紫外线摇动灭菌法建立及在胶原溶液灭菌中的应用

背景：在已有胶原溶液灭菌方法中，微孔膜过滤和紫外线照射可不降低溶液的黏性，但紫外线只能用于表层溶液或透明管道中溶液的灭菌，限制了紫外线的灭菌效率。如果能用紫外线使一个容器中上下各液层都受照射灭菌，就可省去紫外线灭菌的循环管道，可简化操作保持灭菌蛋白特性。 目的：改进紫外线灭菌法，使之适用于胶原溶液的灭菌。 设计、时间及地点：体外观察实验，于2000-02/2008-06在福州市传染病医院肝病研究所完成。 材料：灭菌装置由紫外灯、反光罩(可罩于摇床上)、摇床和容盘组成。 方法：①容盘置于摇床上。将待灭菌的酸性胶原溶液装入容盘，液层深8~12 mm。②开动紫外灯和摇床，使容盘中溶液摇动混合、暴露深层受紫外线照射。③将经照射灭菌的胶原溶液pH调为7.4，使成凝胶。 主要观察指标：①胶原溶液层厚、紫外线照射时间对灭菌效果(菌落计数)的影响。②将紫外照射后的胶原溶液pH调为7.4，形成凝胶时间。 结果：①液层厚8 mm照射30 min的胶原溶液和液层12 mm照射90 min的胶原溶液，经培养均显示细菌和霉菌阴性。②将用本法灭菌的胶原溶液pH变为7.4后可在1 h内形成凝胶。 结论：紫外线摇动灭菌法简便有效，灭菌后不影响胶原溶液成凝胶性，适用于胶原及类似蛋白溶液的灭菌。

Establishment of the sterilization with ultraviolet and rolling and its application in sterilization of collagen solution

BACKGROUND: Among the methods for sterilization of collagen solution, microporous membrane filtration and ultraviolet irradiation would not decrease viscosity of the solution, but ultraviolet ray can only irradiating the surface of solution or solution in hyaline tube, which limits sterile efficiency of ultraviolet ray. If every layers of the solution can be irradiated by ultraviolet ray, as the solution in a circulation pipe, by simply rolling the solution, the efficiency of the sterilization would be increased and the quality of the proteins sterilized may be unchanged. OBJECTIVE: To improve the sterilization method of ultraviolet to be suitable for sterilization of collagen solution. DESIGN, TIME AND SETTING: The in vitro observational study was conducted at the Institute of Liver Diseases, Fuzhou Hospital for Infectious Diseases from February 2000 to June 2008. MATERIALS: Sterilization equipment was composed of ultraviolet lamp, reflector (covered on rolling bed), rolling bed and utensil. METHODS: A utensil was put on the rolling bed. The acidic collagen solution to be sterilized was put in the utensil. The depth of the collagen solution was 8-12 mm. Turn on the ultraviolet lamps and the rolling bed which could roll and mix the solution in the utensil, so the whole collagen solution was irradiated and sterilized. After the irradiation, the collagen solution changed to pH =7.4 could form a gel. MAIN OUTCOME MEASURES: The following parameters were measured: depth of the collagen solution; effects of the irradiation time on the efficiency of sterilization efficiency (colony count); Gel formation time after the ultraviolet irradiation (pH = 7.4). RESULTS: Both the solution of 8-mm depth irradiated for 30 minutes and the solution of 12-mm depth radiated for 90 minutes showed negative bacteria and fungus growth on culture. The collagen solution sterilized by this method could become a gel in 1 hour after changing the pH to 7.4. CONCLUSION: This simple and effective method of sterilization with ultraviolet and rolling would not prevent the collagen solution from becoming a gel and be suitable for sterilization of the protein solution.