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.     

山东省无菌制剂生产质量风险分析与探讨

目的：发现山东省无菌制剂生产企业在生产质量管理方面存在的共性缺陷问题,探讨无菌制剂生产企业存在的主要风险点,为无菌制剂企业更加规范地实施GMP提供参考。方法：对2016-2017年山东省承担的无菌制剂认证检查情况进行统计分析。结果与结论：企业需要关注确认与验证、无菌药品附录、质量控制与质量保证、机构与人员等方面存在的质量风险,有针对性地改进提升。     

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.     

Barrier isolators as an alternative to a cleanroom.

The use of barrier isolators as an alternative to a cleanroom for the preparation of sterile products at ASHP-defined risk levels 2 and 3 and cytotoxic and hazardous drugs is described. The two isolators selected provide enclosed microenvironments. The isolator used for preparing products of risk level 2 has white acrylic and polycarbonate rigid walls that isolate the sterile product from the operator. The isolator used for preparing level 3 products and cytotoxic and hazardous drugs has epoxy-painted steel walls, a front window, and a pair of glove ports; its design theoretically prevents cross contamination between the operator and the product under preparation and between the product and the external environment. An independent contractor familiar with barrier isolators certifies the equipment every six months. A random microbiological sampling is periodically compared with a control. The isolators have been in operation since July 1998 without microbial growth in any sample tested. The barrier isolators cost more than traditional laminar-airflow hoods, but the hospital considers the barrier isolators to be more cost-effective than a cleanroom. A cost saving in supplies for cytotoxic and hazardous drug preparation as a result of a reduction in overall gown and glove use was observed. Barrier isolators offer an alternative to a cleanroom for the preparation of products at risk levels 2 and 3 and offer an aseptic, safe environment for preparing cytotoxic and hazardous agents.     

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.     

注射用尿激酶的质量分析和安全性评价

目的通过注射用尿激酶的全国抽验[1],全面了解国内注射用尿激酶的质量状况及安全隐患。方法采用《中国药典》标准[2]进行质量分析,同时用酶联免疫吸附法（ELISA）和荧光定量法（PCR）检测病毒[3]进行安全性评价。结果在全国共抽取58批样品,涉及9个生产企业,按《中国药典》2005年版检验,合格率为87.9%。不合格批次共7批,涉及2个生产企业,不合格项目为[效价]、[无菌]、[干燥失重],安全性评价合格率为100%。结论现行药典标准能较好地控制产品质量,安全性评价能较好的监控产品的安全性。     

无菌药品生产企业核心区微生物污染调查与分析

目的通过分析上海地区无菌药品生产企业微生物污染情况,评价企业微生物污染控制隔离体系的有效性,探索建立适合于我国实际的药品生产企业环境微生物鉴定和分型技术,为无菌药品生产企业完善微生物监控体系提供参考。方法采用生化鉴定和16SrDNA测序技术,对上海地区13家无菌药品生产企业的核心生产环境（A／B或A／C级洁净区）的微生物污染情况进行调查分析。结果收集到药品生产环境微生物分离株共计265株,经过对不同洁净级别和微生物种类的分析,发现葡萄球菌属、芽孢杆菌属和微球菌属细菌分别占全部分离株的52．7％、11．7％和10．5％。结论空气净化和物理隔离可以有效阻断大部分微生物的传播,但接触和操作引起的污染仍是生产企业面临的主要问题,加强对无菌操作人员的培训和管理是解决这一问题的关键。     

Decision-Support Method for the Choice Between Single-Use and Multi-Use Technologies in Sterile Drug Product Manufacturing

Purpose

Single-use technology has been applied to sterile drug product manufacturing processes as a new technology in contrast with the conventional multi-use technology. This study proposes a decision-support method for choosing between these two technologies in sterile drug product manufacturing.

Methods

The proposed method consists of four steps: create process evaluation models, calculate evaluation results, perform what-if analysis, and interpret results. We created models for evaluating the production cost and life cycle CO₂ emissions of processes using either technology. “What-if” analysis quantifies the effect of the input parameters on the evaluation results, which supports more informed decision-making. Here, it is recommended that the filling speed, which was found to have a significant impact on filling accuracy—a critical quality attribute of sterile drug products—should be investigated.

