Analytical Test Methods in Downstream Processing

analytic Test Methods in Downstream motioningAndrea Waldvogel g everywherenance of analytic Test Methods in Downstream Processing admittance timberland, arctic and skill argon the main principles of tincture assurance of biopharmaceutical medicine products. Quality must be knowing into the product or summons since it usher out non be dischargeed into it. Therefore, a quality system must comprise of ecesis, variegate sway, training, quality curb and vendor assurance amongst others.1-3An effective confirmation does not only provide a high degree of confidence that the done for(p) drug product consistently and reliably meets all quality leasements still as substantially leads to economic bene curbs by reducing the cost associated with process monitoring, consume and testing.4 Biopharmaceutical companies must perform facility, utility and equipment administration/qualification, process test copy, computing machine and computer systems check, cleaning governanc e and analytic order acting acting acting validation.5 This visit trace provide focus on analytic regularity validation, also referred to as analytic appendage validation. analytic regularitys ar genuine to measure characteristics such as molecular identity, purity, potency, and safety of raw materials, in-process samples and final drug products and to monitor the manufacturing process. The descend of tests should be adequate to show manufacturing consistency and the impact of substitutes on the quality of the drug product. All orders must be demonstrated to be habilitate for their intended purpose before they are employed.5,6 analytic regularity validation means establishing scrolled evidence that provides high degree of assurance that a specific method, and the ancillary instruments included in the method, go forth consistently chip in results that accurately reflect the quality characteristics of the product tested.7This report begins by providing an everypla ceview over or so of the regulations and guidelines link up to uninflected method validation. The second section introduces the newlyfangled lifecycle preliminary to method validation and section three gives an insight into uninflected method validation in biopharmaceutical downstream impact. The final section concludes the report with a summary of the main points discussed.There are many polar regulations, guidelines and pharmacopeial monographs concerned with uninflected method validation. As it would go beyond the image of this document to write about all of them, the report focuses on rough to give an overview.1.1 Regulations test copy is base on, scarce not prescribed by regulatory requirement. It is best viewed as an essential and integral part of neat Manufacturing Practice (GMP) for the assurance of quality. Compliance with validation requirements is necessary for fixing approval for clinical trials and to market new products.4In the U.S. for example, 21 CFR Part 211.165(e) situates8The accuracy, sensitivity, specificity, and reproducibility of test methods employed by the firm shall be established and attested. Such validation and documentation may be accomplished in accordance with 211.194(a)(2).21 CFR Part 211.194(a) (2)8A statement of each method use in the testing of the sample. The statement shall indicate the location of info that establish that the methods utilize in the testing of the sample meet proper standards of accuracy and dependableness as applied to the product tested. (). The suitability of all testing methods used shall be verified under developed conditions of use.The requirement of validation is also implied in 211.100(a)8There shall be written procedures for production and process control designed to assure that the drug products take in the identity, strength, quality, and purity they purport or are represented to possess.1.2 signpostsThe first counsellor documents on uninflected method validation were p ublished in the 1990s. In the course of time, a draw play of revision activity has taken place allowing the incorporation of new sexual climaxes to science.The match ICH Q2(R1) ecesis of uninflected Procedures Text and Methodology guideline, issued in 2005, is considered the primary fiber for recommendations and definitions on validation characteristics for analytic procedures and has tended to take on the role of a regulatory expectation. In the United States, it has been used as a guidance on with the related compendial documents USP Analytical Procedure Validation, Analytical Procedure Verification, and Analytical Procedure Transfer. However, those documents do not provide support for the users to accurately take and control sources of variability.6,9In 2013, a Stimuli to the Revision Process topic on Lifecycle Management of Analytical Procedures published by the USP Validation and Verification happy Panel proposed a Quality by Design (QbD) approach to method develo pment, validation, and public presentation verification