HZ-100型臭氧灭菌机对实验动物屏障设施熏蒸灭菌效果观察

目的:对日本产HZ-100型臭氧熏蒸灭菌机在屏障环境内杀菌效果进行验证,为屏障环境内控制微生物污染提供参考。方法:使用日本产HZ-100型臭氧熏蒸灭菌机对密闭屏障房间进行熏蒸消毒处理,通过熏蒸前、后房间内菌落检测比较,观察其灭菌以及使用效果。结果:熏蒸前,房间内落下菌>59CFU/皿,墙壁拭子菌落1 CFU/皿,枯草杆菌试纸培养阳性;熏蒸后,房间内落下菌2.5 CFU/皿,墙壁拭子菌落未检出,枯草杆菌试纸培养结果阴性。结论:HZ-100型臭氧机熏蒸灭菌效果良好,全程自动化,操作便捷,使用安全。     

静脉用药调配中心调配环境微生物动态监测数据的统计分析

目的为静脉用药调配中心( PIVAS)调配环境微生物动态监测提供控制限度制订方法和合适的限度值。方法使用复旦大学附属中山医院东院区静脉用药调配中心 2018年 4月至 2021年 3月调配环境微生物监测数据建立数据库,利用参数模型、阈值法及公差限制法等统计分析方法分别构建控制限度。通过应用效果的覆盖率误差及警报频率,评价限度制订的合理性以确定最佳方法。结果除参数模型外,阈值法与公差限制法成功构建了 PIVAS调配环境微生物动态监测的控制限度。阈值法: A级操作台沉降菌警戒限为 0.67、纠偏限为 <1;A级操作台浮游菌警戒限为 0.5、纠偏限为 <1;C级调配间沉降菌警戒限为2.75、纠偏限为 3.51;C级调配间浮游菌警戒限为 5.18、纠偏限为 6.29;在应用中警报 13次(频率为 5.47%)覆盖率误差为1.88%。公差限制法: A级操作台沉降菌警戒限为 <0.95、纠偏限为 <0.99;A级操作台浮游菌警戒限为 <0.95、限为 <0.99;C级调配间沉降菌警戒限为 47.5、纠偏限为 49.5;C级调配间浮游菌警戒限为 95、纠偏限为 99;在应用中警报 1次(频率为 纠偏,0.47%),覆盖率误差为 2.58%。结论阈值法确定的控制限度可行性较强、灵敏度高,可作为 PIVAS调配环境动态微生物控制限度的制订方法。     

PIVAS洁净区空气微生物动态变化的两种监测方法应用比较

目的： 评估沉降菌法和定量空气浮游菌采样法对PIVAS洁净区空气微生物动态变化的监测效果。方法： 参照《医药工业洁净区（室）沉降菌的测试方法》（GB/T 16294-2010）、《医药工业洁净区（室）浮游菌的测试方法》（GB/T 16293-2010），对洁净区空气微生物进行动态监测，对数据进行统计学分析。结果： 4个洁净区的空气微生物浓度均在合格范围内；两种方法对普通药/营养调配洁净区空气微生物的检出效果具有统计学差异（P<0.05），沉降菌法的检出率高于定量空气浮游菌采样法；抗生素/抗肿瘤药调配洁净区，两种方法的检出率无统计学差异（P>0.05）。结论： PIVAS洁净区空气微生物的动态监测，应同时采取沉降菌法和定量空气浮游菌采样法，两种方法相辅相成，互为补充。     

