Working Safely in the Cleanroom

Jan Eudy has written an interesting article for Controlled Environments about personnel in cleanrooms. In the article, Eudy writes:

A cleanroom safety program would define how the PPE (personal protective equipment) or cleanroom garments are worn to enhance operator safety. Examples of safety criteria are:

• Hoods are worn securely on the head to minimize movement of the hood and maximize operator visibility.
• Masks are fixed securely over mouth and nose to minimize movement of the mask and maximize operator visibility.
• Goggles or safety glasses are worn to protect the eyes and should be worn to maximize operator visibility.
• Coveralls are chosen based on size of the operator for comfort and safety. The bottom of the coverall legs must be secure at the ankles to reduce risk of trips or falls from walking on the fabric of the coverall legs.
• Boots and/or shoe covers are chosen based on size of the operator’s shoes and specific operations performed in the cleanroom. Boots and/or shoe covers should fit snugly over shoes and be secured with strap(s) at the top of the foot. The top fabric of the boots should be secured at the knee to avoid slips or falls.

To access the article, go to Controlled Environments



Posted by Tim Sandle

Antibiotics trigger super-spreaders

In a new study, Salmonella-infected mice that were given antibiotics became sicker and began shedding far more bacteria in their feces than they had before.

When the scientists gave oral antibiotics to mice infected with Salmonella typhimurium, a small minority -- so called "superspreaders" that had been shedding high numbers of salmonella in their feces for weeks -- remained healthy; they were unaffected by either the disease or the antibiotic. The rest of the mice got sicker instead of better and, oddly, started shedding like superspreaders. The findings point to a reason for superspreaders' ability to remain asymptomatic. They also pose ominous questions about the widespread, routine use of sub-therapeutic doses of antibiotics in livestock.

For further details, see:

Smita Gopinath, Joshua S. Lichtman, Donna M. Bouley, Joshua E. Elias, and Denise M. Monack. Role of disease-associated tolerance in infectious superspreaders. PNAS, October 20, 2014 DOI: 10.1073/pnas.1409968111



Posted by Tim Sandle

Bladderwrack and bacterial resistance

The bladderwrack Fucus vesiculosus is a species of brown algae, found along the North Atlantic coasts. The algae has an interesting defence mechanism against bacterial infections.

Bacteria generally play a crucial role in the life of seaweeds. Also the bladderwrack lives in symbiosis with many types of bacteria that feed it with certain growth factors and nutrients. On the other hand, some other bacterial species can harm the seaweed. To deter them, Fucus produces different chemical compounds.

In terms of climate change, under changed light or temperature conditions the production of single defensive compounds decreased in comparison to unchanged conditions.

For further details, refer to:

Mahasweta Saha, Martin Rempt, Stephanie B. Stratil, Martin Wahl, Georg Pohnert, Florian Weinberger. Defence Chemistry Modulation by Light and Temperature Shifts and the Resulting Effects on Associated Epibacteria of Fucus vesiculosus. PLoS ONE, 2014; 9 (10): e105333 DOI: 10.1371/journal.pone.0105333

Posted by Tim Sandle

Biosimilars market rises

A new report suggests that specialty pharmaceuticals are growing in proportion of total pharmaceutical spend in the United States as well as in cost to the patient and healthcare system at an unsustainable rate.

Biosimilars also known as follow-on biologics are biologic medical products whose active drug substance is made by a living organism or derived from a living organism by means of recombinant DNA or controlled gene expression methods. Biosimilars (or follow-on biologics) are terms used to describe officially approved subsequent versions of innovator biopharmaceutical products made by a different sponsor following patent and exclusivity expiry on the innovator product.

The Biologics Price Competition and Innovation Act (BPCIA) of 2009 was enacted as part of the Patient Protection and Affordable Care Act on March 23, 2010, to alleviate some of those pressures. It amended the Public Health Services Act to include an abbreviated pathway in section 351(k) for biological products shown to be biosimilar to or interchangeable with an FDA licensed reference product. The BPCIA opened the door for biosimilar versions of already approved biologics where “generic” competition did not exist prior.

