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蟾酥复方消毒剂性能的实验观察
http://www.100md.com 《中国消毒学杂志》 1998年第1期
     作者:孟庆增 袁青春 姚元庚 侯秀丽 陈 璐

    单位:山东省卫生防疫站, 济南 250014

    关键词:中药消毒剂;蟾酥;苦参;蛇床子;稳定性;杀菌作用;金黄色葡萄球菌;大肠杆菌;白色念珠菌;毒性

    中国中国消毒学杂志980107

    提要 蟾酥复方消毒剂含蟾酥、苦参、蛇床子等成分。以其含67.5%原液的水溶液对白色念珠菌作用2 min,以其含45.0%者对金黄色葡萄球菌作用5 min,对大肠杆菌作用10 min,杀灭率均达100%。杀菌效果随温度升高而增加,受20%小牛血清影响较小。室温下贮存10个月,杀菌效果无明显变化。无毒,基本无刺激。

    用蟾酥、苦参、蛇床子等中药提取液配制成蟾酥复方消毒剂(pH 6.5~7.2),在实验室对其杀灭微生物效果与毒性等进行了实验观察。现将结果报道如下。
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    1 方法

    1.1 菌悬液的制备

    试验菌为金黄色葡萄球菌(ATCC 6538)、大肠杆菌(8099)、白色念珠菌(ATCC 10231)与枯草杆菌黑色变种(ATCC 9372)芽胞。用含1%蛋白胨的磷酸盐缓冲液,将其分别制成含菌量为10 7 cfu/ml的菌悬液。

    1.2 中和剂选择试验

    试验菌为金黄色葡萄球菌。试验分组为:① (消毒剂+菌液)+中和剂;② 中和剂+菌液;③ (消毒剂+中和剂)+菌液;④ 消毒剂+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2、3、5组组间菌数差不超过10%,第1组长菌,第4组不长菌或菌数远少于第1组时所用中和剂及其浓度适宜。

    1.3 定量杀菌试验
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    试验用悬液法进行。取0.5 ml菌悬液与4.5 ml消毒液混匀。作用至预定时间,取0.5 ml菌药混合液,加入盛有4.5 ml中和剂溶液的试管内,混匀。中和作用10 min,作活菌计数,计算杀灭率。

    1.4 稳定性试验

    将玻璃瓶装消毒液放置于室温下避光处。于放置前与放置后,分别用悬液法进行定量杀菌效果检测。

    1.5 毒性试验

    急性经口毒性试验用昆明小白鼠进行。以消毒液灌胃,测LD 50 值。皮肤刺激和粘膜刺激试验用新西兰标准试验用兔进行。皮肤刺激试验时,将兔背部脊柱两侧毛分别剪掉3 cm×2 cm,不损伤表皮。将染消毒液纱布敷贴于一侧皮肤上,另一侧作对照。眼粘膜刺激试验系将消毒液滴入兔一侧眼结膜囊内,另一侧眼作为对照。试验动物均由山东省实验动物中心提供。
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    2 结果

    2.1 中和剂试验结果

    经对不同复方中和剂进行比较,含0.5%硫代硫酸钠、0.07%卵磷脂与2.0%吐温80的磷酸盐缓冲液可完全中和蟾酥复方消毒剂原液(表1)。

    2.2 杀菌效果

    以蟾酥复方消毒剂原液浓度为100%,用其45.0%浓度对金黄色葡萄球菌作用5 min,对大肠杆菌作用10 min,以其67.5%浓度对白色念珠菌作用2 min,杀灭率均达100%(表2)。而以其原液在20 ℃对枯草杆菌黑色变种芽胞作用2 h,杀灭率仅14.4%。

    表1 蟾酥复方消毒剂中和试验结果

    Table 1 Results of neutralization test of compound toad venom disinfectant 组 别 Group
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    平均菌数 Average bac-terial count (cfu/ml)

