By Jennifer Johnson
Routine microbiological testing carried out by the pharmaceutical, cosmetics and toiletry manufacturing industries is determined by regulatory requirements. This applies not only to what should be tested and when, but also to the methods that should be used. Whether the final product is sterile or non-sterile, the bioburden exists from the raw materials, throughout the process and/or within the product’s environment (e.g. water) to the final product. A critical review of the overall microbiological process will determine whether the final product will meet its acceptance criteria. In addition, any “objectionable” or “specified” microorganisms that may be encountered during the procurement of raw materials and the processing must be considered.
Whether you are testing a raw material, an in-process sample, final product, or the environment, you should be aware of the critical role that microorganisms play throughout the process. You should also be familiar with the various microbiological related documents needed to determine whether the SOPs, validations, and regulatory body documents are being maintained to assure the control required to permit the final product to enter the marketplace as safe.
The Unites States Pharmacopeia (USP) is a non-governmental organization. Their documents are recognized as an official compendia book of methods and standards for Pharmaceutical, nutraceutical, dietary supplements, cosmetics, and toiletries. The USP offers a variety of documents such as monographs (published standards) and general chapters. General chapters with numbers <1000 are mandatory chapters and numbers >1000 are considered informational chapters.
Pharmaceutical USP Chapters
USP offers some standardized test methodologies and material specifications relating to microbiological quality and control so as to ensure product quality and safety.
<1111> Microbiological Examination of Nonsterile Products: Acceptance Criteria for Pharmaceutical Preparations and Substances for Pharmaceutical Use – It is suggested that certain categories of products should be tested routinely for total microbial count and for specified indicator microbial contamination. The relevant tests for determining the total count of viable aerobic microorganisms and the total combined yeast and mold count and for detection and identification of designated species are given under Microbial Limit Tests <61>.
<1115> – Bioburden Control of Nonsterile Drug Substances and Products – This chapter deals with the bioburden of non-sterile drug substances and products. The chapter states that the biggest manufacturing risk for these products is the ingredient water. Process water is the single most important risk factor contributing to the contamination of nonsterile products. The purified waters that are used in manufacturing are deionized and do not contain chlorine that helps control microbial growth. Purified water is capable of supporting growth of gram negative rod shaped bacteria and many different molds.
For more information, click here.
<1116> Microbiological Control and Monitoring of Aseptic Processing Environments – The purpose of this chapter is to review the various issues that relate to aseptic processing of bulk drug substances, dosage forms, and to the establishment, maintenance, and control of the microbiological quality of controlled environments.
For more information, click here.
<1117> Microbiological Best Laboratory Practices - This general information chapter outlines what is needed to improve the effectiveness, efficiency and inspection-readiness of a Microbiology Laboratory in respect to media preparation, microbiological cultures, lab equipment, laboratory layout, lab records, interpretation of results, training, and documentation.
<1223> Validation of Alternative Microbial Methods – The purpose of this chapter is to provide guidance for validating methods for use as alternatives to the official compendial microbiological methods. Validation of microbiological methods is the process by which it is experimentally established that the performance characteristics of the method meet the requirements for the intended application, in comparison to the traditional method. Traditional plate methods for microbiological testing require long time to results, highly trained microbiologists, is labor intensive, and creates a bottleneck for product release. Rapid microbiological methods (RMMs) offer a high degree of automation, significant reduction in time to results, reduced staff training, faster product release, and improved process control.
<1227> Validation of Microbial Recovery from Pharmacopeial Articles – This chapter
provides guidelines for the validation of methods for the estimation of the number of viable microorganisms, for the detection of indicators or objectionable microorganisms, for the validation of microbiological methods used in antimicrobial effectiveness testing, and for the sterility testing of Pharmacopeial articles. It is generally understood that if a product possesses antimicrobial properties because of the presence of a specific preservative or because of its formulation, this antimicrobial property must be neutralized to recover viable microorganisms. This neutralization may be achieved by the use of a specific neutralizer, by dilution, by a combination of washing and dilution, or by any combination of these methods.