The FDA continues to clarify biosimilar guidelines. The most recent draft guidance providing additional details was published on May 2014. In an article for Pharm Pro, Sarfaraz K. Niazi examines current FDA regulations and explores the state of the market.

Then article can be accessed here.



Posted by Tim Sandle

ATCC® Minis - new system for QC strains

ATCC, the premier global biological materials resource and standards organization, announces the release of ATCC® Minis to support quality control (QC) testing in pharmaceutical and industrial labs, during the PDA 9th Annual Global Conference on Pharmaceutical Microbiology in Bethesda, MD, Booth # 304.

Healthcare, personal care product, and cosmetic manufacturers are required to test the bio-burden and sterility of their products and production environments to ensure consumer safety. Global alignment and harmonization of microbial testing requirements among the United States Pharmacopeia (USP), Japanese Pharmacopeia (JP), and European Pharmacopeia (EP), have resulted in the need for consistent and reliable control organisms at less than five passages from the ATCC reference stock for reproducible results.

“Many customers spend considerable time and resources creating cryopreserved stocks from ATCC organisms, acquired either directly from ATCC and our exclusive distributors or through ATCC Licensed Derivative® program partners. With the availability of ATCC® Minis, we now provide our high quality strains in a ready to use mini-cryovial format to meet customers’ requirements,” said Dr. Mindy Goldsborough, ATCC Vice President. “We have created single-use glycerol stocks, so our customers can validate their QC work quickly and efficiently without the risk of cross-contamination during banking techniques.”

Only ATCC Genuine Cultures® are authenticated and supported by ATCC polyphasic testing to ensure both microbial identity and phenotypic characteristics. To meet the needs of QC biologists, ATCC developed ATCC® Minis – with the same high-quality ATCC Genuine Cultures® – provided as a six-pack of ready-to-use QC strains preserved in glass-free “mini” cryovials containing glycerol stock. Each tube has a 2D barcode to allow for easy storage and tracking, and offers a peel-off label for fast and reliable recordkeeping.

Today, ATCC has released the following ATCC® Minis for the top 12 strains used in QC testing, including those that support test methods for non-sterile products, such as USP 61 and USP 62: Pseudomonas aeruginosa (ATCC® 9027-MINI-PACKTM), Staphylococcus aureus (ATCC® 6538-MINI-PACKTM), Candida albicans (ATCC® 10231-MINI-PACKTM), Aspergillus brasiliensis (ATCC® 16404-MINI-PACKTM), Escherichia coli (ATCC® 8739-MINI-PACKTM), Bacillus subtilis (ATCC® 6633-MINI-PACKTM), Salmonella enterica (ATCC® 14028-MINI-PACKTM), Escherichia coli (ATCC® 25922-MINI-PACKTM), Staphylococcus aureus (ATCC® 25923-MINI-PACKTM), Pseudomonas aeruginosa (ATCC® 27853-MIN-PACKTM), Enterobacter aerogenes (ATCC® 13048-MINI-PACKTM), and Enterococcus faecalis (ATCC® 29212-MINI-PACKTM). In addition, ATCC is offering convenient ATCC® Minis accessories and a QC pack of the USP recommended ATCC organisms.

For further details, see ATCC.



Posted by Tim Sandle

What is Microbiology?

The world around us is full of organisms that are too small to be seen with naked eye-bacteria, virus, fungi, algae and protozoa. These microbes live in a wide range of habitats from hot springs to the human body and depth of ocean. They affect each and every aspects of life on earth.

We can all think of a few microbes that make us ill – the viruses that cause cold and flu, or food poisoning bacteria. However, there are many more microbes living harmlessly alongside us playing a vital role in the planet’s nutrients cycles, from fixing nitrogen and carbon dioxide at the beginning of the food chain right through to decomposing and recycling essentials nutrients at the end of it.