    (消毒剂+菌液)+中和剂

    (Disinfectant+bacterial suspension)+ neutralizer

    158

    中和剂+菌液

    Neutralizer+bacterial suspension

    1 650 000

    (消毒剂+中和剂)+菌液

    (Disinfectant +neutralizer) +bacterial suspension
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    1 610 000

    消毒剂+菌液

    Disinfectant+bacterial suspension

    0

    正常菌对照

    Normal bacterial control

    1 740 000

    未接种菌的培养基对照

    Uninoculated culture medium control

    0

    注: 试验菌为金黄色葡萄球菌。试验温度为20℃。消毒剂 为原液,加菌液后作用1 min。加中和剂后作用10 min。结果为3次试验平均值。
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    Note: The test bacterium was Staphylococcus aureus. The temperature was 20℃. The disinfectant was stock solution. The contact time after addition of bacterial suspension was 1 min and that after addi- tion of neutralizer was 10 min. The results were means of 3 tests.

    2.3 温度与有机物对杀菌效果的影响

    2.3.1 温度 试验表明,在15~30℃,浓度45.0%该消毒液对白色念珠菌的杀灭率随温度升高而增加(表3)。

    表2 蟾酥复方消毒剂对微生物的杀灭效果

, 百拇医药     Table 2 Efficacy of compound toad venom disin- fectant in killing microorganisms 消毒液浓度Concentration ofdisinfection solution(%)

    作用不同时间(min)的杀灭率(%)Killing rate (%) after exposure fordifferent periods of time (min)

    2

    5

    10

    金黄色葡萄球菌

    Staphylococcus aureus

    22.5
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    50.464

    54.224

    58.724

    45.0

    99.999

    100.00

    100.00

    90.0

    100.00

    100.00

    100.00

    大肠杆菌

    Escherichia coli
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    45.0

    99.896

    99.999

    100.00

    90.0

    100.00

    100.00

    100.00

    白色念珠菌

    Candida albicans

    45.0

    66.372
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    86.382

    95.593

    67.5

    100.00

    100.00

    100.0

    注: 试验温度为20℃。阳性对照组平均菌数金黄色葡萄 球菌为1 100 000 cfu/ml,大肠杆菌为1 300 000 cfu/ml,白色念珠菌为1 300 000 cfu/ml。结果为 5次试验平均值。

    Note: The temperature was 20℃. The average bacterial counts of Staphylococcus aureus, Escherichia coli and Candida albicans were 1 100 000 cfu/ml, 1 300 000 cfu/ml and 1 300 000 cfu/ml respectively in positive control group. The results were means of 3 tests.
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    2.3.2 有机物

    试验结果,20%小牛血清对浓度45.0%该消毒液对白色念珠菌的杀灭率影响较小(表4)。

    2.4 稳定性

    于室温下避光放置10个月,该消毒液未出现浑浊与沉淀。经检测,对白色念珠菌的杀灭率无明显变化(表5)。

    表3 温度对蟾酥复方消毒剂杀灭白色念珠菌效果的影响

    Table 3 Influence of temperature on efficacy of compound toad venom disinfectant in kill- ing Candida albicans 温 度 Temperature (℃)

    作用不同时间(min)的杀灭率(%) Killing rate (%) after exposure for different periods of time (min)
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    2

    5

    10

    15

    57.81

    67.42

    79.19

    20

    72.39

    80.34

    92.74

    25

    86.68
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    94.41

    98.86

    30

    97.37

    99.74

    99.99

    注: 阳性对照组平均菌数为2 300 000 cfu/ml。消毒液浓 度为45.0%。结果为5次试验平均值。

    Note: Average bacterial count of positive control group was 2 300 000 cfu/ml. The concentration of disin- fectant solution was 45.0%. The results were means of 5 tests. 表4 有机物对蟾酥复方消毒剂对白色念珠菌杀灭 效果的影响
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    Table 4 Influence of organic substance on effi- cacy of compound toad venom disinfectant in killing Candida albicans 小牛血清 浓度Concentrationof calfserum(%)

