Method for preparing nitrosohemoglobin by using porcine blood

• Main Authors: LI YU, FU SHAOHUI, CHENG YAN, CHENG YUMING
• Format: Patent
• Language: English
• Published: 30.09.2015
• Subjects: CHEMISTRY; METALLURGY; ORGANIC CHEMISTRY; PEPTIDES

The invention provides a method for preparing nitrosohemoglobin by using porcine blood. The method comprises the following steps: (1) separating blood cells from fresh porcine blood; (2) extracting a hemoglobin crude extract from blood cells; (3) purifying the hemoglobin and measuring the purity of the hemoglobin, namely adding an antioxidant to the hemoglobin crude extract obtained in the step (2) for vacuum degassing, introducing N2 for protection, heating in a water bath at 60 DEG C for 30 minutes so that impurity proteins can be precipitated and then centrifuging to obtain a supernatant; adding 10mg/ml glycerol to the purified hemoglobin solution and then storing the purified hemoglobin solution in a refrigerator at 4 DEG C for later use; finally, measuring the content of the hemoglobin; (4) synthesizing the nitrosohemoglobin, namely adding NaOH to the purified hemoglobin, and meanwhile, adding sodium nitrite and vitamin C, regulating the pH to acidity, heating to the range of 50-70 DEG C and keeping for 25-45 minutes, and performing vacuum freeze-drying to generate a red powder, namely the nitrosohemoglobin. The method is simple in process, relatively low in equipment requirement and suitable for industrial production.