Microbes are also essential to the production of many foods and medicines – imagine our diet without cheese, bread, yoghurt or a world where the slightest bacterial infection or wound could prove fatal because there were no antibiotics or vaccines.

Microbes have always affected our health, food and environment and they will play an important role in the big issues that face us in the future: climate change, renewable energy resources; healthier lifestyles and controlling diseases.

What do Microbiologist do?

Because microbes have such an effect on our lives, they are a major source of interest and employment to thousands of people. Microbiologists study microbes: where they occur, their survival strategies, how they can affect us and how we can explain them.

All around our planet there are microbiologists making a difference to our lives – maybe ensuring the safety of our food or treating and preventing diseases or developing green technologies or tracking the role of microbes in climate change.

Basic Research

Before Microbiologist can solve the problems caused by microbes, or exploits their amazing powers, they have to find out about the detailed workings of microbial cells. The basic knowledge of genetics, cell structure and function can then be used in applied microbiology as well as in other areas of biology.

Healthcare

Microbiologists are essential in the fight against infectious diseases. Many work as biomedical scientists in hospitals and Health Protection Agency labs, investigating the samples of body tissues and fluids to diagnose infections, monitor treatments or track disease outbreaks. Some microbiologist work as clinical scientists in hospital and medical school laboratories where they carry out research and give scientific advice to medical staff who treat patients. Other microbiologists work on pathogens that cause diseases, such as ‘flu’ or TB, and the information they find is used by their colleagues to develop vaccines and better treatments.

Environment

Some microbiologists study how microbes live alongside other creatures in different habitats such as the oceans, salt lakes and Antarctica. They develop early warning sensors to detect pollution and use microbes to treat industrial waste. Other contributes to the worldwide research on climate change, investigating the effect of microbial processes on the composition of atmosphere and climate. Microbiologists also work with technologists and engineers to develop greener sources of energy produced from urban and industrial waste.

Agriculture

Without agriculture there would be no food for us to eat. Microbiologists investigate the vital role of microbes in soil. Some concentrate on plant pests and diseases, developing ways to control them. Others research the pathogens that cause diseases in farm animals. Microbiologists also use microbes to control insects’ pests and weeds, especially in developing countries.

Business

Microbiologists work in many bioscience and food companies. They carry out research and develop new products or work in quality control to monitor manufacturing processes and check the microbiological safety of goods such as medicines, cosmetics, toiletries, biochemical and food and drink.

Where do they work?

In the lab

Universities, research institutes and industrial companies employ microbiologists to do basic, environmental, healthcare and agricultural research.

Medical Microbiologists also work in hospitals and Health Protection Agency laboratories.
Industrial microbiologist work in a range of companies – from big pharmaceutical, biochemical, biotechnology and food businesses through to smaller firms that develop biopharmaceuticals or specialist products.

Outside the lab

If you still love microbiology but find that lab-based work is not for you, there are still some great options where you can use the scientific knowledge and transferable skill you’ve acquired while studying.
Microbiologists can use their knowledge and skills in a wide range of careers in industry (marketing, technical support and regulatory affairs) education (teaching, museums and science centers), business (patent attorney or accountant) and communications (public relations, journalism and publishing).

Posted by Microbiology World

The advantages of R3A agar for water microorganisms (new paper)

Microbiological quality control of pharmaceutical water systems is of importance in ensuring that trends in contamination are detected and responded to. This is not least because water is a niche environment for many types of microorganisms and a vector for their transfer. Trending relates to actual microbial counts recorded, incidents and the types of species recovered. To facilitate species identification, microorganisms need to be subcultured from the isolation medium (R2A agar in Europe). Transfer onto the wrong media can result in the microorganism not growing. This paper describes research into three different media for subculturing: low nutrient (R2A); highly nutritious (TSA) and medium nutrient (R3A) and concludes that a higher recovery is obtained where R3A agar is used.

This is the abstract to a new research paper by Tim Sandle.

