QC.pdf

Module 1: Introduction to QA/QC QAQC Introduction Page 1 If you have the recommended text, please refer to chapter 2 for basic definitions and an introduction to what encompasses a QA/QC system in a laboratory. QA/QC Introduction Quality assurance (QA) refers to an organization's policy to prevent p...

Module 1: Introduction to QA/QC QAQC Introduction Page 1 If you have the recommended text, please refer to chapter 2 for basic definitions and an introduction to what encompasses a QA/QC system in a laboratory. QA/QC Introduction Quality assurance (QA) refers to an organization's policy to prevent problems from occurring. Quality control (QC) refers to the procedure(s) implemented by the organization to guarantee quality. This is a very broad definition and is applicable to any laboratory conducting testing procedures on almost every conceivable product. QA/QC QA/QC has a different meaning to different people depending upon their particular area of expertise. For example, the Federal Bureau of Investigation (FBI) provides detailed standards for QA/QC of a laboratory conducting forensic DNA testing and DNA databasing. These standards differ from those of the American Association of Blood Bank (AABB) and College of American Pathologists (CAP) used by various biological testing laboratories. To further complicate matters, there are numerous international standards, which must also be adhered to in an effort to standardize quality assurance programs. Every graduate entering a career in an analytical, forensic, clinical, manufacturing, or research facility will have to abide by some type of quality assurance program. This course teaches the key components of QA/QC and helps students to understand the need to produce sound scientific data using appropriate standards and controls, written procedures, and validated methods no matter what field they are employed in. While everyone can understand the need for a QA/QC program, it is not surprising that the process can seem daunting to the inexperienced. The program that best meets the needs of the particular laboratory and its regulatory or accrediting body is often a very difficult question to answer. This course provides a generic description of what is required in the formation of a quality management system in any laboratory. This is achieved by describing the key principles in any QA/QC program with reference to the American National Standards Institute National Accreditation Board (ANAB), FBI, and International Organization for Standardization (aka ISO) standards, together with specific examples from different specializations. Discipline-specific forensic science standards will also be discussed as they too play a critical role in the development of a comprehensive quality management system. Quality Assurance and Accreditation Standards In 1994, the Federal DNA Identification Act charged the FBI Director with creating an advisory board to “develop, and if appropriate, periodically revise, recommended standards for quality assurance, including standards for testing the proficiency of forensic laboratories, and forensic analysts, in conducting analyses of DNA”. (Source: https://uscode.house.gov/view.xhtml?req=granuleid:USC-2000-title42section14131&num=0&edition=2000 (https://uscode.house.gov/view.xhtml?req=granuleid:USC-2000-title42- section14131&num=0&edition=2000) ) This newly appointed board, the DNA Advisory Board (DAB), was responsible for creating the first set of regulating standards. The Quality Assurance Standards for Forensic DNA Testing Laboratories was issued by the FBI Director and made effective on October 1, 1998. The Quality Assurance Standards for DNA Databasing Laboratories was issued by the FBI Director and made effective on April 1, 1999. In 2000, the DAB was dissolved. The Scientific Working Group on DNA Analysis Methods (SWGDAM) inherited their role and were charged with making all future recommendations regarding these QAS documents. The Quality Assurance Standards for Forensic DNA Testing Laboratories can be found here: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746) The Quality Assurance Standards for DNA Databasing Laboratories can be found here: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743) Additional information on the history and evolution of these standards can be found here: https://www.swgdam.org/_files/ugd/4344b0_fe649488fd6940fa8da6eae00ed33875.pdf (https://www.swgdam.org/_files/ugd/4344b0_fe649488fd6940fa8da6eae00ed33875.pdf) ) ISO/IEC 17025, General requirements for the competence of testing and calibration laboratories, is a key standard used by many laboratories. As this standard is applicable to many types of laboratories, additional supplemental standards are needed to clarify what is specifically required of a forensic laboratory. For many years, the American Society of Crime Laboratory Directors/Laboratory Accreditation Board (ASCLD/LAB) was used as an accrediting body by many forensic laboratories to ensure the quality of their laboratories. ASCLD/LAB provided supplemental requirements to ISO/IEC 17025 that were used during the accreditation process. In 2016, ASCLD/LAB merged with ANAB who now takes on that role in addition to many others. ANAB provides accreditation for ISO/IEC 17025 testing and calibration laboratories and forensic testing agencies, ISO/IEC 17020 inspection bodies and forensic inspection agencies, ISO Guide 34 reference material producers, ISO/IEC 17043 proficiency test providers, ISO 15189 medical test laboratories, ISO/IEC 17021 management systems certification bodies, and industry-specific programs”. Source: https://www.theauditoronline.com/anab-merge-forensics-operations-ascldlab/ (https://www.theauditoronline.com/anab-merge-forensics-operations-ascldlab/) Please go to the following websites for more information about the merger and information about accreditation services through ANAB https://anab.ansi.org/latest-news/anab-and-ascldlab-merge-operations (https://anab.ansi.org/latest- news/anab-and-ascldlab-merge-operations) https://anab.ansi.org/en/forensic-accreditation (https://anab.ansi.org/en/forensic-accreditation) All ANAB forensic accreditation programs are based on International Laboratory Accreditation Cooperation (ILAC) guidance document, G19, Modules in a Forensic Science Process. ANAB, like many accrediting bodies, are signatories to a Mutual Recognition Agreement (ILAC MRA) with ILAC and are also peer evaluated against the ISO/IEC 17011 standard to demonstrate their competence in conducting accreditations. ANAB also provides supplemental accreditation requirements that forensic and calibration laboratories must comply with. ANAB’s AR 3125 ISO/IEC 17025:2017 – Forensic Testing and Calibration Laboratories Accreditation Requirements, can be found here: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=12371 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=12371) Following the completion of an accreditation process, a forensic laboratory’s ANAB Certificate of Accreditation can include a statement of compliance with the requirements of ISO/IEC 17025, the FBI QAS and ANAB’s AR 3125. For medical/clinical laboratories, accrediting bodies like ANAB, may also be used. Accreditation with these labs involve the standard, ISO 15189, and any additional corresponding supplemental requirements. Accrediting bodies, in this instance, utilize the ILAC guidance document, G26, Guidance for the Implementation of a Medical Accreditation Scheme. With all of this in mind, a laboratory must ensure that their quality management system is wellorganized and integrates proof of conformance with all respective requirements and standards based on the services they provide. References: https://anab.ansi.org/en/forensic-accreditation https://anab.ansi.org/iso-15189-medical-labs (https://anab.ansi.org/en/forensic-accreditation) (https://anab.ansi.org/iso-15189-medical-labs) https://ilac.org/publications-and-resources/ilac-guidance-series/ (https://ilac.org/publications-and- resources/ilac-guidance-series/) Forensic Science Standards In 2014, the National Institute of Standards and Technology (NIST) Organization of Scientific Area Committees (OSAC) for Forensic Science was created. The establishment of the OSAC stemmed from a lack of standardized practices in the forensic science community brought to light publicly by the National Academy of Sciences’ (NAS) National Research Council (NRC) in a highly critical 2009 report entitled “Strengthening Forensic Science in the United States: A Path Forward”. This report was presented to Congress and called for several improvements to forensics, including the development of valid and reliable standardized practices. Reference: https://www.ojp.gov/pdffiles1/nij/grants/228091.pdf (https://www.ojp.gov/pdffiles1/nij/grants/228091.pdf) The OSAC consists of a Forensic Science Standards Board (FSSB), Scientific Area Committees (SACs), discipline-specific subcommittees (SCs) and FSSB Resource Task Groups. The OSAC is responsible for drafting, evaluating, and submitting science-based standards to a standards developing organization (SDO), like the Academy Standards Board (ASB), who are responsible for publishing them following their own review process requirements. The ASB, accredited by American National Standards Institute (ANSI), is responsible for forensic science standards development and publication. Part of the approval process involves asking for public commentary to ensure all parties, forensic, legal, or other stakeholders, have the opportunity to play a role in their development. To view the approval process in greater detail, see: https://www.nist.gov/osac/registry-approval-process#OSAC%20PROPOSED (https://www.nist.gov/osac/registry-approval-process#OSAC%20PROPOSED) Once published, these freely accessible standards are available from the OSAC Registry for integration into quality systems thereby creating discipline-specific standardized best practices for forensic laboratories. As the OSAC website states, “The OSAC Registry is a repository of highquality, technically sound published and proposed standards for forensic science. These written documents define minimum requirements, best practices, standard protocols and other guidance to help ensure that the results of forensic analysis are valid, reliable and reproducible. All the standards on this registry have passed a rigorous technical and quality review by OSAC members, including forensic science practitioners, research scientists, statisticians and legal experts. OSAC encourages the forensic science community to implement these published and proposed standards”. (Source: https://www.nist.gov/organization-scientific-area-committees-forensic-science/osac-registry (https://www.nist.gov/organization-scientific-area-committees-forensic-science/osac-registry) ) For more information regarding the development of forensic standards, see https://ndaa.org/wp-content/uploads/BTL-Vol29-No10-Oct21-Forensic-Science-Standards.pdf (https://ndaa.org/wp-content/uploads/BTL-Vol29-No10-Oct21-Forensic-Science-Standards.pdf) For published forensic science standards and those still open for public comments see: https://www.aafs.org/academy-standards-board (https://www.aafs.org/academy-standards-board). Note that each standard is listed with the “ANSI/ASB” acronyms as the approval process includes both organizations. These standards are referred to as American National Standards. Module 1: Introduction to QA/QC Page 2 Brief History of Laboratory Quality Assurance and Control Brief History of Laboratory Quality Assurance and Control Although the concept of a quality management system may sound fairly recent accompanying the advancements of technology, it is indeed at least a century old. The physicist, engineer, and statistician Walter A. Shewhart was the first to recognize the importance of process quality control to both increase the output as well as the quality of the finished product. He established the first industrial quality guideline while working for Western Electric Company when providing a control chart in 1924 that significantly expanded on quality control during the manufacturing process instead of only inspecting the finished product, as was custom at the time. His approach was actually based on statistics rather than a simple flow process. Using a simple normal distribution (Gaussian bellshape distribution), Shewhart was able to convince the management that implementation of various in-process quality control steps could greatly improve revenue by eliminating malfunctioning pieces early on instead of having to throw away the end product. In 1926, the International Federation of the National Standardizing Associations (ISA) was initiated which mainly focused on standard implementation in mechanical and electrical engineering. The association was the predecessor of the ISO and was disbanded during the Second World War. Poor quality management can result and be the cause of a number of common laboratory errors as shown in the following diagram. Although many of these issues have been addressed over the past years by implementing various policies and analytical quality control (AQC) in laboratories, each individual laboratory will have to find the best approach to starting and keeping the implementation. One important stepping stone after the initial implementations of process quality controls in certain industries was the practical quality policy adopted by the US military in 1942. The policy was basic in nature but required contractors to adhere to certain requirements for shell, aircraft, and missile suppliers. As a result of the implementation of the policy there were significantly less reports of malfunctions and uncontrolled explosions of ammunition and equipment. In 1947, the International Organization for Standardization (ISO, derived from the Greek word isos meaning equal) was created to provide international standards set by a council of representatives from national standards organizations. These policies developed significantly over the past decades. The organization, although non-governmental in nature, develops industrial and commercial standards that often become law through treaties or are being adopted by national organization as standards for accreditation. Interestingly, the US and British military had a significant influence on the development and implementation of industry standards. The US military further increased the scope of its initial quality management guidelines by implementing the MIL-Q-9858 quality standards in 1959 which was different from the British policies for military contractors known as AvP92 which quickly was adopted across Europe. Based on the discrepancies between the two policies, contractors were discouraged from producing for the US military. Eventually, the US military changed the policies to adopt the more stringent European system and the new system was adopted in 1968 by NATO (North Atlantic Treaty Organization) as the Allied Quality Assurance Publication 1 (AQAP-1). Up until this point, quality management was primarily focused on military contractors and engineering specialties – therefore production businesses. A new British policy expanded the scope of quality management systems by adopting the BS 5750 policy in 1979 which would be the predecessor for quality management guidelines established through ISO. This policy expanded the scope from military contractors to include general quality requirements for industrial, commercial, and governmental products and services. Although it took another 8 years, ISO issued the current basis for quality management systems with the release of ISO 9000 in 1987. This set of standards formed the basis for quality operations in a variety of industries – including the services offered by analytical laboratories. Over the past 20 years, ISO guidelines have significantly evolved. Most accreditation bodies in the US, Europe, Asia, and South America utilize the family of ISO 9000 standards for implementation of quality management. A company who has received and maintains ISO 9000 certification has a valid quality management system in place that guarantees the customer to meet certain expectations and requirements in how a company conducts its business and how its employees perform and are being evaluated. The most important ISO guidelines that are the basis for analytical laboratory accreditation are ISO 17025 (General requirements for competence of testing and calibration laboratories), ISO 9001 (quality management), and ISO 15189 (Medical laboratories — Particular requirements for quality and competence). Module 1: Introduction to QA/QC Page 3 Key Elements of a QA Program Key Elements of a QA Program All quality assurance programs have similar key elements that must be addressed. These are designed to ensure that the sample: was protected from any possible contamination problems was handled by appropriately trained personnel using maintained and calibrated equipment and standard validated procedures that all of the above was appropriately documented These key elements are discussed briefly here and in greater detail throughout the course. Personnel and Training health and safety organization management responsibilities All employees should have a personnel file that documents the qualifications and training for the job. Employees should be comfortable in their knowledge of how to conduct a procedure or use a piece of equipment. Once trained, the employee must show that he or she is proficient in the particular task by performing it an appropriate number of times under the supervision of the section supervisor. Once the supervisor is satisfied that the procedure has been learned, a document is signed and dated certifying training and proficiency. Facilities design of the laboratory security storage Separation of activities that might cause cross contamination is a prime consideration. In addition, the control of environmental factors such as temperature and humidity that could influence the validity of the test results must be documented. Samples should be stored under suitable conditions and the whole laboratory must have a security policy that controls access to areas affecting the quality of the testing. Equipment inspection maintenance calibration The laboratory must have written instructions regarding the use of all pieces of equipment, including calibration and maintenance. Each piece of equipment should also have a logbook or other record keeping method such as a computerized maintenance tracking that documents any maintenance performed on that instrument. Test Article Tracking chain of custody characterization stability computer tracking systems There must be a system in place that enables samples to be tracked through the laboratory. A chain of custody form indicates who received the sample at the laboratory and on what date, the condition of the sample upon receipt by the laboratory and what happened to the sample, thereafter, as the laboratory tested it. If a computer tracking system is used for some or this entire requirement, it must be validated to demonstrate that the system is reliable. Standard Operating Procedures preparation of SOP modification of SOP revision of SOP All procedures in the laboratory, including sample preparation, analysis reporting, etc. must have a written standard operating procedure. Deviations from this procedure for any reason must be documented and approved. Should it be necessary to revise an SOP the reason should also be documented and the original procedure archived. Study Protocols relationship between the study protocol and SOPs The study protocol describes procedures that are specific to a particular study, for example enrollment of subjects in a clinical trial as opposed to SOPs that have general application to the laboratory operations. Method Validation analytical method validation validation of computer generated data All analytical methods must be validated prior to use on real samples. Validation includes such parameters as sensitivity, specificity, linearity, and reproducibility. If computer programs are used to generate this data, then the programs must also be validated to ensure that the data is accurate and reproducible. Final Report key elements for inclusion in the final report statistical considerations The final report should include documentation of the process to which the sample was subjected, the results obtained, appropriate statistical analyses, and any conclusions formed. The report is also subject to quality control review before being signed by the appropriate laboratory personnel. Archiving archivist storage retrieval It may take many years for legal action concerning a particular forensic sample to finally conclude. Similarly, clinical data associated with the submission of a regulatory package make take an extended amount of time to be collated. It is therefore important that all raw data, documentation, records, protocols, specimens, and final reports generated as a result of a study are retained. The retention time depends on the type of sample and the regulations pertaining to it, but typically 2-5 years are usual lengths of time for documents to be kept by the laboratory. These archives are the responsibility of a member of the laboratory who is the designated archivist. Module 2: Key Elements of a QA/QC Program Page 1 Module Overview Introduction Welcome to Module Two. In this module we will learn the organizational requirements for an accredited forensic laboratory as an example of the necessary elements of a QAQC system. Although many of these elements may appear logical, it is important to maintain records and address/enforce all necessary quality assurance and control issues in all analytical laboratories. An accredited forensic laboratory needs: an overall manager for the laboratory a technical director who oversees daily operation of the laboratory a person responsible for quality control/quality assurance Management is responsible for training all employees to accurately perform the required analyses. Just as importantly, management is responsible for the health and safety of all employees. It must provide training about the dangers inherent in a forensic laboratory - chemicals, instrumentation, bloodborne pathogens, radiation. Adequate protection from these dangers must also be provided. Although United States regulations are cited in this module, most industrialized nations have similar laws concerning the health and safety of their workers. Objectives At the end of this module students should: know the personnel required for an accredited forensic laboratory, their qualifications, and function within the organization know what is required to implement the OSHA regulations concerning the health and safety be able to find and read a SDS (previously known as MSDS) know when a bloodborne pathogen program is needed have an understanding of Universal Precautions know the responsibilities of management Module 2: Key Elements of a QA/QC Program Page 2 Education and Training Education and Training ISO (International Standards Organization) states, "the management of the laboratory shall have a training policy and procedures for identifying training needs and providing training of personnel. The training programs shall be relevant to the present and anticipated tasks of the laboratory" (ISO/IEC 17025:2017). The ISO 17025 guidelines are applicable to all testing and calibration laboratories and therefore not limited to forensic laboratories. Similar criteria for the selection of personnel are mandated by the EPA (Environmental Protection Agency) and FDA (Food and Drug Administration). Once a person with the appropriate qualifications has been hired, the laboratory must still provide that person with training on the specific tasks he/she will perform. Furthermore, the employee is expected to demonstrate competency for each analytical procedure he performs. Some accreditation bodies actually specify the education and experience required for the various positions. For instance, in order to be accredited as a horseracing and canine racing laboratory in the State of Florida, "the laboratory technical leader shall have a minimum of either: a 4-year baccalaureate with college credit courses in chemistry, pharmacology and toxicology or related subjects, course work in statistics, and 5 years of experience as an analytical chemist in a laboratory analyzing drugs in body fluids including experience in giving evidence, or a graduate degree with college credit courses in chemistry, pharmacology and toxicology or related subjects, course work in statistics, and 2 years of experience as an analytical chemist in a laboratory analyzing drugs in body fluids, including experience in giving evidence." The quality assurance standards for forensic testing laboratories and DNA databasing laboratories issued by the FBI director specified that the technical manager or leader of the laboratory meet the following degree/educational requirements: 1. A graduate degree in a biology, chemistry, or forensic science related area 2. A minimum of 12 credit hours or its equivalent including a combination of graduate and undergraduate course work or classes covering the subject areas of: Biochemistry Genetics Molecular Biology Statistics and/or population genetics In addition, the technical leader must have a minimum of three years forensic experience. There are also requirements for DNA analysts. Each examiner/analyst must meet the following degree/educational requirements: 1. B.A. / B.S. degree or its equivalent in a biology, chemistry, or forensic science related area 2. College course work or classes covering the subject areas of: Biochemistry Genetics Molecular Biology 3. College course work or training which covers the subject area of statistics and/or population genetics In addition each examiner/analyst must have at least six months experience prior to working on unsupervised cases. Module 2: Key Elements of a QA/QC Program Personnel Page 3 Personnel There should be a sufficient number of personnel to ensure analyses are conducted in a timely and proper manner. Each employee may need to wear a lab coat or other item(s) that will: protect him/her from potential dangers in the laboratory-biological, radiological, or chemical-and/or prevent contamination of the test equipment or the test or control articles themselves There are some instances, as in some drug studies, where the illness of an employee may contaminate or otherwise bring into question the integrity of an analysis. In such a case, the employee must be removed from the study until the condition has been corrected. An employer must maintain: A job description for each position. This description lists the tasks to be performed and the training/experience/education required for the successful performance of these tasks. An employee personnel file for each employee that contains the job description and information on that individual's training and experience for the job. Aside from job specific training, training on pertinent health and safety matters must be documented. All training records must be kept up to date. Management In addition to providing qualified personnel, the management staff must provide sufficient resources for the successful completion of the analyses. The personnel must be free from commercial, financial, or other potential pressures that could compromise the integrity or quality of their work. Conflicts of interest should be avoided. Management should establish policies that will minimize influence exerted either by these external sources or by other groups within the organization itself. For instance, a written policy on the acceptance of gifts from clients or vendors may be helpful. The role of each employee must be clearly defined. In other words, the responsibility, authority and interrelationships of all the employees must be specified in order to optimize every step of the testing procedure-from receipt of the sample, analysis, through the data interpretation and final reporting of the data. Furthermore, the laboratory needs to be organized so that each employee is able to exercise his/her independence of judgment. Should problems arise, an employee should have access not only to his own immediate supervisor but to other members of management. The management can be held legally responsible for the accuracy of reported results. Organization A properly functioning forensic laboratory needs: A laboratory director who has overall responsibility for all aspects of the laboratory operation. This includes the coaching and mentoring of staff as well as the technical performance of the analyses, the interpretation, analysis, documentation and reporting of results. A technical manager (group leader, section leader, or similar designation) who has overall responsibility for the technical operations of the laboratory. He/she should be familiar with the instrumentation, calibration or test methods, procedures, and assessment of the results. A quality manager (quality control officer, or similar designation) who is responsible for the quality system and consequently should have direct access to the technical manager and/or laboratory manager. Depending upon the size of the laboratory, a single person may actually be required to perform the functions of both the technical and quality managers. Safeguards should be established to ensure that decisions made as the quality manager do not influence one's decisions made as the technical manager, or vice versa. It should be made clear to the staff whether one is acting as the technical or the quality control manager. Support personnel (lab technicians, chemists, secretaries, etc.) who receive and log in the forensic samples, perform the actual analyses, etc. Some employees who may be asked to serve in various training and informational capacities such as safety officer, hazard communication coordinator, etc. The title of a person filling one of these positions may vary between organizations and one individual may be responsible for several different functions within a small laboratory. Nevertheless, the functions exercised by the laboratory director, technical manager, quality manager, safety officer, and other support personnel are all important for a properly operated forensic laboratory. Module 2: Key Elements of a QA/QC Program Page 4 Health and Safety Health and Safety OSHA (Occupational Safety and Health Administration in the Department of Labor) has issued numerous standards (found in 29 CFR [Code of Federal Regulations] Part 1910 Occupational Safety and Health Standards) for the purpose of safeguarding the health and safety of industrial workers: https://www.osha.gov/pls/oshaweb/owasrch.search_form?p_doc_type= STANDARDS&p_toc_level=1&p_keyvalue=1910 (https://www.osha.gov/pls/oshaweb/owasrch.search_form? p_doc_type=STANDARDS&p_toc_level=1&p_keyvalue=1910) Many of these standards cover areas that do not directly impact laboratory quality assurance work such as shipyard employment, the construction of ladders, etc. There are several OSHA standards that are of direct concern to forensic laboratories in the United States. Hazard Communication (29 CFR 1910.1200) Bloodborne Pathogens (29 CFR 1910.1030) Personal Protective Equipment (29 CFR 1910.132) Eye and Face Protection (29 CFR 1910.133) Respiratory Protection (29 CFR 1910.134) Hand Protection (29 CFR 1910.138) Occupational Exposure to Hazardous Chemicals in Laboratories (29 CFR 1910.1450) Reference: https://www.osha.gov/sites/default/files/publications/OSHA3404laboratory-safety-guidance.pdf (https://www.osha.gov/sites/default/files/publications/OSHA3404laboratory-safety-guidance.pdf) 1910.1200 Hazard Communication Standard (HCS) This standard is frequently referred to as "Right-to-Know" training. (The HCS pre-empts all state and local laws in those states that do not have an OSHA-approved job safety and health program. State right-to-know laws are authorized in those states with OSHA-approved state programs). HCS mandates that employers and employees must be informed of the hazards of all chemicals used (whether purchased or produced) within the laboratory. There are several aspects of this program. The employer must: Maintain a written hazard communication program within the workplace Example of a written hazard communication program: For laboratory personnel: ( https://www.ehs.ufl.edu/about/policies/hazard-communication-policy/ (https://www.ehs.ufl.edu/about/policies/hazard-communication-policy/) ) For non-laboratory personnel: ( https://www.ehs.ufl.edu/departments/research-safety-services/chemical-and-labsafety/chemical-safety/chemical-safety-information/ (https://www.ehs.ufl.edu/departments/research-safety-services/chemical-and-lab-safety/chemicalsafety/chemical-safety-information/) ) Implement an employee training program that provides information about the hazards of the chemicals used within that workplace and the use of protective procedures, clothing, and equipment. Maintain a chemical inventory. The list should at least include: the name (and possible common alternative names) of the chemical approximate quantity storage location date of acquisition Depending upon the function of the lab, further details would include: the manufacturer quantity used (and date of use) frequency of use expiration date hazards CAS number (glossary: CAS number, Chemical Abstracts Services number. This is a unique number that is used to identify a specific known compound.), etc. may be desired and/or required. Maintain a complete collection of pertinent Safety Data Sheets (SDS). Please note, that SDSs were previously known as MSDSs (Material Safety Data Sheets). Some websites may still refer to these documents as such. Both employees and emergency personnel must have ready access to a SDS for each chemical present in the workplace. This means that every employee must be informed as to the physical location of the SDSs within the workplace and be able to access them at all times. Alternatively, many SDSs are internet accessible; and if all employees are instructed how to obtain SDSs on the web and always have access to a computer, the internet SDS can be used in lieu of a hard copy (it is still advisable to have hard copies of all SDSs). The SDSs are primarily intended for use by persons who regularly work with a hazardous substance rather than by the consumer who has limited contact with a product. Under the assumption that some individuals have extensive contact with janitorial chemicals, pesticides, antiodorants, biological products such as gels, and so on, SDSs are available for these products. In some instances, a single chemical may be obtained from a variety of different suppliers. It is best to save a SDS from each manufacturer; however, a single SDS can be saved as long as the emlpoyees understand that one SDS is representative of all the SDSs for that compound. Furthermore, the user must be able to cross reference the single SDS for all bottles of that chemical regardless of manufacturer or supplier. That is, for example, it must be clear that dichloromethane is identical to methylene chloride and that the SDS for dichloromethane is the same as the SDS for methylene chloride. Laws mandating the availability of SDS in the workplace have been enacted by many countries. Where can one obtain a SDS? Chemical manufacturers and importers are required to impart hazard information about their product to the downstream users by means of the SDS and appropriate labeling of the container containing the chemical. Frequently, for one reason or another, a paper copy of the SDS that arrives with the new chemical or the SDS has been lost. A SDS is to be shipped with the first order of a chemical; however, in a large organization, this means that only the first laboratory to order the chemical actually gets a SDS. Other laboratories that order later do not. A SDS for each chemical can be requested from the manufacturer or distributor at the time of purchase. Some companies only provide SDSs via their extensive Web-accessible SDS collections. SDSs are also available on some general sites, governmental and other non-profit sites, and some other industrial sites (pharmaceutical, paint, janitorial products, etc). Other commercial sites have SDSs for sale. Module 2: Key Elements of a QA/QC Program Page 5 Health and Safety (continued) Health and Safety (continued) SDS Information and Sources The following site lists many Internet sources for SDS and also contains other information about SDS: http://www.ilpi.com/msds (http://www.ilpi.com/msds). Some sites having a large number of SDS are: Internet Site URL Chemical Safety https://chemicalsafety.com/sds-search VelocityEHS MilliporeSigma Fisher Scientific Number of (M)SDS Comment 300,000+ (M)SDS, Must register (but free) 90,000 Access via catalog 61,000 Access via catalog (https://chemicalsafety.com/sds-search) https://www.ehs.com/resources/sds (https://www.ehs.com/resources/sds) www.sigma-aldrich.com (http://www.sigmaaldrich.com/) https://www.fishersci.com/us/en/catalog/search/sdshome.html (https://www.fishersci.com/us/en/catalog/search/sdshome.html) What