中药饮片虎掌南星微生物污染状况分析

目的:建立虎掌南星的微生物限度检查方法，考察该饮片微生物污染的程度，并评估其潜在的风险。方法:依据《中国药典》2020年版四部通则1105、1106、1108收载中药饮片微生物限度检查法对虎掌南星进行方法适用性试验，对20个批次药品的需氧菌总数(TAMC)、霉菌和酵母菌总数(TYMC)、耐热菌总数(NAIRE)、耐胆盐革兰阴性菌、大肠埃希菌、沙门菌、金黄色葡萄球菌、铜绿假单胞菌、白念珠菌进行检查，同时对耐热菌做探索性检验。结果:需氧菌总数在10²～10⁷之间；霉菌和酵母菌总数在10¹～10⁵之间；12批次检出耐热菌，检出的耐热菌数在10¹～10³之间；15批次检出耐胆盐革兰阴性菌，N均小于10⁴;13批次检出大肠埃希菌；0批次检出沙门菌；20批次均检出金黄色葡萄球菌；7批次检出铜绿假单胞菌；6批次检出白念珠菌；耐热菌数随水浴时间增加，菌数下降明显。结论:通过对微生物的污染调查分析，给虎掌南星饮片的合理使用提供了一些数据上的保障。建议根...     

医用高压氧舱气道保护盒的临床应用研究

目的研究医用高压氧舱气道保护盒在高压氧治疗中的作用。方法对进行高压氧治疗的76例患者完全随机分为使用医用高压氧舱气道保护盒治疗的保护组（45例）和使用传统高压氧治疗方法的对照组（31例）。保护组患者采用医用高压氧舱气道保护盒进行治疗，对照组采用传统人工防护方法进行治疗。观察治疗效果及患者气道情况。结果保护组痊愈13例（28．9％），显效18例（40．0％），有效8例（17．8％），无效6例（13．3％），总有效率86．7％；对照组痊愈8例（25．8％），显效11例（35．5％），有效5例（16．1％），无效7例（22．6％），总有效率77．4％。2组总有效率比较，差异有统计学意义（P〈0．01）。对照组样本菌落为11500～13260CFU／m^3，平均12850CFU／m^3。保护组样本菌落为2700～5100CFU／m^3，平均3420CFU／m^3。保护组样本菌落数明显低于对照组，差异具有统计学意义（P〈0．05）。结论医用高压氧舱气道保护盒解决了医用高压氧舱气道异物、灰尘污染及感染问题，可以保护使用高压氧舱的患者，有助于患者早日康复。医用高压氧舱气道保护盒制作简单，经济实用，值得推广应用。     

基于激光诱导荧光技术原理的微生物实时监测技术在USP<1223>标准下的替代验证及其在PIVAS洁净室的应用研究

目的 为实现给静脉用药集中调配中心（pharmacy intravenous admixture services，PIVAS）对浮游菌及尘埃粒子的实时监测及预警提供技术支持，依据美国药典（USP）微生物替代方法验证的要求，对微生物实时监测技术进行微生物替代方法可行性验证，并研究该技术与传统浮游菌及尘埃粒子测试方法的效果差异。方法 通过对激光诱导荧光技术的生物气溶胶实时监测系统（biological aerosol real-time monitoring system，BAMS）与浮游菌采样器在线性相关性、准确度、精密度、专属性、定量限、重现性和耐用性等关键参数的数据比对，验证替代方案的可行性。在PIVAS洁净室分别对BAMS、浮游菌采样器及尘埃粒子检测仪的实际应用效果进行比较评估。结果 BAMS相较于传统的浮游菌采样器具有显著的灵敏度优势，回收率、精密度、专属性、线性、重现性及耐用性等验证指标均符合USP<1223>项下的相关要求，BAMS在PIVAS洁净区的尘埃粒子检测效率与传统方法无明显差异。结论 BAMS与传统洁净区环境监测方法无差异，且比传统的浮游菌采样器具备更高的检测灵敏度及时效性，可应用于医院PIVAS洁净室环境微生物污染状况的实时监测。     

无菌异丙醇开瓶后有效期的初步研究

 目的 通过研究3个批次无菌异丙醇在3种不同洁净度的环境中开瓶后，瓶内外观、无菌状态及异丙醇含量是否受到环境影响，初步确定无菌异丙醇开瓶后有效期。方法  从3个批次无菌异丙醇中每个批次随机选取162瓶，每个批次分别放置在A级、D级无菌室，无级别库房各54瓶。喷瓶的喷嘴始终处于打开状态，每个工作日进行3次喷雾操作。在第0、1、7、14、21、30天，检测异丙醇溶液的外观性状；采用薄膜过滤法测定异丙醇溶液中微生物的污染情况；采用气相色谱法测定异丙醇含量，并用独立样本t检验方法进行统计学分析。结果  实验期间所有异丙醇溶液外观无变色、无沉淀、无悬浮物，气味无变化，未出现微生物污染；异丙醇含量为体积分数（69.98~70.17）%，达到标示含量。通过统计学分析，异丙醇含量差异无统计学意义（t值0.07~2.18，p值均＞0.05）。结论  异丙醇在A级、D级无菌室和无级别库房开瓶后使用时间建议不超过30 d。     

