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Rapid Sterility Testing

Rapid Sterility Testing

By: Derek Prince

In June of 1936 the United States Pharmacopeia (USP) implemented the sterility test to better ensure the sterility of parenteral drug products. The test utilized one type of media and applied only to sterile liquids. Since then slight modifications have been made to include a broader range of product types and to increase the overall sensitivity of the method. However, 80 years later the test remains virtually unchanged. Due to advances in microbial detection technologies and a strong initiative from regulatory authorities, alternative sterility testing methods are beginning to take flight.

The addition of chapter <1223> validation of alternative microbiological methods to the USP provides stakeholders with a clear path to validate alternative testing methods. This chapter is clearly the result of the accelerated pace at which alternative detection technologies are advancing and allows manufacturers and testing laboratories alike to validate alternative methodologies in accordance with the outlined criteria. As long as the alternative method yields results equivalent to or better than the compendial method, the proposed alternative method may be validated and used for routine product release.

The current incubation requirements outlined in USP <71> mandate a 14-day incubation period. As many manufacturers contract out their sterility testing, it’s possible that up to 16-17 days (testing + quality assurance time) may pass prior to the release of a final report.  Although this time to result has little impact on some industries/product types, certain industries that manufacture short shelf life products require a more rapid release test.

The Need for Rapid Methods

Pharmacy compounding is a practice where licensed personnel combine drug ingredients to create a personalized medicine.  Unlike commercially manufactured products that have multi-year expiration dates, compounded preparations typically have beyond use dates (BUD) in days or months. This makes it difficult for manufacturers to maintain an inventory of high shelf life products. To further complicate things, the recent proposed changes to the California Code of Regulations states that sterile compounded drug products may no longer be able to be dispensed until an acceptable sterility test result is in hand. The inherent short shelf life of the products and proposed California regulatory changes further highlight the necessity for a more rapid method.

A similar issue exists for products that need to be administered to patients shortly after manufacturing. Cell therapy manufacturers continue to struggle with comprehending the possibility of obtaining a 14-day sterility test result prior to drug administration. Although some companies are able to freeze their product prior to obtaining batch release results, certain manufacturers greatly benefit from the ability to administer fresh, non-frozen drug product. To achieve this preferred product delivery, obtaining a 14-day sterility test result prior to drug administration is not possible. As per the Food and Drug Administration (FDA) Chemistry Manufacturing and Controls (CMC) guidance document “Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs)”, manufacturers are given the task to develop an approach to demonstrate sterility assurance for products that must be administered to patients prior to the results of a 14-day sterility test being available. The document recommends an in-process sterility test to be performed on samples collected 48 to 72 hours prior to final harvest or after the last re-feeding of cultures. Gibraltar has developed several of these in-process tests utilizing our rapid sterility technology to provide a high level of sterility assurance for the full panel of USP <71> organisms in only 48 hours in order to help our customers meet the required criteria.

About the Technology

The Milliflex® Rapid Test System by EMD Millipore is a rapid microbial detection instrument capable of detecting low levels of microbial contamination. The innovative system utilizes a novel membrane filtration system where microbial adenosine triphosphate (ATP) is detected in the form of a bioluminescent signal. Following sample filtration, a media cassette is combined with the filter and incubated for a set period of time. Once incubation is complete, an automated system is used to detect ATP bioluminescence. If the level of ATP detected supersedes the calibrated background threshold, the luminescence is recorded as a CFU.

Validation Strategy and Findings

The Milliflex® instrument underwent an extensive validation plan to demonstrate equivalence to the compendial USP <71> and EP 2.6.1 sterility test. Ruggedness, robustness, specificity, limit of detection and other criteria were assessed to demonstrate the effectiveness of the system. Ultimately, a five-day detection plan was administered as that was the minimum amount of time required to recover growth of a wide variety of environmental and clinical isolates including the notoriously slow growing anaerobe Propionibacterium acnes (4-days + 1-day for assurance). We performed a similar validation at Gibraltar and achieved comparable results. To truly understand the detection power of the instrument we attempted to recover <10 CFUs of all USP <71> organisms at varying incubation times. We were able to detect the presence of all bacteria (Staphylococcus aureus ATCC# 6538, Pseudomonas aeruginosa ATCC# 9027, Bacillus subtilis ATCC# 6633 and Clostridium sporogenes ATCC# 11437 and yeast (Candida albicans ATCC# 10231) in as few as 12 hours and mold (Aspergillus brasiliensis ATCC# 16404) in as few as 24 hours when utilizing the instrument. Despite the powerful results, we generally use a five-day incubation period to allow for any slow growing or stressed contaminants to grow. However, as indicated by our early detection data, the Milliflex® rapid system remains a viable option to provide reliable sterility assurance data for short shelf life products that need to be administered or released before a 5- or 14-day sterility test result is available.


