抗生素微生物检定法

抗生素微生物检定法 一、目 的:阐述抗生素微生物的检定法。 二、适用范围:适用于抗生素微生物检定法。 三、责 任 者:品控部。 四、正 文: 1 简述 抗生素微生物检定法，依试验设计原理不同，可分为稀释法、比浊法和琼脂扩散法。后二者被列为抗生素微生物检定的国际通用方法。中国兽药典采用管碟琼脂扩散法。 抗生素管碟检定法是利用抗生素在摊布特定试验菌的固体培养基内球面形扩散，形成含一定浓度抗生素球形区，抑制了试验菌的繁殖而呈现出透明的抑菌圈。此法系根据抗生素在一定浓度范围内，对数剂量与抑菌圈直径(面积)呈线性关系，通过比较标准品与供试品产生抑菌圈的大小，计算出供试品的效价。 抗生素效价以“单位(u)”或“微克(ug)”示。 2 仪器与用具 2.1 操作室 光线明亮，操作室应分为两部分，彼此分开;一部分为一般操作间，一部分为半无菌操作间。半无菌操作间，设有紫外线灯达到空气消毒。应装有空调设备，控制室温在20-25?。操作台可用稳固的水泥台，台面用玻璃板垫平，用水平仪校准，保持水平。室内注意抗生素的污染。 2.2 双碟 为硬质玻璃或塑料培养平皿，内径约90mm,高16-17mm,碟底厚薄均匀水平,无气泡.碟底平度检查,可将双碟放在水平台上,下垫一张白纸,碟内加2-3ml,再滴加蓝墨水,观察蓝色深汪是否一致.用过的双碟经高压灭菌倒出培养基后,置玻璃仪器专用洗液或其他清洗液中浸泡过夜,冲洗,沥干,置150-160?干热灭菌2小时或高压121?蒸气灭菌30分钟，备用。 2.3 陶瓦盖 内径约为103mm，外径108 mm，平坦，吸水性强，应定期干燥、清洗。 2.4 钢管 内径(6.0?0.1)mm,高(10.0?0.1) mm，外径(8.0?0.1)或(7.8?0.1)mm，每套钢管重量差异不超过?0.05g，内外壁及两端面光洁平坦，管壁厚薄一致。每次使用后应置1:1000新洁尔灭溶液内，浸泡2小时以上，进行灭菌后再行洗涤，先用水洗涤，超声波超声30分钟或用沾有去污粉的纱布条擦内外壁，用水冲洗，淋干，再用蒸馏水冲洗3次后，置带盖的溶器内，在150-160?干热灭菌2小时备用。 pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 2.5 钢管放置器 置于半无菌操作间内，有6孔和4孔两种，钢管下落时应垂直平稳、位置正确，双碟升降平稳。应保持清洁，防止抗生素污染。可定期用75%乙醇棉擦试，并用乙醇棉炎焰烧小孔。置钢管的玻璃管定期干烤灭菌。 2.6 恒温培养箱 隔水式为宜，35-37?及24-26?，隔板上可备有带孔的玻璃板并垫平。 2.7 灭菌刻度吸管 吸取菌液及培养基用。试验用后应立即置5%石炭酸或者0.2%新洁尔灭溶液中消毒后再按玻璃容器常规洗涤。吸口处塞入脱棉(应松动，透气)，置适宜容器中，120?以上干热灭菌2小时或121?蒸气灭菌30分钟，烘干备用。 2.8 玻璃容器包括滴定管、移液管、刻度吸管、容量瓶等，按“玻璃器皿检定规程”进行标定，要符合一等品规定，每次应用前用清洁液浸泡，水冲洗、蒸馏水冲洗3次，淋干。 pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 德信诚培训网 2.9 称量瓶 具塞，重量在10 g以下。用毕先用水冲洗、淋干，置清洁液浸泡2小时以上，水先、蒸馏水冲洗3次，置洁净的大平皿中，在120?干热3小时，待冷至70-60?时，取出置于干燥器中备用。 2.10 毛细滴管 用玻璃管拉制，管口光滑。应用前清洁液浸泡、水冲洗、蒸馏水冲洗3次，置适宜容器中，在120?干燥3小时备用。 2.11 天平 分析天平，感量0.1mg。 2.12 抑菌圈直径(面积)测量仪。 2.12.1 可用CHB型抑菌圈面积测量仪及适当的测试仪。仪器必须按抑菌圈测量仪检验规程检验，符合规定者方能应用。 2.12.2 游标卡尺 精度0.05 mm，长度125 mm。 2.13 超净工作台 用于菌种的接各或传代。超净工作台须置洁净工作室或半无菌室内。 3 试液 3.1 灭菌缓冲液 制备缓冲液的试剂应为分析纯，配制后的缓冲液应澄明，分装于玻璃容器内，经121?