The reference is:

Sandle, T. (2014) Assessment of the suitability of R3A agar for the subculture of microorganisms isolated from pharmaceutical water systems, European Journal of Parenteral and Pharmaceutical Sciences, 19 (3): 85-94

If you are interested in a copy, please contact Tim Sandle



 Posted by Tim Sandle

Real-time tracking system developed to monitor dangerous bacteria

Combining a PET scanner with a new chemical tracer that selectively tags specific types of bacteria, researchers working with mice report they have devised a way to detect and monitor in real time infections with pathogenic Gram-negative bacteria.

The new model emerged from a combination of existing PET scan technology -- a sophisticated 3-D visualization system for tumor imaging -- with an ingredient commonly used in sugar-free foods known as sorbitol. The model capitalizes on Gram-negative bacteria's fondness for sorbitol, which they readily soak up. By contrast, other classes of bacteria and other microorganisms, cancer, and human cells do not absorb sorbitol. The researchers hypothesized that converting an already available PET imaging tracer into radio-labeled sorbitol would selectively tag and light up clusters of Gram-negative bacteria inside the body.

For further details, see:

Edward A. Weinstein, Alvaro A. Ordonez, Vincent P. Demarco, Allison M. Murawski, Supriya Pokkali, Elizabeth M. Macdonald, Mariah Klunk, Ronnie C. Mease, Martin G. Pomper, and Sanjay K. Jain. Imaging Enterobacteriaceae infection in vivo with 18F-fluorodeoxysorbitol positron emission tomography. Science Translational Medicine, October 2014 DOI: 10.1126/scitranslmed.3009815

Posted by Tim Sandle

USP 2014, 2nd supplement

With: United States Pharmacopiea and National Formulary, 2nd Supplement to USP37-NF32. Official 01 Dec 2014, there are some items of interest:

a)      General Chapters

Listed are items in which changes have been made to existing official text:

Physical Tests and Determinations

Chapter 791 pH

Chapter 1196 Pharmacopeial Harmonization

Chapter 1229.6 Liquid Phase Sterilization

Chapter 1240  Virus Testing of Human Plasma for Further Manufacture [NEW]

b)      USP and NF Excipients

Updates to the following

• Polysorbate 80: Identification – Test A and Infrared Absorption, Test B (added), Assay – Composition of Fatty Acids, Impurities – Heavy Metals, Method II, Specific Tests – Specific Gravity and Viscosity – Capillary Viscometer Methods or Rotational Rheometer Methods, Additional Requirements - USP Reference Standards.
• Cupric Sulfate: Specific Tests – Loss on Drying, Additional Requirements – Labelling (added)

Posted by Tim Sandle

Endotoxin testing – European Pharmacopeia changes

A proposal has been made to amend the European Pharmacopeia chapter on guidance for the bacterial endotoxin test – chapter 5.1.10.

The proposed changes are:

In the context of the new bacterial endotoxins Ph. Eur. policy, a new section (2-4) includes aspects to be considered when establishing an endotoxin limit for a specific substance or product; also, reference is made to the fact that an endotoxin limit is not always provided in a specific monograph.

– Reference is made to general chapter 2.6.30. Monocyte-activation test as an alternative to the rabbit pyrogen test, and a recommendation is given to perform a risk assessment when using the bacterial endotoxin test as a pyrogenicity test, due to the potential for contamination by non-endotoxin pyrogens. In this respect, the previous section 11 concerning the replacement of the rabbit pyrogen test by a test for bacterial endotoxins is substituted with a new text in agreement with a strategy to be applied for testing of bacterial endotoxins or non-endotoxin pyrogens. A distinction is made between replacement methods already described in the Ph. Eur. and other alternative methods.

– In the context of the 3R’s, a test for bacterial endotoxins or a monocyte-activation test is preferred to the rabbit pyrogen test. Therefore, implementation of the rabbit pyrogen test has to be justified and authorised and is appropriate only when the bacterial endotoxin test or the monocyte-activation test cannot be validated.

– Reference is made to the use of alternative reagents to the Limulus amoebocyte lysate, such as recombinant factor C: this practice avoids the use of endangered animal species and can be considered in the context of the use of an alternative method as described in the General Notices.