注射剂瓶容器密封完整性:微生物挑战法与色水法的关联比较分析

考察了微生物挑战法和色水法对中性硼硅玻璃管制注射剂瓶容器密封完整性的关联并进行比较分析.微生物挑战法是将密封、灭菌后的空包装系统浸入高浓度铜绿假单胞菌的菌悬液中,施加特定压力驱动并保持4 h,取出后于33℃培养7 d;色水法是将密封后的空包装系统浸没于10％亚甲基蓝水溶液中,放入真空箱中抽至75 kPa并保持30 mi...     

我院自制中药制剂的质量控制

中药制剂的生产 ,由于使用的原料中草药一般都带有细菌、虫卵 ,在生产过程中又受到环境、工具等污染 ,故其成品常含有大量细菌 ,在药品染菌中可能有致病性微生物 ,病人服用后极为不利。因此 ,中药制剂的质量控制极为重要。笔者 1998～ 2 0 0 2年对我院 30 0余批次自制中药制剂进行了微生物限度检查 ,其情况如下 :颗粒剂12 5批 ,细菌数最高为每克 170 0个 ,大部分在每克2 0 0个以内 ,霉菌数每克 <10个 ,12 0批符合规定 ,有 5批细菌数超标 ,灭菌后符合规定。片剂 12 9批 ,工艺改革前 ,细菌数最高为每克 15 0 0 0个 ,霉菌数每克 2 5个。大部分…     

直立式聚丙烯输液袋装输液灭菌工艺研究

目的通过输液灭菌工艺优选、大输液水浴灭菌器性能确认,确定灭菌程序及装载方式的可靠性;通过对产品进行3个批次的热穿透、生物指示剂试验,确定灭菌工艺的灭菌效果。方法使用大输液水浴灭菌器,采用可编程自动控制系统进行灭菌全过程控制,并采用T32-32温度验证系统进行性能验证,在此基础上,使用嗜热脂肪杆菌芽孢生物指示剂实施灭菌工艺验证,确定灭菌效果。结果采用121℃灭菌12 min,F0值>12,无菌保证水平(SAL)≤10-6。袋身无破裂、无变形,可直立放置;制剂质量按标准检测符合《中国药典》2010年版第二增补本标准。结论方法实用,结果准确可靠。     

Comparison of hydrogen peroxide and peracetic acid as isolator sterilization agents in a hospital pharmacy.

PURPOSE: The efficacy of hydrogen peroxide and peracetic acid as isolator sterilization agents was compared. METHODS: Sterilization and efficacy tests were conducted in a flexible 0.8-m3 transfer isolator using a standard load of glass bottles and sterile medical devices in their packing paper. Bacillus stearothermophilus spores were placed in six critical locations of the isolator and incubated at 55 degrees C in a culture medium for 14 days. Sterilization by 4.25 mL/m3 of 33% vapor-phase hydrogen peroxide and 12.5 mL/m3 of 3.5% peracetic acid was tested in triplicate. Sterility was validated for hydrogen peroxide and peracetic acid at 60, 90, 120, and 180 minutes and at 90, 120, 150, 180, 210, and 240 minutes, respectively. RESULTS: In an efficacy test conducted with an empty isolator, the sterilization time required to destroy B. stearothermophilus spores was 90 minutes for both sterilants, indicating that they have comparable bactericidal properties. During the validation test with a standard load, the sterilization time using hydrogen peroxide was 150 minutes versus 120 minutes with peracetic acid. The glove cuff was particularly difficult for hydrogen peroxide to sterilize, likely due to its slower diffusion time than that of peracetic acid. Hydrogen peroxide is an environmentally safer agent than peracetic acid; however, its bacteriostatic properties, lack of odor, and poor diffusion time may limit its use in sterilizing some materials. CONCLUSION: Hydrogen peroxide is a useful alternative to peracetic acid for isolator sterilization in a hospital pharmacy or parenteral nutrition preparation unit.     