蒸气灭菌30分钟备用。 3.1.1 磷酸盐缓冲液(PH6.0)取磷酸氢二钾2g与磷酸二氢钾8g，加水使成1000ml，滤过。 3.1.2 磷酸盐缓冲液(PH7.0)取磷酸氢二钾9.39g与磷酸二氢钾3.5g，加水使成1000ml，滤过。 3.1.3 磷酸盐缓冲液(PH7.8)取磷酸氢二钾5.59g与磷酸二氢钾0.41g，加水使成1000ml，滤过。 3.1.4 磷酸盐缓冲液(PH0.5)取磷酸氢二钾35g，加氢氧化钾液(10mol/L)2ml，加水使成1000ml，滤过。 4 培养基 培养基中原料的质量对抑菌圈边缘清晰度及试验结果的精确度影响较大，因此应对原材料进行预试验，挑选适当的品牌使用。制成的培养基不应有沉淀，如产生沉淀，可在配制培养基后，置115?，20分钟溶化，趁热用纸浆减压或适宜方法过滤，调整PH值，分装灭菌备用。 pipe inspection) shower faucet deviation shall not exceed 更多免费资料下载请进:? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair sl好好学习社区ot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 根据中国兽药典2005年版，抗生素微生物检定法中选用不同处方的培养基。目前市场已有干燥培养基商品生产、供应。临用时按照使用说明书进行配制，但注意培养基的PH，应符合规定，否则必须校正后，灭菌备用。 5 试验用菌种 检定用标准菌种，由中国药品生物制品检定所提供，为冷冻干燥菌种。 5.1 菌种的复苏与保存 在无菌室或超净工作台内进行。 5.1.1 菌种的复苏 5.1.1.1 把冻干菌种管、灭菌1 ml毛细滴管、双碟、镊子、营养琼脂斜面数支，移入接种室或超净工作台。 5.1.1.2 将冻干菌种管外壁用磺酒擦洗消毒、稍干，用75%乙醇棉擦净，放在灭菌双碟内，待干。点燃酒精灯，将菌种管的封口一端，使骤冷而炸裂。 5.1.1.3 取灭菌镊子，在火焰旁，将炸裂的管口打开，放入灭菌双碟内，另取1支灭菌毛细滴管，在火焰旁吸取营养肉汤少许，加至菌种管底部，将冻干菌块搅动促使溶解，随即吸出管内菌液，分别接种至营养琼脂斜面及普通肉汤内，并将毛细滴管及菌种管投入消毒液内，将已接种的营养肉汤及营养琼脂斜面置35-37?培养22-24小时。 5.1.1.4 取出培养物，仔细观察菌苔形态、有无杂菌、涂片、革兰氏染色镜检，呈典型菌落后，转种植代即可应用。如发现菌形不典型，可进行平板分离单菌落。 5.1.2 菌种的接种与保存 5.1.2.1 准备需用的培养基，培养基应新鲜制备，如斜面已无冷凝水者，不宜再使用，标签一注明菌名用接种日期。自冰箱取出的菌种斜面，应在室温放置约30分钟，待温度平衡后再移入接种室或超净工作台。 5.1.2.2 点燃酒精或其他灯，在左手握住菌种斜面，将管口靠近火焰旁，右手拿接种棒后端，将接种环烧红30秒种，随后将全部接种棒金属部分在火焰上烧灼，往返通过3次。右手用无名指、小指及掌部夹住棉塞，左手将管口在火焰上旋转烧灼，右手再轻轻拨开棉塞，将接种环伸入管内先在近壁的琼脂斜面上靠一下，稍冷却再移至菌苔上，刮取少量菌苔，随即取出接种棒，并将菌种管口移至火焰旁。堵上棉塞，左手将菌种管放下，取营养琼脂斜面1支，照上述操作打开棉塞，将接种环但入管内至琼脂斜面的底部，由底向上，将接种环轻贴斜面的表面曲折移动，使细菌划在斜面的表面上。 5.1.2.3 取出接种棒，在火焰旁将培养基管用棉塞堵上，然后将接种过细菌的接种棒在火焰上烧灼灭菌。 5.1.2.4 将已接种毕的细菌管置35-37?培养22-24小时，霉菌管一般置24-25?霉菌pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 德信诚培训网 培养箱内培养7日。取出后放入冰箱保存，一般1-3个月转种一次。 5.2 菌悬液的制备 5.2.1 枯草芽孢杆菌(Bacillus subtilis CMCC(B)63501)悬液取枯草芽杆菌工作用菌种营养琼脂斜面培养物加灭菌水1-2ml将菌苔洗下，制成悬液，用吸管将此悬液接种至盛有营养琼脂培养基的扁培养瓶内，均匀摊布，在35-37?