Good Laboratory Practices for Bioanalytical Laboratories
Conceptually understanding bioanalysis can be difficult, but good introductory resources enable a basic understanding. In simple words bioanalysis, is a testing tool used in drug discovery and development to determine the concentration of drugs and metabolites in biological fluids. Scientists can use a set of rules, regulations, and guidelines to efficiently and effectively develop various methods using instrumentation and process biological samples.
What is GLP?
The FDA requires that each laboratory follow a standard set of laboratory practices widely referred to as good laboratory practices, GLP for this article.
GLP guidelines set out the requirements for the appropriate management of non-clinical safety studies. GLP Principles help define and standardize research institutions’ planning, performance, recording, reporting, monitoring, and archiving processes.
The goal of GLP is to ensure:
Standard operating procedures (SOPs) are in place to cover all activities in the laboratory.
Independent Quality assurance unit is in place.
The staff is well trained, follows protocol and standard procedures.
The equipment used in GLP testing is calibrated and maintained.
Records generated during testing are kept secure.
Results of each test or process are backed up and checked by a quality assurance unit or audited according to current guidelines.
Bioanalytical results provide great insight on the usefulness of the drug under development and generates pharmacokinetic data for FDA to evaluate. FDA look closely and frequently audits laboraotires before drug approval.
Bioanalytical laboratories must follow predetermined rules and regulations to ensure the validity, integrity, and reliability of non-clinical and clinical data submitted for FDA evaluation and approval.
NorthEast BioLab (NEBA) is a GLP compliant laboratory that offers a wide range of bioanalysis services and works to provide the most accurate and complete test results possible. NEBA conducts GLP and Non-GLP method development, validation, and bioanalysis of large and small molecules using various state of the art technologies. NEBA has a team of highly experienced and dedicated professionals who can provide accurate and reliable results that fully comply with current regulatory guidelines and standards.
Pre-clinical development consists of several different in-vitro and in-vivo processes, tests, and study types used to help start the drug development process. When drugs are in the pre-clinical development stage, it is important to document each step of the process thoroughly and follow proper rules and regulations. The primary early-stage development work provides reliable results for the success of the drug through Phase I-III resulting in a marketable drug.
A Pharmacokinetic (PK) study relates to drug absorption and its after-effects. Post drug administration, the body goes through a four-stage process to utilize the medication. The first phase is the initial absorption of the drug into the tissues of the body. The next phase is metabolism, where the body breaks down the drug to be used. After metabolization, the body then distributes the drug through the bloodstream. The last phase entails the excretion of the drug or its unused portion. During the PK study, blood samples are drawn and carefully analyzed to help understand how the body uses and gets affected by the drug.
Toxicology tests are performed at scheduled intervals to help measure the concentration of the drug in the sample, determine how the body uses the drug, how long it takes to distribute in the body, and measure the peak effectiveness of the drug. During this time, the lab will also note any side effects and collect data on the efficacy to determine therapeutic and non-therapeutic doses, including thresholds for safety.
The above studies are performed in pre-clinical and clinical settings to determine how a drug works. Generally, these studies involve a test article studied under GLP conditions to assess the efficacy and safety of any given drug. During these studies, the sponsor provides an approved protocol to bioanalytical laboratories.
The primary agency for the regulation of laboratory practices and for maintaining the guidelines for GLP in the United States is the Food and Drug Administration (FDA). This agency carries the force of the law and audits those laboratories performing such tests. According to the FDA regulations, the practice and guidelines for GLP consist of several different subsets and categories. The GLP definition FDA standards put forth can best be understood by looking at each component of GLP that the FDA expects.
The FDA GLP requires a well-defined structure of the research organization and the responsibilities of each research personnel. The facility management will help with the setup and overall conduct of the study. Some roles of the management are–
To designate Study Director (SD) before the study begins and replace them promptly, if necessary.