A number of additional specific revisions have been made.

– In line with current knowledge, Method A is no longer declared as the reference method, and all methods A to F of general chapter 2.6.14. Bacterial endotoxins can be used. Where the method is stated in the monograph, the use of another method must be supported by evidence of validation.

–The expression ‘threshold endotoxin concentration’ has been replaced by the more appropriate expression ‘endotoxin limit concentration’ to harmonise with general chapter 2.6.30. Monocyte-activation test.

– A new entry has been included in Table 5.1.10.-1 for formulations administered per square metre of body surface.

Finally, the structure of the general chapter has been modified to improve its clarity.

The proposal has been added to Pharmeuropa 26.4.

Posted by Tim Sandle

Pharmig News #57

A new edition of Pharmig News has been published. The current issue features:

• An article by Tony Cundell on bioburden control of non-sterile drug substances and products, based on a critique of the new USP chapter 1115
• A review article, presenting research on Streptococcus anginas, by Antonella Chesca from the Transilvania University in Romania
• The regular update on standards, monographs and articles on interest, by Tim Sandle.

Copies have been sent to Pharmig members. If you are interested in reading a copy, please contact the Pharmig office.

Posted by Tim Sandle

Medicines and the Microbiome

We now know more than ever before about the complex ecosystem of bacteria, fungi, and viruses within our bodies. How can that knowledge be best applied in drug development and manufacturing?

This is the basis of a short opinion piece written by Tim Sandle for a new magazine called The Medicine Maker.

The issue can be accessed here.

The reference is:

Sandle, T. (2014) Medicines and the Microbiome, The Medicine Maker, Issue 1, p18

Exclusion of Objectionable Microorganisms

The PDA has issued a new publication of interest:

PDA Technical Report No. 67 'Exclusion of Objectionable Microorganisms from Nonsterile Pharmaceuticals, Medical Devices, and Cosmetics'.

The purpose of the publication is:

“The purpose of this technical report is to provide guidance to the nonsterile producmanufacturing industry on how to manage the microbial risks associated with manufacturing and storage and how to determine what isolates would be deemed an objectionable microorganism in nonsterile products that is in alignment with the microbial limits requirements for releasing these products into the marketplace. Nonsterile products exceeding the microbial count limit and/or containing specified microorganisms for their product type would be expected to be rejected. Specified microorganisms include microorganisms with compendial requirements to be absent in a particular dosage form, and/or required by a national board of health to be excluded from a registered non-sterile product. The contamination of marketed products by potentially objectionable microorganisms continues to be an infrequent but chronic problem. A U.S. survey of reported microbiologically related recalls between 2004 and 2011 found that 72% of recalls of nonsterile products were associated with objectionable microorganisms rather than exceeding microbial enumeration limits (1). Of the 144 recalls for nonsterile products, 5% involved nonsterile pharmaceutical drug products, 42% were for OTC drug products, 31% were for cosmetics, 14% were for medical devices and 8% were for dietary supplements. The average rate of reported recalls is 20 per year.”

For details see: PDA

Posted by Tim Sandle

Pyrogen testing – European Pharmacopeia

The Ph. Eur. is proposing an amendment for the pyrogen test monograph (2.6.8). The only change is in directing users to consider using an alternative to the rabbit model.

The change states:

“In accordance with the provisions of the European Convention for the Protection of Vertebrate Animals used for Experimental and Other Scientific Purposes, tests must be carried out in such a way as to use the minimum number of animals and to cause the least pain, suffering, distress or lasting harm. Wherever possible and after product-specific validation, the pyrogen test is replaced by the monocyte-activation test (2.6.30).”

Interestingly, the recommendation is towards the MAT rather than LAL testing.

The proposal has been added to Pharmeuropa 26.4.



 Posted by Tim Sandle

Fungicidal activity of biocides against fungal isolates

A new paper of interest has been published: “In vitro fungicidal activity of biocides against pharmaceutical environmental fungal isolates”.