<1231> Water for Pharmaceutical Purposes – According to USP <1231>, although there are no absolute microbial standards for water (other than water intended to be sterile), the cGMP regulations require the establishment of appropriate specifications. The specification must take into account the intended use of the water; i.e., water used to formulate a product should contain no organisms capable of growing in the product. Action or alert limits should be established based upon validation data and must be set low enough to signal significant changes from normal operating conditions.
BioLumix Rapid Microbiology Methods offer innovative new testing solutions that are designed specifically for the challenges of the pharmaceutical industry, including a multitude of microbial tests for raw materials, in-process and finished products, as well as, the capacity to perform environmental and water testing. This simplifies, expedites, and reduces the cost of performing microbiological assays.
The BioLumix Drug Master File (DMF) entitled “Microbiological Rapid Method for the Detection and Enumeration of Microorganisms in Pharmaceutical Products” has been accepted by the US Food and Drug Administration (FDA). The DMF provides specific technical and regulatory information to the FDA which allows companies planning to use the BioLumix Rapid Microbiological System to obtain regulatory approvals for prescription drugs. The DMF can be referenced by drug manufacturers, reducing FDA review times and accelerating the regulatory approval process for use of the BioLumix System. In addition, Over the Counter (OTC) products can be tested in conjunction with the BioLumix Validation Package.
For more information, click here and here.
Posted by Caron Ockerman
Many pharmaceutical, cosmetic, and toiletry products contain preservatives that help extend the product’s shelf life, keeping spoilage at bay for a certain period of time, and also act with antimicrobial properties if microbes are introduced. A consumer dipping a finger into a jar of facial cream has introduced microbes into the cream, and the preservative should effectively kill or reduce the number of microorganisms (for more information on preservative effectiveness, please read the blog “Preservative Efficacy Testing: Guidelines to an Automated Simplified Testing System”).
Why we need suitability testing? The measurement of preservative test requires the ability to count the number of surviving microorganisms after exposure to the antimicrobial agent in the product. Bioburden (also called microbial content) testing has the same requirement because the assay depends on the ability to recover viable microorganisms in the presence of potentially antimicrobial in the product or raw materials. Carryover of residual inhibitors from the product could inhibit growth in the recovery medium, leading to poor microbial recovery. This potential residual activity must be neutralized and it is necessary to demonstrate the adequacy of neutralization for the test. This demonstration of neutralization in compendial microbiological tests is known as demonstration of method suitability.
Therefore, before conducting any microbiological test, it is important to find the most effective neutralizing agent or dilution that will create a suitable environment for growth, allowing for accurate results during microbial content and preservative efficacy testing.
How to test for Suitability- Suitability in the presence of product is the ability of microorganisms to grow when introduced in low numbers. Following the plate count method for suitability in USP Chapter <61> MICROBIOLOGICAL EXAMINATION OF NONSTERILE PRODUCTS: MICROBIAL ENUMERATION TESTS, the appropriate growth media alone (control) and the product (test) are inoculated with 10-100 cfu. The inoculated product is then diluted 1 to 10. If growth occurs on control plates, but does not grow on test plates, or counts are half that of the control plates, neutralization or further dilution may be necessary to show suitability of product.
How to Neutralize Inhibitors- Depending upon the type of product and the inhibitory system being tested, different diluents should be used as some products may require neutralization of a strong preservative that would suppress growth of naturally occurring microbes. Table 1 below summarizes some neutralizing agents used in common neutralizing broths such as Modified Letheen Tween (MLT) broth, D/E Neutralizer, and Tryptone Azolectin Tween (TAT) broth among others.
