1. Cleaning Validation
Eun-Sook Gi
April 12, 2004
㈜ 삼양제넥스 생명공학 연구소

2. Regulatory and Requirements
FDA, July 1993
Guide to inspection for validation of cleaning process
Application regulation and requirements: 21CFR , 21CFR
Written cleaning procedure,21 CFR

3. Reference
Cleaning and cleaning validation: A biotechnology perspective, pub PDA, 1996
www. cleaningvalidation.com
Guidance on aspects of cleaning validation in active pharmaceutical ingredient plants, APIC, 2000
1) Cleaning validation an exclusive publication,
2) J of validation technology: Cleaning validation II pub by Inst. of Validation Technology

4. Cleaning Validation Overview
목적:
Product: purity, safety, efficacy, quality 유지
Product:cross contamination, previous residual product, microbial residue, detergent, degradant 잔류 방지
언제 Cleaning validation 실시?
New equipment/product
Changed : 생산 process, cleaning process, 세척제
Changed : major component

5. Cleaning validation Overview
Good system design
Master validation plan
Preliminary study
- Coupon study
- Cycle Development study
- Continuous data monitoring
- Justifiable acceptance criteria
Effective cleaning process development
Adequate analytical technique

6. Master Validation Plan
Appropriate cleaning procedure
Identification of cleaning agent
Description of sampling procedure
Acceptance criteria
Analytical method
A copy of protocol and final report
A list of equipment
Manufacturing process( a flow diagram)

7. Master Validation Plan
CIP or COP
Equipment matrix for CV
Description of equipment and its location
Surface area of equipment
Average batch size of each equipment
Training program for production and analytical personnel
Reference to the company’s change control program

8. Sampling Procedure Sampling SOP Rinse or swab(surface) sampling
Rinse sampling: volume, valve define
Swab sampling: map of each equipment
the most-difficult-to-clean, easy-to-clean
“Easy-to-clean”: cleaning failure확인

9. Sampling Location for Swab

10. Product/Cleaning Agent 관계
Molecular structure (Bio product or small molecule)
Prod related compound
Solubility: in water or in org. solvent?
Reactivity
Contaminant: Fluid or Solid?
Cleaning agent selection

11. Coupon Study
Coupon: Equipment와 동일surface type SUS or Glass (5 x 5 cm, 10 x10 cm)
Swab: polyester
Characterization of residue
Worst case of cleaning condition
Selection of cleaner visually clean
Cleaning condition 설정: temp. range, cleaning agent conc., pressure(agitation, shaking), rinse volume visual inspection

12. Swab Test / Swab Recovery
Swab method: sample, control and blank test
Spiking of known quantity of analyte/residue
Swab recovery test (70~130%)  recovery factor CV 실시 시 반영
Estimation of swab sample solution stability
Estimation of swab extraction time
Residue limit TOC/prod specific

13. Cycle Development Study
Performed prior to process validation
Characteristic of residuals
Cleaning agent select and concentration
Rinse cycle time and volume
Cleaning agent temperature
CIP-Pressure (Turbulence)
Cleaning procedure development
Cleaning-SOP change or improve
Continuous data monitoring Acceptance limit
Operator training 및 training report

14. Cleaning SOP Cleaning procedure development (SOP정량화)
Pre-rinse: volume, pressure
Soaking: alkaline/acid, organic solvent or other detergent volume, temperature, soaking time
Rinse recycle: time, temperature
Rinse volume, pressure end point
Final rinse volume, pressureend point
End point 측정: conductivity, pH

15. Equipment Validation CIP 설비의 validation status 확인
IQ: requirement of safety, utility, installation and documentation, accuracy of P&ID etc.
OQ: Test of flow rate, volume of washes and rinses, temperature(inlet/outlet), turbulence, heating time of cleaning solution, gas flow, purges
Spray ball: coverage study

16. Coverage Study (CIP) Milk powder visual Rivoflavin UV detection
Pressure
Visual detection
Photo documentation

17. Equipment System Design
Consideration of CIP system for effective cleaning
Piping size 와 구조(slope)
Potential dead leg
Turbulence of CIP solution
Nozzle design: locate seal near vessel wall
Branch piping

18. Instrument Tees  Adequate turbulence (Flow rate) for CIP
Instrument Tee for CIP: L/D <1.5 L D
Bad
Good
Best
 Adequate turbulence (Flow rate) for CIP
5 ft/sec
½ ft/sec
5 ft/sec
<Cleaning and cleaning validation: A biotechnology perspective, pub PDA, 1996>

19. Dead Leg Orientation Branch piping : horizontal Vertical up Bad design
Vertical down
horizontal
Good Design
<Cleaning and cleaning validation: A biotechnology perspective, pub PDA, 1996>