The paper has been written by T. Sandle, R. Vijayakumar, M. Saleh Al Aboody and S. Saravanakumar and it has been published in the Journal of Applied Microbiology.

The abstract states:

Aims: To determine the minimum inhibitory concentrations (MIC) of a range of cleanroom fungi against three disinfectants common to the pharmaceutical and healthcare sectors: biguanide (chlorhexidine) and two quaternary ammonium compounds (benzalkonium chloride and cetrimide).

Methods and Results: The in vitro fungicidal activities of the three biocides were studied against 112 cleanroom fungal isolates using broth microdilution technique (CLSI M38-A2 standard).

Conclusions: Minimum inhibitory concentration (MIC) for all three biocides against hyaline fungi showed results of not more than 16 lg ml^-1. Alternaria showed <32 lg ml^-1 and other dematiaceous fungi reported that 8–16 lg ml^-1 for biguanides and QACs. This study clearly demonstrates that the most frequently isolated micro-organisms from an environmental monitoring programme may be periodically subjected to broth microdilution testing with cleanroom disinfectant agents used in the disinfection programme confirm their sensitivity profile.

Significance and Impact of the Study: No large collection of data exists on the activity of biocides on pharmaceutical cleanroom fungal isolates. This is the first study report with large collection of cleanroom fungal isolates tested against common biocides using the broth microdilution antifungal susceptibility testing to determine the MIC value. The data presented support a quality control procedure for cleanroom disinfection.

The reference is:

Sandle, T., Vijayakumar, R., Saleh Al Aboody, M. and Saravanakumar, S. (2014) In vitro fungicidal activity of biocides against pharmaceutical environmental fungal isolates, Journal of Applied Microbiology, 117 (5): 1267 – 1273

For further details, please contact Tim Sandle

Posted by Tim Sandle

Fighting a pathogen with gut microbes

Clostridium difficile infections can be prevented through the rebalancing of bile acids in the gut by introducing certain commensal microbes, according to a study published in Nature. Eric Pamer of the Memorial Sloan Kettering Cancer Center in New York City and his colleagues have demonstrated the efficacy of this approach in both mice and humans.

While antibiotics work wonders fighting myriad infections, they can also wipe out the large contingent of beneficial bacteria that populate the human gut. Such widespread, indiscriminate destruction paves the way for infectious pathogens, including C. difficile—which can cause extreme diarrhea, abdominal pain, and if left untreated, even death.

The researchers sequenced the gut microbiomes of both mice and humans in search of individual species capable of eradicating infection. They discovered that in antibiotic-treated mice, CDI resistance was strongly correlated with the abundances of C. scindens and, to a lesser extent, 10 other bacterial taxa. A similar exploration of the intestinal microbiota of human patients undergoing a stem-cell transplantation procedure that left them vulnerable to CDI also pointed to C. scindens as the most potent source of infection resistance.

Administering C. scindens to antibiotic-treated mice infected with C. difficile resulted in significantly increased survival rates—80 percent, versus 50 percent for a control substance. A cocktail of C. scindens plus three other C. difficile-inhibiting bacteria was even more effective, resulting in 100 percent survival. In addition, the abundance of both C. scindens and the bacterial cocktail were strongly correlated with resistance.

Next, Pamer’s group investigated the mechanism underlying C. scindens-mediated inhibition of infection, focusing on the ability of the microbe to express the enzyme 7a-hydroxysteroid dehydrogenase. This enzyme, rare among intestinal bacteria, is critical for converting primary bile acids to secondary bile acids in the colon. C. difficile spores interpret the presence of primary bile acids as a signal that they are in the gut and should start germinating. The researchers hypothesized that C. scindens may prevent C. difficile growth by blocking this signal. To confirm this hypothesis, they loaded Petri dishes with the intestinal contents of antibiotic-exposed mice. As expected, C. scindens inhibited C. difficile. But when they added a chemical that binds bile acids, C. difficile flourished, suggesting that C. scindens-mediated inhibition of C. difficile is dependent upon modifying endogenous bile acids.

