冻干蟾皮凝胶剂质量标准及初步药效学的研究.doc

冻干蟾皮凝胶剂质量标准及初步药效学的研究 　　[摘要] 目的 建立冻干蟾皮凝胶剂质量标准和初步药效学研究。方法 采用薄层色谱（TLC）法对制剂中的华蟾酥毒基和酯蟾毒配基进行定性鉴别，高效液相色谱（HPLC）法测定华蟾酥毒基和酯蟾毒配基含量，色谱柱kromasil C18（4.6 mm×250 mm，5 μm）；流动相：乙腈（A）-水（B）（48?U52）；流速：1 mL/min；检测波长：296 nm；柱温：30℃；进样量：10 μL。采用S180腹水型实体瘤小鼠进行初步抑瘤药效学研究。结果 硅胶G薄层板上，在与对照品色谱对应的位置上，显示相同颜色的荧光斑点，且斑点大小适中，分离度良好。华蟾酥毒基和酯蟾毒配基分别在0.056～4.48 μg/mL，0.256～10.24 μg/mL范围内，峰面积与浓度存在良好的线性关系（r=0.9987、0.9996）。加样回收率（n=6）为100.27%、101.68%，相对标准偏差为1.56%、1.67%。初步药效学结果显示冻干蟾皮凝胶剂不同剂量组均有显著抗肿瘤作用，且高剂量组效果最佳，抑瘤率为48.03%。结论 建立了冻干蟾皮凝胶剂质量标准，初步药效学表明冻干蟾皮凝胶剂具有显著抗肿瘤作用。 　　[关键词] 冻干蟾皮凝胶剂；质量?俗迹豢怪琢觯换?蟾酥毒基；酯蟾毒配基 　　[中图分类号] R285.5 [文献标识码] A [文章编号] 1673-7210（2018）04（a）-0029-05 　　[Abstract] Objective To establish the quality standard and preliminary pharmacodynamic study of Freeze-dried Toad Skin Gel. Methods TLC was used for the qualitative identification of Cinobufagin and Resibufogenin， HPLC was adopted to determine content of Cinobufagin and Resibufogenin. Chromatographic column： kromasil C18 （4.6 mm×250 mm， 5 μm）； flowing phase： acetonitrile （A）- water （B） （48∶52）； flow rate： 1 mL/min； UV detection wavelength： 296 nm； column temperature： 30℃； the volume： 10 μL. S180 ascites solid tumor mice were used to study the pharmacodynamics of inhibition of tumor growth. Results The fluorescence spots of the same color were shown in the corresponding position of the Silica gel G thin layer， and the spot size was moderate and the separation degree was good. Cinobufagin and Resibufogenin showed good linearity between the peak area and the concentration in the range of 0.056-4.48 μg/mL （r=0.9987） and 0.256-10.24 μg/mL （0.9996）， respectively. The sample recovery rate（n=6） was 100.27%， 101.68%， RSD was 1.56% and 1.67%. The Preliminary pharmacodynamics study showed that all groups of Freeze-dried Toad Skin Gel had significant anti-tumor effect， and the high-dose group had the best effect， with the inhibitory rate of 48.03%. Conclusion The quality standard of Freeze-dried Toad Skin Gel is established， and the preliminary pharmacodynami