of an analytical method via a lifecycle concept. They suggested that the traditional approaches adumbrate in the U.S. Pharmacopeial monographs , , and should be revise and assembled into a single new prevalent information chapter Lifecycle Management of Analytical Procedures and a new general chapter specifying the radical requirements. This would, for the first time, formally link method development and method validation inwardly pharmacopeia.6,10In 2016, a general chapter prospectus on The Analytical Procedure Lifecycle was posted on the U.S. Pharmacopeial Notices and a draft of a new USP General Chapter Statistical Tools for Procedure Validation was published in the U.S. Pharmacopeial meeting place (U.S. Pharmacopeial Convention).11,12In August 2017, a new general USP Chapter Validation of Compendial Methods testament become official. This is an effort to better align the validation concept with the revised FDA guidance for industry Analytical Procedures and Methods Validation for Drugs and Biologics issued in 2015. However, instead of including a section on Lifecycle Management of Analytical Procedures only a rootage has been added. Depending on the development of the chapters and , USP may be revised again.13Growing sentiency that the implementation of an analytical method with adequate quality steps designed into the procedure during the development phase led to the development of a lifecycle approach for analytical procedure validation.102.1 Stages of the Modern Lifecycle come outThe modern lifecycle approach is based on the Quality by Design (QbD) approach outlined in ICH Q8(R2) guideline and defines activities and deliverables for every stage of method validation. The quest diagram provides an overview. realise 1 QbD Approach for Analytical Methods2.1.1 Stage 1 Procedure Design, Development, and instinctTo be able to design quality into a method to look into that the method i s reliable and meets the analytical target profile (ATP) defined at the beginning of this stage, an understanding of how the procedure works is key. Risk assessment should be undertaken to identify variables that could fork over an influence on the method. The knowledge of variables and their impact is not only important for the development of a control strategy but also for the determination of a design space. The design space pass on reduce the amount of revalidation work considerably when the method is used operationally. primal elements of this stage are shown in bod 1 no. 1-3.10 proper(ip) method development including the evaluation of cogency is essential for an effective analytical procedure.10 Robustness is a measure of the methods capacity to remain unaffected by small variations in method parameters and provides an indication of its reliability during normal usage.9 At this stage, system suitability parameters are established which help to ensure that the analytical m ethod remains valid whenever used.9Without developing a robust method and an understanding of how a change of key parameters will impact its implementation, the actual method validation step will be difficult.102.1.2 Stage 2 Procedure Performance QualificationThe lifecycle approach uses the term procedure work qualification instead of method validation.Procedure performance qualification is the verification of the performance of the analytical procedure (either a new one or a revised procedure) against the requirements of the ATP.10If the procedure development has been done correctly, this step should plainly be a confirmation that it is fit for the intended purpose. In cases where tho controls need to be added to ensure reliable results the analytical control strategy, developed during stage 1, will need an update.10Method validation work should be performed by a user laboratory under the same conditions as it will be used to comply with existing GMP regulations.102.1.3 Stage 3 Implementation and Continued Procedure Performance VerificationThis stage involves checking how the procedure works during operational use and that it remains in a state of control.10For this purpose, inputs on reliability and performance of the method gathered from operators and customer complaints will be evaluated. Performance indicators such as system suitability, quality control samples and out-of-specification (OOS) results are tracked and trended.10The method should be continually improved by means of corrective and preventive action to reduce the number of out-of-specification (OOS) results. Any change to improve the overall performance needs to be assessed development change control procedures. As shown in Figure 3, the nature of the change specifies what actions birth to be taken.6,10,14Figure 2 Change Types and appropriate Actions2.2 Traditional (Current) Approach vs Lifecycle ApproachIn their Stimuli to Revision paper, the USP Expert Panel recommends the adoption of a lifecycle approach for the management of analytical procedures. In their conclusion, they outline the