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OK器械消毒液杀灭微生物效果CJ[100]与腐蚀性的试验观察
http://www.100md.com 《中国消毒学杂志》 1999年第1期
     作者:陈越英 顾健 吴小成 周士新 张晓东

    单位:江苏省卫生防疫站,南京 210009

    关键词:戊二醛复方消毒液;杀菌作用;HBsAg;大肠杆菌;金黄色葡萄球菌;细菌芽胞;枯草杆菌黑色变种;腐蚀性;稳定性

    中国消毒学杂志9901003 提要 OK器械消毒液为含2%戊二醛的复方消毒剂。对枯草杆菌黑色变种芽胞作用3h,杀灭率为100%,作用10min可将HBsAg抗原性破坏。杀菌效果随温度升高而增强,随pH下降、悬液中含有机物、复方中去除亚硝酸钠而减弱。所试金属,除对铝有轻度腐蚀外均基本无腐蚀。存放于54~57℃ 14d,戊二醛浓度下降2.97%。

    EXPERIMENTAL OBSERVATION ON GERMICIDAL EFFICACY AND CORROSIVENESS OF OK INSTRUMENTdISINFECTION SOLUTION
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    Chen Yueying Gu Jian Wu Xiaocheng Zhou Shixin Zhang Xiaodong

    (Jiangsu Provincial Sanitary and Anti-epidemic Station,Nanjing 210009)

    Abstract The OK Instrumentdisinfection Solution is a compounddisinfectant (pH 7.0) containing 2% glutaraldehyde,0.1% compound alcohol,0.4% sodium nitrite and surfactant. The killing rate of the spores of Bacillus subtilis var. niger in suspension exposed to thisdisinfectant for 3h was 100%. Exposure to thisdisinfectant for 3h was 100%. Exposure to thisdisinfectant for 10min coulddestroy the antigenicity ofhBsAg in 5% calf serum. The killing rate of both Escherichia coli Fsexposed to 100-folddilute solution of thisdisinfectant for 15min and Staphylococcus aureus exposed to the same solution for 30min was 100%. Its germicidal efficacy increased with increasing the temperature anddecreased withdecreasing pH,presence of organic substances and removal of sodium nitrite in the compounddisinfectant.It was slightly corrosive to aluminum and essentially non-corrosive to copper,stainless steel and carbon steel. The concentration of glutaraldehyde in its stock solutiondecreased by 2.97% after storing at 54~57℃ for 14days.
, 百拇医药
    Key words compound glutaraldehydedisinfection solution germicidal effect HBsAg Escherichia coli Staphylococcus aureus bacterial spore Bacillus subtilis var. niger&corrosiveness stability

    OK器械消毒液为含2%戊二醛、0.1%复合醇、0.4%亚硝酸钠与表面活性剂的中性(pH 7.0)复方消毒剂。在实验室对其杀灭微生物效果及对金属腐蚀性等进行了试验观察。现将结果报道如下。

    1 方法

    1.1 中和剂选择试验

    试验菌为大肠杆菌(8099)与枯草杆菌黑色变种(ATCC 9372)芽胞。试验分组为:①消毒剂+菌液;②(消毒剂+菌液)+中和剂;③(消毒剂+中和剂)+菌液;④ 中和剂+菌液;⑤ 正常菌对照;⑥ 未接种菌的培养基对照。试验结果,当第6组不长菌,第3、4、5组组间菌数差不超过10%,第1组不长菌或长菌量远低于第2组,第2组长菌量超过100cfu/ml时,表明所用中和剂及其浓度适宜〔1〕
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    1.2定量杀菌试验

    试验采用悬液法。试验菌为枯草杆菌黑色变种芽胞、大肠杆菌(ATCC 25922)、金黄色葡萄球菌(ATCC 6538),均制成含1%蛋白胨的菌悬液。试验时,将0.5ml菌悬液与4.5ml消毒液(对照为磷酸盐缓冲液)混匀。作用至规定时间,取菌药混合液0.5ml,加入盛有4.5ml中和剂的试管中,混匀。中和作用10min,作活菌计数,计算杀灭率。

    1.3乙型肝炎表面抗原破坏试验

    试验用悬液法进行。试验时,以含5%小牛血清的磷酸盐缓冲液将纯化乙型肝炎表面抗原(HBsAg)稀释至浓度为10μg/ml后,取0.1ml与0.4ml消毒液混匀。作用至规定时间,再加0.5ml中和剂。中和作用10min,用ELISA法测OD值。同时设HBsAg阳性与阴性对照,以试验组OD值(S)与阴性对照组OD值(N)的比值(S/N)< 2.1为HBsAg抗原性破坏。
, 百拇医药
    1.4稳定性试验