Sometimes, even the use of a neutralizing agent may not be enough, and dilution further than 1 to 10 may be necessary. It is important to keep in mind that five different organisms are being tested in one product, and what allows one organism to grow might not work as well for another. Essentially, it is important to find any and all different parameters that make a product suitable for inoculation.
Once a proper neutralizer or dilution is found for each product and organism, microbial content testing and PET testing can proceed. Without performing suitability testing first, microbial content testing and PET may yield false negative results; bacteria, yeast or mold may be present, but simply remain dormant until the optimal growth conditions are introduced.
The suitability test can be performed very simply and quickly with the BioLumix system.
Use of BioLumix System for suitability testing- The BioLumix system is capable of determining whether a product meets suitability standards. Products and controls are inoculated, then 1.0 mL is added to the BioLumix vials, entered into the instruments, and initial results may be observed within 18-24 hours. The figure shows examples of curves obtained from the BioLumix system using Staphylococcus aureus with various preservative systems. The top picture shows a product that is properly neutralized. While as the bottom picture demonstrates that without proper neutralization, product can hinder the growth of organisms.
In the BioLumix instrument, if the detection time (DT) of the media without product is within a couple of hours of the product containing media it means that enough neutralization is achieved. If there is a DT without the product in the positive control; but the sample containing vial does not have a DT it means that the product prevents the growth of bacteria and a different neutralization scheme needs to be employed.
What are the advantages of the BioLumix system?
Time Saving: The results are available much faster, for example, the results of the Yeast and Mold vials occurred in less than 48 hours, instead of five days for countable colonies. All products tested with bacteria using the automated BioLumix assay yielded results typically within 10-18 hours, instead of 48 hours for the plate count methodology. The advantage using the BioLumix system is that you can see results an entire day earlier if the product is exhibiting any sort of inhibition.
Labor Saving: The setup of the assay can be done much faster using the BioLumix system as opposed to traditional plating methods, saving significant hands-on labor due to its automation and simplicity of use.
High Correlation with USP: The BioLumix System showed a high correlation between the instrument results and the USP methodology.
Neogen Corporation (NASDAQ: NEOG) announced today that, effective Sept. 30, it has acquired all the outstanding stock of BioLumix, Inc., an Ann Arbor, Mich.-based manufacturer and marketer of automated systems for the detection of microbial contaminants.
The BioLumix business will be consolidated with Neogen’s closely related Soleris® technology, which is widely used for the detection of spoilage organisms in several food industries, as well as the nutraceutical market. The Soleris system was the first in the industry used for the detection of microbial contamination based upon an innovative application of well-accepted classical microbiology.
“Combining of the Soleris and BioLumix technologies, market bases, and outstanding technical staffs will greatly enhance both businesses and offer significant labor saving rapid solutions for the food, pharmaceutical, and personal care businesses,” said James Herbert, Neogen’s chief executive officer and chairman. “The combination also settles litigation of seven years between the two companies, and benefits existing customers of both companies moving forward.”
The BioLumix test platform includes an instrument where test vials are incubated and automatically read for results, and an automatic system to alert users to sample results. The basic unit accommodates 32 different test vials at a time and can be combined in modules to accommodate up to 1,000 different samples simultaneously.
Test vials are the consumable portion of the platform and contain growth media and indicators for a number of different microorganisms. A sample is simply added to the vial, and the vial inserted into the instrument. BioLumix currently has 19 different microorganism tests. Specialized software shows test results as soon as detections occur, and avoids the need for involvement from a hands-on operator. The software also creates and maintains all the necessary audit trails to comply with various government regulations.
Terms of the agreement were not disclosed.
For the full press release, click here.
Web-related information about microbiology is plentiful. There are a variety of social media resources available to a microbiologist, from scientific information, to opportunities to benchmark practices, obtain training opportunities, and to keep current with the latest news and developments in virtually any area of interest .All it takes is a little research and an open mind.
Available Information Sources
News and Company Sites
In recent years, there have been more and more websites dedicated to microbiology on the Internet. There are also sites dedicated to providing news, as well as, websites dedicated to providing microbiology supplies.
