Verification of applicability of the validated/compendial API analytical method for the final formulation

Assay, dissolution test and degradants
Dar es Salaam, August, 21-25, 2006 Dr. Birgit Schmauser, BfArM, Bonn

Guidelines
• ICH Q2A
– Validation of Analytical Methods: Definitions and Terminology (CPMP/ICH/381/95)

• ICH Q2B
– Validation of Analytical Procedures: Methodology (CPMP/ICH/281/95)

• ICH Q6A
– Specifications: Test Procedures and Acceptance Criteria for New Drug Substances and New Drug Products: Chemical Substances (CPMP/ICH/367/96 corr)
Dar es Salaam, August, 21-25, 2006 Dr. Birgit Schmauser, BfArM, Bonn

API
• Assay
– Validation with respect to:
• Specificity, linearity/range, accuracy, precision, robustness

• Impurities
– Validation with respect to:
• Specificity, linearity/range, accuracy, precision, limit of detection (LOD), limit of quantitation (LOQ), robustness
Dar es Salaam, August, 21-25, 2006 Dr. Birgit Schmauser, BfArM, Bonn

21-25. Birgit Schmauser. BfArM. 2006 Dr. August.FPP • Formulation of the drug product – Presence of further APIs – Presence of excipients (individual formulation) – Presence of known impurities/degradants of all APIs and potential new degradants or incompatibility products Dar es Salaam. Bonn .

Birgit Schmauser. Bonn . August. 21-25. BfArM.Requirements • Capability of the analytical method(s): – Assay of each API in the presence of the other APIs and all impurities/degradants – Assay of each degradant in the presence of all APIs and all other degradants/impurities – Influence of formulation components should be excluded/controlled Dar es Salaam. 2006 Dr.

August. Birgit Schmauser. column parameters.Revalidation I • Revalidation of analytical methods with respect to: – Specificity • presence of new API(s) and impurities/degradants/formulation components – Range • test concentrations of API(s) versus FPP – Accuracy • influence of formulation components – Precision • influence of formulation and sample preparation – LOD/LOQ • test concentrations of API(s) versus FPP) – Robustness • change of column material. solvents) Dar es Salaam. Bonn . BfArM. 21-25. 2006 Dr.

Bonn . Birgit Schmauser. robustness) Dar es Salaam. BfArM. 2006 Dr. 21-25.g.Revalidation II • Revalidation reflected by ICH Q2A: – Revalidation may be necessary in the following circumstances: • Changes in the synthesis of the drug substance • Changes in the composition of the finished product • Changes in the analytical procedure – (e. August.

Bonn . 2006 Dr. August. 21-25. BfArM.Specificity • Identification – Discrimination between compounds of closely related structures • positive results (from samples containing the analyte) • negative results (from samples that do not contain the analyte) • components structurally similar to the analyte do not give positive results Dar es Salaam. Birgit Schmauser.

2006 Dr. Birgit Schmauser. August. BfArM. 21-25.Specificity II • Assay and impurities – Chromatographic procedures • Representative chromatograms with appropriate labelling of individual components • Investigation at an appropriate level Dar es Salaam. Bonn .

Specificity III • Chromatogram with retention times and chemical structures of: • (1) arteannuin B • (2) artemisitene • (3) artemisinin • (4) artemisinic acid • (5) artemether (IS) • Analytical standard containing 1.W. August. BfArM.4 µg/ml IS From: F. Bonn .C. A 1118 (2006) 180-187 Dar es Salaam. Birgit Schmauser. J Chromatogr. 2006 Dr.2µg/ml of each analyte and 0. Van Nieuverburgh et al. 21-25..

Birgit Schmauser. 2006 Dr. Bonn .Injection of pure API . BfArM.Specificity IV • Assay and impurities/degradants – Discrimination of analytes where impurities/degradants are available • Assay – Demonstration of discrimination of the analyte in the presence of all impurities/degradants and/or excipients » f. August. assay result unaffected by presence of spiked impurities/degradants: . ex.Injection of API plus impurities/degradants Dar es Salaam. 21-25.

Bonn . Birgit Schmauser. ex. BfArM. August. spiking of API with appropriate levels of impurities/degradants and/or excipients: Chromatographic profiles of API with and without impurities/degradants/excipients Dar es Salaam. 2006 Dr.Specificity V • Assay and impurities/degradants – Discrimination of analytes where impurities/degradants are available • Impurities/Degradants – Demonstration of separation of impurities/degradants individually and/or from excipients » f. 21-25.

Specificity VI • Assay and impurities/degradants – Discrimination of analytes where impurities/degradants are not available • Comparison of the test procedure to a second well-characterized (independent) procedure – Samples » Test samples containing impurities/degradants » Test samples stored under relevant stress conditions (potential degradants arising during shelf life) Dar es Salaam. 2006 Dr. Birgit Schmauser. August. 21-25. Bonn . BfArM.

2006 Dr. 21-25.Specificity VII • Assay and impurities/degradants – Discrimination of analytes where impurities/degradants are not available • Assay – Comparison of test results by the two independent procedures • Impurities/Degradants – Comparison of impurity profiles • Peak purity assessment – Demonstration that the analyte peak is attributable to only one component Dar es Salaam. Bonn . August. Birgit Schmauser. BfArM.

August. 2006 Dr. 21-25. in: Stavros Kromidas. Birgit Schmauser. Validierung in der Analytik. University of Wuerzburg.Specificity VIII • Peak purity • Overlapping peaks in HPLC (simulation) A B C D From: Prof. Wiley-VCH Dar es Salaam. Siegfried Ebel. Bonn . BfArM.

in: Stavros Kromidas.Specificity IX • Peak purity Fatty acids were reacted with ethylene oxide and separated by HPLC (Fractions 1-6) Fraction 5 was analysed by MALDI From: Dr. BfArM. Düsseldorf. Validierung in der Analytik. Bonn . August. Henkel KGaA. Birgit Schmauser. Michael Schmitt. Wiley-VCH Dar es Salaam. 21-25. 2006 Dr.

21-25. Bonn . 2006 Dr.g.Specificity with FDCs • FDC (e. August. Birgit Schmauser. artesunate and amodiaquine) – One analytical method for both APIs • Capability of one method to quantify both APIs and to separate/discriminate one API and its impurities/degradants and potential incompatibility products from the other API and its impurities/degradants/incompatibility products – Some reference material for impurities/degradants will be available (spiking experiments applicable) – Other degradants are not available as reference material (stress testing necessary to generate in situ degradants) Dar es Salaam. BfArM.

Bonn . BfArM. Birgit Schmauser.120% – Impurities/Degradants • Reporting level to 120% of specification limit • Revalidation is necessary. 21-25. 2006 Dr.Range • Minimum specified ranges – Assay • 80 – 120% of the test concentration – Content uniformity » 70 – 130% of the test concentration – Dissolution » Q-20% . if the ranges covered during validation of the API-methods are different from those of the FPP-methods (different test concentrations) Dar es Salaam. August.

21-25. 2006 Dr. Bonn . August. BfArM. Birgit Schmauser.Accuracy • Assay – Application of the analytical procedure to synthetic mixtures of the product components (placebo mixture) to which known quantities of the analyte have been added – In case certain product components are unavailable: • Application of the analytical procedure to the product to which known quantities of the analyte have been added • Comparison of results obtained by a second (independent) procedure with defined accuracy Dar es Salaam.

BfArM. 21-25. Bonn . Birgit Schmauser. August. 2006 Dr.Accuracy II • Impurities/Degradants – Assessment of samples spiked with known amounts of impurities/degradants – In case certain impurities/degradation products are unavailable • Comparison of results obtained by a second (independent) procedure with defined accuracy Dar es Salaam.

