Microbiology & Environmental Monitoring

Avista Pharma’s microbiological testing labs are FDA-registered laboratories, fully compliant with all GMP and GLP testing services and standards. The microbiology labs are equipped with knowledgeable, experienced analysts who understand that, with release testing, fast turnaround time is important –  but quality is essential. Avista Pharma helps pharmaceutical manufacturers provide safe, high-quality products – fast, with maximum cost-effectiveness.  Our experienced team offers a number of services that are listed below.  If you have a question about these or other services, please contact us.

 

Cleanroom Services & Routine Environmental Monitoring

 

Avista Pharma offers a full range of Cleanroom Services to provide our clients with as little or as much assistance as they need to support their programs.  For more information, please see our best-in-class SPECIALIZED AND GENERAL CLEANROOM SERVICES.

 

Avista Pharma offers an all-inclusive support service to provide ongoing, routine environmental monitoring for the intended purpose of the products being manufactured and the existing resources you have in place. Alternatively, we can provide trained staffing to supplement your existing program during start-ups or qualification activities. Finally, we have a plate analysis-only program in which we provide quality control-tested media for your use which can be returned for incubation and analysis by our trained staffing. For more information, please see our full range of ROUTINE ENVIRONMENTAL MONITORING SERVICES.

 

Cleanroom Certification

Avista Pharma Solutions will certify your controlled environment with unmatched quality and integrity.  Learn more about our GMP program led by our CETA-Accredited Cleanroom Services Supervisor.  For more information, please see our leading CLEANROOM CERTIFICATION SERVICES.

 

Facility Qualification Support

The experienced staff at Avista Pharma Solutions brings over 10 years of quality, validation, and contamination control experience to provide superior services during the execution of facility and critical utility qualifications.  For more information, please see our reliable FACILITY QUALIFICATION SERVICES.

 

Microbiological Testing

Avista Pharma provides expert microbial identification services to identify bacteria, fungi and yeasts for the pharmaceutical, medical device, cosmetics, personal care, food and beverage manufacturers and other industries that require identification of microbial isolates. With over 30 years of experience, the Avista Pharma team uses state-of-the-art gene sequencing technologies to complete the identifications and create accurate and reliable GMP-compliant reports. Our microbial identification service utilizes DNA sequencing for identification of microbial organisms, including bacteria, fungi and yeasts – comparing to an up-to-date library containing thousands of identifications based on 500 base pairs of the 16S ribosomal RNA gene.

 

Microbial Identification – Unique Service in the Market

Our unique identification laboratory has two 16 capillary Microseq™ genetic analyzers that allow increased throughput. We are experts in both fungi and environmental contaminants. Genetic analysis of the ribosomal RNA is accurate and validated. Libraries are maintained and periodically updated by the manufacturer. Determining the correct genus and species is essential for developing a robust product and environmental surveillance program.

Container Closure Integrity Testing

Container Closure Integrity (CCI) testing is performed in order to determine whether sterile product packaging forms an integral seal during the manufacturing process and maintains that seal over time. In fact, the FDA Guidance (Guidance for Industry: Container and Closure System Integrity Testing in Lieu of Sterility Testing as a Component of the Stability Protocol for Sterile Products, U.S. Food and Drug Administration, February 2008) states CCI tests “may be more useful than sterility testing in demonstrating the potential for product contamination over the product’s shelf life or dating period.” Scientists at Avista Pharma have been performing Container Closure Integrity testing for over 15 years, and in the process, validating methods for dozens of drug products utilizing traditional methods as well the newest technologies available.

 

Avista Pharma has experience with the following CCI methods:

 

Laser-based Headspace Analysis

Avista utilizes a Lighthouse FMS-III Headspace Oxygen Analyzer for determining the oxygen content in optically clear vials and syringes. The laser used in the system is optically tuned to be able to detect and accurately measure oxygen. Since the method is non-destructive, changes in oxygen levels over time can be monitored and correlated to leak size. Headspace analysis can easily detect defects smaller than 1 micron in size in products packaged in a low-oxygen headspace (for example, lyophilized samples or those containing nitrogen or argon overlays.)

