1,RSD均不大于1.94%。结论该方法简便、快速、准确、灵敏度高、专属性强,可用于灵杞黄斑颗粒剂的质量控制。AIM To establish a reverse-phase high performance liquid chromatography （RP-HPLC） gradient elution method for the simultaneous determination of verbaseoside and salvianolic acid B in Lingqihuangban granules. METHODS The samples were separated on a Agilent Eclipse XDB-C18 （250 mm × 4.6 mm, 5μm） column with mobile gradient elution. The processes of 3 phase of acetonitrile （A） and 0 gradient elutions were as follows： 1% phosphonic acid solution （B） for the first process with volume ratio of mobile phase A ： B （20 ： 80, V/V） was carried out between 0 rain and 8 rain; the second process with volume ratio of mobile phase A ： B （30 ： 70, V/V） was performed between 8 min and 20 min; the third process with volume ratio of mobile phase A ： B （20 ： 80, V/V） was done between 20 rain and 23 min. All processes were taken under flow rate of 1.0 mL·min-1, the UV absorbance detection at 334 nm, column temperature at 30 ℃ and the sample volume of 10 μL. RESULTS The concentration ranges of calibration curves for verbascoside （1.50 - 14.97 mg·L-1） and salvianolic acid B （18.92 - 189.23mg·L-1） showed good linear correlation with peak areas, outcoming with correlative coeffients of 0.999 8 （n = 6） and 0.999 9 in = 6） , respectively. The average recoveries for verbascoside and salvianolie acid B were （99.7 ＋ s 1.1 ）% with RSD = 1.12% and （99.5 ＋ 1.0）% with RSD = 1.02%, respectively. The average contents of the three batches of samples for verbascoside and salvianolic acid B were more than 36.9μg·g-1 with RSD less than 1.54% and more than 344μg·g-1 with RSD less than 1.94%, respectively. CONCLUSION The RP-HPLC gradient elution method is simple, rapid, precise, high sensitive, and strongly specific, providing the quality control of Lingqihuangban granules.

关 键 词：中成药 黄斑变性 色谱法 高压液相 梯度洗脱 灵杞黄斑颗粒 毛蕊花糖苷 丹酚酸B 

分 类 号：R774.5[医药卫生—眼科] R927.2[医药卫生—临床医学]