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 failure 확인

Sampling Location for Swab

No Description Sampling time 1 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

6

P 01

1

2 3

4

5

7 8

9

10

11

12

6

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: Equipment 와 동일 surface 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 (SOP 정량화 )

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, pressureend point End point 측정 : conductivity, pH

Equipment ValidationCIP 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)

P 01

Milk powder visual

Rivoflavin UV detection

Pressure

Visual detection

Photo documentation

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

DL

½ ft/sec 5 ft/sec 5 ft/sec

Adequate turbulence (Flow rate) for CIPAdequate turbulence (Flow rate) for CIP

Bad Good Best

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

Dead Leg Orientation

Branch piping : horizontal

Good Design

Vertical up

Vertical down

horizontal

Bad 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 product 에 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

Identity No 계산 단위 EL- 101 EL101 ET- 101

Capacity 350L ~ET101 150L

① 처리량/Batch [kg/B] 350 3.08 150

② 처리 Batch수 [B/Lot] 6 6 6

③ 처리량/Lot ① x ② [ug/Lot] 2.10E+12 1.85E+10 9.00E+11

④ Surface area [cm2] 53128 10977 41716

⑤ Carryover 10x10- 6[ppm] 1.00E- 05 1.00E- 05 1.00E- 05 1.00E- 05

⑥ Coupon area 25[cm2] 25 25 25

⑦ MACO/swab ⑤x(③/④)x⑥ ug/swab 9882 421 5394

⑧ MACO/equip ⑦x④/(25 x1000) [mg] 21000 185 9000

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

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) CO2NDIR 측정 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 AssaysProtein 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 Contaminants

TOC HPLC ELISA PAGE Lowry Protein

LAL Ion- 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.