Precision • Assay and impurities/degradants – Repeatability • 9 determinations (3 x 3) covering the specified range or • 6 determinations at 100% of the test concentration – Intermediate precision • Effects of random events on the precision of the procedure. August. 21-25. Bonn .g. e. – Days – Analysts – Equipment • To be performed with a test solution prepared from the drug product Dar es Salaam. BfArM. 2006 Dr. Birgit Schmauser.

Birgit Schmauser. 21-25. Bonn . BfArM.3s/S – Estimation of S » from the calibration curve of the analyte – Estimation of s » from the standard deviation of the blank » from the standard deviation (regression line or y-intercept) of a calibration curve in the range of the DL Dar es Salaam. 2006 Dr.Detection Limit • Determination based on – Visual evaluation (non-instrumental and instrumental methods) – Signal to Noise (baseline noise) – Standard deviation of response (s) and slope (S) • DL=3. August.

2006 Dr. BfArM.Quantitation Limit • Determination based on – Visual evaluation (non-instrumental and instrumental methods) – Signal to Noise (baseline noise) – Standard deviation of response (s) and slope (S) • QL=10s/S – Estimation of S » from the calibration curve of the analyte – Estimation of s » from the standard deviation of the blank » from the standard deviation (regression line or y-intercept) of a calibration curve in the range of the QL Dar es Salaam. Bonn . 21-25. August. Birgit Schmauser.

Bonn . Birgit Schmauser.Robustness I • Reliability of an analysis with respect to deliberate variations in method parameters – Susceptibility to variations in analytical conditions? • control of analytical conditions or precautionary statement • establishment of system suitability parameters Dar es Salaam. 2006 Dr. BfArM. August. 21-25.

Birgit Schmauser. 2006 Dr. Bonn .Robustness II • Examples of variations – Stability of analytical solutions – Extraction time • In the case of liquid chromatography – – – – – Influence of variations of pH in a mobile phase Influence of variations in mobile phase composition Influence of columns (different lots and/or suppliers) Influence of temperature Influence of flow rate • In the case of gas chromatography – Influence of columns (different lots and/or suppliers) – Influence of temperature – Influence of flow rate Dar es Salaam. August. BfArM. 21-25.

Bonn .Robustness III • Influence of pH of elution on separation of amino acids by RP-HPLC From: Waters. August. 2006 Dr. in: Stavros Kromidas. Birgit Schmauser. Wiley-VCH Dar es Salaam. 21-25. Validierung in der Analytik. BfArM.

Michael Krämer. Wiley-VCH Dar es Salaam. Birgit Schmauser. in: Stavros Kromidas. 21-25. NOVARTIS.Robustness • Electropherograms under identical conditions by different analytical equipment From: Dr. August. Basel. 2006 Dr. Bonn . BfArM. Validierung in der Analytik.

Birgit Schmauser. BfArM. 2006 Dr.Dissolution • Applicability of the analytical method used for assay and impurities/degradants – Sample preparation – Range • Applicability of the dissolution method – Appropriateness of drug release acceptance criteria • Solubility criteria of the APIs – Appropriateness of test conditions and acceptance criteria • Dissolution affecting bioavailability • Changes in formulation or manufacturing variables affecting dissolution Dar es Salaam. 21-25. August. Bonn .

Dissolution II • Applicability of the analytical method used for assay and impurities/degradants – Potential parameters for revalidation • Sample preparation – Stability of analytes in the dissolution medium? – Preparation of an injectable sample volume according to the analytical method? – Precision of analysis of the prepared dissolution sample? • Range of test concentrations of API / impurities / degradants according to the validated ranges? – Test concentration of prepared dissolution sample versus test concentration of FPP sample Dar es Salaam. 21-25. Bonn . August. BfArM. Birgit Schmauser. 2006 Dr.