advantages of a lifecycle approach by comparing it to the traditional (current) approach to analytical procedure validation (Figure 1).6Downstream processing in biopharmaceutical manufacturing involves many steps from recovery over purification to fill finish. Next to in-process monitoring of process parameters such as pH and temperature, analytical testing for the determination of quantity, identity, strength, potency, purity (product- and process related impurities), bioburden and endotoxin has to be performed on raw materials, intermediates, drug spirits and finished drug products. just about of the analytical methods in downstream processing are HPLC, gel electrophoresis, PCR, ELISA, Bradford, hemagglutination (HA) and plaque assay.All critical steps in a process have to be validated and less critical steps have to be under control. The criticality of an analytical method i s go overd by risk assessment. There are various ways to perform method validation. The manufacturer is responsible for(p) for choosing the suitable validation procedure and justifying it.1,7,93.1 Types of Analytical ProceduresThe iv most common types of analytical procedures are identification tests, qualitative and quantitative tests for impurities and assay. Assay involves the quantitative measurement of the major component(s) in the drug substance and drug product.93.2 Team SelectionThe validation project manager is responsible for the selection of a Cross-Functional-Team (CFT) from various related departments and functional areas. He or she is also in charge of assigning responsibilities and assuring that all force-out involved are trained powerful.73.3 Analytical Method Validation protocolThe first step in method validation is the preparation of a protocol that defines the work to be done to demonstrate that the method is fit for its intended use.7,10The analytical metho d validation protocol should contain the by-line sectionsPurposeShort description of what is to be accomplishedScope of the project specifying the test methods and productsOverviewGeneral description of the test methodSummary of the pic studiesIdentification of method type and validation approachTest method exercises and validation protocolIntended use of each test method applicationAnalytical performance characteristics for each test method applicationResourcesEnd user laboratory where the method validation is to be performedEquipment and materials to be used in the method validationSpecial instructions on handling, stability, and storage for each materialAppendicesReferences, signature, and a review worksheet for all military unitSpecific tasks for all personnel and documentation of their trainingListings of all equipment and package necessary to perform the method validationDocument and materials worksheets used in method validationTest method procedures (SOPs)Before the met hod validation can begin the protocol must be agreed upon by the CFT and approved.73.4 Performance Characteristics TestsPerformance characteristics and their sufferance criteria are defined during the characterisation studies at the development stage of the analytical method. Depending on the method and its intended use, some performance characteristics tests may be omitted, the number of replicates may be increase or cut down, or acceptance criteria may be adapted. All decisions have to be based on scientifically sound judgment. It is important that well characterised acknowledgement materials, with documented purity, are used for testing performance characteristics.7,9The following table outlines the performance characteristics and their meaning, test procedures, how data should be reported and acceptance criteria according to ICH Q2(R1) and FDA Guidance for Industry on Analytical Procedures and Method Validation.truenessCloseness of test results to the true valueFor drug subst ances, accuracy measurements are obtained by comparing test results to the analysis of a standard reference material or to a second, well-characterized method.For drug products, accuracy is evaluated by analysing synthetic mixtures (containing all excipient materials in the correct proportions) spiked with known quantities of analyte.Guidelines recommend that data be collected from a marginal of nine determinations over at least three absorption levels covering the specified range.The data should be reported as the pct recovery of the known, added amount, or as the deflexion between the mean and true value with confidence intervals (such as 1 SD).Acceptability criteria are defined by end users but seldom fall outside 97-103% of the nominal value. Statistical analysis can be applied using a one sample t-test.Precision phase of obligation among test results when the method is applied repeatedly to multiple samplings of a homogeneous samplePrecision is commonly described in moneta ry value of repeatability, intermediate precision, and reproducibilityRepeatability is investigated by analysing a nominal of nine determinations