Principles of HPLC Validation

Transcription

1 Principles of HPLC Validation A short course in the systematic validation of HPLC methods COURSE HANDOUT: [SECTION 2] Method Development and Pre-Validation The Plan Pre-Validation System Suitability

2 Section 1. Introduction to Validation Section 2. Method Development and Pre-Validation The Plan Pre-Validation System Suitability Section 3. Validation and Documentation Section 4. Quality by Design and Robustness Section 5. Data Analysis Principles of HPLC Method Validation 2-1 Without a Plan If you don t know where you are going, any road will take you there. --Lewis Carroll 2-2 Some Things Before You Start Development Who is on the team? Existing methods Regulatory documents SOPs End-user information How will method be used? Validation requirements Timeline Budget 2-3

3 The Development Team Team leader Lab staff End user representative Validation team representative QA/QC? 2-4 Method History Existing methods internal Method documents Method development reports Validation reports Literature methods Ancillary information C of A MSDS Physical properties 2-5 Regulatory Documents & SOPs ICH, FDA, EP, USP, JP, BP SOPs Method development? Validation? Documentation New SOPs? 2-6

4 End-User Requirements Who will use the method? Internal or external client? What equipment is available? Equipment limitations? Skill limitations? Geographic, language barriers? What USP/ICH category is the method? 2-7 Method Use & Required Validation One-off Walk-up Once a month Daily 10 s, 100 s, or 1000 s of samples Dedicated or shared equipment What stage of development will it support? Level of validation required? 2-8 Timeline and Budget When is a final method needed? When is validation to start? Are sufficient resources available? Equipment Lab space Personnel Budget Beware of less than average thinking 2-9

5 HPLC Method Development Advanced HPLC Method Development Using Quality by Design Snyder, Glajch, & Kirkland, Practical HPLC Method Development, 2 nd ed. (Wiley, 1997) Snyder, Kirkland, & Dolan, Introduction to Modern Liquid Chromatography, 3 rd ed. (Wiley, 2010) Snyder & Dolan, High-Performance Gradient Elution, (Wiley, 2007), Chapter Section 1. Introduction to Validation Section 2. Method Development and Pre-Validation The Plan Pre-Validation System Suitability Section 3. Validation and Documentation Section 4. Quality by Design and Robustness Section 5. Data Analysis Principles of HPLC Method Validation 2-11 Pre-Validation Don t gamble! Perform pre-validation experiments first. 2-12

6 Some Possible Pre-Validation Activities Draft method document Draft validation protocol Confirm method performance Linearity, range Precision, accuracy Robustness Durability Method tweaking Method hand-off 2-13 Document Preparation Draft method Transfer from notebook Add details Put in formal method format Draft validation protocol May have blanks to fill in Acceptance criteria (for pre-validation) 2-14 Confirm Method Performance Linearity and range Method limits Precision and accuracy Repeatability Some intermediate precision Robustness Durability Equivalent column? 2-15

7 Finding an Equivalent Column: The Hydrophobic-Subtraction Model log k/k EB = η H - σ S* + β A + α B + κ C η H O O O O O σ S* hydrophobic steric B: H O O O O H O X O BH + O O - O hydrogen bonding cation exchange β A α B κ C 2-16 Comparing Column Selectivity measure H, S*, A, B, and C for each column how to compare? for neutral samples, only H, S*, A important (H 1, S* 1, A 1 ) (H 2, S* 2, A 2 ) difference in selectivity = F s = distance between two points * F s = {(H 2 -H 1 ) 2 + (S* 2 -S* 1 ) 2 + (A 2 -A 1 ) 2 } 1/2 so: small F s, similar columns large F s, different columns 2-17 Characterizing Column Selectivity

8 Identifying Equivalent Columns Ace C18 F s = 0.0, R s = Develosil ODS-UG-5 F s = 1.7, R s = 1.6 Hypersil BetaBasic 18 F s = 2.2, R s = Selecting an Orthogonal Column Original method 5 * Orthogonal method F s = (min) Method and Document Tweaking Sample preparation Instrument settings Mobile phase adjustments Method instructions Validation protocol changes then. Method Hand-Off 2-21

9 Section 1. Introduction to Validation Section 2. Method Development and Pre-Validation The Plan Pre-Validation System Suitability Section 3. Validation and Documentation Section 4. Quality by Design and Robustness Section 5. Data Analysis Principles of HPLC Method Validation 2-22 System Suitability Dress for Success Good System Suitability Test advance assurance that the sample batch will run successfully fails early Bad System Suitability Test passes, but does not really forecast problems 2-23 System Suitability The evaluation of the components of an analytical system to show that the performance of a system meets the standards required by a method. 2-24

10 System Suitability (ICH) System suitability test parameters to be established for a particular procedure depend on the type of procedure being validated. Validation of Analytical Procedures: Text and Methodology Q2(R1) 2-25 Required System Suitability Parameters 2-26 Common System Suitability Parameters Plate number, N Resolution, R s Peak tailing, TF Retention time, t R Retention factor, k Pressure, ΔP Peak area or height Precision (n = 3-6) 2-27

11 System Suitability (FDA) Parameter Recommendation* k k > 2 R s R s > 2 T f T f 2 N N > 2000 Precision RSD 1% (n = 5) * Reviewer Guidance: Validation of Chromatographic Methods (FDA) 2-28 System Suitability: Importance of k finished product drug standard 2-29 System Suitability: Importance of N (and TF) TF = 1.0 TF = 1.5 N = 7500 R s = 2.06 N = 6750 R s = 1.96 N = 5000 R s =

12 System Suitability: Tailing and Resolution R s = 1.5 R s = 1.5 R s = 1.2 R s = 1.5 R s = 1.0 R s = 1.0 R s = 1.2 R s = R s Resolution and Peak Height 1 / 1 10 / / / System Suitability: What Makes Sense? TF = 1.0 vs. 2.0 Area = 1:10 vs. 10:1 2-33

13 Section 1. Introduction to Validation Section 2. Method Development and Pre-Validation The Plan Pre-Validation System Suitability Section 3. Validation and Documentation Section 4. Quality by Design and Robustness Section 5. Data Analysis Principles of HPLC Method Validation 2-34

14 For more information and to discuss your training requirements, contact: FOR NORTH AMERICA & EUROPE Dean Graimes T: M: FOR ASIA PACIFIC Jeroen Reiniers T: Frederick House, Princes Court Nantwich, CW5 6PQ, UK 20 Maxwell Road, Maxwell House Singapore