Results

As a case study, the method was applied in two cases of technology selection: (i) single-batch production of a product with different batch sizes and (ii) single-batch production with different production patterns. The single-use technology showed its economic superiority in producing small batches and in producing multiple small-scale products, whereas in the environmental evaluation, it was always better than multi-use technology. What-if analyses revealed the impact of changing input parameters on the economy, environment, and quality.

Conclusions

Our method can support the choice of single-use and multi-use technologies in plants having both technologies independently. In the case study, economic evaluation showed a critical point in each design case, whereas the environmental evaluation result was always better in single-use technology.

     

Plastic isolators for treatment of acute leukaemia patients under "germ-free" conditions.

A gnotobiotic isolation system based on those developed in veterinary research has been constructed for hospital use. Fifteen patients with leukaemia and neutropenia spent a total of 110 weeks in plastic isolators, and none acquired any infection. Endogenous flora was effectively suppressed by topical antiseptics and gastrointestinal decontamination effected with nonabsorbable antibiotics. The isolator system was acceptable to patients and staff and much cheaper than the use of sterile rooms. Other advantages of the system are portability, easy storage, and use on ordinary open wards without prejudice to the microbiological protection afforded. It is as yet uncertain whether protective environments of this type will substantially improve the outcome of treatment for the acute leukaemias.     

Isolators and barrier devices in pharmaceutical manufacturing

The use of isolators and barrier technology in pharmaceutical manufacturing may cause design problems, some of them unrecognised. These design problems are highlighted in this paper and, where possible, solutions are given.     

A novel approach to the statistical evaluation of media fill tests by the difference from no contamination data

It is obvious that, in the media fill test and process simulation test, positive numbers in total fills should not have any significant difference from zero or asepsis. There are many reports concerning the definition of "sterility" or "asepsis." However, any scientific and practical methods to demonstrate "no significant difference from zero" have not been reported up to now. The existing criteria, such as "less than 0.1%," "less than 0.05%," and "less than two positives" are not appropriate to assure the integrity of processes, and sometimes lead to erroneous results. The purpose of this report is to demonstrate novel, reasonable and practical methods and criteria based on scientific and statistical consideration. According to the ISO 13408-1 Aseptic Processing of Health Care Products, Part 1 (1998), General Requirement for Aseptic Processing, the action level for the number of positive units in media fill tests is specified as 0.1%, and the alert level is 0.05%. In this paper it is shown that the existing ISO standard and other official methods are inappropriate in that zero contaminated units (sterile product) is outside the confidence range of probable distribution of contaminated units, even though the contaminated units are less than 0.1% in larger numbers of fills, and even less than 0.05%. This indicates that the limit of 0.1% or 0.05% is inappropriate in cases of larger numbers of fills. For sterile products, the number of contaminated units other than "zero" at the statistical confidence range must be judged to be contaminated units in process and as non-sterile. In order to harmonize this criteria-"no significant difference from zero"-with the existing criteria, the new criteria may be combined with only the existing criteria of 0.05% in smaller number of fills.     

Near-infrared reflectance spectroscopy as a process analytical technology tool in Ginkgo biloba extract qualification

Here, we describe the use of near-infrared diffuse reflectance spectroscopy for qualification of Ginkgo biloba extract as raw material for use in pharmaceutical products. G. biloba extract shows unpredicted and uncontrolled variability in some of its quality specifications, intrinsic to its natural origin, which have influence on the manufacturing process of solid dosage forms (viz. granulation and compression). Some of these properties could not be determined by conventional quality control tests, so we investigated the use of NIR to qualify the batches of Ginkgo extract accordingly to its different features and establish a relationship with some of the manufacturing steps behaviour based on their qualification. Several approaches were evaluated, and the NIR method developed demonstrated to be sensitive to changes in important quality specifications and therefore adequate to qualify incoming batches of G. biloba extract. This could be considered a process analytical technology (PAT) application since it: (1) establishes the source of variability in a qualitative way, (2) explains its propagation to the final product quality attributes and (3) lays the basis for a control strategy to be applied in the manufacturing process.     

血塞通泡腾片的处方工艺研究及稳定性考察

目的：探讨制备血塞通泡腾片并考察其稳定性。方法：以乳糖、甜菊素、酒石酸、碳酸氢钠、聚乙二醇（PEG6000）等为辅料,优化处方组成与制备工艺,高效液相色谱梯度洗脱法测定该片中人参皂苷Rb1、人参皂苷Rg1、三七皂苷R,的含量．通过室温留样观察试验对其稳定性进行研究。结果：优化出的处方与工艺基本合理、稳定、可行,与放大生产设备相适应,药品质量可控,可工业化生产。三批血塞通泡腾片经室温留样观察试验18个月后,其试验前后的性状和含量无明显变化,鉴别试验均呈正反应。崩解时限、重量差异、微生物限度检查均符合规定。结论：血塞通泡腾片处方组成合理,制备方法简便可行,稳定性良好。     

预灌封注射器装灭菌注射用水作为水痘减毒活疫苗稀释剂的质量和稳定性研究

目的 研究预灌封注射器装灭菌注射用水(注射器装灭菌水)作为水痘减毒活疫苗(水痘疫苗)稀释剂的质量及稳定性,用以取代原有的安瓿瓶装灭菌注射用水.方法 连续生产并分装3批注射器装灭菌水,依据《中华人民共和国药典》2010年版二部(药典二部)附录XIXC“原料药与药物制剂稳定性试验指导原则”,对注射器装灭菌水的质量和稳定性进行研究;同时将注射器装灭菌水作为水痘疫苗的稀释剂,观察水痘疫苗的质量及稳定性.结果 3批注射器装灭菌水在相对湿度(60±10)％的条件下分别于(40±2)℃放置6个月和(25±5)℃放置42个月,各项检测结果均符合药典二部“灭菌注射用水”标准.3批以注射器装灭菌水作为稀释剂的水痘疫苗的加速和长期稳定性试验结果均符合“水痘减毒活疫苗注册标准”的要求,疫苗的病毒滴度≥3.3 lg噬班形成单位/0.5 ml,牛血清白蛋白残留量＜50 ng/ml,抗生素残留量＜50 ng/剂.结论 注射器装灭菌水可作为水痘疫苗的稀释剂.     

Baxa MicroMacrocompounder for parenteral nutrition solutions in a pediatric hospital

The constant increase of parenteral nutrition (PN) manufacturing in our pediatric hospital led us to look for an automatic filling system. The aim of this study was to evaluate the MicroMacrocompounder (MM23) for compounding pediatric PN solutions. MM23 volumetric accuracy was tested with its different inlets (S, D, V) for volumes of sterile water for injection from 0.2 to 2900 mL. The influence of the solution viscosity and the source solution bottle replacements during the filling operation was also investigated. Manufacturing pediatric PN solutions was eventually assessed. Time to set up the system was 30 minutes. Maximum filling speeds with sterile water for injection were 860, 330, 154 mL/min for Vx, V and D inlets, respectively. Inlet S was not tested for MM23 filling speed. Minimal flush volume of 40 mL of sterile water for injection is necessary to clear the tube of residual ions. Average MM23 volumetric accuracy was < 5% for volumes > or = 0.5 mL for S and D inlets, and for volumes > or = 20 mL for V inlet. The volumetric accuracy was equal to 6.25% for 0.2 mL. In all experiments, volumetric accuracy was < 5%. The accuracy of electrolyte measurements performed on bag samples was less than 5% for 150 (74%) samples, between 5 and 10% for 27 (13%) samples and greater than 10% for 27 (13%) samples. Microbiological analysis showed no positive culture. The average manufacturing times were 56.8 +/- 4.5, 188.2 +/- 7.7 and 447.2 +/- 13.8 seconds for 130, 660 and 1800 mL bags, respectively. The MM23 compounder is suitable for compounding pediatric admixtures with source solutions volumes > or = 0.5 mL. This system has been used daily for five months in our department.     

Preformulation studies for the development of a parenteral liquid formulation of the immunomodulator, peldesine