The Milliflex® rapid sterility system is a non-destructive instrument that provides fully automated 21 CFR part 11 compliant data interpretation. Results are dependable and reproducible with a LOD of 1 CFU. The instrument is compatible with a wide variety of membrane filterable products and provides growth-based results consistent with compendial methodology in five days or less. Therefore, this is a quantitative or qualitative growth based method where identification of a positive result is possible. It is important to note that other rapid sterility test systems available do not allow for the microbial identification of a positive result but instead rely on non-growth methods. This means that in the event of a sterility failure, a repeat of the test with the same number of units from the same lot is most likely the only option and the identity of that particular failure will never be known. Gibraltar advises potential users to inquire about the ability of the proposed system to be used for microbial identifications prior to signing on with a rapid sterility test system. We invite any potential users to contact Gibraltar Laboratories to learn more about rapid sterility testing.

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Rapid Sterility Testing – The Why’s And When’s Answered!

Rapid Sterility Testing uses the latest in bioburden scanning and detection technology in order to accurately predict the growth of bacterium, debris and other microorganisms over the course of time. If there is an issue with bacterial colonies growing on a sample or product, rapid sterility testing will quickly detect that the process used to sterilize that product is in fact, not efficient enough for the FDA to accept and authorize said product for public consumption. The FDA is only half of your problem though, the other half falls upon the integrity and respect of your patients, doctors and clientele. Effective sterilization services protect your business, protect your patients and protect your reputation; getting approval from the FDA is an added bonus.

When do we think Rapid Sterility Testing is necessary? Well, any time when something is going to be in direct contact with a sensitive part of your body or inside your body, we believe that sterility testing should be done rapidly to further curb the development of bioburden and debris on any medical or pharmaceutical device or product. With that being said, let’s focus on some things that we think rapid sterility testing is perfect for. You might just agree with us.

Water for Injection (WFI)

WFI gets overlooked by many, but not us. When you make water for injection, that water is going to be infused with some kind of medicine at some point in time. Some kind of vaccination or medication, sometimes with a biological product within will be used in that product, and and presence of microorganisms will greatly hinder that process. If someone’s immune system is already compromised and they need injectable medication, it’s of utmost importance that medical professionals eliminate any bioburden from that WFI.

WFI is quite clean and can be perfect for microorganisms to grow. For that reason, rapid sterility testing is more important than ever. WFI should be bought by medical facilities, used as quickly as possible and then disposed of. Prolonging the sterility testing and shipping processes just decreases the amount of time that product is actually good to be used for.

Liquid Parenteral Products

Liquid parenteral products such as dextrose solution, sodium chloride solution, etc. are among the most frequently used medicines in the acute and post-acute care settings. These products can be released to the marketplace much faster when rapid sterility testing and bioburden testing are performed.

These are just a few items that our rapid sterility testing greatly benefits. Rapid sterility testing provides a great return on investment for anyone investing within the medical and pharmaceutical industries. At Gibraltar Labs we provide comprehensive contract research services from the ground up, working transparently every step of the way. For more information on our services and how our rapid sterility testing services and depyrogenation services can help your business contact us today at 877.315.5847


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Challenges to Validation of a Complex Non-Sterile Medical Device Tray

BITThis scientific publication is an important case study. The challenges associated with demonstrating to a very high level of assurance of sterility are discussed in the context of case design, instruments, density, F0, maximum load configuration, minimum load configuration. In addition, the biological testing using qualitative and quantitative methodologies are illustrated.

