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目的:建立单胺氧化酶(MAO)活性筛选模型,以该模型指导鳖甲活性肽的分离,并检验分离单体活性。方法:参照MAO试剂盒方法,以优降宁为阳性药进行MAO活性筛选模型;对鳖甲水提液依次调节乙醇体积分数为20%,40%,60%,80%,90%,得到各醇沉部位,再以该模型筛选各醇沉部位,选取活性最优部分继续分离,然后将分得的单体以该模型检验活性。结果:该MAO活性筛选模型中,阳性药优降宁质量浓度在1.0 g·L-1时,抑制率为98.30%~103.05%;各部位质量浓度为1.0 g·L-1时,鳖甲水提液部位MAO抑制率为(23.34±9.66)%,20%醇沉部位(28.16±5.78)%,40%醇沉部位(30.69±7.17)%,60%醇沉部位(41.60±8.03)%,80%醇沉部位(64.91±2.94)%,90%醇沉部位(53.34±7.76)%;在80%醇沉部位分得鳖甲七肽(GAGPHGG),质量浓度在0.1~1.6 g·L-1时,鳖甲七肽MAO抑制率为15.30%~54.84%。结论:发现MAO活性筛选模型优降宁抑制效果显著,提示该模型成功;以该模型成功指导了鳖甲七肽的分离,且鳖甲七肽具有较明显MAO抑制活性。
OBJECTIVE: To establish a monoamine oxidase (MAO) activity screening model to guide the separation of active peptides from turtle shell, and to test the activity of isolated monomers. METHODS: The MAO activity screening model was optimized by the method of MAO kit with the method of ginsenoside. The volume fraction of ethanol was 20%, 40%, 60%, 80% and 90% Alcohol precipitation sites, and then the model selection of each alcohol precipitation sites, select the most active part of the continued separation, and then the monomer was separated to test the activity of the model. Results: In the MAO activity screening model, when the concentration of the optimal drug, Nygonin, was 1.0 g · L-1, the inhibitory rates were 98.30% -103.05%. When the mass concentration of each component was 1.0 g · L-1, The MAO inhibition rate was23.34 ± 9.66%, 20% alcohol precipitation (28.16 ± 5.78)%, 40% alcohol precipitation (30.69 ± 7.17)% and60% alcohol precipitation (41.60 ± 8.03)%, 80% alcohol precipitation (64.91 ± 2.94)%, 90% alcohol precipitation (53.34 ± 7.76)%; in the 80% alcohol deposition part of the grading of GAGPHGG, mass concentration of 0.1 ~ 1.6 g · L- 1, the turtle-a-peptide MAO inhibition rate of 15.30% ~ 54.84%. CONCLUSION: MAO activity screening model was found to be significantly inhibitory effect of UNPHA, suggesting that the model was successful. The model successfully instructed the separation of turtle-captapeptide, and the chelerythrine showed obvious MAO inhibitory activity.