黑木耳多糖螯合钙的制备及活性评价

Preparation and Activity Evaluation of Black Fungus Polysaccharide-Calcium Chelate

目的:以黑木耳多糖(AAP)为原料,探究黑木耳多糖螯合钙(AAP-Ca)的最佳制备工艺,并对其理化性质、安全性以及抗氧化、抗炎等生物活性进行评价。方法:采用原子吸收光谱测定钙含量,以螯合率为指标,采用单因素优化AAP-Ca的制备工艺,通过测量AAP-Ca的Zeta电位以分析其在低温、血清以及不同pH值条件下的稳定性;采用溶血实验以及细胞毒性实验以探究其安全性,以DPPH·的清除率分析其体外抗氧化性,同时通过脂多糖(LPS)诱导的炎症模型中NO释放量考察抗炎活性。结果:反应温度30℃、CaCl_(2)质量浓度为4 mg/mL、糖钙质量比4:1、反应时间7 h为制备AAP-Ca的最佳工艺条件,在最佳状态下含钙率高达(69.2±0.36)%。黑木耳多糖与钙螯合之后可见纤维状多糖表面附着球形颗粒结构,AAP-Ca可以在低温、血清以及不同pH值条件下稳定存在,且具有良好的血液相容性。相较于黑木耳多糖,黑木耳多糖螯合钙对DPPH·显示出更强的清除作用,且对脂多糖LPS诱导的炎症因子NO的释放有抑制作用。结论:制备得到的AAP-Ca含钙率较高、稳定性较好,具有良好的生物安全性,体外抗氧化性和抗炎活性良好,为开发新型多糖补钙剂以及提高黑木耳资源的综合利用提供了依据。

Objective:To investigate the optimal preparation process of chelated calcium(AAP-Ca)of black fungus(Auricularia auricular)polysaccharide(AAP),black fungus polysaccharide was used as raw material,and its physical and chemical properties,safety,antioxidant and anti-inflammatory biological activities were evaluated.Methods:Calcium content was determined by atomic absorption spectroscopy,chelation rate was indexed,the preparation process of AAP-Ca was optimized by univariate factor,and its stability at low temperature,serum and different pH conditions was analyzed by measuring the zeta potential of AAP-Ca.Hemolysis experiments and cytotoxicity experiments were used to explore its safety,the in vitro antioxidant properties were analyzed by the clearance of DPPH.,and the anti-inflammatory activity was investigated by the NO-release amount in the lipopolysaccharide(LPS)-induced inflammatory model.Results:The reaction temperature was 30℃,the CaCl_(2)mass concentration was 4 mg/mL,the sugar-calcium mass ratio was 4:1,and the reaction time was 7 h,and the calcium content was as high as(69.2±0.36)%under the best state.After chelation with calcium,the surface of the fibrous polysaccharide was attached to the spherical granular structure,and AAP-Ca can be stable at low temperature,serum and different pH conditions,and had good hematocompatibility.Compared with black fungus polysaccharides,black fungus polysaccharide chelated calcium showed a stronger clearance effect on DPPH.,and inhibited the NO release of the LPS-induced inflammatory factor.Conclusion:The prepared AAP-Ca has a high calcium content,good stability,good biosafety,good antioxidant and anti-inflammatory activity in vitro,which provides a basis for the development of new polysaccharide calcium supplements and the comprehensive utilization of black fungus resources.