using the same equipment and sample, covering the specified range of the procedure, or a minimum of six determinations at 100% of the test niggardness and reported as percent relative standard deviation (RSD).Intermediate precision refers to the agreement among the results from a single laboratory, despite potential variations in sample preparation, analysts, or equipment.Reproducibility refers to the agreement among the results from different laboratories. Results are reported as % RSD, and the percent difference in the mean values between the analysts must be within specifications. Less than 2% RSD is often recommended, but less than 5% RSD can be acceptable for minor components.Specifity ability to measure accurately and specifically the analyte of interest in the presence of other componentsIn drug assays, specificity takes into acc ount the degree of interference from other active ingredients, excipients, impurities, degradation products, or matrices.In chromatography, it ensures that a chromatographic peak corresponds to a single component. Specificity can be demonstrated by the resolution between peaks of interest.Limit of catching(LOD)Lowest concentration of an analyte in a sample that can be detectedIn a chromatography laboratory, the most common way to determine both the LOD and the LOQ is using signal-to-noise ratios (S/N), commonly 31 for LOD and 101 for LOQ.An appropriate number of samples must be analyzed to richly validate the method performance at the limit.Limit of quantitation(LOQ)Lowest concentration of an analyte in a sample that can be quantified with acceptable precision and accuracy under the stated operational conditions of the methodLinearityAbility of a method to provide results that are directly proportional to analyte concentration within a given rangeGuidelines specify that a minimum of five concentration levels be used to determine the range and linearity, along with certain minimum specified ranges depending on the type of method.The range is usually expressed in the same units as the test results obtained by the method (for example, nanograms per millilitre).Data to be reported generally include the equation for the calibration curve line, the coefficient of determination (r 2), residuals, and the curve itself.RangeInterval between the fastness and lower concentrations of an analyte that have been demonstrated to be determined with acceptable precision, accuracy, and linearity using the methodRobustnessMeasure of a methods capacity to obtain comparable and acceptable results when perturbed by small but deliberate variations in procedural parametersIt provides an indication of the methods suitability and reliability during normal use.During a robustness learning, method parameters (such as eluent report card, gradient, and detector settings) are intention ally varied to study the effects on analytical results. Common chromatography parameters used to measure and document robustness include critical peak pair resolution (R s), crustal plate number (N) or peak width in gradient elution, safekeeping time (t R), tailing factor (T F), peak area (and height) and concentration.Robustness studies are expected to be done during method development.Table 7 gives an overview of the performance characteristic tests that have to be performed on different types of analytical procedures.9Figure 6 Performance Characteristic tests performed on different Types of Analytical ProceduresThe performance characteristics are evaluated by comparing the results to the specifications defined at the development stage.An analytical method is considered to be validated when it meets the specifications defined at the development stage. erst an analytical method has been made a formal part of the manufacturing process, it is passing difficult to remove it. In th e event of changes in the drug substance, the composition of the finished product and in the analytical procedure, revalidation may be necessary.5,7,93.5 Validation DocumentationEvery validation step needs to be documented to be able to provide written evidence to the regulatory governing that a specific method is fit for its purpose. Documentation associated with method validation are validation protocols, standard operating procedures (SOPs), specifications and validation reports.Downstream processing in biopharmaceutical manufacturing involves many analytical methods which help to ensure quality, safety and efficacy of the final drug product. Development, validation and control of a robust analytical method is a lengthy and difficult task. However, without written evidence that an analytical method is fit for its intended use the company will not obtain a marketing authorisation.Over time, many guidelines and pharmacopeial monographs have been issued and a lot of revision activi ty has happened especially following the Stimuli to Revision paper published in 2013. Even though, no comprehensive guideline or monograph incorporating the modern lifecycle