肝水解肽体外活性测定方法的探讨研究

目的: 建立肝水解肽体外生物学活性测定方法。 方法: 将不同浓度肝水解肽作用于正常人肝L02细胞,WST-8比色法测定L02细胞增殖情况。同时对实验条件,包括稀释液、细胞接种密度、药物作用时间等进行探讨,建立肝水解肽体外活性测定方法,并用该方法对2个厂家生产的2批产品进行活性检测。 结果: 获得了比较稳定可靠的实验参数:稀释液为RPMI-1640培养液,细胞接种密度为4.0×10⁴ 个·mL^-1,药物作用时间为48~72 h,肝水解肽浓度为0.5,0.25,0.12,0.06,0.03,0.015 mg·mL^-1。由这些实验参数建立了肝水解肽活性测定方法。用该方法检测的2批产品活性均合格,重复实验3次,每次实验结果均一致,RSD均低于10%。 结论: 该方法可用于肝水解肽体外生物学活性测定。

Objective: To establish the method for determining bioactivity of liver hydrolysates in vitro. Methods: Treated L02 cell with different concentration of liver hydrolysates,cell proliferation was determined by WST-8 colorimetric assay.Experiment conditions are explored,including dilution,cell density,cell culture time. Results: Stable and reliable method parameters were derived:RPMI-1640 culture is more suitable dilution,cell density was 4.0×10⁴ mL^-1,culture time was 48-72 hours.As a result,we established the method for determining bioactivity of liver hydrolysates in vitro.Using the method,bioactivities of 2 bathches' products from different companies were determined 3 times,the results were consistent and the RSD was lower than 10%. Conclusion: The method was suitable for determining bioactivity of liver hydrolysates in vitro.