目的从生物检定角度,探索并建立表征注射用双黄连冻干粉针质量波动性的方法。方法采用微量量热法,建立基于痢疾杆菌的注射用双黄连冻干粉针质量波动评价方法,制备避光室温、高温(60℃)及光照(4 500±500)lx样品S1～S9。以生物热活性图谱的相似度和微量量热学参数如指数生长期速率常数(k1),最大发热功率(P1)、达峰时间(T1)、发热量(Q1),稳定期最大发热功率(P2)、达峰时间(T2)、发热量(Q2)为评价指标,进行定性定量分析,以生物热活性图谱相似度S、参数T'm、P'm为指标进行聚类分析。结果相似度分析结果显示,与避光室温(22℃)样品S1～S4相比,光照(4 500±500)lx样品S7～S9相似度较高(R>0.85),而高温(60℃)样品S5～S6的生物热活性图谱相似度较低(R≤0.700)。结论以相似度及微量量热学特征参数定性定量表征不同样品的抑菌活性,所建立的生物热活性图谱全面反映模型生物的热生物学信息,提供了专属性较强的二维信息,可作为化学指纹图谱的有益补充,可以用于注射用双黄连冻干粉针的质量波动性检测。

Objective To provide the methods for quality fluctuation of Shuanghuanglian Freeze-dried Powder for Injection(SFPI) including bioassay method.Methods Biological profiles of SFPI on Shigella dysenteriae had been established by microcalorimetry.Samples 1—4 were kept at room temperature;Samples 5—6 were kept at storage at 60 ℃;Samples 7—9 were kept under the condition of illumination intensity(4 500±500) lx.All samples were analyzed qualitatively and quantitatively by similarity values of biological profiles and thermodynamics parameters,such as T1m,P1m,Q1,k1,T2m,and I.The normal samples and special samples could be distinguished by cluster analysis with similar values(S),the main thermal activity parameters(T1m,P1m).Results Compared with samples 1—4,the similarity tendency of samples 7—9 was similar,while similar values of S5—S6 was less than or equal to 0.700.Conclusion All the results show that the biotic thermal activity of different samples could be evaluated qualitatively and quantitatively by similarity values of biological profiles and thermodynamics parameters.In Conclusion,characterized by two-dimension,microcalorimetry could supply thermograms as biological profiles characterized to describe the bioactivity of drugs,which were beneficial supplement to chemical analysis.Biological profiles are proposed as an efficient strategy for quality fluctuation of SFPIs.

国家重大新药创制科技重大专项(2009ZX09502-003,2009ZX09502-022);国家自然科学基金资助项目(30701109)