A few examples of major sites containing information and news related specifically to microbiology are:
- American Society for Microbiology
- ScienceDaily – Microbiology News
- The Scientist
- The Independent
BioLumix News and Information:
The BioLumix website is updated frequently with information about the technology, new development and current events. Click on the icon to connect to BioLumix website
Blogs can be viewed as online journals that are created by individuals, groups, or companies. Blogs has exploded in recent years due to the ease of blogging and the multitude of hosting services available. The growth of the blogosphere has become so pervasive that some question the viability of print magazines.
Many blogs, of interest to a microbiologist, are published frequently and may well serve to replace printed publications in terms of timely information. Below are a few examples:
- Small Things Considered
- Microbiology Network Blog
- Pharmaceutical Microbiology
- The RMM Blog
BioLumix posts bi-weekly blogs on a variety of topics in rapid microbiology, about regulations, and about the BioLumix technology. To stay current on our new developments click on the icon to connect to BioLumix blog.
E Mail discussion Groups
Email discussion groups are created to encourage discussion around a unifying topic. Users subscribe to the group. The basic mechanism is to have a central Email address to which all mails are sent. If a subscriber has a question or comment it is sent by Email to that address. The email is then resent to all of the participants. If one (or more) participant wishes to comment on the Email, that reply is then sent to the central address for redistribution and the cycle repeats.
The Email discussion group is the oldest form of social media participation. There are two Email discussion groups of use to the microbiologist – the PMFList and the PSDGList. Both focus on Pharmaceutical production and validation.
LinkedIn is a business-oriented social network and it provides a variety of opportunities to connect with other microbiologists and join discussions. LinkedIn also supports the formation of interest groups, and many people use the groups on LinkedIn as a kind of think tank or open brainstorming session. Others use it to disseminate information. It is an essential business resource of information, LinkedIn can help you stay up-to-date with industry trends and share information with others who do similar work to you. Many companies have a company page on LinkedIn.
Examples of relevant discussion groups:
- Rapid Microbiology Testing and Methods
- Rapid Microbiology
- Microbiology Professionals
- Food Microbiology- HACCP
- Pharmaceutical Microbiology
BioLumix on LinkedIn:
BioLumix has a company page on LinkedIn that contains relevant information about the products as well as corporate news. To view our LinkedIn page click on the icon. BioLumix Linked In page:
BioLumix also sponsors the discussion group Rapid Microbiology Testing and Methods. To join the group and read the content of this informative group click on the icon. Rapid Microbiology Testing and Methods
Facebook defines social media today and is more of a pure social media, than it is used for personal purposes. However, in recent years it started serving also in the promotion of companies and services. Many companies have Company Pages that can promote their service or products with information and special offers.
BioLumix on Facebook: To view the BioLumix Facebook page click on the icon.
Twitter is a messaging service that allows users to post short (140 character) “Tweets” to their followers. Its main use is not in science, but it can offer a good way to rapidly disseminate information. Many Companies and organizations are using this service to get information out quickly and to direct people to web-based information sources. The very brevity of the tweet format can encourage concise expression of ideas and concepts.
A few examples:
BioLumix on Twitter:
To view the BioLumix Twitter page click on the icon.
Google Plus was established in the beginning of 2013 and is currently the second largest social network in the world. It was launched by Google, in order to evolve the way we relate to one another, and how we communicate with the world. At the moment its presence in the microbiology world is limited, but with the Google power behind it is expected to be a major source of information in the near future. Some companies have Google+ pages.
BioLumix on Google+:
To view the BioLumix Google+ page click on the icon.
YouTube is a video sharing service. Video provides an excellent opportunity to explain and teach scientific concepts. In the current impatient, busy world, many people would prefer to watch a video instead of trying to digest massive pieces of written information. Since more consumers are spending time online instead of watching TV, the popularity of watching videos online is growing exponentially.