The web address is,

and in print

Challenges to Validation Of a Complex Nonsterile Medical Device Tray
Daniel Prince, Jozef Mastej, Isabel Hoverman, Raja Chatterjee, Diana Easton, and Daniela Behzad
Biomedical Instrumentation & Technology, Vol. 48, No. 4, July/August 2014: 306-311.
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“Microbiological Testing and Contamination Control – an opinion on the microbial limit test with reference to harmonization”

contractpharmaDear All,

Gibraltar Laboratories has published its 90th paper. The current issue of Contract Pharma has our article entitled “Microbiological Testing and Contamination Control – an opinion on the microbial limit test with reference to harmonization”. We are proud of the contributions of our technical services team to this effort: Christopher Waskewich, Kristah Kohan and Danina Rinaldi.  Our paper provides an opinion on the harmonization of USP microbial limits testing. It provocatively analyzes the changes that were made to USP Chapter <62> in order to win consent from the Europeans. You will find a copy  at

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Dear Friends

Governor ChristieProcalamation

Gibraltar was AGAIN awarded with another prestigious award – the 2013 New Jersey Family Business of the Year Award!

This award is presented by the Rothman Institute of Entrepreneurial Studies at Fairleigh Dickinson University’s Silberman College of Business. The award was presented to Gibraltar Laboratories by New Jersey’s Governor Chris Christie at a ceremony on October 16, 2013.

Thank you for all of your continued support of Gibraltar Laboratories!

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Burkholderia cepacia and BCC – the next indicator organism?


The main interest in the presence of pseudomonads of any type (including Burkholderia cepacia complex (BCC) – the former Pseudomonas cepacia) is that they are strictly water-borne organisms (deionized water) which is used as the main ingredient in all of the medicaments listed in this report. Gram negative bacilli of this type thrive heartily in deionized water because of the sparse nutritional requirements.  Pseudomonas cepacia was first discovered by Walter Burkholder in 1949 while studying the cause of onion rot and gained more recognition in the 1980’s after it was found to cause a 35% death rate in cystic fibrosis patients that became infected, and subsequently was renamed in his honor.

Burkholderia cepacia is from a group of catalase-positive, lactose-negative, gram negative aerobic bacteria. Gram negative bacilli are divided into three biochemical groups based on their metabolism of glucose fermenters, non-fermenters, oxidizers.  E. coli, acaligenes and Pseudomonas/cepacia are examples of these three groups, respectively. They are motile due to a polar flagellum. Sometimes referred to as the Burkholderia cepacia complex (BCC) the group is comprised of B. ambifaria, B. anthina, B. cenocepacia, B. cepacia, B. dolosa, B. multivorans, B. pyrrocinia, B. stabilis, B. vietnamiensis and several others. BCC organisms are typically isolated from water, soil and agricultural products.

BCC organisms are naturally multi-drug resistant to many common antibiotics such as polymyxin B and aminoglycosides. There is a special agar called “Oxidation-fermentation polymyxin-bacitracin-lactose (OFPBL) that contains polymyxin (which inhibits most gram negative bacteria, including Pseudomonas aeruginosa) and bacitracin (which inhibits most gram positive bacteria as well as Neisseria sp.).  The presence of lactose distinguishes between fermenting and non-fermenting gram negative bacilli by rendering the medium yellow in the presence of lactic acid and bromthymol blue.

The microbe is able to remain viable for many months in harsh conditions such as low and high temperature extremes (12C to 48C) and in the presence of organic solvents, low nutrients, antiseptics, preservatives, antibiotics and preservatives. It has been found to survive for months in water (low nutrient) but for only maybe 1 week on a dry surface. Its ability to survive such conditions is due to its ability to produce a biofilm.

Over the years B. cepacia has become notorious for its ability to grow in the presence of the substances it is supposed to inhibit e.g., benzalkonium chloride in the barber shop and povidone-idoine antiseptics. A famous paradoxical case of antiseptic drug resistance occurred several years ago with the incident of pseudobacteremia where blood cultures grew cepacia but the organism did not come from blood – it came from contaminated PVP-I that was used to decontaminate the blood culture bottle.

Gibraltar recommends adding this organism to preservative challenge testing, disinfectant validations for facilities and USP <61/62> as our experience has indicated that it can be more resistant to standard preservatives than organisms in the current battery of USP methods. It can be noted though that finished product testing can yield false-negative results due to “nutrient shock” when using nutrient rich media. Special cultivation is necessary (pre-enrichment step) confirmatory identification.

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Instrument Sterilization versus Disinfection

In the medical field, every step of a doctor has to be taken for the betterment of his/her patients. Medical instruments are really delicate and they can cause problems no one would have ever thought of. These medical instruments need to be cleaned after being used on one patient and before being used on another in order to prevent the chances of potentially transferring diseases. Instrument sterilization is one of the most important responsibilities of a good doctor.  There are many instruments that need to be cleaned and taken care of. Surgical sterilization can either be done in the clinic/hospital or this procedure can also be outsourced to a reliable laboratory.

What most people fail to realize is the difference between disinfection and sterilization. Instrument sterilization results in the thorough cleaning of all microbial life present on surgical instruments such as surgical blades. On the contrary, simple disinfection can only reduce the percentage of certain types of microorganisms present. You may probably think that extensive disinfection can be used to clear all microorganism, but this is not so. High level disinfection will eliminate vegetative microorganisms and TB but resistant bacteria spores will not be killed using this process. Common disinfectant chemicals used by most hospitals include phenolic compounds, alcohol, quaternary ammonium and iodophors. However, these should not be confused as effective sterilants and therefore cannot be used as substitutes for proper surgical sterilization.

On the other hand, preferred methods of surgical sterilization are steam/high temperatures in autoclaves used for instruments which can withstand extreme temperatures. Other methods of effective instrument sterilization include ethylene oxide gas and cold chemical sterilants.

Medical institutes have used this process to provide medical staff with clean and sterilized instruments to use on patients. Instrument sterilization has to be done in order to make sure that the medical tools used are germ free and do not pose the risk of transferring a disease from one patient to another. This process doesn’t take more than a day and after that the instruments are as new as ever. Surgical sterilization is an effective way to keep infections away and keep patients stress free about the sanitary conditions of the surgical tools. It is advised that instrument sterilization should be done by every medical institute in order to control the potential spread of diseases. This is the most effective way of preventing the transferring of diseases that can occur because of using non-sterilized medical instruments.


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Gibraltar Laboratories Equipment and Instrument Sterilization Services – Why Should You Choose Us?

Providing state of the art services using heat and steam equipment and instrument sterilization methods, Gibraltar Laboratories is a reliable name in the sterilization industry today. As people are gaining awareness about health and safety, a drastic need in sterile processing has been experienced from biology, chemistry, dentistry, cosmetics and other major industries of the world. This is exactly where our services come into play. Catering to the said industries and more, we take pride in bringing about modern methods of sterilizing medical equipment as well as reprocessing the implements used during aseptic manufacturing. With over 43 years of impeccable service, Gibraltar Laboratories has been successful in creating a reputation based on customer satisfaction and quality testing?.

Headed by Dr. Daniel Prince we are an independent, passionate team of professional scientists and technicians who put forth our best efforts to comply with government regulations and customer requirements intended to protect patient safety. The company is the brainchild of Dr. Herbert Prince. Gibraltar Laboratories was formed in 1970 and has increased in size and market share in the last 43 years. We use only the best equipment sterilizationand instrument sterilization practices. Be it validation, lab testing, and instrument sterilization or sterilizing medical equipment, at Gibraltar we are a strong and focused family catering to your needs in the best possible manner.

Previously there was no concept of the reprocessing such as equipment sterilization.As a result too many people treated in the hospital acquired some kind of infection. These acquired infections can be fatal in many cases. Due to this reason it is important to realize that only by sterilizing medical equipment and instrument sterilization, the lives of innocent individuals can be saved. In addition to sterilization of surgical instruments, the management should also work on proper cleaning of floors, doors, windows, operating tables manufacturing lines and other sensitive areas which are associated with patient safety.

Just as sterilized bottles are important for a baby’s safety, reprocessing and sterilizing instruments and medical equipment by validated procedures is essential. At Gibraltar we offervalidated services which are in the best interests of our clients which are also a major reason why large and small customers turn to us in their hour of need. Using top quality autoclaves, chemical liquids and sterilization pouches is only one of our features, Gibraltar Laboratories is overall a stable outsourcing partner who understands the importance of instrument and equipment sterilization, reprocessing  and therefore leaves no stone unturned in serving its clients with a service that is hard to find elsewhere


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Understanding the Basics of Sterilization and Liquid Chemical Sterilization

When it comes to sterilization, you can never be too careful. The main objective of proper sterilization of medical devices, supplies and surgical instruments is to remove and destroy all forms of micro life to achieve an acceptable level of sterility.