Peldesine (BCX-34) is a potent inhibitor of the enzyme purine nucleoside phosphorylase (PNP). The object of this study was to determine the preformulation parameters of BCX-34 and subsequently prepare a sterile liquid formulation for intravenous infusion. Ionization and solubility parameters were assessed prior to formulation development. Three pKa values of approximately 4.0, 5.2 and 10.3 were assigned for BCX-34. The pH-solubility profile showed an increase in solubility when the pH of the solution was less than the pKa1 value of 4.0 and when the pH was more than the pKa3 value of 10.3. BCX-34 has an octanol-water partition coefficient of approximately 7:1 (Log P = 0.8). The Van't Hoff temperature dependent solubility when measured over a range of 4-40 degrees C produced a heat of solution (delta Hc) of 7.68 kcal/mole. Based on these preformulation observations, two parenteral formulations containing 1 and 5 mg/mL of BCX-34 were prepared in an acidified saline solution and filled into 30 cc and 100 cc glass vials. Both formulation batches were sterile filtered, while only one 1 mg/mL batch was terminally autoclaved. The long-term stability of both solutions was performed and compared. Results of this study show that the drug product was stable. Following a screening study, Lactated Ringer's injection USP was selected as a suitable infusion medium for the dilution of the drug concentrate solutions.     

消毒供应中心集中化管理效果分析

目的探讨消毒供应中心集中化管理的效果。方法通过对消毒供应中心集中化管理前、后回顾性总结与分析及问卷调查,分析消毒供应中心集中化管理的效果。结果消毒供应中心集中化管理后治疗包和消毒包、手术器械包、各种管道类物品、外来医疗器械、外院消毒包、总收入均多于管理前,且满意度高于管理前。结论消毒供应中心集中化管理,可以保证物品的消毒灭菌质量,有效保障医疗护理工作质量,降低医院感染发生率,而且可以使医院人力、设备等达到资源共享,从而减少投入,减少资源的浪费。     

Syringe Filling of High-Concentration mAb Products Using Peristaltic Pump-Based Mechanism: Challenges and Mitigation Strategies

Syringe filling of high-concentration mAb formulation during manufacturing of large-scale drug product batches may present challenges such as product deposition onto the area of the syringe barrel where the stopper is inserted, product splashing or dripping, droplets left after the fill cycle, filling needle clogging, product build-up inside the needle during line stoppages, variation in fill weight/volume, and potential impact on product quality attributes. In this article, a summary of these issues and approaches to overcome them are summarized. Potential failure modes of the syringe filling process and appropriate in-process controls are provided. In addition to developing the filling process or resolving manufacturing issues, the pharmaceutical company developing the product and associated drug product manufacturing process may want to implement long-term strategic approaches to support the portfolio progression. Potential long-term approaches such as use of a viscosity reducing formulation development approach, improving peristaltic filling technology performance, building small-scale filling capability and establishing a streamlined filling process management cycle are also summarized. The aspects summarized in this article may be used to develop a robust filling process and control strategy for high-concentration mAb products and implement long-term strategic approaches to support the portfolio progression.     

Sterile Filtration of a Parenteral Emulsion

The Syntex adjuvant formulation (SAP) containing [thr¹]-muramyldipeptide in an oil-in-water emulsion has proven to be an effective adjuvant eliciting both cell-mediated and humoral immune response. As a parenteral emulsion, sterility of the final product was a concern, and various methods of achieving sterility were considered. For emulsions, most conventional sterilization methods are not viable, requiring the more cumbersome technique of sterilizing individual components and assembling/manufacturing under sterile conditions. Emulsion vehicles were manufactured with various models in the Microfluidizer M110 series. All equipment examined was capable of reducing the average dispersed oil droplet size to approximately 160 nm, with varying size ranges. Operating at an internal equipment pressure of greater than 16,000 psi, with at least five cycles through the interaction chamber, the resulting emulsion had a narrow droplet size range distribution, with the largest droplets being small enough to enable sterile filtration. Under specific-manufacturing conditions, the adjuvant emulsion becomes easily filtered through a 0.22-µm cartridge filter, thus yielding a sterile end product. This is the first published example of emulsion sterilization being achieved by terminal filtration.     

盐酸西替利嗪片的溶出度考察

目的 :考察国产与进口盐酸西替利嗪片的体外溶出度 ,为其质量控制提供参考。方法 :采用转篮法测定溶出度 ,用威布尔 (Weibull)分布模型拟合溶出曲线 ,并对溶出参数m、T50 、Td 等采用方差分析及t检验进行统计。结果 :国产与进口盐酸西替利嗪片的溶出参数差异有显著性 (P <0 .0 5 ) ;国产片 3批号间差异无显著性 (P >0 .0 5 ) ,进口片 2批号间差异无显著性(P >0 .0 5 )。结论 :进口片的体外溶出度较国产片为优。国产片及进口片制剂工艺均较稳定