21-25.2 to 6. pH 4.Dissolution III • Applicability of the dissolution method – Appropriateness of drug release acceptance criteria • Solubility of the APIs (ICH Q6A Definitions) – Rapidly dissolving products » Not less than 80% of the label amount of the drug substance dissolves within 15 minutes in each of the following media: pH 1.8 – Low solubility drugs » Drugs with a dose/solubility volume of more than 250 ml Dar es Salaam.2. August. Bonn . BfArM. Birgit Schmauser. pH 6.0. 2006 Dr.8 – Highly water soluble drugs » Drugs with a dose/solubility volume of less than or equal to 250 ml over a pH range of 1.

Bonn .Dissolution IV • Appropriateness of drug release acceptance criteria – Solubility of the APIs • Problem with low solubility drugs: – Solution of the drugs may become a time-limiting step » Dissolution also dependent on the strength of the drug product » Dissolution test cannot reflect batch to batch consistency • Possible solution of the problem – Extending the dissolution volume and – Validation of the dissolution procedure with extended volume (applicability of the pharmacopoeial procedure) Dar es Salaam. 21-25. August. 2006 Dr. Birgit Schmauser. BfArM.

. BfArM.3: . Bonn .Dissolution V • Sink conditions – Ph. Birgit Schmauser.9. 21-25. August.the material already in solution does not exert a significant modifying effect on the rate of dissolution of the remainder… – „Sink conditions normally occur in a volume of dissolution medium that is at least 3 to 10 times the saturation medium – Consequently: the amount of API contained in the dosage form should be soluble in NMT 300 ml of dissolution medium Dar es Salaam. Eur 2. 2006 Dr.

Birgit Schmauser. 21-25. August. Bonn .Dissolution V • Applicability of the dissolution method – Appropriateness of test conditions and acceptance criteria (ICH Q6A) • Dissolution significantly affecting bioavailability – Have relevant developmental batches exhibited unacceptable bioavailability? » Development of test conditions and acceptance criteria which can distinguish batches with unacceptable bioavailability • Changes in formulation or manufacturing variables affecting dissolution – Control of these changes by another procedure and acceptance criterion or – Development of test conditions and acceptance criteria which can distinguish these changes Dar es Salaam. BfArM. 2006 Dr.

Major problems • Solubility of Artemisinins – Sink condition cannot be established • (+) Addition of solubilizers could help establish a (dis)solution test • (-) The test would disconnect dissolution and bioavalability and could only serve as parameter for batch to batch consistency • Disintegration could be considered as additional parameter • Stability of Artemisinins – Artesunate decomposes (to DHA) in buffers required for dissolution testing (e.g. 2006 Dr. August. Bonn . 21-25. BfArM. pH 4.0) Dar es Salaam. Birgit Schmauser.5) • Dissolution could only be performed at a neutral pH (~ 7. pH 1.2.

BfArM. Birgit Schmauser. Bonn . 21-25. 2006 Dr.Deficiencies from PQ • Validation of precision – Precision of the drug substance solution lower than precision of the drug product solution – Acceptance criteria for precision of the drug substance solution wider than for precision of the drug product solution – Acceptance criteria much wider than real values assessed – Acceptance criteria of assay specifications and precision do not match • (3 x RSD outside the specification range) Dar es Salaam. August.

Deficiencies from PQ II • Assay of API and impurities/degradants – No acceptable mass balance found between assay of API and impurities/degradants – Quantitation limit of impurities too high • ICH requirement on threshold for identification and qualification of unknown impurities cannot be fulfilled Dar es Salaam. Bonn . 21-25. August. 2006 Dr. Birgit Schmauser. BfArM.

Deficiencies from PQ III • Dissolution – Necessary information on development of dissolution test not presented – Dissolution method (pharmacopoeial) not presented along with development of dissolution test and/or validation of applicability of analytical methods – Test conditions and acceptance criteria of the dissolution test not justified Dar es Salaam. 2006 Dr. August. 21-25. Bonn . Birgit Schmauser. BfArM.

THANK YOU FOR YOUR ATTENTION Dar es Salaam. Bonn . August. Birgit Schmauser. 21-25. BfArM. 2006 Dr.