1. 4/26/2018
Richard R. Goodin
Senior Analytical Chemist,
CRD –Early Development Team
Pharmaceutical Development Services
Rapid Chromatographic Method Development for Pre-Clinical Drug Programs
Pharmaceutical Development Services

2. Agenda Research phase Experimental set-up Run scouting trials
4/26/2018
Agenda
Research phase
Experimental set-up
Run scouting trials
Qualify your methods
Commercial Capacity Swindon

3. 4/26/2018
Analytical Development is an Essential Element to Support Early Drug Development
All active pharmaceutical ingredients must be tested for identity, purity and stability
Most purity indicating methods are chromatographic
You will need an “assay only” fast method and a “stability indicating” method for both your API and your dosage form
When do you need to validate your methods What is “phase appropriate” ?
Commercial Capacity Swindon

4. Early Drug Development Timeline

5. Rapid Chromatographic Method Development for Pre-Clinical Drug Programs
Research Phase

6. THE BEGINNING, If your product is an NCE
4/26/2018
THE BEGINNING, If your product is an NCE
Consult your synthetic chemistry group
They already have an HPLC method! Is there a UV chromophore? Other detection methods (MS, ELSD, fluorescence, refractive index, etc.) ?
They may already know the API process impurities! Can they provide them?
They may even know the stability requirements of your drug (this will require verification)
Consult your formulation group
What excipients do they anticipate?
What are the release characteristics?
Commercial Capacity Swindon

7. Phase I Quality System Attributes
4/26/2018
Phase I Quality System Attributes
Methods need to show your product is:
Free of organic contamination
Synthetic by-products (or unintended chiral products, epimers)
Degraded API products
Free of inorganic contaminants
Stable enough to meet the shelf life needs for the clinical study (24 hrs - 1yr)
Chromatography can determine API label claim content and release (soluble?)
Commercial Capacity Swindon

8. Use All Your Resources to Develop a Method
4/26/2018
Use All Your Resources to Develop a Method
Determine synonyms for your API (IUPAC, CAS, PubChem, DrugBank)
Conduct literature searches, National Library of Medicine, USP, NF, EP, plus structure searches
Look for HPLC methods, columns, mobile phases, GC methods, UV spectroscopy
Commercial Capacity Swindon

9. What Are Your Impurities?
4/26/2018
What Are Your Impurities?
Assemble data on all API lots
Create a chromatographic “history” of the analysis results for different lots
Fully characterize your active ingredients including preservatives (parabens etc.)
Conduct forced degradation studies of your API and your formulations
Conduct high stress excipient compatibility studies.
Commercial Capacity Swindon

10. An Example of an Active Ingredient Specification
4/26/2018
An Example of an Active Ingredient Specification
Test
Method
Specification ID
FTIR
matches
Purity
HPLC
95%-103%
Impurities
Total NMT 5 %
Any one NMT 1%
Residual Solvents
Headspace GC
ICH Class 1-3
Salt form
(dihydrochloride)
Chloride electrode
6-8%
Residue on Ignition
Furnace
<2%
Micro testing
Test for pathogens
CFU or Pyrogens
< 10 CFU /gram
Appearance
Observation
White solid
Commercial Capacity Swindon

11. Rapid Chromatographic Method Development for Pre-Clinical Drug Programs
Scouting Experiments

12. HPLC Method Development Initial Surveys for Reverse Phase
4/26/2018
HPLC Method Development Initial Surveys for Reverse Phase
Begin with one to six different columns
Start with a strong eluting condition and later you may be able to narrow the gradient or even shift to isocratic
For UV absorbance detection, use a diode array instrument to establish UV maximums
Set up many versions of your method for your best test mixture. Spike impurities into your product. Let the instrument do the work
Commercial Capacity Swindon

13. HPLC Method Development Best Configuration for Method development
4/26/2018
HPLC Method Development Best Configuration for Method development
Automated control
Multiple buffers
Multiple columns
Select a test mix
Agilent supports 4 buffers and >6 columns (two column modules)
Waters supports up to 10 solvents and 6 columns in their FUSION software
Both support independent thermal control of each col.
Commercial Capacity Swindon

14. 4/26/2018
HPLC Method Scouting Software “ChemSword” Sergey Galushko, Ralf Honsberg Agilent, Technologies GmbH,
Commercial Capacity Swindon

15. 4/26/2018
HPLC Method Gradient Scouting Software from Agilent Website Sergey Galushko, Ralf Honsberg Agilent, Technologies GmbH,
Commercial Capacity Swindon

16. 4/26/2018
HPLC Method Column Scouting Sergey Galushko , Ralf Honsberg Agilent , Technologies GmbH,
Commercial Capacity Swindon

17. HPLC Method Development UPLC Versus HPLC Advantages and Disadvantages
4/26/2018
HPLC Method Development UPLC Versus HPLC Advantages and Disadvantages
Parameter
HPLC
UPLC
Column Length (cm) 25 5
Column Inside Diameter (mm)
4.6
2.1
Particle Size (µm)
1.7
Injection volume ( µL) 20
Pressure (psi)
1000
10177
Run time (min.) 40
5.44
Equilibration time (min) 10
3.95
UPLC Advantages
Much faster
No loss of performance
UPLC Disadvantages
Instrument cost
Reduced column selection
Less easily transferred contractor may not have capability
Commercial Capacity Swindon

18. UPLC Methods Can be Stability Indicating
4/26/2018
UPLC Methods Can be Stability Indicating
Commercial Capacity Swindon

19. Evaluate Chromatographic Methods During Development
4/26/2018
Evaluate Chromatographic Methods During Development
Parameter
Needed
Resolution
From API, impurities and void volume
Plate Count
Look at peak shape also
Selectivity (polarity, pKa, basic, acidic)
Best column / mobile phase pair
S:N (baseline noise vs peak height)
ASTM E (PK to PK)
Relative Standard Deviation
NMT 2%
Linearity
If non-linear, can it be fitted? quadratic?
Range
NMT 2 AU for UV
LLOQ & ULOQ
LOD for impurities?
Commercial Capacity Swindon

20. Qualify Your Methods for Use
Rapid Chromatographic Method Development for Pre-Clinical Drug Programs
Qualify Your Methods for Use

21. 4/26/2018
Requirements of Your IND Filing Patheon Method Qualification Standard Practices for Phase I
System Suitability
Linearity
Specificity
Range
Accuracy
Repeatability
Solution Stability
Quantitation Limit (if applicable)
Detection Limit (if applicable)
Commercial Capacity Swindon

22. Data Elements Required For Assay, Qualification using USP Categories
Category I: Quantitation of active ingredients (including preservatives) in finished pharmaceutical products
Category II: Purity Methods
Category III: Dissolution methods
Category IV: Identification tests
From Validation of Compendial Methods USP29 <1225>
Also see ICH Q2 (R1)

23. Qualification vs. Validation USP guidelines for Marketed Products
4/26/2018
Qualification vs. Validation USP guidelines for Marketed Products
Analytical
Performance
Characteristics
Assay Category I II
III IV
Quantitative
Limit test
Accuracy
YES * NO
Precision
Specificity
Detection Limit
Quantitation Limit
Linearity
Range
Commercial Capacity Swindon

24. Requirements of Your IND Filing FDA Guidance Doc Nov 1995
4/26/2018
Requirements of Your IND Filing FDA Guidance Doc Nov 1995
Validation data and established specifications ordinarily need not be submitted at the initial stage of drug development
Most companies choose to partially validate as part of their “risk assessment”
Ruggedness and robustness can be delayed to phase II or phase III
Commercial Capacity Swindon

25. Preparing for Your IND Filing FDA Guidance Doc Nov 1995
4/26/2018
Preparing for Your IND Filing FDA Guidance Doc Nov 1995
A brief description of each test method should be prepared
API Lot history
Proposed acceptable limits
Characterization of your Product
Characterization of your API
Commercial Capacity Swindon

26. 4/26/2018
Summary Points
Research many different methods and use all your resources
Scout the variables in your lab to optimize GC or LC conditions
Choose your “best” method
Characterize performance using real samples (not just standards)
Qualify your final choices for Phase I studies
Commercial Capacity Swindon

27. Thank you for Listening – Any Questions?
Thank you for Listening – Any Questions?