Medical devices that are not properly sterilized can be life threatening and pose a serious threat to proper patient medical care.

It is imperative then that medical technicians and health workers realize the complex process of sterilization and acquire knowledge about the proper methods and scientific principles of sterilization.

Therefore, it is essential that the standards and principals of sterilization be met in every health care facility, whether it is a dental or a physician’s office, a hospital, a surgical centre, or an outpatient facility.

The Three Key Essentials of the Sterilization Process are:

The conditions should be ideally matched to ensure that the equipment is properly sterilized in a way that would kill any bacterial organism effectively.

The equipment that is sterilized should undergo thorough cleaning to ensure the effectiveness of the sterilization. If the cleaning process is not undergone with strict sterilization parameters, it could damage the whole sterilization process as ineffective.

The sterilized equipment should be thoroughly sterilized with the sterilant so that all surfaces and cracks in the instrument are sterilized.

Achieving proper sterilization is not just based on these three essentials, and proper care should be given to medical device cleaning so that no risk or harm comes to the people that are involved in the sterilization process.

Sterilization involves the use of a chemical or physical procedure to destroy micro bacterial life and organisms. The major agents used to sterilize equipment and instruments are:

  • Saturated Steam
  • Ethylene Oxide gas
  • Hydrogen Peroxide Gas Plasma
  • Liquid Chemicals

Dry heat is used as well but has not been commonly used in recent times. Recently, a new sterilizing agent has emerged, known as Ozone, and it is being used in the US.

Liquid Chemical Sterilization:

Liquid Chemical Sterilization is used to sterilize heat sensitive devices which can be immersed. The method requires the item to be completely immersed in a germicidal solution for a prescribed period of time to kill all the bacterial micro-organisms.

There are two types of liquid chemical sterilization involved in sterilizing equipment:

  • Peracetic Acid
  • Glutaraldehyde

Although it thoroughly accomplishes the task of sterilizing medical devices and equipment, chemical sterility poses a huge risk to the lives of patients and health care workers. Chemicals that come in contact with skin, mucous membrane, tissue, and eyes can have hazardous results. Therefore, it is advised to use protective equipment like goggles, masks, aprons and gloves when performing the process.

Gibraltar Laboratories enjoys an impeccable reputation as a health care service provider. The company boasts a long list of satisfied clients.

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Why Medical Instruments need Sterilization

Surely each one of us, at some point in our lives, must have seen several medical instruments. These simple looking tools play a very significant role in diagnosing and treating different kinds of medical problems. You must have also noticed that medical practitioners use the same set of medical instruments again and again on different people during the course of a day. Have you ever wondered about the safety of these instruments because they are being used on different people with infections or diseases?

There is always a chance of a perfectly healthy person catching the same infections through one of these medical equipments. The danger is increased exponentially since the parts of the body that come into contact with these equipments are most vulnerable to infection and contamination. There is no doubt about the significance and importance of these medical tools, the absence of which can cause the whole health care service to cripple. It is, however, also imperative that these tools are properly taken care of in such a manner that they do not transmit infections to healthy patients. This is where medical instrument sterilization comes in.

There are several kinds of medical equipment in use in hospitals and laboratories. Regardless of type, make or size, sterilizing medical equipment is crucial to the proper usage and maintenance of all the instruments. Instrument sterilization not only extends the life of expensive equipment so that they remain useable for a longer period of time, but also ensures safety against transmissible diseases. There are, of course, a few tools that do not require instrument sterilization since they are meant to be used just once. These are discarded immediately after usage.

Sterilizing medical equipment can be carried out in a simple manner with the help of alcohol and cotton. The equipment is rubbed with cotton that has been immersed in alcohol. Ultrasonic bath is another popular method of sterilizing medical equipment. This process involves placing the equipment in a disinfectant liquid for more than 15 minutes. The ultrasonic waves are able to remove even the tiniest of particles from the said equipment. After the ultrasound bath, the equipment is rinsed in running water and dried with paper towels.

An autoclave is also very useful equipment for sterilizing. Instrument sterilization is carried out with the help of high-pressured steam. The equipment, after being subjected to steaming, is kept in a clean and dry place.

There are several other procedures for sterilizing instruments all of which are designed to carry out the same purpose.

Gibraltar Laboratories enjoys an impeccable reputation as a health care service provider. The company boasts a long list of satisfied clients.