Assure there is a Principal Investigator (PI) and an independent Quality Assurance (QA) unit in place
Ensure test and control articles have been appropriately tested for identity, strength, purity, stability, and uniformity as needed
Has authority to provide resources and implement changes to comply with GLP requirements and, if necessary, to take corrective actions for addressing audit findings
Approves all test facility SOPs
The FDA GLP requires a Study Director (SD) to be appointed, who is the pivotal point of control for the whole study. This director supervises the entire study from start to finish and is a single point of control. The designated SD –
Adheres to study design, conduct, and reporting guidelines, including compliance with protocol and any relevant GLP
Ensures that all circumstances affecting the study’s integrity are documented and assessed for impact and that corrective actions are taken as necessary
Resides, generally, at the testing facility, where the test article is administered, to evaluate the system as effectively as possible
Must be a scientist or other professional with the proper education, training, and experience or combination of these elements
Must be one person and cannot have a Co-Study Director but can have an acting director or deputy when the primary SD is unavailable.
Cannot be the CEO of the company
The SD will assert a GLP Compliance Statement, which is an integral part of the study report. For this, the SD needs to be aware of all the events that may impact the quality and integrity of the study, evaluate their influence and deploy corrective actions as required. Even if certain study phases are delegated to other test sites (as in the case of multisite studies), the study director maintains overall responsibility for the entire research study and the global interpretation of the study data.
The FDA GLP requires a Principal Investigator (PI) to be appointed as well. The PI, or contributing scientist, is designated to oversee the testing performed at each facility and has different responsibilities and authority than the SD. The PI is also selected for non-clinical studies that are conducted across multiple test sites.
The FDA also requires that an Independent Quality Assurance Unit (QAU) be appointed. The QAU is responsible for various duties to help ensure that the study goes smoothly, is documented thoroughly, and follows all GLP guidelines. QUA unit is responsible to multiple audits during course of study and provide full audit reports to SD in timely manner.
GLP compliant laboratories must have adequate facilities and equipment to perform GLP studies. All laboratory equipment used in the generation, measurement, or assessment of data must be of appropriate design and adequate capacity to function as per protocol and maintained according to applicable SOPs. SOPs must be in place for calibration and maintenance of all critical pieces used, impacting the results. Laboratory staff must keep records for all repairs, maintenance, and calibration.
SOPs shall be maintained and followed for the following at a minimum.
Monitoring and Maintenance of Freezers, Refrigerators, and backup generators
Calibration, use, and maintenance of laboratory equipment such as pH meters, balance, pipettes, centrifuges
Use of instruments such as HPLC equipment, Mass Spectrophotometer, Plate reader and others
Environmental Monitoring System
Reference materials and biological sample handling
The GLP laboratories must maintain a complete chain of custody for all test articles (reference materials and biological samples) received in the facility. This is achieved by keeping the entire record from the time of receipt to the disposal of all test articles. All refrigerators and freezers shall have a full history of temperatures and any excursions around the clock and have backup power in case of power failure.
Quality Assurance (QA) – sometimes also known as the Quality Assurance Unit (QAU) – as defined by GLP, is a department or team of persons who, along with management, assure that GLP compliance has been attained in the test facility as a whole and for each study. QA must be independent of the operational aspects of the studies and function as a “witness” to all studies related activities.
Primary responsibilities of the Quality Assurance Unit are-
QAU Is responsible for monitoring each study and ensuring that the facilities, equipment, personnel, practices, records, and controls meet regulatory standards
Maintains and continuously improves the quality system
Is not a scientific evaluator
Must be distinct and independent of the personnel engaged in the conduct of the study
QAU functions can only be performed by an individual uninvolved in the study
QAU cannot report to the Study Director
Maintains a Master Schedule Sheet listing all studies conducted on FDA-regulated products to support a research application or marketing permit. Tracked information includes:
Nature of the study
Date of study initiation
Current status of the study
Sponsor – applies to the FDA for a research or marketing permit
Maintains copies of all approved protocols
Reviews all final reports and amendments
Inspects each study at adequate intervals
Verifies the study is being conducted as per:
Maintains signed records from each inspection with
Date of inspection
Person performing the inspection
Maintains records of the SOPs for the study, including:
Availability, where needed
Master SOP binder
GLP Compliance review
Compliance with working procedures
Reports SOP deviations to the SD and Management
Helps with inspections of the study, the process, and the facility in which the study is performed
Responsible to maintain documents related to Job descriptions, training programs, training plans, maintaining training records,
Increase awareness and commitment to quality and provide GLP related training
All research studies generate raw data, also referred to as source data, which is nothing but the original data recorded during the conduct of a study. Raw data also documents the procedures and circumstances under which the study was conducted. They are, therefore, essential to reconstruct the research study and contribute to the traceability of the events during the study.
The FDA GLP also has guidelines regarding the documentation process of all studies that are conducted. FDA requires that all findings are well documented, and they can be validated.
Some documentation that the FDA requires is as follows:
(Planned/Unplanned) Deviations, or requests to accept nonconformance to a requirement, need to be presented before the work begins and are often risk-based, and Waivers, these are usually filed after work has been completed and noncompliance was found; this can also be root cause investigation papers and preventative and corrective action paperwork as well
Engineering Change and Change Control must be documented in cases where permanent change involves revision.
Laboratory Notebooks are to be kept with Good Documentation Practices (GDP) in mind – researchers should record recreations of the experiment, patent protection, and consider that documents can be audited at any time and therefore should be thorough and complete.
FDA Guidelines regarding Laboratory Information Management Systems (LIMS)
In keeping with GLP, FDA has set the following guidelines regarding Laboratory Information Management Systems (LIMS):
Only authorized individuals should make data entries, password controls should be present, system access should be limited, and electronic signatures should be required
Data entries cannot be deleted but should be modified such that the initial data can still be accessed and seen, leaving an audit trail that can be followed – the database should be as tamper-proof as possible.
SOPs should describe the procedures that are being used for collecting data and ensuring its validity.
The primary goal of LIMS is to ensure that data that is collected is accurate, that no one has been able to tamper with it, and that only authorized individuals can access the data.
It is also vital that the data can only be modified, when necessary, by authorized individuals and that electronic signatures are used to make sure that if changes are made, it can be traced back to the person who recorded it.
Information technology is another essential aspect of documentation and electronic data. Backing up data in a cloud-based storage solution is recommended with proper controls to allow access to only authorized personnel. It is crucial that the IT server room is protected and has restricted access with a complete record of activities. Proper fire suppression systems should also be in place, as well as temperature controls and monitoring.
We live in a day and age where computers are far more prevalent than other recording methods. As such, the FDA also has some stipulations regarding computerized systems that are being used in experiments.
The FDA has strict guidelines for electronic data. Title 21 Code for Federal Regulations (CFR) Part 11 outlines criteria that must be met for electronic records to be considered reliable and equivalent to paper records. Essentially, files must be considered ‘valid’ and contain original recordings, and a user must be recognized and held accountable for the data. Under this code, electronic signatures are the equivalent of handwritten signatures and indicate that specific actions have been approved, reviewed, and verified.
Researchers need to keep accurate and up-to-date records in their computerized systems with administrative and procedural controls to protect the information within the systems.
Data integrity is vital as this is the data that the FDA would review when approving or disapproving a drug. The FDA states that records should be complete, consistent, and accurate. Documents need to be legible, contemporaneously recorded, original or true copy, and accurate. This information also needs to have electronic signatures equivalent to handwritten signatures so that the content can be attributed to one person or other.
A digital signature is equivalent to a handwritten signature which, when present on a document, guarantees that the owner of the signature acknowledges and knowingly signs the document. He is then responsible for all electronic documents containing this electronic signature.
Employees must be accountable for the actions that are taken using their electronic signatures.
The Standard for Exchange of Nonclinical Data or SEND provides the organization, structure, and format of standard non-clinical (animal toxicology studies) tabulations datasets for regulatory submissions and dates in the ISO 8601 format. The Study Tabulation Model or SDTM provides a structure for data for clinical and clinical toxicology studies and a standard structure for human clinical trials tabulation databases. An Analysis Data Model or ADaM should be used for clinical data sets. The Annotated Case Report Form or CRF should be submitted in a PDF document to capture subject data.
The Clinical Data Interchange Standards Consortium or CDISC helps to develop global standards for clinical and non-clinical research. The FDA cannot process a single data set over 5GB and only processes the American Code for Standard Information Interchange text codes.
Study Data Standardization and Terminology is also essential for GLP:
Controlled terminology standards specify critical concepts such as definitions, preferred terms, synonyms, and codes:
Defined in SDRG (Study Data Reviewer’s Guide)
Defined in CDISC at the National Cancer Institute (NCI)
MedDRA is used for coding adverse events
Unique Ingredient Identifier (UNII) is found in FDA’s Substance Registration System.