Validation of microbial recovery from hydrogen peroxide-sterilized air

The use of hydrogen peroxide as a sanitant in isolators and other barrier systems is well documented. To confirm that the isolator maintains a germ-free environment between decontamination cycles, microbiological air monitoring is performed after the sanitation and aeration cycles. In this study, we have shown that residual levels of hydrogen peroxide as low as 1 ppm can remain in the isolator and inhibit the growth of microorganisms after concentration on agar media. This lingering hydrogen peroxide can make accurate microbiological air monitoring difficult and can even cause false negative test results. To solve this issue, we have developed a new media that can mediate the effects of residual peroxide and prevent false negative test results. Initially, catalase was tested as a neutralizing agent but proved not to be efficient enough. Instead, 1% pyruvate was added, which was able to tolerate as much as 15 ppm Vaporous Hydrogen Peroxide (VHP) and ensured growth promotion of Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis, Candida albicans and Aspergillus niger. The 1% pyruvate retained its neutralizing activity for Micrococcus luteus at up to 100 ppm VHP. Raising the pyruvate concentration to 5% pyruvate enabled neutralization of up to 300 ppm VHP, permitting subsequent growth of Micrococcus luteus on agar media.     

Near-Infrared (NIR) Monitoring of H2O2 Vapor Concentration During Vapor Hydrogen Peroxide (VHP) Sterilisation

Purpose. There is an increasing use in the pharmaceutical industry of barrier systems such as transfer isolators, sterilisation tunnels and work station isolators. As Vapor Hydrogen Peroxide (VHP) sterilisation of isolators and lyophilizers becomes an important sterilisation method, there is an acute need for a VHP monitoring system to be used for in-process control and validation. In this study, near infrared (NIR) spectrofotometry was evaluated as a potential technique to monitor hydrogen peroxide. Additionally the H₂O₂ vapor permeability of different packaging materials, commonly used in steam and ethylene oxide sterilisation, was evaluated. Methods. NIR spectrofotometry, using a gas cell connected with optic fibres, was evaluated as a potential technique to monitor hydrogen peroxide vapor and water vapor during VHP sterilisation of an isolator. A NIR spectrum was taken every 30 s during VHP sterilisation of an isolator. The influence of injection rate, air flow rate, working temperature and gas distribution was investigated. The H₂O₂ vapor permeability of different packaging materials was determined by placing the gas cell in the sterilisation bags and sealing the bags hermetically. The sterilisation bag was then subjected to VHP sterilisation. Results. The NIR spectra taken at steady state sterilization conditions showed 4 absorption peaks: at 1364,1378 and 1400 nm attributed to water and at 1420 nm attributed to H₂O₂ vapor. By measuring the absorbance level at these wavelengths, the actual concentration of H₂O and H₂O₂ vapor in the isolator was calculated. The water vapor permeation of the sterilisation bags, measured with NIR, appeared to be equal for all materials tested. Whereas Tyvek^® was the most permeable material for hydrogen peroxide vapor (82.7% of the reference concentration outside the bag), only 30% was found in bags made of medical paper. Sterilisation bags consisting of laminate films and PVC sealed to medical paper showed intermediate permeability. Conclusions. Near-infrared (NIR) spectroscopy using a gascell with optic fibres is a useful technique to monitor VHP sterilisation cycles. There was a difference in H₂O₂ vapor permeability of different packaging materials, commonly used in steam and ethylene oxide sterilisation.     