 

Dye Ingress Testing

Avista utilizes a dye ingress test to evaluate the container closure integrity of vials, syringes and bottles. The dye ingress test involves performing a vacuum and pressure challenge on the test samples while they are submerged in methylene blue dye. After the challenge the samples are visually evaluated and then measured on a UV-Vis spectrophotometer at the specific wavelength of methylene blue dye. The absorbance measured from the samples is compared to a limit standard in order to determine whether a leak is present. The Avista Pharma dye ingress test is able to detect leaks as small as 5 microns in size. Validation is recommended to ensure the drug product is compatible with the methylene blue dye, that the detection limit is adequate and that the vacuum and pressure conditions are suitable for the container configuration.

 

Microbial Immersion Testing

Avista Pharma can perform a microbial immersion CCI test upon client request. The test involves performing a vacuum and pressure challenge on media-filled samples while they are submerged in a microbial broth. Following the challenge the samples are incubated and observed for the presence of growth. If growth is observed, the organism is identified to determine whether it is the challenge organism, and if found to be so, a leak is confirmed.

Bacteriophage Screening

Avista Pharma offers testing to determine the resistance of clothing materials to the passage of blood-borne pathogens using Phi-X 174 bacteriophage penetration as a test system, per ASTM F1671/F1671M. In this test, patient use of protective clothing is simulated by exposing it to Phi-X 174 bacteriophage suspended in a body fluid simulant under conditions of continuous contact. The material is observed throughout the test for liquid penetration, and if none is present, the material is rinsed with sterile fluid. The rinse fluid is then tested to determine whether bacteriophage has penetrated through the material.

 

Avista Pharma also offers a method for barrier testing of ENT Endoscope Sheaths (and Materials) per the FDA Guidance for Manufacturers Seeking Marketing Clearance of Ear, Nose, and Throat Endoscope Sheaths used as Protective Barriers, issued on March 12, 2000. In this test, the sheath is inoculated with a Phi-X 174 bacteriophage suspension and attached to the corresponding scope. The endoscope with the sheath is articulated in submersion buffer to simulate the movement of actual use. At the end of the articulation test, the submersion buffer is tested to determine whether bacteriophage has penetrated through the sheath.  The pass/fail evaluation for both tests is based on the detection of penetrated Phi-X bacteriophage.

 

Avista Pharma offers bacteriophage screening of master and working cell lines with and without induction.

Filter Integrity Testing

Bacterial Filter Integrity

Filters are tested for their ability to retain bacteria that have a diameter equal to or slightly larger than the pore size of the filter being tested. Filters are challenged with ≥ 1.0 x 107 cfu/cm2  of the effective filtration area at a specified pressure. The filtrate obtained from the challenge is filtered through an appropriate assay filter which is plated onto agar. After incubation, the assay filters are observed for the presence of the challenge organism.  Acceptable performance of the filters is demonstrated by the membrane retaining the entire bacterial challenge of ≥ 1.0 x107 cfu/cm2 of the filter area. Bubble points are performed on hydrophilic and hydrophobic filters. A test used to indicate maximum pore size of a filter. Used to test the integrity of filters with a specific, validated, pressure values, wetting liquids and temperatures for a specific pore size. Notes: currently test 0.1, 0.22 and 0.45µm filters – hydrophobic and hydrophobic capsules and membranes

 

Bacterial Filter Retention Studies

These studies test the ability of a filter to retain bacteria after being exposed to product during process conditions.  A smaller filter made from the same material as the process filter can be used.  Three different lots of test filters are exposed to product at specific pressure, temperature, flow rate and membrane fluid contact time. The product is recirculated through the test filters per the process conditions. After re-circulation, the filters are challenged with a bacterial challenge of ≥ 1.0 x107 cfu/cm2 of the effective filtration area. Viability study results determine whether the bacterial challenge is performed in the test product or in a surrogate fluid. The filtrate obtained from the challenge is filtered through an appropriate assay filter which is plated onto agar. After incubation, the assay filters are observed for the presence of the challenge organism.  Acceptable performance of the filters is demonstrated by the membrane retaining the entire bacterial challenge of ≥ 1.0 x107 cfu/cm2 of the filter area.

Mycoplasma Testing

Avista Pharma provides Mycoplasma testing services to meet regulatory requirements in USP <63>, EP2.6.7 and FDA Points to Consider:

 

  • Direct Culture Method per USP <63> and EP2.6.7
  • Indicator Cell Culture Method per USP <63> and EP2.6.7
  • Direct and Indicator Cell Culture Methods per FDA Points to Consider Guidelines

 

Testing for Mycoplasma is a necessary quality control requirement to assure reliably pure biotechnological products and allied materials used to generate these products. The genus Mycoplasma, a common cell/ tissue culture contaminant, represents a group of minute bacteria which have no cell walls. The presence of Mycoplasma species does not always result in turbid growth in cultures or visible alteration of cells, and a cell culture infection may persist for an extended period of time without causing apparent cell damage.

Sterilization Validation Testing

Properly designed and executed sterilization validations are critical to ensuring the sterilization process is capable of achieving the intended sterility assurance level (SAL). An SAL of 10-6, meaning there is a probability of only 1 non-sterile device in every 1 million sterilized, is most common with today’s sterilization processes. With over 30 years of experience, Avista Pharma offers superior expertise and high quality testing to support your radiation (gamma/e-beam) and gaseous (Ethylene Oxide) sterilization validations and monitoring. Avista Pharma’s experts have years of experience in sterility, bioburden and ethylene oxide residual testing. Avista Pharma has also fostered good relationships with leading industrial sterilizers and work closely with them on clients’ sterilization projects. This networking makes the sterilization validation process very streamlined.

 

Our testing services include:

 

  • Bioburden testing: performed per ANSI/AAMI/ISO guidelines. Testing can be performed using pour plate or membrane filtration methods.
  • Bioburden Recovery Factor testing: performed per ANSI/AAMI/ISO guidelines. This testing ensures the validity of the bioburden testing by establishing how much of a control organism can be recovered from the product using the extraction methodology.
  • Sterility testing: per ANSI/AAMI/ISO and USP/EP guidelines. Avista Pharma offers a wide range of vessel sizes that can handle most samples.
  • Bacteriostasis/Fungistasis testing: performed per ANSI/AAMI/ISO or USP/EP guidelines depending on the testing method. This testing ensures the validity of the sterility test by demonstrating that control organisms can be recovered in the presence of the sample.
  • Ethylene Oxide Residuals testing: performed per ANSI/AAMI/ISO guidelines using state of the art gas chromatograph (GC) equipment. Avista also is outfitted with GC/mass spectrometry (MS) equipment, which can be used to test devices that contain compounds such plasticizers. Such compounds can interfere with conventional GC methods.

Cleaning Validation

Validating the cleaning steps of your drug product manufacturing process is a critical component of avoiding an adulterated product. Adulteration can occur if there is any remaining product or cleaning agent in the manufacturing line prior to the start of the next batch. In order to prove your cleaning process is effective the validation of the process and subsequent monitoring are critical.

 

Avista Pharma routinely helps clients validate their cleaning processes by performing the following services:

 

  • Method development and validation of analytical assays (HPLC, TOC)
  • Performing swab recovery studies using the client’s swabbing instructions, drug product, and coupons of materials used in the manufacturing line
  • Performing routine testing of post-cleaning samples (HPLC, TOC)
  • Development of equipment specific cleaning validation protocols and reports
  • Development of protocols and sample testing to support qualifying technicians in swabbing

 

Avista Pharma utilizes Total Organic Carbon (TOC) methodology as well as High Performance Liquid Chromatography (HPLC) in cleaning validation assays. TOC is a great method for cleaning validations because it is a non-specific method that will measure organic compounds. It is useful in cleaning validations because it is able to detect numerous compounds at very low limits in a single measurement. A drawback of TOC is that, because it is a non-specific method, the source of the measured result is not known. It must be assumed that all of the TOC measured is from a single source. This understanding is very important when a cleaning validation is performed, as multiple limits may need to be applied against a single measured TOC result. If a component in the drug product is of particular concern and its presence or absence needs to be known,  a specific assay (such as HPLC) is recommended over the TOC assay. Avista Pharma has the equipment and knowledgeable scientists available to help develop these methods, validate these methods and apply these methods for assessing your cleaning results.