information does a SDS provide? Each chemical company provides its own SDS format. Nevertheless, the company must include in the SDS all the pertinent information it has on: Product identification-including name, synonyms, formula and molecular weight, CAS number Composition/information on ingredients Health hazards/first aid/personal protection Handling and storage Flammability Physical and chemical properties Toxicology Disposal methods Transportation Regulatory Other There are some limitations to the usefulness of the SDSs: A chemical manufacturer is not required to perform exhaustive studies of the health hazards and toxicology, etc. of every chemical sold but must merely report what is known about the compound. Thus, complete information is primarily available for chemicals that are used in relatively large quantities or that are exceedingly toxic. The composition of some chemicals is sometimes considered proprietary knowledge. In case of a medical or other emergency, the manufacturer is supposed to provide privileged information as to the identity of the proprietary compound(s) to the appropriate medical personnel with the understanding that the information will not be made available to anyone else. For an example SDS see: https://www.sigmaaldrich.com/US/en/sds/sigald/e7023 (https://www.sigmaaldrich.com/US/en/sds/sigald/e7023) Provide employees information on how to read chemical labels. All chemicals received by the laboratory should be appropriately labeled with the name of the substance and hazards. The exact manner of labeling is determined by the manufacturer; however, hazard pictograms frequently indicate whether the substance is explosive, an oxidizer, flammable, toxic, harmful or an irritant, corrosive, environmentally toxic, or to be kept away from food. Specific hazards may also be enumerated on the label. Fisher Scientific uses the National Fire Protection Association (NFPA) Hazard Code symbol to depict dangers. The degree of danger-4 being the most severe, 0 being least severe-is found within a section of the symbol. When the symbol is color-coded, blue signifies health, red-- flammability, yellow--reactivity (yellow) dangers. The white section at the bottom may contain special warnings. https://www.bgsu.edu/content/dam/BGSU/envhs/documents/Lab-Safety/NFPA-Labeling-Information.pdf (https://www.bgsu.edu/content/dam/BGSU/envhs/documents/Lab-Safety/NFPA-Labeling-Information.pdf) All chemicals and mixtures that are prepared in the laboratory must also be adequately labeled with the name of the chemical(s). Similarly, all hazardous waste materials must be labeled. The disposal of this waste will be discussed later in the course. Module 2: Key Elements of a QA/QC Program Page 6 Health and Safety (continued) Health and Safety (continued) 1910.1030 Bloodborne Pathogens An annual bloodborne pathogen exposure control program must be implemented if the laboratory personnel has any exposure to human blood, human blood components, products made from human blood, or any other potentially infectious material. Essentially, this means any substance that has been in contact with human blood, tissue, saliva in dental procedures, or any substance contaminated with blood (such as soiled linens, etc.). A bloodborne pathogen is any pathogenic (disease causing) microorganism present in human blood that causes disease and includes the hepatitis B virus (HIB) and the human immunodeficiency virus (HIV). The major points in this training include: It is mandatory that all personnel who come in contact with bloodborne pathogens must receive bloodborne pathogen training once a year. Hepatitis B virus vaccinations must be made available to an employee within 10 days of assignment to a position involving bloodborne pathogens. Universal Precautions must be practiced. This means that all human blood and human body fluids are treated as if known to be infectious for HIV, HBV, and other bloodborne pathogens. Engineering and work practice controls must be implemented. These include use of gloves and handwashing upon their removal. Proper disposal of contaminated sharps and needles, no eating and drinking on the premises, etc. Personal protective equipment such as gloves, lab coats, etc. must be used at all times. Gloves should not be reused and care should be taken to minimize contamination by changing gloves frequently, and always when moving from an area that contains post amplified product. All equipment and laboratory surfaces must be regularly cleaned and disinfected. All waste must be labeled appropriately and must be either incinerated or removed by an authorized biological waste removal service. In addition it is important not to contaminate areas where pre-amplified DNA is analyzed with material (either equipment or waste) from the post-amplified DNA area. All areas contaminated with biological waste (whether it be a room, refrigerator or bag of waste) must have fluorescent orange or orange red labels with the biohazard symbol. A detailed bloodborne pathogen program exposure control plan can be found on the web at: https://webfiles.ehs.ufl.edu/BBP_ECP.pdf (https://webfiles.ehs.ufl.edu/BBP_ECP.pdf). Personal Protective Equipment, Eye and Face Protection, Respiratory Protection and Hand Protection (29 CFR 1910.132, 1910.133, 1910.134, & 1910.138) These straightforward standards discuss the importance of providing the correct protective equipment to laboratory personnel depending on the job they are tasked with. These safety related standards discuss the need for laboratory personnel to be supplied with and don the appropriate protective equipment some of which may require training and testing as a prelude to their use. Occupational Exposure to Hazardous Chemicals in Laboratories (29 CFR 1910.1450) This OSHA standard discusses the importance of a Chemical Hygiene Plan (CHP). The CHP is defined by OSHA as “a written program developed and implemented by the employer which sets forth procedures, equipment, personal protective equipment, and work practices that (i) are capable of protecting employees from the health hazards presented by hazardous chemicals used in that particular workplace…”. Source: https://www.osha.gov/laws-regs/regulations/standardnumber/1910/1910.1450 (https://www.osha.gov/laws-regs/regulations/standardnumber/1910/1910.1450) As discussed further in OSHA’s Laboratory Safety Guidance booklet, “The CHP must include provisions for worker training, chemical exposure monitoring where appropriate, medical consultation when exposure occurs, criteria for the use of personal protective equipment (PPE) and engineering controls, special precautions for particularly hazardous substances, and a requirement for a Chemical Hygiene Officer responsible for implementation of the CHP”. Source: https://www.osha.gov/sites/default/files/publications/OSHA3404laboratory-safety-guidance.pdf (https://www.osha.gov/sites/default/files/publications/OSHA3404laboratory-safety-guidance.pdf) Module 2: Key Elements of a QA/QC Program Page 7 Other Considerations Other Considerations Further stringent guidelines must be followed by all nonclinical, clinical and research facilities that have studies involving animals and/or humans. Good laboratory practices (GLP) must be followed for nonclinical laboratory studies of substances regulated by the FDA (reference: 21 CFR Part 58, http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFR/CFRSearch.cfm?CFRPart=58 (http://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfCFR/CFRSearch.cfm?CFRPart=58) ) and for EPA studies relating to health effects, environmental effects, and chemical fate testing (reference: 40 CFR Part 792, https://www.ecfr.gov/current/title-40/chapter-I/subchapter-R/part-792 (https://www.ecfr.gov/current/title-40/chapter-I/subchapter-R/part-792) ). The EPA also mandates Good Automated Laboratory Practices for laboratories that employ laboratory information management systems (LIMS) to acquire, record, manipulate, store and archive their data. Depending on the study and the end user of the data, different rules and regulations may govern the conduct of the analyses. Data produced using GLP tend to be accurate and reproducible and can be compared to data from other laboratories also practicing GLP. Furthermore, many granting agencies and other organizations will provide funds only to those laboratories observing GLP. Thus, GLP practices and procedures are advisable whether mandated by law or not. The care and use of animals is regulated as well (reference: 9 CFR Animals and Animal Products, Parts 1-199). The FDA and EPA GLP also address this subject. Basically, requirements for feeding, housing, and transporting are specified. To insure that the animals are used as humanely as possible, an Institutional Animal Care and Use Committee (IACUC) must review and approve all studies involving animal. There are several organizations such as the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC, http://www.aaalac.org (http://www.aaalac.org) ) that provide accreditation for facilities that use animals. Frequently, funds are made available only to those laboratories having accredited animal use programs. Studies involving humans are even more stringently regulated than those using animals. See CFR Title 45 Part 46, Protection of Human Subjects https://www.ecfr.gov/current/title-45/part-46 (https://www.ecfr.gov/current/title-45/part-46) ). An Institutional Review Board (IRB) reviews and approves all such studies. More information regarding research involving human subjects can be found at: https://www.ecfr.gov/current/title-21/chapter-I/subchapter-A/part-56 (https://www.ecfr.gov/current/title-21/chapter-I/subchapter-A/part-56) A variety of other training programs may be provided, and attendance possibly required, by the organization itself. These may include: waste management, AIDS prevention, diversity training, and others. Module 2: Key Elements of a QA/QC Program Case Study Page 8 Case Study Safety precautions in a morgue and clinical laboratory A morgue, as the place in a medical laboratory where autopsies are conducted, has personnel safety and health hazards that must be considered. The combination of potential exposure to infectious diseases from bodily fluids, as well as exposure to chemicals that are used to preserve samples puts the pathologist (as well as anybody handling the samples/body) at an increased risk of exposure and contamination. Much has changed in terms of personal protective equipment (PPE) over the past decades. While minimal protective gear has been worn in the 1970s, it is now mandatory to wear masks, face shields, double and triple gloves, gowns, shoe covers, etc. when handling bodies. The use of many different chemicals is extensive and many are hazardous by themselves and even more so in combination with other chemicals and require specific handling and storage. Accidents may cause significant injuries and damages in a laboratory; one example is sodium azide, which was commonly used in the past, as this chemical can combine with metals to form highly explosive intermediates. Please read the full report and description of how safety measures can significantly affect the work conditions in a laboratory. http://hubpages.com/hub/Health-Hazards-and-Safety-Issues-in-a-Medical-Laboratory (http://hubpages.com/hub/Health-Hazards-and-Safety-Issues-in-a-Medical-Laboratory) Module 3: Facility Standards and Safety Measures Page 1 Module Overview Introduction Welcome to Module Three. In order to perform quality work, the facilities housing the laboratory must meet certain minimum standards. Separation of activities that might cause cross-contamination is a prime consideration. The control of environmental factors such as temperature and humidity that could influence the validity of the test results must be documented. Samples should be stored under suitable conditions and the whole laboratory must have a security policy that controls access to areas affecting the quality of the testing. Objectives At the end of this module students should: Know the requirements for handling hazardous chemicals, radiation, and biological materials. Understand the importance of the safety measures presented. Understand the importance of appropriate facility layout and design. Be familiar with the requirements for the use of research animals. Module 3: Facility Standards