推荐回答(3个)
湿热灭菌的原理是使微生物的蛋白质及核酸变形导致其死亡。这种变形首先是分子中的氢键分裂,当氢键断裂时,蛋白质及核酸内部结构被破坏,进而丧失了原有功能。蛋白质及核酸的这种变形可以是可逆的,也可以是不可逆的。虽然无所谓动能行结构被破坏,若氢键破裂的数量未达到微生物死亡的临界值,则其分子很可能恢复到它原有的形式,微生物就没有被杀死。为有效地使蛋白质变形,如采用高压蒸汽灭菌时,就需要水蒸气有足够的温度和持续时间,这对灭菌效果十分重要。高温饱和水蒸气可迅速使蛋白质变形,在规定操作条件下,蛋白质发生变形的过程即微生物死亡的过程,是可预见和重复的。微生物的灭活符合一级动力学方程,微生物死亡速率是微生物耐热参数D和杀灭时间的函数。即在给定的时间下被灭活的微生物与仍然存活数成正比。
LgNt=lgNo-F(T,Z) /DT
Nt:t分钟后微生物计数值;
No:初始微生物计数值;
DT:在T温度下的微生物降低一个对数单位所需要的时间,分钟;
F(T,Z):灭菌程序在确定温度系数Z的T温度的等效灭菌时间。
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湿热灭菌法是指用饱和水蒸气、沸水或流通蒸汽进行灭菌的方法,由于蒸汽潜热大,穿透力强,容易使蛋白质变性或凝固,所以该法的灭菌效率比干热灭菌法高,是药物制剂生产过程中最常用的灭菌方法。湿热灭菌法可分为:煮沸灭菌法、巴氏消毒法、高压蒸汽灭菌法、流通蒸汽灭菌法、和间歇蒸汽灭菌法。
影响湿热灭菌的主要因素有:微生物的种类与数量、蒸汽的性质、药品性质和灭菌时间等。
(1)煮沸灭菌法:将水煮沸至100摄氏度,保持5-10分钟可杀死细菌繁殖体,保持1-3小时可杀死芽胞。在水中加入百分之一至百分之二的碳酸氢钠时沸点可达105摄氏度,能增强杀菌作用,还可去污防锈。此法适用于食具、刀箭、载玻片及注射器等。
(2)巴氏消毒法:一种低温消毒法,因巴斯德首创而得名。有两种具体方法,一是低温维持法:62摄氏度维持30分钟;二是高温瞬时法:75摄氏度作用15-30秒。该法适用于食品的消毒。
(3)流通蒸气灭菌法:利用常压下的流通蒸汽进行灭菌。
(4)间歇蒸汽灭菌法
(5)高压蒸汽灭菌法:103.4千帕蒸汽压温度达121.3摄氏度,维持15-20分钟。
湿热灭菌法 湿热法可在较低的温度下达到与干热法相同的灭菌效果,因为:①湿热中蛋白吸收水份,更易凝固变性;②水分子的穿透力比空气大,更易均匀传递热能;③蒸汽有潜热存在,每1克水由气态变成液态可释放出529卡热能,可迅速提高物体的温度。
湿热灭菌法一般采用121摄氏度,灭菌20-30min,如果是产孢子的微生物则应采用灭菌后适宜温度下培养几小时,再灭菌一次,以用于杀死刚刚萌发的孢子。
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