Water System Vaildation & Monitoring

Process water is the single most important raw material used in pharmaceutical and medical device manufacturing processes. For the last 30 years, Avista Pharma has been among the most experienced and capable laboratories for United States Pharmacopeia (USP) and European Pharmacopeia (EP) purified water testing. Our experts can also advise you on how to set up your pharmaceutical/medical device water testing programs and system validations to ensure they meet specific regulatory requirements. Avista Pharma’s microbiology and analytical testing labs offer a wide range of  methods for water analyses to support your validation and routine monitoring needs.

 

Purified water requires a specific panel of testing be performed as per USP and EP requirements. Avista Pharma is fully outfitted with state of the art equipment and highly trained experts to perform these tests in a timely and accurate manner ensuring the highest quality results.

 

Avista Pharma’s purified water testing services include:

 

  • Bioburden testing: used to establish the microbial load in water. Bioburden testing is offered using mHPC, R2A and mENDO agars offering a wider range of options to suit your specific needs.
  • Conductivity testing: used to establish the sample’s ability to conduct electricity. This measurement relates to the number of dissolved salts (ions) in the sample, which can indicate a potential processing problem.
  • Total Organic Carbon (TOC) testing: used to establish whether the level of carbon present in the sample is below the mandated limit. High TOC levels, which are above the mandated levels, are a reliable indicator of sample contamination.
  • Heavy Metals and Nitrates testing: if your water also needs to meet EP requirements, Avista is fully equipped to perform Heavy Metals and Nitrates testing. These analyses are used to establish the level of metallic impurities and nitrates in the sample, both of which can lead to significant water quality problems.

 

If your water is being used for injection or internal applications, USP and EP requires that testing for endotoxins be added to the purified water testing panel. Endotoxin testing is designed to detect specific toxins (lipopolysaccharide) that are found as a structural component of the cell wall of Gram negative organisms. If these toxins are released from the cells, they can elicit a pyrogenic (fever) response in the patient. Avista Pharma is fully capable and experienced in performing endotoxin testing using various methodologies, including gel clot, chromogenic and turbidimetric methods.

Package Validation Testing & Accelerated Aging Studies

Avista Pharma offers a comprehensive portfolio of package validation testing and accelerated aging studies.

 

Our services include:

 

Accelerated Aging Studies

Accelerated aging studies can be predictive of what will occur to the packaging or product during normal storage conditions. Studies are used to set preliminary shelf life dates (i.e. expiration dates). Shelf life is the period during which the product remains stable for its intended use.

 

Our studies include:

 

  • Standard chamber temperature:
    • (25°C 60% RH), (30°C 65%RH), (40°C 75%RH), (50°C 10-50% RH),( 55°C 10-50%RH), (60°C 10-50%RH), (55°C), (60°C)
  • Custom studies available
  • Electronic monitoring (24-hour / 7-days) of all chambers that alerts Avista Pharma personnel to any out of tolerance conditions
  • Back-up power
  • Backup storage guaranteed
  • Post-stability/aging testing available

 

Package Validation/Flexible Package Testing

Before accelerated aging of the product or package begins, the integrity of the packaging should be determined.

 

Our services include:

 

  • ISO 11607 regimens
  • Package integrity testing
    • Burst test (ASTM F 1140)
    • Dye test (ASTM F 1929)
    • Bubble test (ASTM F 2096)
    • Peel strength (ASTM F 88)

 

Real-Time Aging Studies

Medical Devices are required to have expiration dates based on accelerated and real-time data.  Real-time aging studies are recommended by the FDA.