and Safety Measures Facilities Page 2 Facilities Although facilities need not be palatial, they need to be large enough that one laboratory function cannot adversely affect any of the other laboratory functions. Specifically, this means that the various laboratory operations should be sufficiently removed from one another so that sample contamination is prevented and the health and safety of the personnel is ensured. There are numerous regulations that spell out the minimum requirements for laboratories using biological materials, animals, chemicals and radiation. The reference “Prudent Practices in the Laboratory” by the National Research Council describes the various safety features that should be available in a chemical laboratory facility and the reasons for their inclusion. It also provides the details needed for a chemical hygiene program. Reference: https://nap.nationalacademies.org/catalog/12654/prudent-practices-in-the-laboratory-handling-andmanagement-of-chemical (https://nap.nationalacademies.org/catalog/12654/prudent-practices-in-thelaboratory-handling-and-management-of-chemical) An abbreviated version of "Prudent Practices" is found in 29 CFR 1910.1450 App A, National Research Council Recommendations Concerning Chemical Hygiene in Laboratories, http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=standards&p_id=10106 (http://www.osha.gov/pls/oshaweb/owadisp.show_document?p_table=standards&p_id=10106). A fairly specific description of facilities needed for forensic laboratories is found in Forensic Laboratories: Handbook for Facility Planning, Design, Construction, and Relocation, The US Department of Justice, Office of Justice Programs, National Institute of Standards and Technology, June 2013, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=913987 (https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=913987) In addition, the Federal Bureau of Investigation (FBI) provides detailed guidelines for a laboratory conducting forensic DNA testing and databasing with respect to the separation of pre- and postamplified DNA, equipment and work areas. Reference: The Quality Assurance Standards for Forensic DNA Testing Laboratories can be found here: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746) The Quality Assurance Standards for DNA Databasing Laboratories can be found here: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743) In an ideal situation, facilities will have been specifically constructed to house the laboratory, each room being equipped to adequately handle the function for which it was designed. For instance, rooms having instrumentation may require electrical strips on all walls and more power than some other areas. The "wet chemistry" laboratories will need adequate sinks, hoods, etc. Furthermore, a logical arrangement of rooms will facilitate the transfer of the test sample through the various stages of its analysis. As mentioned above, dedicated rooms for pre- and post-amplified DNA are required. Some laboratories also include the separation of air conditioning systems to ensure no inadvertent contamination between these areas occurs. To ensure the integrity of the samples (and analytical results), the facility must be secure. Doors into the facility should be locked at all times, not left ajar to allow someone temporary access. Laboratory personnel must enter the facility through a checkpoint or must use a cardkey, keypad or similar access at the entry door(s). When visiting the facility, guests must be escorted by the personnel. In larger organizations, identification badges may be desirable. Exit from the facilities in case of fire or other emergency should not be limited. Receipt and Storage of the Test, Control and Reference Substances Presumably, all materials will be received at a central receiving location. The substances, whether a test material, control material or reference material (standard), will be logged in and, when necessary, labeled more completely. Different areas will be set aside for storage based on the substance’s category. This prevents cross-contamination as well as possible human error. Depending upon the materials, refrigeration, freezing, a controlled temperature, or other conditions may be required for storage. Provisions should be made for keeping all "controlled substances" under lock and key and their use must be documented. Reference samples for DNA analysis must be contained so that crosscontamination between reference and evidence samples does not occur. This is generally achieved by packaging samples in leak proof containers sealed with tamper evident seals. The same applies to any other analytical laboratory – especially in quality control for pharmaceutical companies where inprocess controls and final drug products are evaluated for purity, identity, and concentration. Similarly, to prevent contamination or human error, separate areas must be set aside for: preparation of the test sample/unknown preparation of the control and reference materials storage of the test, control, and reference materials It is prudent to perform the functions of the examination of reference samples and the examination of evidence such as clothing, in separate locations or at separate times so that cross-contamination does not occur. Module 3: Facility Standards and Safety Measures Page 3 Facilities (continued) Facilities (continued) Laboratory Operation Areas Ventilation In general, all laboratory areas should have a ventilation system that continually replaces the air in the room with air taken from a non-laboratory area. Assuming that hoods, vents, scrubbers, or similar devices are employed to control the emission of toxic substances into the air, four to twelve air changes per hour should provide adequate air quality in the laboratory. It is vital that the air intake must not be located in close proximity to the air exhaust. The quality and quantity of ventilation should be established initially and be reevaluated at 3-month intervals or whenever changes in the ventilation system occur. The ventilation characteristics in a room in which hoods are operating can differ significantly from the characteristics when the hoods are not in operation. Areas in which volatile chemicals are used should be equipped with fume hoods that are vented separately to the outside. (A toxic substance in one hood may actually spew forth into and out from other hoods that have a common exhaust, especially when the other hoods are not in operation.) Generally 2.5 linear feet of hood per person should be provided for every two persons who work regularly with chemicals. A specially constructed fume hood is required when perchloric acid is used. To assure that a hood is working properly, the hood should have a continuous flow-monitoring device that is checked prior to and during use. When toxic compounds are stored in the hood, it should be operated 24 hours a day. Ventilated storerooms, stockrooms, storage cabinets, canopies or any other ventilation devices should also have their individual exhaust ducts. Glove boxes, isolation units, laminar flow hoods and other devices that exhaust into the room must be equipped with the appropriate filters, chemical scrubbers, etc. Biohazard hoods are necessary for the containment and protection of samples and the operator. When dealing with DNA, it is preferable to use Class II biohazard cabinets that protect both the operator and the sample (see this review of biosafety at https://www.labconco.com/articles/what-are-the-biosafety-cabinet-classes (https://www.labconco.com/articles/what-are-the-biosafety-cabinet-classes) ). The location of biohazard hoods within the laboratory is also important. They should be located away from walkways to minimize external drafts around the hood and also minimize the likelihood of bumping the operator. In addition to biohazard hoods, PCR set up cabinets are also advantageous. They provide a contained clean area to set up PCR reactions without the expense of a biohazard hood. Ideally these cabinets should have their own set of equipment such as dedicated pipettes for use solely in these cabinets. No DNA should ever be introduced into this cabinet so that contamination of reaction mixes is minimized. These cabinets are fitted with a UV light source to aid in cleaning of the cabinet. Sealed systems such as a cold room or a warm room must not only have adequate ventilation, but must also have a way for a person to exit rapidly if the electricity (and hence, the ventilation) fails. Water Usage Every laboratory must be provided with at least one sink that is always available for washing one's hands. The same, or other sinks, can be used for washing glassware, etc. The presence of at least one sink in each hood, although not always necessary, is very useful. Every laboratory should have an eyewash device. This can be either a permanently plumbed fixture that is attached to the wall or sink faucet or can be a portable system employing a plastic water-filled bottle and eye-cup. Two advantages of the permanently fixed eyewashes are that they cannot be misplaced or removed and they are more likely to be periodically flushed (the water in all eyewashes should be changed regularly for purposes of hygiene). A safety shower or a drench shower connected to the sink should be available in case a chemical is spilled on an employee or his/her clothing. Drains under the showers make clean-up easier after usage. Showers are usually not needed in instrumental laboratories and may even present the danger of electrocution. Depending on the use of the room, a sprinkler system may be advisable. Because some chemicals will undoubtedly be poured down the sink, all plumbing should be made of acid-resistant materials. To minimize problems with DNA analysis it is prudent to use pure water of at least 18MΩ-cm for the preparation of reagents used in DNA analysis performed on the pre-amplification side of the laboratory. In addition, deionized water is also required for the post-amplified side of the DNA laboratory. Although water can be carried into the post amplification side of the laboratory, an effective decontamination procedure and monitoring of any potential contamination is then necessary. It is therefore easier and less time consuming to run an additional line into the post-amplification side of the laboratory. Module 3: Facility Standards and Safety Measures Page 4 Other Safety Equipment Other Safety Equipment Sufficient space must be reserved near each outer laboratory door for an all-purpose fire extinguisher. A fire alarm system and emergency phones must be available. Furniture All laboratories need ample bench top space and storage cabinets. Ideally, the furniture and its arrangement within a laboratory are determined by the tasks to be accomplished within a specific laboratory. Just as in the kitchen in your house, the "traffic flow" within each laboratory should be considered in order to maximize efficiency. A few generalizations can be made with regards to furnishing a laboratory. A "wet chemistry" laboratory requires sinks, bench tops, electrical outlets, and hoods. Only a small amount of chemicals can be stored within the individual laboratory in an appropriate cabinet; yet, separate cabinets are needed for acids and bases, volatile liquids, and so on. Incompatible chemicals should not be stored side-by-side in a single cabinet. An instrumentation laboratory requires even more electrical power, but may need fewer sinks and hoods. Free-standing instruments need floor space whereas others stand on bench tops. Frequently, easy access to all four sides of an instrument is desired for purposes of maintenance and repair of the instrumental "think-works". Canopy hoods may be desirable for some of the instruments. Stockrooms and Storerooms The potential toxicity and flammability of chemicals make separate stockrooms and storerooms mandatory. A central stockroom to which all laboratories have access is more efficient and costeffective than having separate storage for each lab. Storerooms and stockrooms must have continuous mechanical ventilation to the outside, appropriate fire extinguishers, and no potential sources of ignition. For instance, explosion-proof refrigerators must be used to store flammable liquids (all explosion-proof refrigerator parts that might ignite a fire are sealed to prevent this from occurring; however the