超声辅助酶解制备猪血血红素及紫外-可见吸收光谱分析

摘要

运用超声辅助酶解进行猪血血红素的制备，并对猪血血红素在不同环境下紫外-可见吸收光谱进行分析。结果表明：超声可以使酶活力提高25.5%，提高了酶解反应效率，缩短了酶解反应所需时间；对酶解产物pH值调节后进行离心分离血红素，并经纯水洗涤后再次离心，对血红素样品进行真空冷冻干燥；通过与血红素标准品比对，所得产品纯度达到83.2%。在碱性条件下，血红素溶液具有明显的特征吸收光谱；在酸性条件下，血红素溶液在300～400 nm和640 nm附近的特征吸收光谱发生了显著变化，吸收值明显降低。%Porcine heme was prepared by ultrasonic-assisted enzymatic hydrolysis of porcine red blood cells with Protamex and its UV-Vis absorption characteristics under acidic and alkaline conditions were analyzed. Results revealed that ultrasonic treatment resulted in a 25.5% increase in the enzyme activity, augmenting the hydrolysis efficiency and shortening the hydrolysis time. After pH adjustment of the hydrolysate, centrifugal separation of heme was carried out, followed by washing with purified water, second centrifugation and vacuum freeze drying. The product purity was determined using heme standard to be 83.2%. Under alkaline condition, the heme exhibited the obvious characteristic absorption, whereas under acidic condition, its characteristic absorption in the region of 300–400 nm and near 640 nm changed noticeably, showing a pronounced decrease in absorbance values.