A few examples are:
- Microbiology video library
- Microbeworld Video
BioLumix on YouTube:
BioLumix has posted many videos on-line explaining the technology and dealing with a variety of topics of interest to microbiologist. To view the BioLumix YouTube videos click on the icon.
A year ago Scott Sutton and the Microbiology Network conducted a survey of the use of social media by microbiologists in a few regulated industries. There has been a significant increase in the use of social media by scientists and microbiologists as these sources provide an opportunities to keep current with the latest in news and developments in virtually any area of interest to the microbiologist.
This survey was conducted online, with a group that is part of an internet email group. It must therefore be assumed that the respondents are more technologically sophisticated than is the norm, and had familiarity with social media on its side.
The survey looked at two main questions – What social media are being used and what is it used for?
As shown in the figure, LinkedIn was the most popular social media used by microbiologists, followed by Facebook. The main reason for using the social media was for personal interest, followed by Benchmarking. This survey also shows that once a technology matures (for example the PMFList and the PSDGList Email discussion groups) participation becomes enthusiastic.
The internet and social media offer a wealth of information and capabilities to the microbiologist; it seems as if more microbiologists can take advantage of the opportunities presented. BioLumix participates in many of the forms of social media and offers a wealth of information to microbiologists
Come to Booth #16091 @ Supply Side West 2014 and Learn More
The implementation of cGMP for all manufacturers (large and small) of dietary supplements and Nutraceutical products means that all manufacturers must test their products for microbiology quality assurance and generate a Certificate of Analysis for each batch. Products must be tested in accordance with the methods described in USP <2021> and <2022>. This includes testing for Total Aerobic Microbial Count, Total Combined Yeast and Mold, Bile-Tolerant-Gram-Negative bacteria, and objectionable organisms (absent in 10 grams of organisms such as Salmonella, E. coli, and sometimes S. aureus). To do these tests companies either send product samples to contract laboratories for microbiological testing or test them internally.
Internal testing of products for microbiology gives the manufacturer much more control over the operation, but some manufacturers are hesitant to bring the microbiology testing in-house for fear of the complexity of testing or the need for trained microbiologist(s). Also daunting is the need for a full validation package which is required for any method that deviates from the USP methodology. Here’s the good news – BioLumix offers a solution that gives faster time to results, quicker product release, cost effective operation, and strengthens key profit drivers. This is all achieved on a single platform that is easy to use, validated to USP requirements, and doesn’t require additional skilled personnel.
At BioLumix, we don’t just offer you a simplified, rapid, automated microbiology system… we provide the Complete BioLumix Experience!
What is the Complete BioLumix Experience? It’s having, from initial contact to the completion of the installation and beyond, our commitment to providing unparalleled support and total customer satisfaction.
In 2006, Ruth & Gideon Eden established BioLumix, Inc. Today, because of their extensive backgrounds in science and technology combined with customer-oriented sales and support staff, we have raised the simplification, automation and speed of microbiology testing to a new level.
- Needs Assessment - Whether you contact us direct or through our website you can count on our sales team to respond quickly. The focus is on your needs and goals. Our BioLumix Technology Presentation provides an in depth look into how and why our system will work for you. In addition, we can quickly and easily show you the value and the ROI the system will bring to your company.
- Expertise – Knowing how the BioLumix system will work with your samples gives you peace of mind. That’s why we test samples – YOUR samples – for you at our laboratory in Ann Arbor, MI. We do this for you at no cost and at no obligation! Once completed, expect a comprehensive report from one of our expert microbiologists. That same microbiologist spends two and a half days on-site for training and installation of the system.
- Validation – If you are concerned about validation, here is more good news. We provide complete SOPs along with a customized Validation book which includes Installation Qualification (IQ), Operations Qualification (OQ) and Performance Qualification (PQ).
