Cleaning Validation

Eun-Sook Gi
April 12, 2004

㈜ 삼양제넥스 생명공학 연구소

Regulatory and Requirements
FDA, July 1993
Guide to inspection for validation of cleaning
process

Applicationregulation and requirements:

21CFR 211.65, 21CFR 211.67

Written cleaning procedure,21 CFR 211.182

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
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
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
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)
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
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
Sampling Location for Swab

No Description Sampling time 12

1 1번 Impeller 상
11
2 2번 Impeller 상
3 2번 Impeller 하 10
4 Shaft
6
5 벽면 1 6
6 벽면 2
4
7 Sparger 하
8 P01 valve 부위 5
3 2

9 Sparger 상
1
10 배양액 경계벽면 9
11 P09 line 옆 벽면 7 8
12 발효조 상부 P01
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
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
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
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
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
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
Coverage Study (CIP)

Milk powder visual

Rivoflavin UV detection
Pressure
Visual detection
Photo documentation

P01
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
Instrument Tees
 Instrument Tee for CIP: L/D <1.5

L D

Bad Good Best

 Adequate turbulence (Flow rate) for CIP

½ ft/sec 5 ft/sec 5 ft/sec

<Cleaning and cleaning validation: A biotechnology perspective, pub PDA, 1996>

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>

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
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
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
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= 0.00001(mg/mg) x 200 000 000 (mg)

= 2000 (mg)

<Ref: APIC 2000: cleaning validation guidance>

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

Identity N o 계산 단위 E L -101 E L 101 E T -101

C apacity 350L ~ E T 101 150L
① 처리량/B atch [kg/B ] 350 3.08 150
② 처리 B atch수 [B /L ot] 6 6 6
③ 처리량/L ot ①x ② [ug/L ot] 2.10E + 12 1.85E + 10 9.00E + 11
④ S urface area [cm 2 ] 53128 10977 41716
⑤ C arryover 10x 10 - 6 [ppm ] 1.00E - 05 1.00E - 05 1.00E - 05 1.00E - 05
⑥ C oupon area 25[cm 2 ] 25 25 25
⑦ M A C O /sw ab ⑤x (③/④)x ⑥ ug/sw ab 9882 421 5394
⑧ M A C O /equip ⑦x ④/(25 x 100 0) [m g] 21000 185 9000
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>
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>

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

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)
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
Commonly used anal. Method for
biopharm.Contaminants and Impurities
Impurities and TOC HPLC ELISA PAGE Lowry LAL Ion-
Contaminants Protein Exchange
Media + + - + + - +
Metabolites + + - - - - +
Endotoxin + - - - - +
DNA/nucleic acid + - + - - - +
Carbohydrates + + + - - - +
Lipids + + + - - - +
Proteins
Native + + + + + - -
Denaturated
+ + - - + - -
Stabilizers + + - - - - +
Cleaning agent
Organic + + - - - - +
Inorganic
+ - - - - - +

J. of Parental Science & Technology, 13-19(45), 1991

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