Cleaning Validation

Eun-Sook Gi April 12, 2004

Regulatory and Requirements

July 1993 Guide to inspection for validation of cleaning process regulation and requirements: 21CFR 211.65, 21CFR 211.67 jWritten cleaning procedure,21 CFR 211.182
Application FDA,

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



: Product: purity, safety, efficacy, quality Product:cross contamination, previous residual product, microbial residue, detergent, degradant

Cleaning validation ? New equipment/product Changed : process, cleaning process, 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 companys 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

Sampling Location for Swab

No 1 2 3 4 5 6 7 8 9 10 11 12 P09 line
7

Description 1 Impeller 2 2 Impeller Impeller Shaft 1 2 Sparger P01 valve Sparger

Sampling time
11 10

12

6 6 4 5 3 1 9 8
P01

Product/Cleaning Agent
 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

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 : 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 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 training report

Cleaning SOP
Cleaning procedure development (SOP
 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 : : conductivity, pH

Equipment Validation
CIP
 IQ:

validation status

requirement of safety, utility, installation and documentation, accuracy of P&ID etc. 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

 OQ:

Coverage Study (CIP)

jMilk powder visual jRivoflavin UV detection jPressure jVisual detection jPhoto documentation
P01

Equipment System Design

Consideration of CIP system for effective cleaning  Piping size (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 D

Good Best

L
Bad

j Adequate turbulence (Flow rate) for CIP

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  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 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 : 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
Ide ntity No Capacity /Batc h Batc h /Lot S urfac e are a Carryove r Coupon are a MACO/s wab MACO/e quip x x( / )x /(25 x100 0) 10 x1 0 - 6 [ ppm] x [ kg/B] [ B/Lot] [ ug/Lot] [ c m2] 1.0 0E- 05 25 [ c m 2 ] ug/s wab [ mg] EL- 1 0 1 35 0L 35 0 6 2.1 0E+ 12 53 128 1.0 0E- 05 25 98 82 21 000 EL101 ~ ET 101 3.0 8 6 1.8 5E+ 10 10 977 1.0 0E- 05 25 42 1 18 5 ET - 1 0 1 15 0L 15 0 6 9.0 0E+ 11 41 716 1.0 0E- 05 25 53 94 90 00

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  Microbial contamination  Endotoxins  Sanitizing agent  Lubricant  Environmental dust  Residual rinse

active ingredient

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 process-/medium component,

 Inorganic comp:

detergent
 Biol

contaminat: mycoplasma, viral, bacterial, non viral host bacteria

Specific Assays
 Cytotoxicity: to

verify the detoxification of bacterial toxin by heat inactivation specific but poor

 Immunoassay: ELISA,

reproducible
 HPLC:

protein, peptide, nucleic acid, small molecules accuracy, reproducibility, recovery rate very good, 1~2% SD specificity is limited to protein size

 PAGE:

Non-Specific Assays
 TOC:

Determination of various compound or compound class (Pt)/ (uv induced) CO2 NDIR: 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 Media Metabolites Endotoxin DNA/nucleic acid Carbohydrates Lipids Proteins Native Denaturated Stabilizers Cleaning agent Organic Inorganic TOC HPLC ELISA PAGE Lowry Protein LAL IonExchange

+ + + + + + + + + + +

+ + + + + + + + -

+ + + + -

+ + -

+ + + -

+ -

+ + + + + + + +

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  Procedure  Acceptance criteria:

method

training, deviation, batch, ..  Work sheet/equipment: cleaning procedure, raw data record, sampling, analytical procedure, etc.

Cleaning Validation Report

 Introduction  Summary:method

table  Conclusion  Recommendation  Appendices: analytical raw data, chromatogram, etc.

 Results: