Commentary
写雾的作文Quantitative Bioanalytical Methods Validation and Implementation:Best Practices for Chromatographic and Ligand Binding Assays
关于感恩的诗歌
C.T.Viswanathan,1Surendra Bansal,2Brian Booth,3Anthony J.DeStefano,4Mark J.Ro,5,11Jeffrey Sailstad,6Vinod P.Shah,7Jerome P.Skelly,8Patrick G.Swann,9and Rusll Weiner 10
Received March 2,2007;accepted March 8,2007;published online April 26,2007
Abstract.The Third AAPS/FDA Bioanalytical Workshop,entitled B Quantitative Bioanalytical Methods Validation and Implementation:Best Practices for Chromatographic and Ligand Binding Assays’’was held on May 1–3,2006in Arlington,VA.The format of this workshop consisted of prentations on bioanalytical topics,followed by discussion ssions where the topics could be debated,with the goal of reaching connsus,or identifying subjects where addition input or clarification was required.The discussion also addresd bioanalytical validation requirements of regulatory agencies,with the purpo of clarifying expectations for regulatory submissions.The proceedings from each day were reviewed and summarized in the evening ssions among the speakers and moderators of the day.The connsus summary was prented back to the workshop on the last day
and was further debated.This communication reprents the distillate of the workshop proceedings and provides the summary of connsus reached and also contains the validation topics where no connsus was reached.KEY WORDS:bioanalytical;bioequivalence;LC-MS/MS;ligand binding;validation.
INTRODUCTION
Bioanalysis,employed for the quantitative determina-tion of drugs and their metabolites in biological fluids,plays a significant role in the evaluation and interpretation of bioequivalence,pharmacokinetic (PK),and toxicoki-netic studies.The quality of the studies,which are often ud to support regulatory filings,is directly related to the quality of the underlying bioanalytical data.It is therefore
important that guiding principles for the validation of the analytical methods be established and disminated to the pharmaceutical community.
The first American Association of Pharmaceutical Scientists (AAPS)/Food and Drug Administration (FDA)Bioanalytical Workshop in 1990focud on key issues relevant to bioanalytical methodology and provided a platform for scientific discussions and deliberations.The workshop and the report (1),raid awareness of the need for validated bioanalytical methods for the regulatory acceptance o
f bioequivalence and pharmacokinetic data.Although the workshop addresd bioanalysis in general,it acknowledged the differences between chromatographic and ligand-binding (non-chromatographic bad)methods.The workshop identified the esntial parameters for bio-analytical method ,accuracy,precision,lec-tivity,nsitivity,reproducibility,limit of detection and stability.The outcome of the first workshop and its report resulted in improved quality of data submissions to regula-tory agencies.
Following the first workshop report (1)and the experi-ence gained at the FDA,the draft Guidance on Bioanalytical Methods Validation was issued by the FDA in January 1999.This draft guidance provided stimulus and opportunity for further discussion at the 2nd AAPS/FDA Bioanalytical Work-shop in January 2000.In addition,newer technology,such as chromatography coupled to tandem mass spectrometry (LC-
0724-8741/07/1000-1962/0#2007Springer Science +Business Media,LLC
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Pharmaceutical Rearch Vol.24,No.10,October 2007(#2007)DOI:10.1007/s11095-007-9291-7
The views expresd in this article are tho of the authors and do not reflect official policy at the FDA.No official endorment by the FDA is intended or should be inferred.
1
Food and Drug Administration,Center for Drug Evaluation and Rearch,Rockville,Maryland 20857,USA.2
Hoffmann-La Roche Inc.,Nutley,New Jery 07110,USA.3
Food and Drug Administration,Center for Drug Evaluation and Rearch,Silver Spring,Maryland 20993,USA.4
Procter &Gamble Pharmaceuticals,Mason,Ohio 45040,USA.5
Amgen Inc.,Thousand Oaks,California 91320,USA.6
Sailstad and Associates,Durham,North Carolina 27707,USA.7
Consultant,Gaithersburg,Maryland 20878,USA.8
Consultant,Alexandria,Virginia 22309,USA.9
Food and Drug Administration,Center for Drug Evaluation and Rearch,Bethesda,Maryland 20892,USA.10
Bristol-Myers Squibb Inc.,Pennington,New Jery 08534,USA.11
To whom correspondence should be addresd.(e-mail:)
MS/MS),was discusd along with an update on ligand binding assays.This workshop resulted in a report B Bioanalytical Method Validation-A Revisit with a Decade of Progress^(2) and formed the basis for the FDA Guidance on Bioanalytical Methods Validation in May2001(3).
The evolution of divergent analytical technologies for conventional small molecules and macromolecules,and the growth in marketing interest in macromolecular therapies, has led to the workshop held in2000to specifically discuss bioanalytical methods validation for macromolecules.Due to the complexity of the issues,the workshop failed to achieve a connsus.To address the need for guiding principles for the validation of bioanalytical methods for macromolecules,the AAPS Ligand Binding Assay Bioanalytical Focus Group developed and published recommendations for the develop-ment and validation of ligand binding assays in2003(4).
As bioanalytical tools and techniques have continued to evolve and significant scientific and regulatory experience has been gained,the bioanalytical community has continued its critical review of the scope,applicability,and success of the prently employed bioanalytical guiding principles.The purpo of this Third AAPS/FDA Bioanalytical Workshop was to identify,review and evaluate the existing practices, white papers,articles and clarify the FDA Guidance.The workshop addresd quantitative bioanalytical methods val-idation and their u in sample analysis,focusing on both chromatographic and ligand binding assays.
GOALS AND OBJECTIVES
The purpo of the Third AAPS/FDA Bioanalytical Workshop was to:
&Review the scope and applicability of bioanalytical principles and procedures for the quantitative analysis of samples from bioequivalence,pharmacokinetic and compa-rability studies in both human and non-human subjects;
&Review current practices for scientific excellence and regulatory compliance,suggesting clarifications and improve-ments where needed;
&Review and evaluate validation and implementation requirements for Chromatographic and Ligand-bad Quan-titative Bioanalytical Assays,covering all types(sizes)of molecules;
&Review recent advances in technology,automation, regulatory and scientific requirements and data archiving on the performance and reporting of quantitative bioanalytical work;and
&Discuss current best approaches for the conduct of quantitative bioanalytical work regardless of the size of the molecule analyzed.
The Third AAPS/FDA Bioanalytical Workshop,held on May1–3,2006in Arlington,VA,concluded with veral recommendations to achieve the above goals and objectives. While the FDA guidance(3)remains valid,the recommen-dations obtained during the workshop were aimed at pro-viding clarification and some recommendations to enhance the quality of bioanalytical work.This publication provides the clarification and recommendations obtained at the workshop with a view to achieve uniformity among the practitioners and urs of quantitative bioanalysis for all types of molecules.
NON-CHROMATOGRAPHIC ASSAY—SPECIFIC ISSUES
Differences between Ligand Binding Assays Supporting Macromolecule PK Analysis and Small Molecule Analysis by Chromatography
Ligand Binding Assays(LBAs)are ud throughout many organizations attempting to discover or develop new chemical entities(NCE).Besides the obvious size difference between small and macromolecule analytes,there are key structural differences.Small molecules typically are organic molecules while macromolecules are complex biopolymers. In addition,small molecules are prepared by organic synthesis while macromolecules are typically formed biolog-ically.As a direct result of how macromolecules are pro-duced,the reference standards tend to be heterogeneous, often due to post-translational modifi,glycosyla-tion or phosphorylation).In contrast,small molecule refer-ence standards are homogeneous with a high degree of purity.Generally,small molecules are often hydrophobic and macromolecules are often hydrophilic.While chemical sta-bility is assd for small molecules with relative ea, macromolecule stability asssment is generally more com-plex,requiring the evaluation of not only chemical and physical properties,but also biological ,is receptor binding affinity maintained?).Macromolecules are endogenous and/or structurally similar to endogenous counter-parts,while small molecules are generally xenobiotics,foreign and not prent in the sample matrix.The catabolism of small molecules is typical
ly well defined while for macromolecules few specifics are known.Macromolecules typically have specific carrier proteins while small molecules can be generically bound to a number of endogenous proteins.Due to the significant differences between small and macromolecule analytes,different technologies,such as LC-MS for small molecules and LBAs for macromolecules,are often employed to determine drug levels for PK asssments.
