Transcript Analytical Method Validation

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
Application
regulation 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
1
Description
1번 Impeller 상
2
2번 Impeller 상
3
2번 Impeller 하
4
Shaft
5
벽면 1
6
벽면 2
7
Sparger 하
8
P01 valve 부위
9
Sparger 상
10
배양액 경계벽면
11
P09 line 옆 벽면
12
발효조 상부
Sampling time
12
11
10
6
6
4
2
3
5
1
9
7
8
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
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
 Coupon:
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 No
계산
단위
Capacity
EL-101
EL101
ET-101
350L
~ET101
150L
① 처리량/Batch
[kg/B]
350
3.08
150
② 처리 Batch수
[B/Lot]
6
6
6
[ug/Lot]
2.10E+12
1.85E+10
9.00E+11
[cm2]
53128
10977
41716
1.00E-05
1.00E-05
1.00E-05
1.00E-05
25[cm2]
25
25
25
③ 처리량/Lot
① x ②
④ Surface area
⑤ Carryover
10x10-6[ppm]
⑥ Coupon area
⑦ MACO/swab
⑤x(③/④)x⑥
ug/swab
9882
421
5394
⑧ MACO/equip
⑦x④/(25 x1000)
[mg]
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
Protein
Org compound
Inorg compound
Biol Systems
Assays
Bioassay, ELISA, HPLC
PAGE, Absorbance, TOC
TOC, HPLC, UV-Abs., TDS
Conductivity, pH,
o-Phosphate, ICP, TDS
Viable cell analysis
Commonly used anal. Method for
biopharm.Contaminants and Impurities
Impurities and
Contaminants
TOC
HPLC
ELISA
PAGE
Lowry
Protein
LAL
IonExchange
Media
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Metabolites
Endotoxin
DNA/nucleic acid
Carbohydrates
Lipids
Proteins
Native
Denaturated
Stabilizers
Cleaning agent
Organic
Inorganic
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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.