培养7日。取菌苔少许涂片，革兰氏染色镜检，应有芽孢85%以上，用灭菌水10ml将芽孢洗下，制成芽孢悬液，合并至灭菌大试管内，在70-75?水浴内加热30分钟将菌体杀死，待冷后放冰箱贮藏为浓菌液。 日常用菌悬液取上述浓菌液，用灭菌水1:3稀释至灭菌试管中，冰箱保存备用。 5.2.2 短小芽孢杆菌[Bacillus pumilus aureus CMCC(B)26003]悬液取短小芽孢杆菌工作用菌种营养琼脂斜面培养物，照上述方法制备芽孢悬液。 5.2.3 金黄色葡萄球菌[Staphylococcus aureus CMCC(B)26003]悬液取金黄色葡萄球菌工作用菌种的营养琼脂斜面培养物，用接种环取菌苔少许接种至营养琼脂斜上，在35-37?培养20-22小时，临用时，用灭菌水将菌苔洗下，制成悬液。此液在冰箱保存，可使用日。 5.2.4 藤黄微球菌[Micrococcus luteus CMCC(B)28001]悬液取藤黄微球菌工作菌种的营养琼脂斜面培养物，用1ml培养基?将菌苔洗下，用吸管移至盛有营养琼脂培养基的扁培养瓶中，将菌液布满培养基表面，将培养瓶反放于培养箱中26-27?培养24小时后，取出用吸管吸取培养基?5ml，至培养瓶中，将菌苔洗下，合并菌液至灭菌大管中备用。该菌液在冰箱中保存，可使用1-2月。 5.2.5 大肠杆菌[Escherichia coli CMCC(B)441033]悬液取大肠杆菌工作用菌种的营养琼脂斜面培养物，照金黄色葡萄球菌悬液制备方法进行，此菌液只供当天使用。 试验菌的菌龄对抑菌圈边缘清晰度有一定影响，保持菌种的新鲜，对易变的菌株如藤黄微球菌等在制备菌液前进行单菌落的分离，选择典型菌落以保持悬液中菌群的一致性，所得抑菌圈边缘清晰、整齐。 6 操作方法 6.1 称量 6.1.1 称量前，将标准品从冰箱取出，使与室温平衡;供试品应放于干燥器内至少30分钟方可称取。 6.1.2 称量供试品与标准品应用同一天平;吸湿性较强的抗生素，在称量前1-2小时更换天平内干燥剂。 6.1.3 标准品与供试品的称量最好一次取样称取，不得将已取出的标准品或供试品倒pipe inspection) shower faucet deviation shall not exceed 更多免费资料下载请进:? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair sl好好学习社区ot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 回听容器内，标准品称量不可少于20mg，取样后立即将称量瓶或适宜的容器及被称物盖好，以免吸水。 称样量的计算: VC W= P 式中 W为需称取标准品或供试品的重量(mg); V为溶解标准品或供试品制成浓溶液时用容量瓶的体积量 C为标准品或供试品的浓度(单位µ/ ml或µg/ml); P为标准品的纯度或供试品的估计效价(单位µ/mg或µg/mg) 6.2 稀释 稀释操作应遵照容量分析的操作规程。 6.2.1 从冰箱中取出的标准品溶液，必须先在室温放置，使其温度达到室温后，方可量取。 6.2.2 标准品与供试品溶液的稀释应采用容量瓶，每步稀释，取样量不得少于2 ml为宜，稀释步骤一般不超过3步。 举例:取浓溶液1000µ/ ml。 第一步:取5ml(1000µ/ ml)?50ml量瓶中?100u/ml。 第二步:取5ml(100µ/ ml)?50ml量瓶中?10u/ml(H)。 取5ml(100µ/ ml)?100 ml量瓶中?5u/ml(L) 6.2.3 每次吸取溶液用密刻度玻璃吸管，量取溶液前要用被量液流洗吸管2,3次，吸取样品溶液后，用滤纸将外壁多余液体擦去，从起始刻度开始放溶液。 6.2.4 稀释标准品与供试品用的缓冲液应同一批和同瓶，以免因PH或浓度不同影响测定结果。 6.2.5 稀释时，每次加液至容量瓶近刻度前，稍放置片刻，待瓶壁的液体完全流下，再准确补加至刻度。 6.2.6 (2.2)法，标准品与供试品高、低溶液浓度之比为2:1或4:1。