For further details, see the following paper published in Nature: "Precision microbiome reconstitution restores bile acid mediated resistance to Clostridium difficile."

Posted by The Scientist

Archaeal origins and bacterial gene acquisitions

A phylogenetic analysis of more than 25,000 archaeal gene families found that the integration of bacterial genes into the genome paralleled the pattern of archaeal-specific genes in each of 13 orders of archaea, suggesting that archaeal lineages picked up groups of bacterial genes at the time of their formation.

About a third of the archaeal genes analyzed had homologs among bacterial genomes, and the phylogenies of these apparent genetic imports closely mirrored those of archaeal-specific genes in each of 13 orders of archaea, suggesting that the birth of each order coincided with the gene acquisitions.

The phylogenies revealed that 83 percent of the bacterial introductions occurred in methanogenic archaea, which are thought by some to be the most ancient groups of archaea and have a simple metabolism—anaerobic reduction of carbon dioxide to methane in the presence of hydrogen gas.

The conclusion that archaeal lineages abruptly picked up large groups of bacterial genes at the time of their formation, though supported by statistical tests, will require further analysis.

For further information, see Nature "Origins of major archaeal clades correspond to gene acquisitions from bacteria."

 Posted by Tim Sandle
(Reference: The Scientist)

European Pharmacopeia: Preparation of sterile products

The European Pharmacopeia has put the following revised chapter out for public comment:

“5.1.1. METHODS OF PREPARATION OF STERILE PRODUCTS”

The chapter states:

Sterility is the absence of viable micro-organisms, as defined by a sterility assurance level of 10^{− 6}. Sterility is a critical quality attribute for a wide variety of human and veterinary preparations, including but not restricted to:

• Preparations administered to normally sterile areas of the body, such as parenteral and ophthalmic preparations, some irrigation and inhalation preparations, intramammary and intrauterine preparations;
• Preparations applied to severely injured skin, such as semi-solid preparations for cutaneous
• application;
• Preparations that are supplied sterile in order to prolong the shelf life of the product.

With the revised chapter, the following revisions are proposed: 

• In the introduction, the reference to GMP has been removed;
• In the section Sterility assurance level, the reference to exponential inactivation has been removed as membrane filtration is not a first order process;
• The sections on the different sterilisation processes, where appropriate, now have the same format: principle, equipment, sterilisation cycle, cycle effectiveness and routine control; where required, relevant information has been added;
• Modern concepts for validation of steam sterilisation have been added;
• A wider description of the equipment suitable for dry heat sterilisation has been provided;
• In the section Ionising radiation sterilisation, the reference to European Notes for Guidance
• has been removed;
• In the section Gas sterilisation, two different types of agents are defined: alkylating agents and oxidising agents. The establishment of the cycle effectiveness has been described in more detail;
• In the section Membrane filtration, the description of the microbial challenge test has been removed as it is proposed for inclusion in the revised chapter on biological indicators;
• In the section Aseptic preparation, freeze-drying under aseptic conditions is added.

The proposed revision is published in Pharmeuropa which is the free online publication from the EDQM.

Posted by Tim Sandle

ISO 14644 Part 1 update

Introduction

As part of the ISO periodic review cycle, ISO 14644-1, Cleanrooms and associated controlled environments — Part 1: Classification of air cleanliness is being revised. ISO TC209 Working Group 01, an international team of subject matter experts, is planning to assess the existing ISO 14644 document and propose changes for enhancements.

After discussing with statisticians, the expert working group is recommending modifications to the classification method. It is also implementing the key elements of ISO 14644-3, Cleanrooms and associated controlled environments — Part 3: Test methods in an effort to improve the document and thus enhance credibility and resulting quality.

Changes in Classification Method

The present ISO 14644-1 document depends on an ad-hoc approach to ascertain the location and number of sampling areas for classifying cleanrooms. This method, however, is not built on statistical sampling technique and also does not consider any risk-based evaluation. At present, the working group is recommending a randomized sampling location selection technique, in tandem with a risk-assessed, fixed-location sampling technique.

