提取诺邓火腿抗氧化肽,经过Sephadex G-25凝胶色谱进行分离纯化,对各组分进行体外抗氧化试验(清除·OH、清除DPPH自由基、螯合Fe2+)研究抗氧化活性;将抗氧化活性最强的组分经葡聚糖凝胶G-15(Sephadex G-15)凝胶色谱再次分离纯化,最后对抗氧化活性最强的组分通过基质辅助激光解析串联飞行时间质谱仪(MALDI-TOF-MS)对抗氧化肽进行测序鉴定。试验结果:诺邓火腿抗氧化肽经Sephadex G-25凝胶色谱进行分离纯化后得到4个组分(A、B、C和D),其中组分C的抗氧化活性最强,质量浓度为1 mg/mL时,·OH清除率、DPPH自由基清除率和Fe2+螯合率分别达到50.01%、48.32%和34.37%,与其他组分(A、B和D)相比,差异极显著(p<0.01);组分C经Sephadex G-15凝胶色谱分离纯化的5个组分(C1、C2、C3、C4和C5),其中C3组分抗氧化活性最强,质量浓度为1 mg/mL时,·OH清除率、DPPH自由基清除率和螯合Fe2+分别达到73.01%、51.21%和65.23%,与其他组分(C1、C2、C4和C5)相比,差异极显著(p<0.01);通过MALDI-TOF-MS对C3组分抗氧化肽进行测序鉴定,得到抗氧化肽序列。
We extracted and purified the anti-oxidized peptide of Nuodeng ham through Sephadex G-25 gel chromatography. The anti-oxidative activity of components was studied in vitro, including ·OH radical scavenging assay, DPPH radical scavenging assay and chelation Combined Fe2+ experiment. The components with the highest antioxidant activity were separated and purified by Sephadex G-15 gel chromatography. Finally, the antioxidant peptides were identified by MALDI-TOF-MS. Results: Noudeng ham antioxidant peptide were separated and purified by Sephadex G-25 gel chromatography to obtain four components, namely A, B, C and D. Component C had the strongest antioxidant activity. The free radical scavenging rate, DPPH radical scavenging rate and Fe2+ chelation rate reached 50.01%, 48.32% and 34.37% when the concentration of component C was 1 mg/mL, respectively. Compared with other components (A, B and D), the difference was significant (p<0.01). The component C was Purified by Sephadex G-15 gel chromatography of the five components (C1, C2, C4 and C5) , wherein the component C3 has the highest antioxidant activity. The free radical scavenging rate, DPPH radical scavenging rate and Fe2+ chelation rate reached 73.01%, 51.21% and 65.23% when the concentration of component C3 was 1 mg/mL, respectively. Compared with other components (C1, C2, C4 and C5), the difference was significant (p<0.01). The antioxidant peptides of C3 were identified by MALDI-TOF-MS.