(3.3)法，标准品与供试品高、中、低溶液浓度之比为1:0.8。但所选用的浓度必须在剂量反应直线范围内。 6.3 双碾的制备在半无菌室内进行，应注意微生物及抗生素的污染，培养基应在水浴中或微波炉中融化，避免直火加热。 6.3.1 底层用灭菌大口吸管(20ml)或其它灭菌分装器,吸取已融化的培养基20ml注入双碟内，等凝固后更换干燥的陶瓦盖，放于35-37?培养箱中保温，使易于摊布菌层。 6.3.2 菌层 取出试验用菌悬液，按已试验妥的菌量[(2.2)法标准品溶液的高浓度pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 德信诚培训网 所致的抑菌圈直径在18-22mm，(3.3)法标准品溶液的中心浓度所致的抑菌直径在15-18mm]，用灭菌吸管吸取菌悬液加入已融化并保温在水浴中(一般细菌48-50?，芽孢可至60?)的培养基内，摇匀作为菌层用。用灭菌大口10ml吸管或其它分装器，吸取菌层培养基5ml，使均匀摊布在底层培养基上，置水平台上，用陶瓦圆盖覆盖，放置20-30分钟，待凝固，备用。 6.3.3 放置钢管用钢管放置器，将干热灭菌的镊子或适宜方法将钢管装于玻璃管中，放于钢管放置器上，将双碟打开，放入双碟台上，托起双碟台，使钢管平稳落在培养基上，注意使各个钢管下落的高度基本一致。钢管放妥后，应使双碟静置5-10分钟，使钢管在琼脂内稍下沉稳定后，再开始滴加抗生素溶液。 6.4 滴加抗生素溶液 6.4.1 每批供试品取4-10个双碟,滴加溶液用毛细滴管或定量加样器,在滴加之前须用滴加液洗2-3次。 6.4.2 滴加标准品与供试品溶液，因实验设计方法不同而异。(2.2)法,在双碟的4个钢管中分别成对角滴加标准品(S)及供试品(T)高(H)、低(L)两种浓度的溶液。(3.3)法的6个钢管中间隔3个钢管分别滴加标准品及供试品高、中(M)、低3种浓度溶液，并使相同浓度成对角。滴加溶液的顺序:(2.2)法SH?TH?SL?TL;(3.3)法SH?TH?SM?TM?SL?TL。滴加溶液至钢管口平满，注意滴加溶液间隔不可过长，因溶液的扩散时间不同影响测定结果。 6.4.3 每份滴加的溶液为1单位，但双碟数每次不超过5个，如1份溶液滴加双碟数目多，可分次滴加。每种溶液各用1支毛细滴管。 6.4.4 滴加完毕，用陶瓦盖覆盖双碟，平稳置于双碟托盘内，双碟叠放不可超过3个，避免受热不均，影响抑菌圈大小，以水平位置平稳移入培养箱中间位置，35-37?或30-32?培养至所需时间。 6.5 抑菌圈测量 6.5.1 将培养妥的双碟取出，打开陶瓦盖，将钢管倒入盛有1:1000新洁尔灭溶液或其它消毒液内，换以玻璃盖，按样品号排妥;测量抑菌圈前应检查抑菌是否圆整，如有破圈或圈不圆整应将该碟弃之，切忌主观挑选抑菌圈及双碟，使结果造成偏倚。 6.5.2 测量换菌圈可用抑菌圈自动测量分析系统，也可用游标卡尺;使用游标卡尺测量时，眼睛视线应与读数刻度垂直，用游标卡尺的尖端与抑菌圈直径的切点成垂直方向测量。 7 记录与计算 7.1 试验记录应包括抗生素的品种、剂型、标示量、生产厂、批号、检验目的、检验pipe inspection) shower faucet deviation shall not exceed 更多免费资料下载请进:? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair sl好好学习社区ot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 依据、检验日期、温度、湿度，标准品与供试品的称量、稀释步骤与核对人，抑菌圈测量结果。当用游标卡尺测量抑菌圈直径时，应将测试数据以框图方式顺双碟数记录清。当用测量仪测量面积或直径时，电脑测试、计算、统计分析的打印纸贴附于上。 7.2 计算 7.2.1 剂量法计算公式 T+T-S-S 2121-1P=log[————————×I]×100% T+S-T-S 2211 式中 P为供试品效价(相当于标示量或估计效价的百分数); S为标准品高浓度溶液所致抑菌圈直径(面积)的总和; 2 S为标准品低浓度溶液所致抑菌圈直径(面积)的总和; 1 T为供试品高浓度溶液所致抑菌圈直径(面积)的总和; 2 T为供试品低浓度溶液所致抑菌圈直径(面积)的总和; 1 I为高、低剂量之比的对数值，2:1时，I=0.301。