Method validations for the divergent methods should consider important differences including the basis of mea-surement,the detection modality and whether a sample is measured directly in the matrix or extracted prior to analysis. The basis of measurement of LC-MS is owed to the chemical properties of the analyte,while for LBAs,the measurement depends on a high affinity biological binding interaction between the macromolecule analyte and another macro-molecule(s)in the form of one or more capture/detection antibodies.Detection in LC-MS methods is direct and typically results in a linear measured respon,where higher concentrations of analyte have a proportional increa in respon.In contrast,the measured respon in LBAs is indirect and this results in a non-linear,often sigmoidal, measured respon.Owing to the characteristics of the assay system,the calibration standard curve range for a LC-MS method is broad,often covering veral orders of magnitude. In contrast,the calibration range for an LBA is typically limited to less than two orders of magnitude.The analyte differences,combined with the unique technologies ud to
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Quantitative Bioanalytical Methods Validation and Implementation
measure analyte concentration,provide a strong rationale as to why consideration should be given to the need of employ-ing some analyte specific(small versus macromolecule) method validation guidelines.
One major point of concern in discussing the method validations for the divergent technologies centers on stan-dards and quality control(QC)acceptance ,the acceptable deviation from a nominal value expresd as a percentage).Current guidance recommends the15/20rule, where thefirst number,in this ca15%,is the acceptance criterion for all standards and quality control samples(QCs) with the exception of the lower limit of quantitation(LLOQ), where the acceptance criterion is incread to a20% deviation.This rule was developed prior to the routine u of LC-MS,where chromatographic methods were employed,but internal standards were analyte analogues and not stable isotopes.When the15/20rule was propod,most PK asss-ments that utilized ,radioimmunoassay)measured small molecules.The typical radioimmunoassay(RIA)ud high affinity polyclonal antibodies that were quite suitable to measure well characterized homogeneo
us organic small mol-ecules.In most of the small molecule RIAs,meeting the15/ 20challenge was achievable and it is recommended that the 15/20rule be continued when LBAs are ud for small molecule analysis.However,nearly all small molecule analysis performed today is by LC-MS,often with the incorporation of a stable isotope internal standard;as a result,assay precision has continued to improve.In fact,the results of a method validation survey conducted for the Third AAPS/FDA Bio-analytical Workshop found that89%of chromatography respondents ud the15/20target.
As a result of small molecule analysis moving to the LC-MS platform,LBAs are now almost exclusively ud to measure macromolecules.While some LBAs continue to be developed and validated to meet the15/20rule,different criteria are sometimes required becau of the heteroge-neous nature of macromolecules,and the fact that other macromolecules(antibodies)are employed in the assay.In fact,the Third AAPS/FDA Bioanalytical Workshop survey found that only23%of the LBA respondents follow the 15/20rule.Instead,53%of respondents ud somewhere between20/25(42%)and30/30(2%)as their acceptance rule,while23%ud B other criteria.^The B other criteria^ could possibly include statistically bad approaches that estimate in-study assay performance bad on pre-study validation results.
Ligand Binding Assays Pre-study Validation
During pre-study validation,method precision and accu-racy are determined through the analysis of QCs(validation samples)prepared in a biological matrix equivalent to that anticipated for study samples.Due to the endogenous nature of some biopharmaceuticals,it may be necessary to deplete the matrix of the analyte or employ a F surrogate_matrix to evaluate method accuracy and precision.One propod validation protocol(4)recommends that matrix be spiked at 5or more validation sample concentrations that span the range of quantifi,the anticipated lower limit of quantitation(LLOQ),õ3times LLOQ,mid(geometric mean),high(õ75%of the upper limit of quantitation [ULOQ],andfinally the anticipated ULOQ.As previously noted,the major sources of variability(imprecision and inaccuracy)differ bad on technology.For LBAs,the inter-batch variance component is usually a greater contrib-utor to the overall variability than the intra-batch variance component.It is recommended that at least two independent determinations be made for each validation sample per assay run across a minimum of six independent assays runs (balance validation design).For example,12reportable values would result from two measurements across six independent assay runs.An appropriate statistical method should then be ud to compute the summary ,each validation sample,the repeated measurements from all runs should be analyzed together).A detailed description of this approach has been described previously(4).
For a method to be considered acceptable,it is recommended that both the inter-batch imprecision(%CV) and the accuracy,expresd as absolute mean bias(%RE)be T20%(25%at LLOQ and ULOQ).As an additional constraint to control method error,it is recommended that the target total error[sum of the absolute value of the%RE (accuracy)and precision(%CV)be less than T30%(T40% at the LLOQ and ULOQ].The additional constraint of total error allows for consistency between the criteria for pre-study method validation and in-study batch acceptance.In asss-ing the acceptability of a method,including total error,it is not appropriate to reject assay runs.All assay runs during the validation should be included in the computation of summary statistics.The only exception would be runs rejected for cau or in cas where errors are obvious and documented. Ligand Binding Assays In-study Acceptance Criteria The recommended standard curve acceptance criteria for macromolecule LBAs are that at least75%of the standard points should be within20%of the nominal concentration(%RE of the back-calculated values),except at the LLOQ and ULOQ where the value should be within 25%.This requirement does not apply to F anchor calibrators_ which are typically outside the anticipated validation range of the assay and ud to facilitate and improve F sigmoidal_ curve-fitting.
The recommended QC acceptance criteria for macro-molecule LBAs includes the u of low,medium
and high (LQC,MQC and HQC)QCs typically run in , 6results=3concentrationsÂ2reportable values per concen-tration),with assays being accepted bad upon a4-6-20rule. Exceptions to this criterion should be justifi,pre-study total error data approaching30%).At least four of the six QCs must be within20%of the nominal value.In addition,at least one QC sample per concentration needs to meet this criterion.If additional ts of QCs are ud in a run,then50%of them need to be F in-range_at each concentration.The following are recommendations for the placement of the controls in relation to the standard curve range.The LQC should be placed above the cond non-anchor standard,approximately3times the LLOQ.The MQC is placed near the mid point(geometric not arithmetic mean)of the standard curve,while the HQC should be placed below the cond non-anchor point high standard and/or about75%of the ULOQ.
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CALIBRATION CURVE AND QC RANGES
QC samples rve to monitor the performance of the methodology throughout the cour of the analysis.They are the basis for demonstrating,as required in21CFR320.29(a), that the analytical method is sufficiently accurate,preci, and nsitive to measure the actual concentrations achieved
in the body.For studies involving pharmacokinetic profiles spanning all or most of the calibration curve,three QC samples run in duplicate(or at least5%of the unknown samples),spaced across the standard curve as per the FDA Guidance(3),are likely sufficient to adequately monitor method performance.For an analysis where the study data fall over a small percentage of the calibration curve,it is possible that none of the QC concentrations is near the concentrations of the unknowns,thus limiting the monitoring power of the QC samples.
If a narrow range of analysis values is known or anticipated prior to the start of sample analysis,it is recommended that either the standard curve be narrowed and new QC concentrations ud as appropriate,or if the original curve is ud,existing QC concentrations be revid or sufficient QC samples at additional concentration(s)added to adequately reflect the concentrations of the study samples. Narrowing of the standard curve and preparation of new QC samples requires only a partial validation to assure adequate performance of the new curve and QCs.A full validation is not required.
统计图表怎么做If a narrow range of analysis values is unanticipated,but obrved after start of the sample analysis,it is recommended that the analysis be stopped and either the standard curve narrowed,existing QC concentrations revid,or QC sam-ples at additional concentrations be added to the original curve pr
ior to continuing with sample analysis.It is not necessary to reanalyze samples analyzed prior to optimizing the standard curve or QC concentrations.
CARRYOVER AND CONTAMINATION EVALUATION
Contamination,carryover,or blank respon from matrix or reagents can affect the accuracy and precision of quantita-tion at all concentrations.However,low concentration sam-ples are most affected as a percentage of concentration.Care should be taken to minimize interference from all contamina-tion factors and the interference should not significantly affect the accuracy and precision of the assay.