    阳性对照组平均菌数Average bac-terial count ofpositive controlgroup (cfu/ml)

    作用不同时间(min)的杀灭率(%) Killing rate (%) afterexposure for different periods of time (min)

    2

    5

    10
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    0

    2240000

    66.37

    86.38

    95.59

    10

    2570000

    62.89

    85.43

    94.28

    20

    1970000

    59.66
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    85.38

    93.44

    注: 试验温度为20℃。消毒液浓度为45.0%。结果 为5次试验平均值。

    Note: The temperature was 20℃. The concentration of disinfectant solution was 45.0%. The results were means of 5 tests.

    2.5 毒性

    经测定,该消毒剂原液的小白鼠经口LD 50 > 10 000 mg/kg,属实际无毒类物质。皮肤刺激试验表明,兔皮肤未出现红肿和水肿。对皮肤属无刺激性物质。兔眼粘膜在滴消毒液后6 h内出现充血、水肿及分泌物,4 d后恢复正常。对粘膜属轻度刺激类物质。
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    表5 贮存前后蟾酥复方消毒剂对白色念珠菌杀灭效果的比较

    Table 5 Comparison of efficacy of compound toad venom disinfectant before and after storage in killing Candida albicans 贮存时间(月)Storingtime(month)

    对照组平均菌数Average bacterialcount ofcontrol group(cfu/ml)

    作用不同时间(min)杀灭率(%)Killing rate (%) after exposure for differentperiods of time (min)

    2

    5
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    10

    0

    224000

    66.37

    86.38

    95.59

    10

    2300000

    65.87

    80.34

    92.74

    注: 试验温度为20℃。消毒液浓度为45.0%。结果 为5次试验平均值。
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    Note: The temperature was 20℃. The concentration of disinfectant solution was 45.0%. The results were means of 5 tests.

    3 结语

    结果表明,浓度45.0%的蟾酥复方消毒剂对金黄色葡萄球菌、大肠杆菌作用5 min,浓度67.5%的溶液对白色念珠菌作用2 min,杀灭率为99.999%~100%;其原液对枯草杆菌黑色变种芽胞作用2 h,仍无明显杀灭效果。在15~30℃,温度对其杀菌效果影响较明显。20%小牛血清对其杀灭白色念珠菌的效果影响较小。于室温下放置10个月,该消毒剂杀菌作用无明显减弱。该消毒剂无毒,对皮肤基本无刺激,对粘膜有轻度刺激。

    EXPERIMENTAL OBSERVATION ON PERFORMANCE OF COMPOUND TOAD VENOM DISINFECTANT
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    Meng Qingzeng Yuan Qingchun Yao Yuangeng Hou Xiuli Chen Lu ( Shangdong Provincial Sanitary and Anti-epidemic Station, Jinan 250014)

    Abstract

    Compound toad venom disinfectant was prepared from Chinese drugs such as toad venom, flavescent sophora root and cnidium fruit. Its pH value is 6.5~7.2. Its efficacy in killing microorganisms, stability and toxicity were observed in laboratory. The killing rates of Candida albicans exposed to its solution containing 67.5% stock solution for 2 min, Staphylococcus aureus exposed to its solution containing 45.0% stock solution for 5 min and Escherichia coli exposed to this solution for 10 min were 100%. Its stock solution with a 2 hr contact time had no significant killing effect on spores of Bacillus subtilis var. niger. At 15~30℃, its germicidal efficacy increased with increasing temperature. 20% calf serum had little influence on its germicidal efficacy. No obvious change in its germicidal efficacy was observed after storage at room temperature for 10 months. This disinfectant is nontoxic, essentially not irritative to skin and slightly irritative to mucous membrane.

    Key words Chinese drug disinfectant toad venom flavescent sophora root cnidium fruit Stabilit germicidal effect Staphylococcus aureus Escherichia coli Candida albicans toxicity

    (1997-02-05收稿 1997-05-28修回), http://www.100md.com


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