    将消毒液原包装置54~57℃水浴中2周。于放置前、后,分别取样测戊二醛含量,计算下降率。同时,对放于室温下70d的消毒液进行对枯草杆菌黑色变种芽胞杀灭效果检测。

    1.5金属腐蚀试验

    将不锈钢、碳钢、铝、铜等金属制成直径2.4cm、厚0.1cm的圆片。试验时,每种金属取3片,将不同金属分别经该消毒液浸泡72h,于浸泡前、后称重,取3片平均值计算腐蚀速率(R)〔2〕

    2 结果

    2.1 中和剂试验结果

    试验以含1.0%甘氨酸、0.5%吐温80的磷酸盐缓冲液中和含2%戊二醛的OK器械消毒液(对细菌芽胞)或含0.02%戊二醛的OK器械消毒液(对大肠杆菌),结果均达到所定指标(表1)。 表1 OK器械消毒液中和试验结果
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    Table 1 Results of neutralization test for OK Instrumentdisinfection Solution 组别

    Group

    不同菌平均菌数(cfu/ml)

    Average bacterial count (cfu/ml)

    枯草杆菌黑色变种芽胞

    Spores of B. subtilis var. niger

    大肠杆菌

    E.coli

    消毒剂+菌液
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    Disinfectant+bacterial suspension

    0

    0

    (消毒剂+菌液)+中和剂

    (Disinfectant+bacterial suspension)+neutralizer

    330

    1750

    (消毒剂+中和剂)+菌液

    (Disinfectant+neutralizer)+bacterial suspension

    1220000
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    2950000

    中和剂+菌液

    Neutralizer+bacterial suspension

    1210000

    2970000

    正常菌对照

    Normal bacteria control

    1220000

    2940000

    未接种菌的培养基对照

    Uninoculated culture medium control
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    0

    0

    注: 试验温度为22℃。作用于细菌芽胞的OK器械消毒液含2%戊二醛,作用于大肠杆菌者为OK器械消毒液的1∶100稀释液。加细菌芽胞与大肠杆菌后作用时间分别为2h与15min。加中和剂后作用10min。结果为3次试验平均值。

    Note:The temperature was 22℃. The OK Instrumentdisinfection solution for killing bacterial spores contained 2% glutaraldehyde and that for killing E.coli was 1∶100dilute solution of OK Instrumentdisinfection solution. The exposure time of bacterial spores and E.coli to thedisinfectant was 2h and 15min respectively. The neutralizing time was 10min. The results are means of 3 tests.
, 百拇医药
    2.2 杀菌效果

    试验表明,该消毒液稀释100倍(戊二醛含量为0.02%)的中性溶液对大肠杆菌作用15min,对金黄色葡萄球菌作用30min,杀灭率均达100%(表2)。其原液对枯草杆菌黑色变种芽胞作用3h,杀灭率亦为100%,但如复方中不含亚硝酸钠则杀灭率仅96.00%(表3)。表2 1∶100稀释OK器械消毒液对细菌繁殖体的杀灭效果

    Table 2 Efficacy of 1∶100dilute solution of OK Instrumentdisinfection Solution in killing bacterial vegetative form 作用时间

    Exposure time

    (min)

    不同菌的杀灭率(%)
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    奒illing rate (%)

    大肠杆菌

    E.coli

    金黄色葡萄球菌

    S.aureus

    5

    98.69

    96.96

    10

    99.95

    99.87

    15

    100.00
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    99.95

    30

    100.00

    100.00

    注:试验温度为22℃。对照组菌数大肠杆菌为2 280 000cfu/ml,金黄色葡萄球菌为3 540 000cfu/ml。结果为5次试验平均值。

    Note:The temperature was 22℃. The bacterial count of the positive control group was 2 280 000cfu/ml forE.coli and 3 540 000cfu/ml for S.aureus.The results are means of 5 tests.

    表3 OK器械消毒液对枯草杆菌黑色变种芽胞的杀灭效果
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    Table 3 Efficacy of OK Instrumentdisinfection Solution in killing spores of B.subtilis var.niger

    组别

    Group

    对照组菌数

    作用不同时间(h)的杀灭率(%)

    Killing rate (%) after exposure for

    different periods of time (h)

    Bacterial count of

    control group
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    (cfu/ml)

    1

    2

    3

    复方含亚硝酸钠

    Compounddisinfectent containing sodium nitrite

    2050000

    93.10

    99.96

    100.00

    复方不含亚硝酸钠

    Compounddisinfectant without sodium nitrite
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    2490000

    22.17

    68.51

    96.00

    注:试验温度为22℃。结果为5次试验平均值。

    Note:The temperature was 22℃.The results are means of 5tests. 2.3 对HBsAg的破坏作用经检测,以该复方原液作用10min,可使5%小牛血清中的HBsAg抗原性破坏(表4)。

    表4OK器械消毒液对悬液中HBsAg的破坏作用

    Table 4 Destructive effect of OK Instrumentdisin fection Solution onhBsAg in suspension
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    作用时间

    Exposure time

    (min)

    S/N值

    S/N value

    5

    2.38

    10

    1.81

    15

    1.75

    30

    1.62
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    注:试验温度为22℃。阳性对照S/N值为35.20。结果为5次试验平均值。

    Note:The temperature was 22℃. S/N value of the positive control was 35.20. The results are means of 5 tests.