美国药典41版汽相灭菌法简述

以汽化过氧化氢表面灭菌为例,介绍了美国药典(USP)41版中汽相灭菌法相关章节(1229.11汽相灭菌法)的内容,主要包括过氧化氢灭菌原理、汽化过氧化氢表面灭菌的特点、灭菌效果的影响因素、灭菌验证用生物指示剂的特点以及使用的注意事项,并将USP第41版1229.11汽相灭菌法的全文翻译。     

Development of a sterilizing in-place application for a production machine using Vaporized Hydrogen Peroxide

The use of steam in sterilization processes is limited by the implementation of heat-sensitive components inside the machines to be sterilized. Alternative low-temperature sterilization methods need to be found and their suitability evaluated. Vaporized Hydrogen Peroxide (VHP) technology was adapted for a production machine consisting of highly sensitive pressure sensors and thermo-labile air tube systems. This new kind of "cold" surface sterilization, known from the Barrier Isolator Technology, is based on the controlled release of hydrogen peroxide vapour into sealed enclosures. A mobile VHP generator was used to generate the hydrogen peroxide vapour. The unit was combined with the air conduction system of the production machine. Terminal vacuum pumps were installed to distribute the gas within the production machine and for its elimination. In order to control the sterilization process, different physical process monitors were incorporated. The validation of the process was based on biological indicators (Geobacillus stearothermophilus). The Limited Spearman Karber Method (LSKM) was used to statistically evaluate the sterilization process. The results show that it is possible to sterilize surfaces in a complex tube system with the use of gaseous hydrogen peroxide. A total microbial reduction of 6 log units was reached.     

Evaluating the Potential,Applicability, and Effectiveness of Ozone Sterilization Process for Medical Devices

Purpose

Ozone (O₃) can be considered the most potent natural germicide against microorganisms (in vegetative and spore forms) with high efficiency and speed, because of its highly oxidizing activity. Despite this, there are a few studies describing the application of ozone as a sterilizing agent of medical devices. The aim of this communication was to describe the development and validation of a sterilization cycle applied to medical devices.

Methods

The sterilization process was challenged using Geobacillus stearothermophilus ATCC 7953 spores, which have shown great resistance. The sterilizing effect of ozone was measured using carriers inoculated with 10⁶ CFU/mL spores, introduced into a 3-mL syringe and lumens of tubes of different sizes and diameters simulating hospital medical products, which have undergone a half-cycle or complete cycle.

Results

The results of sterilization process studied in active vegetative form of microorganisms showed that the ozone sterilization was effective with a bioburden between 10⁵ to 10⁷ CFU/mL with one pulse sterilizing action. The validation of the process was confirmed by the satisfactory results for the half-cycle, corresponding to a treatment with four pulses allowed sterilizing the material with bioburdens <?10⁶ CFU/mL spores which indicate an appropriate sterility assurance level.

Conclusion

The results showed that the ozone may be considered as effective and promising alternative for sterilization of thermosensitive materials and medical devices.

     

Permeation risks with peracetic acid and hydrogen peroxide sterilizing agent inside ambulatory pumps

The sterilizing agent commonly used to sterilize materials for an isolator is a peracetic acid (PA) and hydrogen peroxide (HP) mixture. The permeation of this agent through ambulatory pumps should reveal a potential toxic risk for the patient and a stability modification of the drug by a pH change. Six wrapped and six unwrapped ambulatory pumps from each laboratory were introduced in the transfer chamber for the sterilizing process over 2 h 45 min. The presence of PA and HP were determined by using analytical strips. If the analytical strips of HP were positive, the level of HP was determined by using a specific spectrometric kit. No acid permeation was found in all wrapped pumps. Acid permeation was found in two samples of Ultraflow unwrapped series and in one unwrapped sample of Easypump series by the analytical strips. In other unwrapped samples, no acid permeation was detected. In four unwrapped ambulatory pumps (Accufuser, Infusor, Ultraflow, and Easypump), the analytical strips of HP were positive in the range of 0.5 to 25 mg/L, varying by laboratory. In only one sample (Surefuser), no detection of HP was found. The quantitative dosage of HP by spectrophotometry confirmed the permeation risk inside all pumps except the Surefuser. Our investigation shows that the permeation risk inside ambulatory pumps is real when pumps are unwrapped and exposed at high levels to PA and HP mixture. The results of our study recommend retaining the wrapping for the peracetic acid sterilization of the ambulatory pumps.     