Founded on the hypergeometric statistical model, the randomized sampling technique enhances confidence that the selected sample provides 95% assurance that the results represent 90% of all areas in the cleanroom. Examining the additional risk, the fixed sampling location technique enables cleanroom users to choose the areas where their product is believed to be most at risk. When the cleanroom is classified first, these locations will be sampled, and whenever the room is re-qualified the locations will be sampled again.
When combined together, these two methods provide improved confidence that the cleanroom is operating as expected and ideal for providing the right quality-controlled environment for life science applications. These methods are better than the method specified in the existing ISO 14644-1 document.

Improving Confidence in Quality

In addition to cleanroom monitoring and classification applications in the life sciences and pharmaceutical industries, there are certain particle counters that can be utilized in indoor air quality  and other less critical industrial applications.

Figure 1. Particle counters can be designed for different applications

Although a number of air particle counters available in the market are designed for use in all three application areas, some give incorrect results in low particle concentration environments such as life-critical cleanroom applications, while others are unable to handle high particle population environments found in the industrial applications. Therefore, particle counters developed for the less critical industrial applications may not meet the rigorous requirements specified in ISO 21501-4 and ISO 14644-3.

In an effort to enhance confidence in the quality of cleanroom environments, the amended ISO 14644-1 document will comprise the performance criteria of particle counter specified in ISO 14644-3 and give a normative reference to ISO 21501-4, Determination of particle size distribution — Single particle light interaction methods — Part 4: Light scattering airborne particle counter for clean spaces, to guide the reader through a consistent test method for the calibration of particle counters. The ISO 21501-4 document was initially published in 2007.

Additionally, manufacturers of particle counters and national standards agencies have publicized the normative reference to ISO 21501-4 in the amended document ever since the initial draft of the revised ISO 14644-1 document was published in December 2010. Therefore, the working group believed that, by the estimated publication date of the amended ISO 14644-1 in 2013, the industries using the ISO 14644-1 document would have sufficient notification of the need to conform to the ISO 21501-4 calibration method and to also upgrade any particle counters that are non-compliant.

Prior to the publication of ISO 21501-4 in 2007, no calibration standards were available for airborne particle counters which resulted in fluctuating levels of performance from particle counting systems. Since 2007, many particle counter manufacturers across the globe have modified the designs of their particle counters meant for critical life science ISO 14644 applications, so that these instruments can fulfill the more complex expectations of the latest calibration standard. Now, cleanroom users can have improved confidence that their cleanrooms are providing the required level of quality-controlled environment for their specific processes.

Content Imported from ISO 14644-3

The performance requirements of particle counter have been laid down in the ISO 14644-3 document since its first publication in 2005 and corresponds with the ISO 21501-4.

Figure 2. Specifications for the light-scattering discrete-particle counter from ISO 14644-3

In an effort to enhance clarity, the International Standards Organization Technical Committee ISO TC 209 determined that particle counter performance methods and criteria used to attain classification should be integrated into each ISO 14644 standard. Therefore, the performance criteria of particle counter specified in Table C.1 of ISO 14644-3 will appear in the amended ISO 14644-1 document.

Publication timeline

The initial publication of the Draft International Standard (DIS) DIS ISO 14644-1 in December 2010 triggered a heated debate. After assessing the questions and comments from the international community with regard to the DIS, the expert working group planned to publish an amended draft DIS document in 2013. In the interim, ISO Technical Committee TC209 recommends that users who want to leverage the improvements in the 2010 DIS ISO 14644-1 document can use it, as long as they know that the final published version will be modified.

Conclusion

The improved sampling method in the revised ISO 14644-1 document along with the enhanced guidance for particle counter calibration and performance will provide improved confidence to cleanroom users that their cleanroom is delivering the preferred level of quality-controlled environment for critical life science applications.

Posted by Tim Sandle