refrigerator will not necessarily contain an explosion caused in some other way). The chemicals should not be exposed to direct sunlight, another heat source or stored under a sink. All bottles should be placed below eye-level, preferably in cabinets with see-through doors. Limitations are placed on the quantity of flammable liquids that can be stored in a laboratory or in a storeroom. Care must be taken to separate chemicals that will react if mixed together. A review of the requirements for storage of hazardous chemicals can be found at https://www.jove.com/v/10380/chemical-storage-categories-hazards-and-compatibilities (https://www.jove.com/v/10380/chemical-storage-categories-hazards-and-compatibilities) A more detailed description of standards set by the US Department of Labor can be found at: https://www.osha.gov/laws-regs/regulations/standardnumber/1910/1910.1450AppA (https://www.osha.gov/laws-regs/regulations/standardnumber/1910/1910.1450AppA) Some facilities may actually store chemicals and chemical waste in a separate one-story building that has 2-hour fire-rated exterior walls having no opening within 10 feet of such storage. When not in use, gas cylinders must be stored outside the laboratory. At all times, the cylinders must be stored upright with their valve protection caps on and they must be secured so that they cannot fall over. Module 3: Facility Standards and Safety Measures Page 5 Animal Care Facilities Animal Care Facilities A number of separate rooms are required when live animals are used at a facility. Both the EPA and FDA GLPs briefly outline the facilities required for animal care. Housing AAALAC and other accreditation organizations provide specific requirements for the size of the cage to house an animal, bedding, the number of air exchanges in the room per hour, and so on. Generally, Only one species can be accommodated in a room (aside from the general incompatibility of some species such as cats and dogs, a disease carried by one species may cause serious illness in a second species). The exception to the one species per room requirement occurs in the case of aquatic animals, where each species can be maintained in a different tank within the same room. Special rooms must be set aside for studies involving materials that are either biohazardous, volatile, aerosols, radioactive, or infectious. Different analyses or studies may need separate rooms. Studies requiring special housing may require separate rooms. The animals are not to be housed within the individual laboratories, but in some instances may be used in designated areas of the laboratory. A separate area shall be provided for the diagnosis, treatment and control of laboratory animal diseases. Sick animals must be isolated from well animals. In a large organization, there may be an additional area in which all newly received animals are quarantined until shown free from disease. The environment of the room can be altered according to the study protocol. This includes temperature, humidity, photoperiod (lighting). The AAALAC has outlined their accreditation standards in the “Guide for the Care and Use of Laboratory Animals”, NRC, 1996, http://www.aaalac.org/resources/theguide.cfm (http://www.aaalac.org/resources/theguide.cfm). Other Considerations Separate storage areas for feed and bedding are required. These cannot be the same areas used to store or analyze the test materials. Furthermore, they must be kept sanitary and vermin-free. Adequate provisions must be made for perishable supplies. Provisions must be made for the proper collection and disposal of animal waste, bedding, and animals and/or to their storage prior to removal from the site. Storage must be sanitary, the potential for vermin, odor, disease or environmental contamination being minimized. Access to any animal care facility needs to be limited to those persons using the animals. This helps to prevent the spread of disease and is less disruptive to the animals. Limited access also reduces possible difficulties with "animal rights" activists. Veterinary care must be provided for the animals, either by an "in-house" veterinarian or by one under contract to the facility. Animal care personnel must have received appropriate training. Module 3: Facility Standards and Safety Measures Page 6 Bio- and Radioactive Materials Bio- and Radioactive Materials Bio- Materials Lab Biological experiments are best conducted in a laboratory having restricted access, thus minimizing the number of people exposed to potentially infectious biological materials. Biological laboratories are often equipped with biological safety cabinets having HEPA (high efficiency particulate air) filters. The purpose of these safety cabinets is twofold: 1. to protect the laboratory personnel from possible infections 2. to protect biological material such as cell cultures from infections carried by the personnel. If animal studies are being conducted, other equipment such as operating tables may be needed. The degree of danger associated with the biological material determines the design of the room (or suite of rooms) and the precautions that are observed. For example, studies involving a highly contagious virus would require the personnel to change into other clothing prior to entering the laboratory, wear protective masks, shower upon exiting the lab, etc. Other studies, such as determining the amount of arsenic in tissue, might require nothing more than observation of universal precautions. The Center for Disease Control site provides a manual that describes the facilities and precautions necessary for work with biological materials: https://www.cdc.gov/labs/pdf/CDC-BiosafetyMicrobiologicalBiomedicalLaboratories-2009-P.PDF (https://www.cdc.gov/labs/pdf/CDC-BiosafetyMicrobiologicalBiomedicalLaboratories-2009-P.PDF) Radioactive Materials Studies involving radioactive materials may also be best conducted in a room specifically set aside for such purposes. Analytical work often uses beta-emitters such as tritium and carbon-14 and requires the use of bulky scintillation counters that stand on the floor. Use of the higher energy gamma-emitters may require lead-lined storage drawers, lead bricks to place in front of an experiment, and Geiger counters as well as other counters. In the case of volatile substances such as iodides, a fume hood having leaded glass and a HEPA filter may be required. It is also wise to restrict radioactive experiments to as small an area as practical because all surfaces and equipment have to be periodically checked (by "swiping", wiping the surface and counting the radioactivity) for radioactive contamination. A few micrograms of radioactive material that is not visible to the human eye can still be highly radioactive. Specimen / Data Storage Area There must be an area, separate from the area holding the unanalyzed specimens, in which any remaining specimen can be stored under the appropriate conditions. It is prudent to maintain a tracking system for the location of these remaining specimens particularly if the laboratory processes a large number of samples and there is a likelihood that further analysis will be required by an external agency such as a court of law. Furthermore, in a forensic context, the development of new technology often allows the analysis of samples in the future that cannot be analyzed using current technology. It is therefore essential that these samples are stored appropriately and easily located at a later date. Waste Disposal Facilities Aside from the usual waste generated by any business, a laboratory must dispose of chemical biological radioactive waste Usually, a private waste concern picks up the waste and disposes of it as mandated by law. Thus, the laboratory must have appropriate facilities, preferably away from the main building, in which to store the waste until its removal. Some laboratories have access to incinerators in which some of the chemical and biological wastes can be destroyed. In addition, post-amplified DNA waste should not be stored in the pre-amplified DNA area. Other Facilities Aside from the facilities described above, space must be allocated for staff members' offices, secretarial services and supplies, rest rooms, a "break room", and the building support systems (heating, air conditioning, janitorial closets, etc). Room may also be required for a library, an incinerator, and so on. Corridors must be unobstructed in order to provide easy access in case of emergency. Module 3: Facility Standards and Safety Measures Case Study Page 7 Case Study Handling of flammable liquids – burn death at UCLA and response A research assistant at UCLA died of burn wounds sustained after handling a container with highly reactive and flammable t-butyl lithium. According to the findings of California’s OSHA division the plunger on the syringe of the container became dislodged and resulted in spontaneous self-ignition of the material. The research assistant was not wearing a flame-retardant lab coat which resulted in immediate engulfing of all of her clothing into flames. The university is taking appropriate steps to increase safety measures as well as provide training to personnel to avoid any further incidences. Read the article here: https://www.latimes.com/local/lanow/la-xpm-2009-mar-01-me-uclaburn1-story.html (https://www.latimes.com/local/lanow/la-xpm-2009-mar-01-me-uclaburn1-story.html) Module 4: Validation of Analytical Procedures Page 1 Module Overview Introduction “Validation is confirmation that a method of measurement, through assessment of performance characteristics, is suitable for its intended use”. Source: https://www.nist.gov/mml/csd/organic-chemical-metrology/primary-focus-areas/fundamental-chemicalmetrology/validation (https://www.nist.gov/mml/csd/organic-chemical-metrology/primary-focusareas/fundamental-chemical-metrology/validation) Having decided upon an analytical method that will be used to verify the identity and concentration of a compound or a method to distinguish between different genotypes of a new genetic marker, that method must be developed and in turn also be validated. There are many approaches that may be taken to validate an analytical procedure. The main objective of method validation, however, is to demonstrate that the procedure is adequate for its intended use. This module will include a discussion on some of the parameters utilized for this purpose as well as some general guidelines for each. If you have the recommended text, please refer to chapter 3, sections 3.1, 3.2 and 3.3 for basic definitions and an introduction to internal quality control in an analytical method; additionally, chapter 5 discusses uncertainty of measurement and chapter 8 discusses method validation, which may assist you with the assignment. Objectives At the end of this module students should: Understand the need for validating analytical methods Know the requirements for the validation of an analytical method Understand accuracy in terms of trueness (bias) and precision (repeatability & reproducibility) Understand the limit of detection and lower limit of quantitation Module 4: Validation of Analytical Procedures Page 2 Method Development & Validation Plan Method Development & Validation Plan How do you know that your method is ready for validation? Before we move into the validation of a method, let’s review how we got to this point. Validation may be conducted for many reasons such as when a new method is to be brought online or when the approval of a new standard by a regulatory or governing body requires adjustments be made to a laboratory’s current validated methods. Time and resources are vital to day-to-day operations for any laboratory so time spent in validating a method must be preplanned, well-organized, and effective. Prior to validation, a review of governing requirements, potential training needs of staff and availability of instrumentation/equipment would all play a role in the selection of an appropriate candidate method. In developing the method, instrumental and data analysis parameters should be defined, tested, evaluated, and adjusted, as needed. Additionally, reagents and reference standards should be prepared, to include the appropriate matrix, where needed, and concentration range to ensure all aspects from extraction through detection have also been appropriately assessed based on the laboratory’s needs. Some