In vitro assay for the screening of the plaque-reducing activity of antimicrobial agents

Bacteria grown in biofilms are less susceptible to antimicrobial agents than planktonic bacteria. One of the most common biofilms in humans is dental plaque. To investigate the antimicrobial activity against dental bacteria grown in biofilms, biofilms were generated with Streptococcus sanguis on hydroxyapatite disks to mimic the tooth surface. After 48 h of aerobic incubation at 37 degrees C in a continuous flow culture, a biofilm consisting of Streptococcus sanguis became visible on the surface of the disks. The disks were removed from the growth chamber and placed in different vessels containing either chlorhexidine digluconate (CAS 55-56-1, 0.1% or 1.0%), polyvinylpyrrolidone iodine (1.5% or 7.5%), or octenidine dihydrochloride (CAS 70775-75-6, 0.05% or 0.1%) for 5 or 30 min. In addition, the antiseptics were applied to the bacterial suspension in the growth chamber. A significant difference was observed in the antimicrobial activity against bacteria in the suspension liquid compared to the bacteria grown in biofilms. The best reduction factors were obtained with chlorhexidine (1.0%, 30 min) for either sessile (3.97 Ig) or planktonic bacteria (> or = 5.58 Ig). Clear relationships between the doses and times of action were found for the assessed agents. Therefore, the authors conclude that the present in vitro assay is a quick and cost-effective model to screen the activity of antimicrobial agents against bacteria grown in biofilms.     

The Influence of Humidity, Hydrogen Peroxide Concentration, and Condensation on the Inactivation of Geobacillus stearothermophilus Spores with Hydrogen Peroxide Vapor

The study presented here examined the factors influencing the effectiveness of surface decontamination with hydrogen peroxide vapor. The impact of relative humidity and hydrogen peroxide gas concentrations was investigated and compared to a dew point analysis of these various sterilant atmospheres. For this purpose, a series of different H₂O₂ decontamination cycles were developed and tested for antimicrobial effectiveness using biological indicators inoculated with greater than 10⁶ spores of Geobacillus stearothermophilus. The results indicate that an increasing concentration of hydrogen peroxide in the gas phase and higher humidity levels result in a faster inactivation of the test organisms. The higher the H₂O₂ gas phase concentration was, the more independent the inactivation effect from the humidity level. At lower H₂O₂ concentrations, the same kill was achieved with higher humidity. Subvisible condensation was found to be necessary for short inactivation times, but condensation in the visible range did not further enhance the sporicidal activity. The molecular deposition of water and hydrogen peroxide on the target surface represents the determining factor for microbial inactivation, whereas the hydrogen peroxide concentration in the gas phase is of secondary importance.     

Antimicrobial properties of porphyrins

A large number of natural and synthetic porphyrins of diverse chemical compositions and characteristics can be isolated from nature or synthesised in the laboratory. Antimicrobial and antiviral activities of porphyrins are based on their ability to catalyse peroxidase and oxidase reactions, absorb photons and generate reactive oxygen species (ROS) and partition into lipids of bacterial membranes. Light-dependent, photodynamic activity of natural and synthetic porphyrins and pthalocyanines against Gram-positive and Gram-negative bacteria has been well demonstrated. Some non-iron metalloporphyrins (MPs) possess a powerful light-independent antimicrobial activity that is based on the ability of these compounds to increase the sensitivity of bacteria to ROS or directly produce ROS. MPs mimic haem in their molecular structure and are actively accumulated by bacteria via high affinity haem-uptake systems. The same uptake systems can be used to deliver antibiotic-porphyrin and antibacterial peptide-porphyrin conjugates. Haemin, the most well known natural porphyrin, possesses a significant antibacterial activity that is augmented by the presence of physiological concentrations of hydrogen peroxide or a reducing agent. Natural and synthetic porphyrins have relatively low toxicity in vitro and in vivo. The ability for numerous chemical modifications and the large number of different mechanisms by which porphyrins affect microbial and viral pathogens place porphyrins into a group of compounds with an outstanding potential for discovery of novel agents, procedures and materials active against pathogenic microorganisms.