There are specific GLP guidelines for archiving or storage of records. Sometimes it is required to reconstruct a study many years after it has ended. Thus, storage of records must enable their safekeeping for long periods without loss or deterioration and, preferably, in a way that allows quick retrieval. For safe storage of precious data, it is usual to restrict access to archive facilities to a limited number of staff and record the documents logged in and out. Although there is restricted access to specific staff, records are also kept of the people entering and leaving the archives.
There should also be controlled procedures for adding and removing items from the archive and a standard for a retention time of at least two years following the date of the application for research marketing permit and five years following the submitted results to the FDA.
The study director oversees numerous archival duties and ensures that the data is transferred appropriately during archival efforts.
Study Records are as follows:
Sample Preparation and Dilution
Mobile Phase Preparation
Conditions of Chromatographic System
All raw data generated during conduct of study and final results including final study reports
Dated signature of Analyst who performed tests
Non-study items can be filed separately from the study, including pest control, instrument qualification, and calibration efforts.
Other records to be archived regarding the study are as follows:
QAU Records and Inspection Records
Data that Supports Method Validation
The archival room must have adequate fire suppression safeguard and an environmental monitoring system to take proper precautions to protect data integrity and prevent deterioration.
Those laboratories that go out of business must transfer the GLP records to the sponsor of the study. Proper disposal methods should also be in place for documents that are no longer going to be archived. A disaster recovery plan also needs to be in place that relates explicitly to equipment and engineering control, facility and cabinet designs, document restoration, and client contact for notification in emergencies that can compromise the archived data.
There are good laboratory practices for sample analysis as well. The first is the chain of custody of samples that are used for biological testing. Such samples must be accounted for from the time they are collected to the time they are stored and after being used. This includes every movement that the samples make while they are being used. Tissue slides can be labeled with accession numbers only if these numbers get translated to all required sample label information.
As the samples move through different stages in the analysis life cycle, it becomes essential to conduct a stability study on the freeze and thaw cycles of samples to ensure that they do not deteriorate and are still viable after freezing and subsequent thawing.
It is crucial that samples are stored safely to avoid any contamination and deterioration and that the samples are persevered for the study’s duration and beyond. Regarding controlled substances, these must be stored per DEA (Drug Enforcement Administration) standards.
When archiving and storing samples after they have been used for the study, samples must be cataloged and stored per regulations and maintain integrity for reevaluation if necessary. Once the samples have deteriorated past the point of being useful, they should be disposed of accordingly. When storing reagents and solutions in the lab, these items must be labeled to identify the composition, concentration, storage requirements, and any pertinent expiration dates.
Reference Standards is a critical piece of the process and must have valid Certificate of analysis (COA). Reference standards must be kept at proper temperature as per COA . When it comes to controlled substances, these items should be stored in a secure location in keeping with the DEA regulations, and compounds should only be accessible to the appropriate staff.
High potency compounds are also something to keep in mind. Often these compounds can be dangerous. Proper mixing instructions, potency information, and any safety information (MSDS) should be clearly labeled to protect those in the lab and anyone else who can access these substances.
Another element of GLP is the presence of up-to-date standard operating procedures (SOPs). It is challenging to include all the technical details of study conduct in the protocol, and so the details of all routine procedures are described in SOPs, which are part of the documentation system of the institution.
SOPs reduce bias in studies by standardizing frequently performed techniques. Thus, SOPs are basically a list of rules and procedures put in place to ensure data quality and integrity at the facility. These rules are intended to minimize the introduction of systemic error as all staff uses the same testing practices.
It is necessary to have SOPs in place to cover important tasks in a laboratory. .
Following are examples of content that one may find in an SOP which can facilitate understanding better what they are and why they are critical for any GLP lab or study:
SOPs writing and approval
Documentation practices and procedures and control
Managing reference compounds and reagents
Equipment calibration and maintenance
Sample storage, shipment and disposal
Managing and monitoring sample storage units
Quality management system
QAU role and responsibilities
You may also find supplemental information to the SOPs to use that is in adherence to current GLPs and current documents that are approved for use.