parameters, such as the robustness and ruggedness of the method, may be evaluated during this development phase. Taking steps to ensure the method works, finetune instrumental parameters, and evaluate factors external to the method prior to validation should help prevent the need for starting the process over or the need for the selection of a new method all together once validation is underway. Overall, the completion of this development step ensures the method has been optimized for use, seems capable of meeting the needs of the lab, and is ready for validation. A validation plan should then be prepared and approved by quality management. This plan might include a discussion on why the method and validation is needed, information from the method’s development to include the corresponding optimized conditions to be used, extraction procedures, and the validation parameters that will need to be included as well as the laboratory’s designated acceptance criteria. A Standard Operating Procedure (SOP) would also be generated to provide the components of the procedure, quality control measures, and written instructions for the use of this method as well as other required elements. This document would also require approval prior to the commencement of the validation. The creation and content of SOPs will be discussed in greater detail later in the course. Components of Analytical Procedures A quantitative analytical procedure can generally be considered to include the following components: extraction of the test compound from a matrix construction of a calibration curve quantitation of the test article inclusion of quality control measures evaluation of method performance The following are key parameters that should be considered when validating such a process: Linearity and Working Range Sensitivity (Limit of Detection and Lower Limit of Quantitation) Specificity Accuracy (Trueness and Precision) Stability Quality control In the case of validation of procedures for DNA laboratories, the validation is split into developmental and internal validation. The FBI's Quality Assurance Standards for Forensic DNA Testing Laboratories states, that developmental validation “is the acquisition of test data and determination of conditions and limitations of a new or novel DNA method for use on forensic samples” and that internal validation “is an accumulation of test data within the laboratory to demonstrate that established methods and procedures perform as expected in the laboratory". (Source: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746) Developmental Validation Developmental validation must include, where applicable: Characterization of the genetic marker Species specificity Sensitivity studies Stability studies Case-type samples Population studies Mixture studies Precision and accuracy studies PCR-based studies Internal Validation Internal validation must be conducted by any laboratory when developmental validation has been performed by another laboratory or at another site of a multi-site laboratory system. The FBI Quality Assurance Standards for Forensic DNA Testing Laboratories and DNA Databasing Laboratories require that internal validation include, where applicable: known and non-probative evidence samples or mock evidence samples, known database-type samples (for database validation studies) precision and accuracy studies sensitivity and stochastic studies mixture studies contamination assessment studies References: The FBI Quality Assurance Standards for Forensic DNA Testing Laboratories: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15746) The FBI Quality Assurance Standards for DNA Databasing Laboratories: https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743 (https://anab.qualtraxcloud.com/ShowDocument.aspx?ID=15743) Additional reading: The FDA’s Bioanalytical Method Validation, Guidance for Industry: https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf (https://www.fda.gov/files/drugs/published/Bioanalytical-Method-Validation-Guidance-for-Industry.pdf) Module 4: Validation of Analytical Procedures Page 3 Linearity and Working Range Linearity and Working Range The acceptable working concentration range of the assay is normally derived from linearity studies and depends on the intended application of the procedure. It is established by confirming that the analytical procedure provides an acceptable degree of linearity, bias, and precision when applied to samples containing amounts of analyte within or at the extremes of the specified range of the analytical procedure. Once the working range of the analytical procedure is determined, the linear relationship between concentration and response should be demonstrated across this range. If there is a linear relationship, test results should be evaluated by appropriate statistical methods, for example, by calculation of a least squares regression line. Linearity is assessed by analyzing a series of calibrators and observing the relationship between the instrument’s response versus their respective concentrations. These studies begin with the preparation of the calibrators using reference materials, such as certified reference material (CRM), which contain the analyte(s) of interest. Several calibrators should be used to construct a standard curve. For example, the ANSI/ASB standard for toxicology calls for a minimum of six non-zero calibrators with five replicates prepared for each concentration. The concentrations for these calibrators should provide an even distribution across the working range of interest. This series of calibrators is then analyzed and a standard curve is constructed. This standard curve provides a graphical representation of the relationship between the calibrators’ concentrations and the associated instrumental responses, which should reveal that they are directly proportional. As an indicator of the strength of the linear relationship, a correlation coefficient (r) and/or a coefficient of determination (r2) is normally calculated. A correlation coefficient of 0.99, for example, might be used as acceptance criteria in evaluating the curve. The closer this value is to one (1), the better the fit of the points to the line or calibration curve. A visual check for any outliers is also recommended such as with the creation of a residuals plot. For more on creating a residuals plot, see: https://www.statisticshowto.com/residual-plot/ (https://www.statisticshowto.com/residual-plot/) Additionally, as a quality control check of the calibration curve, control samples prepared in an appropriate matrix are also run. These should also be prepared from reference material such as CRMs. Typically, these are purchased from a different source or made from a different preparation than that of the calibrators. The end result of this phase of the validation process is to demonstrate that the working range is capable of meeting the demands of the laboratory when quantitating analytes at levels expected to be typically encountered and to select the appropriate calibration model, most commonly a linear model. If these studies reveal a non-linear relationship, other calibration models may be sought to serve as a viable alternative. For more information on calibration models see the ANSI/ASB standard for toxicology: https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf (https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf) Module 4: Validation of Analytical Procedures Sensitivity Page 4 Limit of Detection Limit of Detection (LOD) can be defined as, “An estimate of the lowest concentration of an analyte in a sample that can be reliably differentiated from blank matrix and identified by the analytical method”. (Source: https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf (https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf) ) Determination of the LOD can be accomplished by several means depending on the method. For this discussion, the focus will be on LOD determination utilizing signal-to-noise ratio. This is performed by comparing measured instrumental response/signals from samples with known low concentrations of analyte with those of blank samples and establishing the minimum concentration at which the analyte can be reliably detected. It should be remembered, however, that this approach is only applicable to instrumentation that exhibits enough noise to be applicable. LOD is determined with samples prepared for analyses using the appropriate matrix fortified with the analyte(s) of interest and should undergo all the method’s procedural steps such as extraction. The use of multiple blank matrix samples, obtained from different sources, are important to include. These would be fortified or spiked with decreasing concentrations of the analyte of interest and analyzed multiple times in assessing LOD. The ANSI/ASB standard for toxicology, for example, suggests the use of at least three different sources for blanks, the preparation of duplicates for each chosen concentration and analyzing each a minimum of three times. Once analyzed, the lowest concentration suitable for selection as the LOD would need to demonstrate reproducibility, meeting all criteria required for identification, and be at least three times the noise produced from the instrument’s background/baseline signal for every run made. This calculation may be completed through the use of the instrument’s software. The detection limit and the method used for determining the detection limit should be presented in the method validation report, together with suitable data to justify its determination. Lower Limit of Quantitation (LLOQ) The lower limit of quantitation (LLOQ) can be defined as, “An estimate of the lowest concentration of an analyte in a sample that can be reliably measured with acceptable bias and precision”. (Source: https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf (https://www.aafs.org/sites/default/files/media/documents/036_Std_e1.pdf) ) The upper limit of quantitation is generally established through the linearity and working range studies discussed previously. As with the LOD, the LLOQ can also be determined in more than one manner. The lowest concentration calibrator from the standard curve could be chosen to be the LLOQ or it may be defined in terms of an administratively defined decision point based on what concentrations the lab may choose to report even if the method can identify much smaller quantities. LLOQ can also be determined by reference to signal-to-noise ratio and establishing the minimum concentration at which the analyte can be reliably quantified. These studies would also be conducted by analyzing matrix blanks fortified with the target analyte(s). A typical signal-to-noise ratio is 10:1. LLOQ may also be chosen as the lowest non-zero calibrator concentration found during the working range studies. Studies would still be required by fortifying blanks with the target analyte and analyzing each multiple times. The ANSI/ASB standard for toxicology, for example, suggests the use of at least three different sources for blanks, the preparation of duplicates for the lowest non-zero calibrator concentration and analyzing each a minimum of three times. Once analyzed, this concentration would be deemed the LLOQ if it were found to demonstrate reproducibility and met all criteria required for identification, including bias and precision acceptance criteria. The quantitation limit and the method used for determining the quantitation limit should be presented in the validation report and should show that the requirements set for detection and identification were met. Specificity During validation, it is critical to confirm that the method can discriminate between the analyte of interest and interferents such as compounds with closely related chemical structures during the analyses. The choice of materials to include for these studies should be made after reasonable consideration of the potential interferences that could be present in the sample have been assessed. In seized drug analyses, for example, interference may originate from sources like cutting agents, impurities, by-products, or precursors. Reference materials/standards of closely related compounds and adulterants/diluents should be analyzed to determine whether they affect the target analyte. For toxicology, these interferences might arise from the matrix, isotopically-labeled