- Simplicity and Ease-of-Use- The easy to master BioLumix system offers a new level of productivity. Sample preparation takes less than 2 minutes; just add the sample to the diluent and mix. Pipette the appropriate amount into the vial and place it into the instrument and record the sample and assay information into the computer. The rest is totally automated. The Certificates of Analysis will automatically be generated in as little as 24-48 hours!
- Support – Our jobs center on satisfying our customers and providing unparalleled support. At BioLumix you receive 24/7 support plus a microbiologist dedicated to service you and answer your questions. What happens if you encounter a problem? Troubleshooting can be done via the internet and in most cases problems are resolved. If needed we provide a loaner instrument while your instrument is being fixed to minimize downtime. Our website provides instant access to information and resources about our technology, instruments, assays, and more. It’s also mobile-enabled to format to your handheld devices. Links to social media, white papers, blogs, videos, and other informative contact information puts everything you need at your fingertips.
- Continuous Product Improvement – At BioLumix we have an ongoing commitment to evolving and adding to our platform. One way this is achieved is through our partnership with our customers. Customer feedback, along with a desire to increase the breadth and depth of our offerings, helps us to make improvements to our media and our assays. We have recently introduced two new assays – Listeria and Bacillus cereus.
With a simplified method such as the BioLumix System we can provide a simple cost effective method to test products for microbiology. Come visit our booth (# 16091) and take the first step into your own personal BioLumix Experience. This year the show is being held at the beautiful Mandalay Bay Convention Resort, in Las Vegas, October 8th and 9th.
BioLumix is now part of Neogen
For Food Safety Diagnostic solutions including allergens and toxins, visit Neogen.com
Simplified, Rapid, Automated Microbiology – BioLumix
The validated BioLumix System for Simplified, Rapid, Automated Microbiology Testing
BioLumix was established in 2006 and has developed a novel, patented system for microbiological testing of nutraceutical, food, meat, dairy, beverage, cosmetics, toiletry, and pharmaceutical products.
Customer support and innovation are the cornerstones of the BioLumix business. Our mission is to assist our customers in their quality assurance by offering simplified, rapid, automated, reliable solutions for microbiological quality control and assurance. We offer individualized customer and industry specific validation packages with IQ, OQ, and PQ.
BioLumix allows for easy compliance with cGMP, will streamline your microbiological testing saving significant time labor and money, reducing cost, increasing efficiencies and showing a rapid return on investment. BioLumix can save multiple days versus current testing methods and dramatically impact key profit drivers. BioLumix delivers shorter sample preparation, automated real-time communication, fast product release and early warning of contamination. It is paperless and efficient, saving on disposables, time and space. The system eliminates product interference, delivering accurate results.
"With over 500 systems in the field, it was gratifying to learn that all our customers said that they would recommend the BioLumix system to their vendors, suppliers and customers! "- Ruth Eden, President, BioLumix
The BioLumix System Drives Profit Ability!
With BioLumix Rapid, Automated Microbiology you can view microbiology product testing from a position of company strength and competitive advantage! The BioLumix system is fully automated, validated against USP methodology and 21 CFR Part 11 Compliant. The system delivers complete microbiology testing results within 24-48 hours, allowing for faster product release time, shorter sample prep time – saving time, money and other resources.
The BioLumix System Advantage:
- Single platform testing for all assays: Total aerobic count, yeast and molds, coliforms, Enterobacteriaceae, E. coli, P. aeruginosa, S. aureus, Salmonella, Listeria, Lactic acid bacteria Probiotic, etc.
- Real-time communication – early warning of contamination
- Automated Certificate of Analysis in 48 hours
- No product interference with ready-to-use vials with organism specific enrichment medium
- Stackable compact instruments take up little bench-top space
- Technical service available 24/7
- Ready-to-use assay vials are easy to use
- Vial structure eliminate product interference
- Capable of performing: Microbial limits, Suitability testing, and Preservative efficacy testing