GLP requires that approved written protocols indicate the objective and all the methods for the conduct of the study. The protocol must also list the type and frequency of tests, analyses, and measurements to be made in the study. A proper protocol needs to include a statement about the regulatory standards under which the study will be conducted. The approved written protocol supersedes the SOPs. All changes and revisions or amendments must be approved by the study director and signed. If a systematic change occurs, it must be entered as an amendment; if it is a one-time deviation, it should be listed as such.
The study report or final report, must describe all relevant details of the study accurately. The Study Director is responsible for the scientific interpretation included in the study report and for declaring to what extent the study is conducted in compliance with the GLP Principles.
GLP guidelines require that final reports or study reports follow a specific format and include all pertinent information of the study. These must include:
Name and address of facility performing study
Date when the study was initiated (date Management signed/approved the protocol) and completed (the date the Study Director signed the final report)
All circumstances potentially affecting data quality; deviations
There are also specific GLP requirements for equipment, mainly whether the equipment has been approved before installation and the installer is qualified to do the installation. This process must be documented and verified. It should be confirmed that the individual operating the machine is doing so under the predefined specifications, and these operations should be periodically checked during actual use.
All types of equipment must be routinely calibrated and maintained in working order. A program of validation, calibration, and maintenance can help achieve this. Documenting use and maintenance is essential to know the precise status of the equipment and its history
GLP guidelines for equipment used during a GLP study are as follows:
Design Qualification (DQ, Pre-installation)
Installation Qualification (IQ)
Verify procedure and documentation of installation
Operation Qualification (OQ)
Verify operating according to predefined specifications
Performance Qualification (PQ)
Verify consistently performing as intended
Performance during “Actual” use
System Suitability Testing
Calibration and Maintenance
GLP testing laboratories require that the equipment status should be documented in a notebook or logbook. Equipment such as refrigerators and freezers should be routinely monitored to ensure the performance and integrity of the items stored inside. Password or controlled equipment use is deemed necessary for an audit trail during inspections.
The facility chosen to conduct testing must live up to the GLP guidelines. Facilities must fit the design, capacity, and maintenance requirements for the study being done.
The facility should be spacious in size to avoid overcrowding, confusion between projects, or cross-contamination. Utilities (water, electricity, etc.) must be adequate, sufficient, and stable.
The floor plan and maps of the building must fit the study being conducted, and proper security measures, monitoring, and access controls are also necessary. The site needs to be monitored regularly to ensure that only authorized personnel have access to the facility and its contents. The building needs to be secured with proper monitoring so that staff and security can always see the facility to prevent tampering.
The facility must be free of pests that could contaminate the study, have adequate space for the process and flow of the study and have proper environmental controls to help prevent contamination of the study.
The method for testing is as important as the lab, the procedures, and the protocol. Study Directors must possess the ability of the researchers to demonstrate that the method is reliable and reproducible and has been validated as per FDA guidance on Bioanalytical method validation.
There are several levels of method validation under GLP. The first one proves the method is fit-for-purpose intended. Then, there is full validation, partial validation, and cross-validation.
Inspections are an essential part of meeting any GLP guidelines. The Quality Assurance Unit typically conducts most internal audits to make sure that standards are being met. They will perform critical phase audits, in-process study audits, data audits, and report audits and will make a detailed report of findings from any audits conducted.
Internal audits are the most common and will occur at various stages through the process of the study. These audits occur on a schedule and can include inspections of facilities, equipment, training, adherence to documents, GLP guidelines, sample management, and more. These audits are performed to keep the study on track and make sure that all regulations are being met so that the findings of the study will be considered valid. The results are then sent to the site management to handle any issues that need to be corrected.
As far as the FDA goes, four different types of regulatory audits can take place. These audits are in place to regulate studies of all kinds across the board. The four types of FDA Inspections for GLP are as follows:
GLP inspection: Announced. Scheduled once every two years. Periodic, routine assessment of a laboratory compliance
Data Audit: This may not be announced in advance and is scheduled as needed.
Conducted to verify the information submitted to the FDA is accurate in the final report.
Directed inspection: May not be announced in advance. Scheduled as needed. Any series of inspections for a specified reason, such as questionable data in the final report or tip from an informer.
Follow-up inspection: May not be announced in advance and scheduled as needed. This kind of inspection is made after a previous GLP inspection has revealed objectionable practices and conditions. These inspections assure proper corrective actions have been taken.
