Stress studies. ⺠Identification of impurities. ⺠Validation of analytical methods. Content. Presentation: Purity Testing â regulatory needs and analytical method ...
Purity Testing – regulatory needs and analytical method requirements RPD 5th Year Anniversary 04.05.2012 Dr. Stefanie Seitz
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Purity Testing – regulatory needs and analytical method requirements Content › › › › ›
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Regulatory needs Method development Stress studies Identification of impurities Validation of analytical methods
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Classification of impurities › Organic impurities (process-, drug-, excipient-related) › › › › › › ›
Starting materials By-products Intermediates Degradation products Residual solvents Reaction products with excipients Leachables from container closure systems
› Inorganic impurities › Reagents, ligands and catalysts › Heavy metals or other residual metals › Inorganic salts
› Other materials (e.g., filter aids, charcoal)
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Regulatory needs – ICH Guidelines › CPMP/ICH/2737/99: NOTE FOR GUIDANCE ON IMPURITIES TESTING: IMPURITIES IN NEW DRUG SUBSTANCES › CPMP/ICH/2738/99: NOTE FOR GUIDANCE ON IMPURITIES IN NEW DRUG PRODUCTS › =>Distinct limits:
› Reporting limit (= Ph. Eur. Disregard limit) › Identification limit › Qualification limit ›
Action limits!
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Specification of: › Identified impurities › Specified unidentified impurities › Unspecified identified impurities › Sum of impurities
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Regulatory needs – ICH Guidelines CPMP/ICH/2737/99 & 2738/99
›The CHMP/CVMP QWP has prepared a series of guidelines, which provide recommendation on the information which has to be submitted in an application file for marketing authorisation (MA). It is assumed that these guidelines represent the current knowledge on technical and scientific progress. ›In Europe, the pharmaceutical legislation makes no distinction between the quality requirements for new active substances and the quality requirements for existing generic active substances (Directives 2001/83 and 2001/82 as amended). Therefore in principle (V)ICH guidelines adopted as EU guidelines may also equally apply to new products containing existing/known active substances.
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Presentation: Purity Testing – regulatory needs and analytical method requirements
CPMP/ICH/2737/99 - IMPURITIES IN NEW DRUG SUBSTANCES › This document is intended to provide guidance for registration applications on the content and qualification of impurities in new drug substances produced by chemical syntheses and not previously registered in a region or member state. › It is not intended to apply to new drug substances used during the clinical research stage of development. › The following types of drug substances are not covered in this guideline: biological/biotechnological, peptide, oligonucleotide, radiopharmaceutical, fermentation product and semi-synthetic products derived there from, herbal products, and crude products of animal or plant origin.
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Presentation: Purity Testing – regulatory needs and analytical method requirements
CPMP/ICH/2738/99 - IMPURITIES IN NEW DRUG PRODUCTS › This guideline addresses only those impurities in new drug products classified as degradation products of the drug substance or reaction products of the drug substance with an excipient and/or immediate container closure system (collectively referred to as "degradation products" in this guideline). › Generally, impurities present in the new drug substance need not be monitored or specified in the new drug product unless they are also degradation products (see ICH Q6A guideline on specifications). › Impurities arising from excipients present in the new drug product or extracted or leached from the container closure system are not covered by this guideline.
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Method development for purity testing › Performance of stress studies › Selectivity of the used methods › Integration of impurities › Identification of impurities
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Stress studies in method development for purity testing › are of fundamental importance › for the development of "stability-indicating methods“ › for the evaluation of potential degradation products (-pathways) › to discriminate between different types of impurities › to determine the stability of drug substance and drug product
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Stress testing (CPMP/QWP/122/02, corr.) › Stress testing is likely to be carried out on a single batch of the active
substance. › It should include the effect of temperatures (in 10°C increments (e.g., 50°C, 60°C, etc.) above that for accelerated testing, humidity (e.g., 75 % RH or greater) where appropriate, oxidation, and photolysis on the active substance. › The testing should also evaluate the susceptibility of the active substance to hydrolysis across a wide range of pH values when in solution or suspension. › Photostability testing should be an integral part of stress testing. The standard conditions for photostability testing are described in the Note for Guidance on Photostability Testing of New Active Substances and Medicinal Products (CPMP/ICH/279/95)
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Properties of a molecule
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Performance of stress testing
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Hydrolysis
Acid
Oxidation
Base
Isomerisation
Oxidant
Racemisation
pH
Epimerisation
Temperature
Rearrangement
Humidity
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Performance of stress testing › Acid: 1 % and 5% of sample solution, pH = 1, 60 °C , closed container, test period: 2-3 days › Base: 1 % and 5% of sample solution, pH 10, 60°C, closed container, test period: 2-3 days › Humidity: 25°C/75% r. h. and 40°C/75% r. h. , storage in open containers, testing period: one week
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Performance of stress testing › Humidified samples, different temperatures (50 °C - 90°C) in closed container, testing period: one week › Temperature: dry, different temperatures (50 °C – 90 °C), closed containers, testing period: one week › Oxidation: 1% and 5% sample solution, 0,03 % H2O2, different temperatures (30°C - 50°C), closed containers, testing period: some days
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Method development 40min, 30°C 0
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Determination of integration parameters
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Determination of integration parameters
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Strategies for identification › › › › ›
Literature search HPLC/MS Data?? Evaluation of suitable conditions for forced degradation In-situ generation of target impurity by forced degradation of the API (Selective) Extraction Purification, e.g. › (Flash) Chromatography on silica gel › Further purification on Sephadex®/C-18 › (Synthesis of the impurity) › Structure elucidation (NMR, X-Ray,…)
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Strategy of structure elucidation › 1H-NMR with signal assignment › 13C-NMR with signal assignment › 2D-NMR-spectra for an unambiguous assignment › Mass spectra, e.g. EI, ESI, CI, etc.. › interpretation of spectra (molecule ion & fragmentation) › high resolution mass spectra (?) › IR spectroscopy with assignment of characteristic absorption bands › UV/VIS spectroscopy › Literature search with recorded data!