Carryover does not necessarily involve only the next sample in the quence.In fact,carryover from late eluting residues on columns may affect chromatograms veral samples later.Carryover from residues in rotary sampling/ switching valves often appears later in the samples.Precau-tions should be taken to avoid contamination during sample collection and preparation.Carryover should be assd during validation by injecting one or more blank samples, after a high concentration sample or standard.The injector should beflushed with appropriate solvents to minimize carryover.If carryover is unavoidable for a highly retained compound,specific procedures should be provided in t
he method to handle known carryover.This could include injection of blanks after certain samples.Randomization of samples should be avoided,since it may interfere with the asssment of carryover problems.Contamination can be assd by monitoring blank respon in the prence of high concentration samples or standards.The assay platform (manual or automated),configuration of sampling and extraction method(manual,automated,on-line or solid pha,etc.)in the assay should be taken into consideration when ascertaining contamination.There is no standard acceptable magnitude of carryover for a passing bioanalytical run.Carryover should be addresd in validation and minimized,and an objective determination should be made in the evaluation of analytical runs.
During validation,the operator should asss the analyte respon due to blank matrix while eliminating or minimizing other contaminations.The analyte respon at the LLOQ should be at least5times the respon due to blank matrix. For immunoassays,and if the analyte is prent endogenous-ly in the matrix,the blank respon can exceed20%of LLOQ,but the contribution should not interfere with the required accuracy in the measurement of the LLOQ.In such cas,specific procedures should be provided in the method to handle blank matrix respon.
DETERMINATION OF METABOLITES
DURING DRUG DEVELOPMENT
A draft FDA Guidance for Industry,entitled
B Safety Testing of Drug Metabolites^was issued in June2005by the Center for Drug Evaluation and Rearch(CDER)(5).There is general support from the pharmaceutical community for the idea that a more extensive characterization of the pharmaco-kinetics of unique and/or major human metabolites(UMMs) would provide greater insight into the connection between metabolites and toxicological obrvations.This information would be best generated by the u of rugged,bioanalytical methods applied at appropriate times in drug development.
Characterization of UMMs should proceed using a flexible,B tiered^approach to bioanalytical methods valida-tion.This tiered approach would allow metabolite screening studies to be performed in early drug development using bioanalytical methods with limited validation,with validation criteria increasing as a product moves into clinical trials.A tiered validation approach to metabolite determination would defer bioanalytical resource allocation to later in the drug development timeline when there is a greater likelihood of drug success.As a minimum,the specifics of this tiered validation process should be driven by scientifically-appropriate criteria,established a priori.
INCURRED SAMPLE REANALYSIS
There are a number of situations where the performance of standards and QCs may not adequately mimic that of study samples from dod subjects(incurred samples). Examples include,metabolites converting to the parent species,protein binding differences in patient samples, recovery issues,sample inhomogeneity,and mass spectro-metric ionization matrix effects.The factors can affect both the reproducibility and accuracy of the concentration deter-mined in incurred samples.While the effects are often
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Quantitative Bioanalytical Methods Validation and Implementation
T a b l e I .D e t a i l s o f D o c u m e n t a t i o n D e s i r a b l e a t t h e A n a l y t i c a l S i t e a n d i n V a l i d a t i o n a n d A n a l y t i c a l R e p o r t s
I t e m s
