高效液相色谱测定六味地黄丸的含量

  六味地黄丸处方为:熟地黄160g,山茱萸(制)80g,牡丹皮60g,山药80g,茯苓60g,泽泻60g。2005版《中国药典》采用高效液相色谱法测定了山茱萸中马钱苷和牡丹皮中丹皮酚的含量。有关六味地黄丸制剂的含量测定方法总结如下。
  
  一、2005《中国药典》六味地黄丸含量测定项下:
  
  山茱萸:
  
  色谱条件与系统适用性试验:十八烷基硅烷键合硅胶为填充剂;以四氢呋喃-乙腈-甲醇-0.05%磷酸溶液(1:8:4:87)为流动相;检测波长为236nm,柱温40℃。理论板数按马钱苷峰计应不低于4000。
  
  供试品溶液的制备:取本品水蜜丸或小蜜丸,切碎,取约0.7g,精密称定;或取重量差异检查项下的大蜜丸,剪碎,取约0.1g,精密称定,置具塞锥形瓶中,精密加入50%甲醇25ml,密塞,称定重量,超声提取(功率250W,频率33kHZ)15分钟,放冷,再称定重量,用50%甲醇补足减失的重量,摇匀,滤过。精密量取续滤液10ml,置中性氧化铝柱(100~200目,4g,内径1cm,干法装柱)上,用40%甲醇适量使溶解,并转移至10ml量瓶中,加50%甲醇稀释至刻度,摇匀,即得。
  
  牡丹皮
  
  色谱条件与系统适用性试验:以十八烷基硅烷键合硅胶为填充剂;以甲醇-水(70:30)为流动相;检测波长为274nm。理论板数按丹皮酚峰计应不低于3500。
  
  供试品溶液的制备:取本品水蜜丸或小蜜丸,切碎,取约0.3g,精密称定;或取重量差异检查项下的大蜜丸,剪碎,取约0.4g,精密称定,置具塞锥形瓶中,精密加入50%甲醇50ml,密塞,称定重量,超声提取(功率250W,频率33kHZ)45分钟,放冷,再称定重量,用50%甲醇补足减失的重量,摇匀,滤过,取续滤液,即得。
  
  二、以单皮酚为指标
  
  李玮玲等采用SPE-HPLC法测定六味地黄丸中丹皮酚的含量。高效液相色谱仪为Waters2695;色谱柱为HypersilODS2柱(205mm×4.6mm,5μm);流动相为甲醇-2%冰醋酸(55:45);流速为1.0mL/min;检测波长为275nm;柱温25℃。样品用甲醇超声提取后经Sep-PakC18微柱处理,收集第12~14mL流出液。
  
  仲英等采用薄层扫描测定了六味地黄丸中丹皮酚的含量,采用日本岛津CS-910双波长薄层扫描仪。层析条件:硅胶G板;展开溶剂:环乙烷-氯仿-无水乙醇7:3:1。薄层扫描条件:反射法锯齿扫描,波长s=274nm,R=365nm;扫描速度20mm/min;狭缝1.25

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