4:1时，I=0.602。 举例 乳糖酸红霉素效价测定结果计算(见表1)。 估计效价(Ar)=6.03u/mg 剂距(I)=0.301 15532+12528-15491-12472 -1P=log[——————————————×0.301]×100%=101.12% 15491+15532-12472-12528 效价(Pr)=P×Ar=603 u/mg×101.12%=609.75 u/mg pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 德信诚培训网 表1 抑菌圈面积结果 SH SL TH TL 1 1571 1276 1581 1268 2 1481 1199 1523 1173 3 1523 1302 1497 1263 4 1518 1214 1533 1226 5 1573 1237 1543 1268 6 1546 1219 1580 1224 7 1495 1260 1526 1240 8 1530 1237 1548 1263 9 1605 1256 1593 1293 10 1649 1272 1608 1320 ? 15491 12472 15532 12528 8 结果判断 8.1 可靠性测验 该法的设计是根据量反应平行线原理，在实验所用的剂量范围内，对数剂量和反应呈直线关系，供试品和标准品的直线应平行。当用卡尺测量后的结果，将参数输入电脑，有专用(2.2)法和(3.3)法的软件程序进行统计学处理。用测量仪测量抑菌圈时，测量与统计学处理一次完成，统计学分析按药典生物检定统计规定进行F的显著性测验。(2.2) 此法要求直线回归、剂间P,0.01，偏离平行P,0.05;(3.3)法除(2.2)法的规定外,尚考查二次曲线和反二次曲线P,0.05，试验结果认为可靠，方可进行效价和可信限率计算。 8.2 可信限率 考核实验的精密度，除兽药典各论另有规定外，本法的可信限率杆菌不得超过7%，球菌不得超过8%。 上述各项规定都能符合者，试验结果成立。 8.3 实验计算所得效价低于估计效价的90%或高于估计效价的110%，则检验结果仅作为初试，应调整供试品估计效价，予以重试。 8.4 原料药效价测定一般需双份样品，平行实验以便核对。对不符合规定的样品应至少有2次符合规定的结果，才能发出。 8.5 效价结果的数字按药学规定取舍小数位。 pipe inspection) shower faucet deviation shall not exceed 更多免费资料下载请进:? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair sl好好学习社区ot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 9 供试品测定操作要点 9.1 原料药品 是指大包装或半成品干燥粉末或结晶性粉末，不含辅料，一般测定纯度(u/mg或μg/mg)，根据抗生素品种及厂方提供的效价进行估计单位，称取样品，估计效价尽量接近真实效价，如估计效价与真实效价距离较远时，可先作初测试验，然后按初测结果来估计效价，再作精密测定。 原料药品一般皆以干燥品或无水物计算效价，先测其含水的供试品效价，再根据供试品的水分或干燥失重的结果折算成干燥品或无水物的效价。 湿品效价(u/mg) 干燥品效价(u/mg)= —————————— 1-供试品干燥失重量% 9.2 水针剂 水针剂系抗生素无菌水溶液，标示量以每亳升含效价单位数计，效价测定时启开安瓿或小瓶塞后，用干燥的标准吸管吸取一定量供试品，将吸管外壁用滤纸擦净，再弃去过多的供试品，沿着容量瓶口内壁缓缓放入已盛有一定溶剂的容量瓶内，以免抗生素结晶析出，振摇，继续加溶液至刻度，摇匀，再稀释至规定的浓度。 pipe inspection) shower faucet deviation shall not exceed ? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair slot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread 德信诚培训网 pipe inspection) shower faucet deviation shall not exceed 更多免费资料下载请进:? 3 mm spacing; 3.4 construction for water after the first test, then check 1.00MPA hydraulic test, pipe joint is no seepage and leakage to grade, time after pressure relief plug nozzle; 3.5 construction after the completion of 10# cement mortar repair sl好好学习社区ot and fixed water supply reserve heads; 4.2.4.2 lining plastic pipe installation project construction of vertical water supply pipe lining plastic pipe, pipe diameter for DN20~DN100, with threaded connections, installed inside the pipe shaft. In charge of water supply network pressure-tube diameter DN100 above by machinery slots clamp connection, clean the pipe shaft 1, in accordance with design drawings, to guarantee the pipeline's convenient hanging tube simultaneously, determine bracket mounting location and height; 2, pipe L40*4 steel processed into "men" type, holding cards finished galvanized material, 0.70 meters installed on each 1 set of brackets; 3, can only be used on an electric pipe threader pipe cutting machine cutting pipe, high temperature cutting so as not to damage the plastic inner lining; completion of cutting with a blade to remove pipe-end deburring plastics, and file to remove burrs and Flash of steel pipe end; 4, according to the different pipe diameters pipe Threading 2~3 system is complete, it tube lines should be clear, broken chains or lack of deduction shall not be greater than the length of thread length 10%; after processing applications standard thread gauge check; 5, steel pipes ribbed connection using flax oil or PTFE thread