A n a l y t i c a l S i t e V a l i d a t i o n R e p o r t V a l i d a t i o n R e p o r t A p p e n d i x A n a l y t i c a l R e p o r t
A n a l y t i c a l R e p o r t A p p e n d i x
S t a n d a r d
C e r t i fic a t e o f a n a l y s i s ,p u r i t y ,s t a b i l i t y f o r a n a l y t e B a t c h /L o t #,p u r i t y a n d m a n u f a c t u r e r
B a t c h /L o t #,p u r i t y a n d m a n u f a c t u r e r
R e c o r d o f r e c e i p t a n d s t o r a g e S t a b i l i t y a t t i m e o f u s e S t a b i l i t y a t t i m e o f u s e
L a c k o f i n t e r f e r e n c e b e t w e e n I S a n d a n a l y t e S t o c k s o l u t i o n p r e p a r a t i o n
R e c o r d s o f p r e p a r a t i o n S t o r a g e l o c a t i o n &c o n d i t i o n C a l i b r a t o r s a n d Q C s p r e p a r a t i o n
R e c o r d s o f p r e p a r a t i o n
P r e p a r a t i o n d a t e s
S t o r a g e c o n d i t i o n s
安踏广告词是什么
F r e e z e r l o g (s a m p l e i n g r e s s /e g r e s s ,t e m p e r a t u r e )
S t o r a g e c o n d i t i o n s
R u n a c c e p t a n c e c r i t e r i a S O P f o r c a l i b r a t o r s ,Q C s &c h r o m a t o g r a p h i c i n t e r f e r e n c e s S h o r t d e s c r i p t i o n S O P (o p t i o n a l )S h o r t d e s c r i p t i o n
雪地胎S O P (o p t i o n a l )
A s s a y p r o c e d u r e
S O P f o r t h e m e t h o d
B r i e f d e s c r i p t i o n o f m e t h o d o f e x t r a c t i o n ,a n d a n a l y s i s S O P (o p t i o n a l )B r i e f d e s c r i p t i o n S O P (o p t i o n a l )
S a m p l e t r a c k i n g
S t u d y s a m p l e r e c e i p t ,c o n d i t i o n o n r e c e i p t &l o c a t i o n o f s t o r a g e S t o r a g e c o n d i t i o n &l o c a t i o n
D a t e s o f r e c e i p t o f s h i p m e n t s a n d c o n t e n t s T r a c k i n g o f Q C ,c a l i b r a t o r s a n d s t u d y s a m p l e s S a m p l e c o n d i t i o n o n r e c e i p t F r e e z e r l o g s S t o r a g e l o c a t i o n &c o n d i t i o n A n a l y s i s
D a t e s o f e x t r a c t i o n a n d a n a l y s i s a n d i n s t r u m e n t I D f o r e a c h r u n T a b l e o f r u n s ,i n s t r u m e n t I D &a n a l y s i s d a t e s
R e p r e s e n t a t i v e c h r o m a t o g r a m s
T a b l e o f a l l r u n s ,&a n a l y s i s d a t e s
C h r o m a t o g r a m s f r o m 5–20%o f s u b j e c t s f o r
I d e n t i t y o f Q C s ,c a l i b r a t o r s &s t u d y s a m p l e s
T a b l e o f c a l i b r a t o r r e s u l t s o f a l l r u n s w i t h a c c u r a c y a n d p r e c i s i o n C r o s s -v a l i d a t i o n ,i f a p p l i c a b l e
T a b l e o f c a l i b r a t o r r e s u l t s o f a l l p a s s e d r u n s w i t h m e a n &%C V A N D A a n d
D o c u m e n t a t i o n o f p r o c e s s i n g o f c a l i b r a t o r s ,Q C s a n d s t u d y s a m p l e s f o r e a c h r u n
T a b l e s o f w i t h i n a n d b e t w e e n r u n Q C r e s u l t s (a c c u r a c y a n d p r e c i s i o n )
风的造句A d d i t i o n a l v a l i d a t i o n ,i f a n y
T a b l e s o f Q C r e s u l t s o f a l l p a s s e d r u n s w i t h a c c u r a c y a n d p r e c i s i o n .O .K t o i n c l u d e Q C r e s u l t s o f t h e f a i l e d r u n s .
R e p r e s e n t a t i v e c h r o m a t o g r a m s f o r N D A s u b m i s s i o n s
D o c u m e n t a t i o n o f i n s t r u m e n t s e t t i n g s a n d m a i n t e n a n c e
B e n c h -t o p ,f r e e z e -t h a w ,l o n g -t e r m a n d p o s t -p r e p a r a t i v e &s t o c k s o l u t i o n s t a b i l i t y d a t a L o n g -t e r m s t a b i l i t y a p p e n d e d o r w r i t t e n i n a s e p a r a t e r e p o r t
R u n s u m m a r y s h e e t s
E x t r a c t i o n r e c o v e r y &m a t r i x e f f e c t
100%c h r o m a t o g r a m s L I M S a n d M o d e o f i n t e g r a t i o n E x t r a c t i o n d a t e s
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Viswanathan et al.
characterized and minimized during method development using QC samples,it is important to assure that they are under control when the method is applied to the analysis of incurred samples.