 

  • Side-by-side real-time aging studies conducted in conjunction with accelerated aging available

 

Ship Testing

The purpose of a ship test is to challenge the strength and robustness of the product and package when faced with transport hazards.

 

  • ISTA 1A/2A: Follows ISTA guidelines

 

Medical Device Package Testing

In addition to strengths in biological and chemical testing for pharmaceutical and medical device clients, Avista Pharma offers medical device package testing. It’s all part of the scope that makes Avista Pharma your ideal single source for laboratory testing.  Materials for all package validation and integrity tests are quality-controlled via our extensive Good Manufacturing Practices (GMP) program. Tests are performed by trained, experienced package integrity technicians.

 

Our wide range of medical device package testing includes the following fast, reliable, expertly administered tests:

 

Peel Strength Test (ASTM F88)
This test measures the seal strength of flexible barrier materials.  Seal strength is a quantitative measure for use in process validation, process control, and capability. The test measures and records the maximum peel strength — of a sample 1 inch wide, in pounds per square inch.

 

Dye Test (ASTM F1929)
This testing provides clearly distinguished visual evidence of package integrity.  This test method evaluates the integrity of porous medical packaging seals by exposure to a dye solution penetrant followed by a visual examination.

 

Bubble Test (ASTM F2096)
This testing is used to detect gross leaks in packaging by internal pressurization of a package while submerged in water.  The package is inflated underwater to a predetermined pressure and then observed for a steady stream of air bubbles which indicates an area of failure.  This test method may be used for tray or pouch packages.

 

Burst Test (ASTM F1140)
This test measures package seal strength of items such as TyvekTM pouches and lid trays. Air pressure on the package and seal is increased until the material separates – the highest pressure attained before bursting is noted. This test method provides a rapid means of evaluating tendencies for package failure when the package is exposed to a pressure differential. Pressure differentials may occur during processes such as sterilization and transportation. Burst testing determines the average burst pressure in force per unit area of a given package design. The test measures and records the burst pressure in inches of water, with mean and standard deviation.

Disinfectant Qualification

A controlled manufacturing cleanroom environment requires a sound disinfectant qualification program. Disinfectants utilized within these environments must be validated to confirm their effectiveness for use. Proper rotation of the disinfectants can be determined based on the data collected in this study.  Avista Pharma is your capable, dependable partner for performing disinfectant qualifications.

 

Some common questions and issues regarding disinfectant qualification include:

 

Why should I choose Avista Pharma to perform my disinfectant qualification study?

  • Avista Pharma has been performing microbiological work for more than 25 years
  • Our specialized microbiology group possesses years of experience performing qualification work on many different surface types using a wide range of disinfectants
  • Avista has an internal collection of ATCC organisms ready for use onsite, plus the ability to identify and cryopreserve client environmental isolates

How do I know which method to follow for my disinfectant testing study? Tube Dilution Method

  • For clients with new facilities with little or no environmental trending history
  • Utilizes ATCC strains and some environmental isolates, where applicable
  • Isolates are directly inoculated into the desired concentration of disinfectant being tested
  • Studies of this sort allow a facility to begin production with some assurance that their disinfectants can kill specific microorganisms of concern
  • Initial screening of disinfectants does not validate that they are effective on hard surfaces

Coupon Testing Using the Swab Recovery Method

  • For established facilities with environmental programs and trending
  • Coupons are prepared from representative surfaces present in the client’s clean room environment
  • The disinfectants are sprayed on the surfaces using the method and contact time captured in the client’s cleaning procedures
  • To ensure implementation of a proper disinfection program, the FDA recommends the use of the facility’s environmental isolates. However ATCC organisms may be used as well

Avista Pharma can create a proposal for your disinfectant qualification study with the following information:

  • Quantity of surface materials found in your facility to be used in the study
  • Quantity of environmental isolates/ATCC strains to be used in the study
  • Quantity of disinfectants to be challenged in study
  • Quantity of exposure time-points to be challenged in the study (time disinfectant sits on a given surface)