internal standards or metabolites. In determining selectivity, the evaluation of several matrix-based blank samples from multiple sources would be important during these studies. Matrix samples fortified with the analyte, previously tested case samples and/or samples made directly from reference materials that contain these potential interferents might also be analyzed. Additionally, any interference related to the internal standard would also be important to assess. For example, isotopically-labeled internal standards may contain non-labeled analyte as an impurity, which should be considered as well. Analyzing blank samples fortified with just the internal standard would provide information as to whether there were any interferents, such as the non-labeled analyte, above the limit of detection. During both seized drug and toxicology analyses, the selection of techniques that can separate a sample’s components, like chromatography, can assist with the method being capable of greater selectivity. Overall, any method being validated must be evaluated for selectivity. Efforts must be made to identify and eliminate any potential sources of interference that could directly impact the analysis of the desired target. Module 4: Validation of Analytical Procedures Page 5 Accuracy (Trueness and Precision) Accuracy (Trueness and Precision) The Joint Committee for Guides in Metrology’s (JCGM), International Vocabulary of Metrology (VIM), defines each in the following manner: Accuracy is defined as, “closeness of agreement between a measured value and a true value of a measurand”. The term “true value” has also been substituted with “reference value” (ex. certified reference material) in defining accuracy. Trueness is defined as “closeness of agreement between the average of an infinite number of replicate measured values and a reference value”. Precision is defined as “closeness of agreement between indications or measured values obtained by replicate measurements on the same or similar objects under specified conditions”. (Source: https://www.nist.gov/system/files/documents/pml/div688/grp40/International-Vocabulary-ofMetrology.pdf (https://www.nist.gov/system/files/documents/pml/div688/grp40/International-Vocabulary-ofMetrology.pdf) Accuracy can be affected by both systematic and random error. As stated in ISO 5725, accuracy “should imply the total displacement of a result from a reference value, due to random as well as systematic effects”. (Source: https://www.iso.org/standard/11833.html (https://www.iso.org/standard/11833.html) ) All measurements have some form of error associated with them. The two components of error are systematic and random error. Accuracy incorporates both trueness and precision, which are related to systematic and random error. In calculating these, we are determining the magnitude of the error and in doing things like incorporating large numbers of runs we are attempting to minimize it wherever possible. When both trueness and precision improve so does accuracy. Trueness is a measure of systematic error and is measured in terms of bias. Bias can be caused by several factors such as loss of analyte during the sample’s preparation, stability issues with the analyte, matrix effects and the reference standard’s purity. In determining bias, a large set of replicate reference standard measurements are made, typically prepared in three concentrations, low medium and high, encompassing the range of interest. Bias is generally reported as a percentage of the difference between the mean of the measured concentrations and the accepted reference concentration divided by the reference concentration times 100%. This calculation is made for each of the three. Reference values can be obtained from CRMs, which are supplied with a Certificate of Analysis provided by the vendor. These certificates provide the certified standard’s concentration. See an example Certificate of Analysis for cocaine at: https://www.cerilliant.com/shoponline/COA.aspx?itemno=a8fffeab-5377-4e07-8eda40fe976722d1&lotno=FE09091901 (https://www.cerilliant.com/shoponline/COA.aspx?itemno=a8fffeab-5377- 4e07-8eda-40fe976722d1&lotno=FE09091901) Precision is the measure of random error present in a data set. Repeatability and reproducibility studies are typically conducted when determining precision. Repeatability is in reference to analyzing samples using the same operator in the same location, using the same procedure and instrument within a short timeframe. Reproducibility is assessed by changing those conditions such as testing conducted by a different analyst, using a different instrument and/or a different location. The conditions that are changed and those that remain unchanged should be documented. These studies should be conducted by preparing samples over the concentration range and run multiple times. Precision is generally reported in terms of standard deviation or relative standard deviation so as the population or number of measurements increases, the precision would improve and the standard deviation would decrease as would the random error. In saving time and resources, bias and precision studies may be run concurrently by utilizing the same set of prepared standards. As discussed previously, the validation plan would indicate the preset minimum acceptance criteria for bias and precision. The ANSI/ASB standard for toxicology, for example, states these parameters should be no more than ±20% at each concentration. In calculating these, the magnitude of error is being determined and by making some adjustments, such as increasing the number of samples included in the calculation of precision, we are attempting to minimize it wherever possible. When both trueness and precision improve so does accuracy. The terminology used for validation parameters may sometimes differ depending upon the field/discipline in question. While this module discusses accuracy in terms of trueness (bias) and precision, there are other instances in which accuracy is defined as being the closeness of agreement between the measured value (or even the mean) and reference value. In those instances, the recommended validation parameters may be listed just in terms of accuracy and precision. A laboratory preparing for validation must be cognizant of the particulars of the validation they are charged with utilizing, how the respective parameters are defined as well as the requirements for determining each. Stability During validation, the stability of the targeted analyte(s) must also be evaluated. Any conditions that could be detrimental to the samples using the validated method need to be evaluated through experimental studies. Stability variables that samples could be subjected to such as temperature and pH changes or conditions that might arise during the extraction process or instrumental analysis need to be included. In considering blood samples, for example, several freeze/thaw cycles may occur during the time spent in the laboratory, which can affect the analyte’s stability. Long-term storage may also need to be evaluated as many labs are backlogged and may need to store samples for longer periods prior to and following analyses. Additionally, if a sample undergoes an extraction as a part of the method, the extract containing the analyte of interest may have to await analysis. The time the extract may have to spend on an instrument’s autosampler or time spent waiting for an instrument to be available for testing, for example, should be studied. Consideration may also be made regarding potential issues that could arise with the intended instrumentation, such as when an electrical outage or software problem occurs. In these instances, a longer-term stability study may be considered to assess the sample’s viability. In conducting these studies, quality control samples should be prepared, typically matrix-matched blanks fortified with the target analyte(s) from CRM, and subjected to the stress conditions of concern. Once the samples have undergone the stress conditions, these samples should be analyzed with calibrators and controls made fresh for this purpose. Stock solutions that are prepared by the laboratory should also be evaluated for stability. Expiration dates for anything purchased or prepared should also be reviewed. As an example, in completing a stability study, fortified matrix-matched samples containing the analyte could be prepared at low and high concentrations and analyzed several times to establish the initial instrumental response. These prepared samples could then undergo stability studies to include long-term storage, and cycles of freezing and thawing. The resulting data following each study would then be used to compare to the initial runs in determining stability. Stability factors, such as the type of container, the temperature, the period of time samples might be stored should all factor into these studies. Overall, stability studies should provide the laboratory with a clear understanding of the timeframe and/or conditions in which an extract should remain stable and when reprocessing may be needed. Quality Assurance & Quality Control Quality control (QC) measures taken during validation are vital in ensuring a quality product. Numerous QC measures are implemented throughout the validation process as has been discussed by the use CRMs and the appropriate matrices/blanks, establishing linearity through calibration curves while also running controls as a crosscheck, incorporating an internal standard, including replicate control samples throughout each batch to monitor the analytical run, evaluating acceptance criteria, and implementing appropriate corrective measures where needed. It is important that the standards used come from traceable sources like the NIST CRM. Traceability to national standards, in general, is vital to quality control related measures. Measurement or metrological traceability is defined as, “property of a measurement result whereby the result can be related to a reference through a documented unbroken chain of calibrations, each contributing to the measurement uncertainty”. (Source: https://www.bipm.org/documents/20126/2071204/JCGM_200_2012.pdf/f0e1ad45-d337-bbeb-53a615fe649d0ff1?version=1.15&t=1641292389029&download=true (https://www.bipm.org/documents/20126/2071204/JCGM_200_2012.pdf/f0e1ad45-d337-bbeb-53a6-15fe649d0ff1? version=1.15&t=1641292389029&download=true) ) As discussed previously, Certificates of Analysis that vendors provide with CRM’s contain traceability information. In general, laboratories must evaluate vendors they choose and ensure the uncertainty associated with all purchased reference materials and equipment is considered prior to purchase and/or use. These vendors should be appropriately accredited, for example to ISO/IEC 17025 or 17034 standards. In addition, many analytical techniques now incorporate a means of testing system suitability. System suitability is based on the concept that the equipment, electronics, analytical operations, and samples to be analyzed constitute an integral system that can be evaluated as such. System suitability test parameters to be established for a particular procedure depend on the type of procedure being validated. Upon completion of the validation all related information and data should be submitted for comprehensive review by the laboratory’s quality management staff. Once this review has been completed and the validated method and corresponding Standard Operating Procedure has been approved, any laboratory personnel who will utilize this procedure will need to successfully show competency in its use prior to routine analyses. Successful completion of this step should also be documented. Web Resources For more information on validation of analytical procedures see the following websites: FDA, “Q2(R1) Validation of Analytical Procedures: Text and Methodology Guidance for Industry”: https://www.fda.gov/media/152208/download (https://www.fda.gov/media/152208/download) FDA, “Bioanalytical Method Validation, Guidance for Industry”: https://www.fda.gov/media/70858/download (https://www.fda.gov/media/70858/download) Scientific Working Group on DNA Analysis Methods, “Validation Guidelines for DNA Analysis Methods”: https://www.swgdam.org/_files/ugd/4344b0_813b241e8944497e99b9c45b163b76bd.pdf (https://www.swgdam.org/_files/ugd/4344b0_813b241e8944497e99b9c45b163b76bd.pdf) ANSI/