A proper evaluation of incurred sample reproducibility and accuracy needs to be performed on each species ud for Good Laboratory Practice (GLP)toxicology experiments.It is not necessary for additional incurred sample investigations to be performed in toxicology species once the initial asssment has been performed.Incurred sample evaluations performed using samples from one study would be sufficient for all other studies using that same species.
It is generally accepted that the chance of incurred sample variability is greater in humans than in animals,so the following discussion pertains primarily to clinical studies.The final decision as to the extent and nature of the incurred sample testing is left to the analytical investigator,and should be bad on an in-depth understanding of the method,the behavior of the drug,metabolites,and any con
comitant medications in the matrices of interest.There should be some asssment of both reproducibility and accuracy of the reported concentration.Sufficient data should be generated to demonstrate that the current matrix produces results similar to tho previously validated.It is recognized that accuracy of the result generated from incurred samples can be more difficult to asss.It requires evaluation of any additional factors besides reproducibility upon storage,which could perturb the reported concentration.The could include metabolites converted to parent during sample preparation or LC-MS/MS analysis,matrix effects from high concentrations of metabolites,or variable recovery between analyte and internal standard.If a lack of accuracy is not due to assay performance (i.e.,analyte instability or interconver-sion)then the reason for the lack of accuracy should be investigated and its impact on the study assd.The extent and nature of the experiments is dependent on the specific sample being addresd and should provide sufficient confi-dence that the concentration being reported is accurate.
The results of incurred sample reanalysis studies may be documented in the final bioanalytical or clinical report for the study,and/or as an addendum to the method validation report.
In lecting samples to be reassayed,it is encouraged that issues such as concentration,patient population and special populations (e.g.,renally impaired)be considered,depending on what is known
about the drug,its metabolism and its clearance.First-in-human,proof-of-concept in patients,spe-cial population and bioequivalence studies are examples of studies that should be considered for incurred-sample con-centration verification.The study sample results obtained for establishing incurred sample reproducibility may be ud for comparison purpos,and do not necessarily have to be ud in calculating reported sample concentrations.DOCUMENTATION ISSUES
Although the current guidance for the documentation ction remains valid,further issues are now addresd and details are provided herein (Table I )to facilitate effective documentation.Records generated during the cour of method validation and study sample analysis are source
F a i l e d r u n s
S a m e a s i n B A n a l y s i s ^
I d e n t i f y r u n s ,a s s a y d a t e a n d r e a s o n f o r f a i l u r e
I d e n t i f y r u n s ,a s s a y d a t e s ,a n d r e a s o n f o r f a i l u r e
R e i n t e g r a t i o n太阳能的应用
A u d i t t r a i l :o r i g i n a l &r e i n t e g r a t i o n S O P (o p t i o n a l )
R e a s o n f o r r e i n t e g r a t i o n M o d e o f r e i n t e g r a t i o n D e v i a t i o n s f r o m S O P s /m e t h o d
D o c u m e n t a t i o n o f d e v i a t i o n s a n d u n e x p e c t e d e v e n t s D e s c r i p t i o n o f d e v i a t i o n s D e s c r i p t i o n o f d e v i a t i o n s
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S O P f o r r e a s s a y c r i t e r i a C o m m u n i c a t i o n
B e t w e e n a n a l y t i c a l s i t e a n d c l i n i c a l s i t e /s p o n s o r
S O P s t a n d a r d o p e r a t i n g p r o c e d u r e s ,I S i n t e r n a l s t a n d a r d ,Q C q u a l i t y c o n t r o l ,Q C s q u a l i t y c o n t r o l s a m p l e s ,%C V i n t e r b a t c h i m p r e c i s i o n ,A N D A A b b r e v i a t e d N e w D r u g A p p l i c a t i o n ,N D A N e w D r u g A p p l i c a t i o n ,L I M S L a b o r a t o r y I n f o r m a t i o n M a n a g e m e n t S y s t e m s
1967
Quantitative Bioanalytical Methods Validation and Implementation
