1. Damiana Gentili Qualified Person/QU Director 09/05/2019
L’importanza della scelta del metodo analitico nella cleaning validation
Damiana Gentili
Qualified Person/QU Director
09/05/2019

2. Agenda Specific vs non-specific Visual inspection
Regulatory framework
Specific vs non-specific
Visual inspection
Selection criteria for analytical methods & case history
Analytical Validation overview
Opportunities

3. FDA outcomes on analytical methods for CLEANING VALIDATION
Failure to adequately validate written procedures for the cleaning and maintenance of equipment

4. Italy

5. Analytical tecniques overview
Product
Active ingredient
Specific
HPLC
Mass Spectrometry
Gas Chromatography
Non-specific
Total Organic Carbon
UV-VIS
Conductivity/pH
Excipients

6. Analytical tecniques overview
Detergent
Specific
HPLC
Mass Spectrometry
Gas Chromtography
Non-specific
Total Organic Carbon
UV-VIS
Conductivity/pH
Microbial
Live microbes
Bioburden
Microbe breakdown
Endotoxin

7. Detection of residues: from 1992 up to date… Specific vs. Non-specific

8. Specific vs. Non-specific Testing
Variety of residues
Only one specific analyte
Worst case results
Expected by Regulatory Authority
Highly sensitive
Highly sensitive (sometimes)
Less costly
Costly
Little sample preparation
Sample preparation required
Easy to manage
Required technical skills
Monitoring purposes
Validation purposes

9. Characteristics Visible residue
This testing is designed to make observations directly from the clean equipment for quick turnaround
Visual inspection using a visible residue limit (VRL) can monitor equipment cleaning efficiency
The VRL is the level below which the residue of interest is not visible to the equipment
Characteristics
A VRL inspection is non-specific
Any visible residue is considered unacceptable
Several of its applications and associated risks have been demonstrate
Since this is a limit test, the detection limit is the primary validation parameter.
Precision should be also determined by using multiple observers in order to minimise subjectivity of the observers
Subjectivity and training of the inspectors is the primary concern with this process
Results from validation activity can be used to establish the difference between the
Analytical Residue Limit and the Validation Residue Limit?

10. Analytical Methods - main criteria selection
The key factors in selecting a method are:
Number of components to be analyzed
Sampling technique
Sample preparation processes
Frequency and number of samples
Data handling
… besides sensitivity and selectivity

11. Case history : UPLC-MS Selection criteria:
High level of sensitivity requested due to a low LOD/ LOQ requested. LOQ in HPLC too high!
High level of specificity requested due to 3 different contaminants to be detected
Evaluation of sample preparation:
Organic vs inorganic/soluble in water or other solvents
Avoid sample manipulation (i.e. sample was in Pyridine/DMSO which would affect HPLC sensitivity)
Swab recovery verification
Is Analyte stable in the cleaning environment? Or does it degrade due to temperature during cleaning?
Speed of analysis (throughput) as requirement  5 minutes analysis time
Available instrumentation  yes, UPLC-MS (more than one)
Operational cost: Time of QC development (pre-linearity, LOD and LOQ verification, swab recovery at 3 concentrations)  ab. 2 weeks

12. Final output Chromatographic conditions
The chromatographic procedure may be carried out using:
- column: ACQUITY UPLC BEH C18, 100 × 2.1 mm, 1.7 μm or equivalent;
- column temperature: 30°C;
- Mobile phases:
Mobile phase A: 0.1% formic acid in water (for example transfer 1.0 ml of formic acid 98% into 1000 ml water);
Mobile phase B: 0.1% formic acid in acetonitrile (for example transfer 1.0 ml of formic acid 98% into 1000 ml water);
- Detection: MS (settings are optimized for Waters Acquity QDa Detector), Single Ion Recording (SIR)
Ionization Mode: ESI +; Probe Temp. 600 °C; Capillary 0.8 kV; Cone: 15 V;
Acquisition Type: Selected Ion Recording (SIR);
MASS: amu M984 MASS: amu N5.1 MASS: amu N5
- Injection volume: 1 µl;
- Flow rate: 0.5 ml/min;
- Run time: 5 min

13. Test method validation for CV samples
Testing cleaning validation samples requires a validated method
The extent of validation is dependent upon
the type of method employed
the capabilities of the method
the scientific and regulatory needs of the resulting data
the anticipated outcome of the testing

14. Validation of Analytical Methods
It is essential to validate the analytical methods chosen for cleaning validation and routine monitoring in the ranges encountered during cleaning
The minimum requirement for method validation must include the following tests:
LOQ. In practice it may not be necessary to determine the LOQ but to show that it sufficiently low for the current purposes when sensitive techniques are being used
LOD
Specificity for chromatographic or UV methods
Accuracy
Repeatability
Recovery from spiked surfaces
Stability of the analyte in the cleaning solvent

15. Where we are as API manufacturer
We use specific analytical method rather than non-specific
We use solvents as cleaning agents and very low use of detergents
We use high level of sophisticated equipment
NEW OPPORTUNITIES TO EXPLORE
Use non-specific methods/easiest analytical tecniques for monitoring purposes
Use detergents to reduce safety issues in handling solvents as cleaning agents
Correlate monitoring results with validation data
Increase the level of control of cleaning

16. Thank you very much gentili@procos.it
Tempio del padiglione d'oro
Thank you very much