    2.4 影响杀菌效果因素

    2.4.1pH值经比较,在pH值为6.0~7.8范围内,OK器械消毒液对枯草杆菌黑色变种芽胞的杀灭效果,随溶液pH值上升而增强(表5) 表5 溶液pH对OK器械消毒液杀灭枯草杆菌黑色变种芽胞效果的影响

    Table 5 Influence of pH of the solution on the efficacy of OK Instrumentdisinfection Solution in killing spores of B.subtilis var.niger pH值
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    pHvalue

    对照组菌数

    Bacterialcount

    of control

    group

    (cfu/ml)

    作用不同时间(h)

    的杀灭率(%)

    Killing rate (%) after

    exposure fordifferent periods

    of time (h)
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    1

    2

    3

    6.0

    2490000

    23.69

    66.37

    95.37

    7.0

    2050000

    93.10

    99.96

    100.00
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    7.8

    930000

    99.86

    99.99

    100.00

    注:消毒液戊二醛浓度为2%。试验温度为22℃。结果为5次试验平均值。

    Note:The concentration of glutaraldehyde in the disinfectant was 2%. The temperature was 22℃. The results are means of 5 tests. 2.4.2有机物以代表有机物,当枯草杆菌黑色变种芽胞悬液中小牛血清含量为10%~20%时,对该消毒液作用2h的杀菌效果有所影响,但对作用3h者则无(表6)。
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    表6 有机物对OK器械消毒液杀灭枯草杆菌黑色变种芽胞效果的影响

    Table 6 Influence of organic aubstance on the efficacy of OK Instrumentdisinfection Solution in killing spores of B.subtilis var.niger

    小牛血清含量

    Calf serum content(%)

    对照组

    菌数

    Bacterial count

    of positive
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    control group(cfu/ml)

    作用不同时间的杀灭率(%)

    Killing rate (%) after

    exposure fordifferent

    periods of time(h)

    2

    3

    0

    3140000

    99.99

    100.00
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    10

    553000

    99.49

    100.00

    20

    603000

    99.16

    100.00

    注:消毒液戊二醛浓度为2%。试验温度为22℃。结果为5次试验平均值。

    Note:The concentration of glutaraldehyde in thedisin fecfant was 2%. The temperature was 22℃. The results are means of 5 tests. 2.4.3温度试验表明,在温度分别为22℃、24~26℃、30℃下,该复方原液对菌数为2680000cfu/ml的枯草杆菌黑色变种芽胞作用2h,杀灭率分别为99.96%、99.99%与99.99%。
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    2.5戊二醛复方消毒液稳定性

    经检测,该消毒液以原包装存放于54~57℃下14d,戊二醛含量由2.02%减至1.96%,下降率为2.97%。

    将该消毒液于室温(18~38℃)下存放70d,对枯草杆菌黑色变种芽胞杀灭率达100%的作用时间需由原3h延长至5h。

    2.6 对金属的腐蚀性

    经试验,该消毒液除对铝有轻度腐蚀(R=0.0138mm/a)外,对碳钢、不锈钢和铜均为基本无腐蚀(R≤0.0037mm/a)。

    3 结语

    悬液试验结果,OK 器械消毒液对枯草杆菌黑色变种芽胞作用3h,杀灭率可达100%;对5%小牛血清中HBsAg 作用10min,可将其抗原性破坏。其100 倍稀释液(含0.02%戊二醛)对大肠杆菌作用15min,对金黄色葡萄球菌作用30min,杀灭率均为100%。在试验范围内,该消毒剂的杀菌效果随作用时间延长、溶液pH值上升、温度升高而增强,有机物存在与复方中不含亚硝酸钠均可使杀菌作用减弱。OK 器械消毒液存放于54~57℃下14d,戊二醛浓度下降2.97%。该液对铝有轻度腐蚀,对铜、碳钢、不锈钢基本无腐蚀。

    参考文献

    〔1〕 陈胤忠,袁朝森,胡琳,等. 过氧化氢复方消毒剂杀灭微生物效果的实验观察. 中国消毒学杂志,1998; 15 (2): 65.

    〔2〕李秋丽,袁洽. 戊二醛对金属腐蚀性的实验观察. 中国消毒学杂志,1997;14 (2):91.

    (1997-12-23收稿 1989-09-16修回), http://www.100md.com