approach has been issued yet.Although proper development of robust and effective analytical methods is more time-consuming and expensive, it has many advantages. It leads to more efficient validation, variability is reduced and controlled and analytical method-related out-of-specification results and failure investigation are minimised. Additionally, changing method parameters within the design space facilitates continual improvement as it does not require regulatory re-approval.Validation is a team effort. Members of the CFT need to be properly trained. Their first and most demanding task is the preparation of a protocol which defines the scene of the validation project and provides all details necessary for a undefeated validation. It also defines, depending on the type of the analytical procedure, which per formance characteristics need to be tested. The use of well characterised reference materials with known purity is important. Analytical method validation is considered to be complete when all acceptance criteria are met and a validation report has been written. BibliographyReferences1 Choudhary, A. (2009). Validation in pharmaceutic Manufacturing. Pharmaceutical Guideline. Accessed on 1 March 2017. acquirable on network http//www.pharmaguideline.com/2010/12/validation.html2 International Conference on Harmonization (2009). Harmonised three-party Guideline ICH Q8(R2) Pharmaceutical Development. Accessed on 1 March 2017. open on net profit https//www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q8_R1/Step4/Q8_R2_Guideline.pdf3 Stockbridge, P. (2008). Biopharmaceutical Fill and Finish Technical and operational Challenges for the Latest Formulations and Devices. BioProcess International. Accessed on 7 March 2017. Available on meshwork http//www.bioprocessin tl.com/2008/biopharmaceutical-quality-assurance-184041/4 Nandhakumar, L. Dharmamoorthy, G. Rameshkumar, S. Chandrasekaran, S. (2011). An Overview of Pharmaceutical Validation Quality Assurance View Point. IJRPC, 1(4). Accessed on 1 March 2017. Available on cyberspace http//www.caidat.org/m4atomp3/2561456335400862.pdf5 Lutz, H. (2005). Introduction to Validation of Biopharmaceuticals. BioPharm International. Accessed on 1 March 2017. Available on meshing http//www.biopharminternational.com/introduction-validation-biopharmaceuticals6 USP Validation and Verification Expert Panel (2013). Lifecycle Management of Analytical Procedures Method Development, Procedure Performance Qualification, and Procedure Performance Verification. Stimuli to the Revision Process Article. Accessed on 1 March 2017. Available on Internet https//www.usp.org/sites/ default/files/usp_pdf/EN/USPNF/revisions/lifecycle_pdf.pdf7 Shabir, G. A. (2004). Step-by-Step Analytical Methods Validation and Protocol in the Q uality System Compliance Industry. IVT Network Analytical Method Validation, pp. 4-14. Accessed on 2 March 2017. Available on Internet http//www.ivtnetwork.com/sites/default/files/Analytical%20Method%20Validation.pdf8 U.S. Food and Drug Administration. Code of Federal Regulations, Title 21, split 211.165(e), 211.194(a) and 211.100(a). Accessed on 2 March 2017. Available on Internet http//www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfcfr/cfrsearch.cfm?cfrpart=2119 International Conference on Harmonization (2005). Harmonised Tripartite Guideline ICH Q2(R1) Validation of Analytical Procedures, Text and Methodology. Accessed on 1 March 2017. Available on Internet http//www.ich.org/products/guidelines/quality/article/quality-guidelines.html10 McDowall, R. D. (2014). GLP and GMP Approaches to Method Validation Going the same Way?. Spectroscopy, 29(4). Accessed on 1 March 2017. Available on Internet http//www.spectroscopyonline.com/glp-and-gmp-approaches-method-validation-going-same-way11 U.S. Pharmacopeial Convention (2016). General Chapter Prospectus The Analytical Procedure Lifecycle. USP-NF, Notices. Accessed on 2 March 2017. Available on Internet http//www.usp.org/usp-nf/notices/1220-analytical-procedure-lifecycle12 U.S. Pharmacopeial Convention (2014). New USP requirements for Analytical Method Validation. USP-NF, Notices. Accessed on 2 March 2017. Available on Internet http//www.usp.org/usp-nf/pharmacopeial-forum13 ECA Academy (2017). rewrite USP Chapter Validation of Compendial Methods approved. Accessed on 2 March 2017. Available on Internet http//www.gmp-compliance.org/gmp-news/revised-usp-chapter-1225-validation-of-compendial-methods-approved14 Huber, L. (2015). Recent Updates and Trends in Analytical Method Validation. PPP of The Agilent critical Compliance Seminar. Accessed on 7 March 2017. Available on Internet http//www.agilent.com/cs/library/flyers/Public/Recent_regulatory_updates_and_trends_in_analytical_method_validation.pdfIllustrationsFigure 1 Huber, L. (2015). Recent Updates and Trends in Analytical Method Validation. PPP of The Agilent Critical Compliance Seminar. Accessed on 7 March 2017. Available on Internet http//www.agilent.com/cs/library/fl