Analytical Method Validation

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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

1

Impeller



2

2

Impeller



3

2

Impeller



4

Shaft


5

벽면
1


6

벽면
2


7

Sparger



8

P01 valve
부위


9

Sparger



10

배양액

경계벽면


11

P09 line


벽면


12

발효조

상부



6

P 0 1
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


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



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


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, 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)

P01

Milk powder


visual


Rivoflavin


UV detection


Pressure


Visual detection


Photo documentation


Equipment System Design

Consideration of CIP system for effective
cleaning


Piping size


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

D

L

½ ft/sec

5 ft/sec

5 ft/sec



Adequate 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


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


: 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

EL
1
01

E
T
-
101

Capacity



350L

~ET101

150L



처리량
/Batch


[kg/B]

350

3.08

150



처리
Batch



[B/Lot]

6

6

6



처리량
/Lot


x


[ug/Lot]

2.10E+12

1.85E+10

9.00E+11



Surface area


[cm
2
]

53128

10977

41716


C
arryover

10x10
-
6
[ppm]

1.00E
-
0
5

1.00E
-
05

1.00E
-
05

1.00E
-
05



Coupon area


25[cm
2
]

25

25

25



MACO/swab


x
(

/

)x


ug/swab

9882

421

5394



MACO/equip


x

/
(
25

x100
0)

[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)


CO
2

NDIR
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


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

Contaminants

TOC

HPLC

ELISA

PAGE

Lowry

Protein

LAL

Ion
-

Exchange

Media

+

+

-

+

+

-

+

Metabolites

+

+

-

-

-

-

+

Endotoxin

+

-

-

-

-

+


DNA/nucleic acid

+

-

+

-

-

-

+

Carbohydrates

+

+

+

-

-

-

+

Lipids

+

+

+

-

-

-

+

Proteins

Native

Den
aturated


+

+


+

+


+

-


+

-


+

+


-

-


-

-

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.