Microtox中药注射剂微毒测试体系明亮发光杆菌502和青海弧菌Q67发光反应条件的比较研究  

Reaction conditions comparison of Photosbacterium phosphoreum 502 and Vibrio qinghaiensis Q67 in Microtox-based fast testing system for traditional Chinese medicine injection

作　　者：熊静悦[1,2] 李孝容 鄢良春[2] 赵军宁[2] 

机构地区：[1]成都中医药大学,成都610072 [2]四川省中医药科学院.国家中医药管理局中药质量生物评价重点研究室.四川省道地药材系统开发工程技术研究中心.中药品质评价与创新中药研究四川省重点实验室,成都610041

出　　处：《中药药理与临床》2016年第2期227-230,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金："重大新药创制"科技重大专项项目(2015ZX09501004-001-005);国家自然科学基金项目(81470180);四川省科技计划项目(2016SZ0031)

摘　　要：目的:基于明亮发光杆菌502和青海弧菌Q67的Microtox微毒测试体系比较研究,明确两者适宜的反应条件。方法:1考察明亮发光杆菌502和青海弧菌Q67在15℃±1℃、20℃±1℃、25℃±1℃下的相对发光情况;2采用Plackett-Burman法优化设计影响明亮发光杆菌502和青海弧菌Q67发光的因素;3采用Plackett-Burman法优化得到的反应体系,考察不同p H复苏液对上述两种细菌相对发光强度的影响。结果:1明亮发光杆菌502的相对发光强度随温度的升高而增强,而青海弧菌Q67的相对发光强度随温度的升高先增强后减弱。2Plackett-Burman法得到最优发光条件为:明亮发光杆菌502冻干粉平衡10min后加入复苏液2ml,混匀15min,取100μl菌液测试初始发光度,随后立即加入1ml复苏液或待测样品,反应15min后再次测试相应发光强度;青海弧菌Q67冻干粉平衡10min后加入复苏液2ml,混匀10min,取100μl菌液测试初始发光度,随后立即加入2ml复苏液或待测样品,反应10min后再次测试相应发光强度。3复苏液p H 3.6-p H 3.8时对明亮发光杆菌502发光的影响由抑制转为增强;复苏液p H 4.5-p H 5.0时对青海弧菌Q67发光强度的影响<±15%。结论:与明亮发光杆菌502比较,青海弧菌Q67具有更宽的p H耐受范围并且在检测时无需调节待测样品渗透压。Objective： Microtox assay system based on Photosbacterium phosphoreum 502（ P. p. 502） and Vibrio qinghaiensis Q67（ V. q. Q67）were compared to definite suitable reaction conditions. Method： 1 The relative luminous intensity of P. p. 502 and V. q. Q67 were investigated under 15℃ ± 1℃,20℃ ± 1℃ and 25℃ ± 1℃. 2The Plackett- Burman trials were carried out to optimize the agents which affected P. p. 502 and V. q. Q67. 3 The p H ranges suitable for each assay system was also investigated. Results： 1 With increased temperature,relative luminous intensity enhanced in P. p. 502 while enhanced first and then fell in V. q. Q67. 2 At a temperature of 15℃ ± 1℃,P. p.502 vials were placed at a thermostat for 10 min,then,2ml rehydration solution（ 3% Na Cl solution） was pour quickly into each vial and blended them into suspensions. After a waiting time of 15 min,added 100 μl bacteria suspension into the test tube to measure the luminescence intensity,immediately before the addition of 1 ml rehydration solution or test samples. The same time intervals（ 10s） were used for later intensity measurement. After a contact time of 15 min,the luminescence intensity of each vial was measured again. At a temperature of15℃ ± 1℃,V. q. Q67 vials were placed at a thermostat for 10 min,then,2 ml rehydration solution（ 0. 85% Na Cl solution） was pour quickly into each vial and blended them into suspensions. After a waiting time of 10 min,added 100 μl bacteria suspension into the test tube to measure the luminescence intensity,immediately before the addition of 2 ml rehydration solution or test samples. The same time intervals（ 10s） were used for later intensity measurement. After a contact time of 10 min,the luminescence intensity of each vial was measured again.3 p H 3. 6- p H 3. 8 reversed luminescence intensity of P. p. 502 from inhibition to enhancement. p H 4. 5- p H 5. 0 inhibited lower than ±15% luminescence intensity of V. q. Q67. The suitable reaction conditions of the two Microtox as

关 键 词：明亮发光杆菌502 青海弧菌Q67 微毒测试 

分 类 号：R285[医药卫生—中药学]