20. Recommended Drain Line Size
Drain line: locate vessel drain high enough to slope down to CIP return pump
- 100L tank: 1.0 inch
- 1,000L tank: 1.5 inch
- 10,000L tank: 2.0 inch
Sampling valve: Diaphram valve
Design: CIP visually 확인 가능한 설계
Cleaning and cleaning validation: A

21. Acceptance Criteria Visually clean Cleaning capability
Sample test time limit
Cleaning time limit: DEHT, CEHT
Number of batches: 3consecutive batch
Deviation handling
Allowed contaminant limit
- General limit
- Maximum daily dose
- Toxicity based carry over

22. Allowable Contaminant Limit
General ppm Limit:toxicological data for intermediate are not known, API product에 적용
MACOppm= MAXCONC x MBS
MACOppm: maximum allowable carry over from previous product, calculated from general ppm limit
MAXCONC:general limit for maximum allowed concentration of “previous” substance to next batch
MBS: Minimum batch size for the next product

23. Allowable Contaminant Limit
General ppm limit
MAXCONC is often set to 5~100ppm depending on toxicity and pharmacological activity
MAXCONC for API : 10ppm is very frequent
계산 예: MBS of next product: 200kg
MACOppm= (mg/mg) x (mg)
= 2000 (mg)
<Ref: APIC 2000: cleaning validation guidance>

24. Allowable Contaminant Limit
General limit (10ppm carry over) for swab area
10mg/kg x MBS(kg)/equipment surface area x Swab area

25. Allowable Contaminant Limit
Based on Therpeutic Daily Dose
MACO= TDDprev x MBS/SF x TDDnext
- MACO: Maximum allowable carry over
- TDDprev: Std therapeutic dose of inv. prod
- TDDnext: Std therapeutic daily dose for next prod
- MBS: Minimum batch size for next prod
- SF: Safety factor (normally 1000 in calculations based on TDD)
<Ref: APIC 2000: cleaning validation guidance>

26. Allowable Contaminant Limit
Based on Toxicological Data
NOEL= LD50(g/kg) x 70kg/2000
MACO= NOEL x MBS/SF x TDSnext
- NOEL: no observed effect level
- 2000: an empirical constant
- TDSnext: Largest normal daily dose for next prod
- Safety factor : parental: 1000~10000
oral prod: 100~1000
topicals: 10~100
<Ref: APIC 2000: cleaning validation guidance>

27. What is being removed Active ingredient
Decomposition product of active ingredient
Microbial contamination
Endotoxins
Sanitizing agent
Lubricant
Environmental dust
Residual rinse water

28. Type of Analytes
Proteins: active, inactive but intact, fragmented protein, as contaminating intra-/extra cellular protein
Organic comp: cellular comp. DNA, RNA, endotoxin, carbohydrate, lipid, other org compound
Inorganic comp: process-/medium component, detergent
Biol contaminat: mycoplasma, viral, bacterial, non viral host bacteria

29. Specific Assays
Cytotoxicity: to verify the detoxification of bacterial toxin by heat inactivation
Immunoassay: ELISA, specific but poor reproducible
HPLC: protein, peptide, nucleic acid, small molecules accuracy, reproducibility, recovery rate very good, 1~2% SD
PAGE: specificity is limited to protein size

30. Non-Specific Assays
TOC: Determination of various compound or compound class 연소산화(Pt)/습식산화(uv induced) CO2NDIR측정 by 1700cm-1
Colorimetric protein assay: binding to dye(Blue G250), determine spectrophotometrically by 595nm
Coductivity: simple and effective for measurement of residual inorganic material
pH, UV/VIS
TDS(Total dissolved solids)

31. Category of Analysis Type of analyte Assays
Protein Bioassay, ELISA, HPLC PAGE, Absorbance, TOC
Org compound TOC, HPLC, UV-Abs., TDS
Inorg compound Conductivity, pH,
o-Phosphate, ICP, TDS
Biol Systems Viable cell analysis

32. Commonly used anal. Method for biopharm.Contaminants and Impurities
J. of Parental Science & Technology, 13-19(45), 1991

33. Analytical Method Validation
ICH Q2A/Q2B
Accuracy
Precision
Linearity and Range
Specificity (no need to perform by TOC-method)
LOD/LOQ
Intermediate precision
Sample solution stability
Allowable acceptance limit > LOQ

34. Cleaning Validation Protocol
Objective
Scope
Reference inclusive SOP
Responsibility
Material and method
Procedure
Acceptance criteria: training, deviation, batch, …..
Work sheet/equipment: cleaning procedure, raw data record, sampling, analytical procedure, etc.

35. Cleaning Validation Report
Introduction
Summary:method
Results: table
Conclusion
Recommendation
Appendices: analytical raw data, chromatogram, etc.