5种大黄蒽醌类衍生物的同时测定及应用论文.doc

5种大黄蒽醌类衍生物的同时测定及应用 【摘要】? 　　目的建立同时测定大黄药材中5种蒽醌类衍生物（芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚）含量的方法，优化大黄蒽醌的超声提取工艺。方法采用反相高效液相法，色谱柱：Waters C18柱（250 mm×4.6 mm, 5 μm）；流动相：甲醇-乙腈-0.01%磷酸溶液 (3∶5∶2)；柱温：25 ℃；流速：1.0 ml/min；检测波长：225 nm。结果5种游离蒽醌均具有较宽的线性范围且呈良好的线性关系（R2=0.998 7～0.999 7），平均回收率为92.79%～99.28%，日内精密度为0.65%～1.71%，日间精密度为2.52%～3.46%。结论该方法成功应用于大黄活性物质的提取工艺优化，根据所建立的方法测得的5种游离蒽醌含量，确定了大黄蒽醌的最佳超声提取工艺为：以70％乙醇为溶剂、料液比为1∶30 (g∶ml)、超声提取2次、每次提取20 min。 【关键词】? 超声提取； 大黄； RP-HPLC； 蒽醌 　　Abstract：ObjectiveTo develop a method for the simultaneous determination of five anthraquinone derivatives (aloe-emodin, rhein, emodin, chrysophanol and physcion) in rhubarb, and to optimize the ultrasonic extraction conditions.MethodsRP-HPLC on a Waters C18 column (250 mm×4.6 mm, 5? μm) was used. The mobile phase was methanol-acetonitrile-0.01% phosphoric acid solution (3:5:2), at a flow rate of 1.0 ml/min, and the detection wavelength was 225 nm. ResultsThe linear ranges of the five free anthraquinones were wide, showing good linear correlations with correlation coefficients (R2) of 0.9987 to 0.9997. The average recoveries were from 92.79% to 99.28%. While the intra-day and inter-day precision were 0.65%～1.71% and 2.52%～3.46%, respectively.ConclusionThrough the contents of five anthraquinone derivatives obtained by the developed RP-HPLC method, the optimum ultrasonic extraction conditions are determined as follows: using 70% EtOH as extracting solvent, the sample is extracted two times and each time for 20 min by a ultrasonic processor, when the solid to liquid ratio is 1:30 (g/ml). 　　Key words：Ultrasonic extraction;? Rhubarb;? RP-HPLC;? Anthraquinones????　　大黄Radix et Rhizoma Rhei为蓼科植物掌叶大黄Rheum Palmatum L.，唐古特大黄Rheum tanguticum Maxim. ex Balf 或药用大黄Rheum officinale Baill的干燥根和根茎，味苦、性寒。临床研究证实，大黄具有抗衰老、降血脂、抗肿瘤和抗炎、泻下作用及抗菌性、止血性等多种生物学活性［1］。其主要有效成分为蒽醌类衍生物,包括芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚以及与葡萄糖结合成苷。5种蒽醌是大黄以及衍生中药大黄属［2］质量控制的基础。?????　　大黄中蒽醌的分离和测定方法有薄层色谱法(TLC)［3］，高速逆流色谱法(HSCCC)［4，5］，胶束电动毛细管色谱法(MEKC)［6］，毛细管区带电泳法(CZE)［7］，高效液相色谱法(HPLC)［8］，加压毛细管电