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Strategy of structure elucidation › 3D structure › X-ray diffraction (single crystals?) › Polymorphic forms / Solid state characterisation › X-ray diffraction (powder / single crystals?) › Raman spectrometry › IR › Absolute configuration / Enantiomeric purity › X-ray structure analysis › Polarimetry
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation of analytical methods › Q2 (R1): Validation of Analytical Procedures: Text and Methodology › Note for Guidance on Validation of Analytical Methods: Definitions and Terminology (CPMP/ICH/381/95) › Note for Guidance on Validation of Analytical Procedures: Methodology (CPMP/ICH/281/95) › „The objective of validation of an analytical procedure is to demonstrate that it is suitable for its intended purpose.“ › „It is the responsibility of the applicant to choose the validation procedure and protocol most suitable for their product.“
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation of analytical methods – ICH guideline
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation protocol – validation report › Introduction › Principle of the method (assay, purity, ...) › Reference to the analytical method › Sample and Reference standards › Composition › Specification
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation protocol – validation report › Validation parameter › Specificity › Linearity / Range › Accuracy › Repeatability / intermediate precision › Detection limit / Quantitation limit › Robustness
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation protocol – validation report › Summary, table › Statistic evaluation of the data › Evaluation of the results with repect to the acceptance criteria › Deviations › Chromatograms
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Specificity › Proof of the identity of the drug substance by comparison with a reference standard › Retention time and spectrum (UV, MS) › Co-injection of drug substance and reference standard › Acceptance criteria › No interference with Drug substance › Proof of peak purity › Chromatograms of › Solvent › Reference standard › Drug substance › Drug substance + Reference standard › Matrix › Drug product 27
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Specificity › Chromatograms of › Identified impurities, individually › Drug product, spiked with impurity › Stress testing (acidic, basic, oxidative, reductive, thermal) › Reference samples of „aged“ batches › Reference solution at specification limit and reporting level
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Linearity › „A linear relationship should be evaluated across the range of the analytical procedure.“ › „The correlation coefficient, y-intercept, slope of the regression line and residual sum of squares should be submitted. A plot of the data should be included.“ › Minimum specified ranges to be considered › purity: reporting level - 120 % of specification › Purity 100 % method: reporting level – 120 % of the assay specification › Validation is performed using the drug substance as well as identified impurities
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Linearity
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Linearity › Evaluation of the residuals
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Range › „The specified range is normally derived from the 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, accuracy and precision when applied to samples containing amounts of analyte within or at the extremes of the specified range of the analytical procedure.“
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Accuracy › „application of the analytical procedure to synthetic mixtures of the product components to which known quantities of the substance to be analysed have been added“ › „in cases where it is impossible to obtain samples of all product components, it may be acceptable to add known quantities of the analyte to the product or to compare the results from a second, well characterised procedure, the accuracy of which is stated and/or defined (independent procedure)“
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Accuracy › Proof of accuracy by determination of recovery › Impurity is spiked to drug product: concentration of the impurity at n = 3 levels (reporting level, specification, 200 % of the specification, each n = 3 determinations) › accuracy for unknown impurities on the drug substance or identified impurity › Acceptance criteria › recovery = 90 - 110 % › COV < 10 %
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Precision
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation - Repeatability › Precision with minimum of n = 6 › Drug product, if impurities are present › Reference samples of „aged“ batches › Stressed samples › Acceptance criteria: COV < 5 - 10 %
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Presentation: Purity Testing – regulatory needs and analytical method requirements
Validation – Intermediate precision › according to the repeatability › Acceptance criteria: › precision: COV < 5 - 10 % › if necessary, compare the mean values: deviation of the mean values with respect to the higher value of