The FDA will also inspect international studies based on having submitted the results of significant studies on essential products to the FDA.
Before inspections, materials such as the GLP regulations, management briefings on post-conference report, miscellaneous memoranda and policy issuance, GLP compliance program, the protocol of an ongoing study, the final report of a completed study, and report from the most recent inspection are usually reviewed. During the investigation, the agency may take photos. FDA may issue a range of responses after an inspection that can be anything from a warning letter to a notice of adverse findings.
FDA Inspection Reports
There are also several reports issued that detail the findings of the inspection. These forms are the EIR, Establishment Inspection Report, and the Form FDA-483, the FDA inspection observations. Sometimes, it may be necessary to re-inspect a site, hold a conference on findings, or disqualify a facility. In cases where the facility is invalidated, all information and data that is collected there becomes unacceptable and cannot be submitted.
Some agency responses that the FDA may administer are:
Form FDA-483 (FDA inspectional observations): written notice of objectionable practices or deviations from the regulations compiled by the FDA investigator at inspection end. These items serve as the basis for the exit discussion with laboratory management. Management acknowledges the issues and offers possible corrective actions to be taken. They may also respond to the district office in writing after reviewing the FDA-483.
Major Deviations: Response requested within ten days, describing corrective actions that the laboratory has taken or plans to undertake and the timeline for completion (this should be ASAP). The response should include items listed on the 483 and considered in the exit discussion with laboratory management.
Notice of Adverse Findings letter: for less significant deviations. Response requested within 30 days.
EIR: “Establishment Inspection Report.” Internal agency document, which reflects the observations of the inspector. Summarizes the remarks made at the laboratory and contains exhibits concerning the audited studies (Protocols, SOPs, CVs). The EIR is reviewed by District and Headquarters personnel, potentially resulting in additional GLP violations by the laboratory management.
The United States is not the only country that is held to good laboratory practices. Several agencies help to regulate laboratory practices in laboratories and studies across the globe.
The International Council for Harmonization (ICH)
The ICH is observed by the EU, Japan, the US, and some parts of Canada and Switzerland. Its goal is to facilitate the mutual acceptance of data. ICH Q9 Guidance is in place to assess the risk to the quality of a drug across the span of its lifetime. ICH Q8 is designed to make sure the design product and manufacturing process are consistent and deliver the product’s intended performance.
World Health Organization (WHO)
The World Health Organization puts forth the need for quality standards in drug research, development, and testing in developing countries.
Organization for Economic Co-operation and Development (OECD)
The OECD is an Intergovernmental organization of 30 industrialized countries in North America, the Pacific, Europe that work together to manage chemicals concerning human health and the environment. OECD GLPs were developed to promote data quality and validity and establish a single monitoring authority for all member countries.
Good Laboratory Practice is defined in the OECD Principles as “a quality system concerned with the organizational process and the conditions under which non-clinical health and environmental safety studies are planned, performed, monitored, recorded, archived and reported.”
European Medicines Agency (EMEA)
EMEA is a European Commission that is designed to create and enforce the GLP practices in Europe.
Japan has its agencies, including the Ministry MHLW, Pharmaceutical and Medical Devices Agency, and Japan Pharmaceuticals Manufacturer Association (JPMA), that enforce the GLP in Japan.
ISO, or the International Organization for Standardization Standards, has two main clauses, management requirements, and technical requirements. Management pertains to the operation and effectiveness of the Quality Management System (QMS) within the laboratory. Technical specifications address the competence of the staff, sampling and testing methodology, equipment, and the quality and reporting of test and calibration results.
GxP is the general abbreviation for quality guidelines and regulations, with the “x” standing in various fields (Clinical, Manufacturing, Laboratory, etc.).
Maintaining good laboratory practices is not advised but rather something essential to maintaining a laboratory that will meet all the standards and regulations required by the FDA and other regulatory authorities.
Principles of good laboratory practices allow GLP laboratories to improvise their present method of conducting studies and fully understand the GLP guidelines that are meant to be followed.
It can make a world of difference in how your study gets approved, whether it passes regulatory inspections and if their findings are valid.
Good laboratory practices are essential across the board and globe, with more and more companies and laboratories striving to meet these standards to make sure their findings will be accepted and approved.