validation general

7/23/2019 Validation General

1/59

1

International Federation forConsulting GmbH IFC

ValidationValidation

by

Dr. Michael Scheutwinkel


2/59

2

eferences

eferences

EURACHEM Guide

The Fitness for Purpose of Analytical Methods -

A Laboratory Guide to Method alidation

and Related Topics !"#$"%%&'((()eurache*)co*

C+TAC $ EURACHEM GuideGuide to ,uality in Analytical Che*istry

An Aid to Accreditation !##'

((()eurache*)co*


3/59

3

alidation is the confir*ation by e.a*ination

and the pro/ision of ob0ecti/e e/idence that

the particular re1uire*ents for a specific

intended use are fulfilled)

alidation !+23$+EC "4#5'

Definition in clause 5.4.5.1 of ISO/IEC 17025

6)fit for purpose


4/59

4

Validation of an analytical test method

Which analytes can be dete!ined in "hich !ati#in the $esence of "hich intefeences%

Within these conditions "hat le&els of $ecision

and accuacy can be achie&ed%

'alidation is the $ocess of establishin( the

) *efo!ance chaacteistics

) +i!itations of a !ethod

) Identification of the influences "hich !aychan(e these chaacteistics and to "hat e#tent.


5/59

5

E,ui$!ent and !ethod &aldiation

6 different ite*s

Method /alidation

2oft and hard(are

E1uip*ent /alidation

+,P,

3,

7,


6/59

6

'alidation in ode to analytical !ethod

alidation

Modified standardi8ed and

in-house *ethods

erification

2tandardi8ed-*ethods


7/597

'eification of an analytical test !ethod

-he tools ae the sa!e as fo &alidation

the laboatoy needs only to &eify that

the docu!ented $efo!ance chaacteistics

can be !et e.(. accuacy.

'eification is the $ocess to de!onstate the

co!$etence in test $efo!ance

of an aleady &alidated standadied test !ethod.


8/598

When should be &alidated%

) Chan(es in established !ethods

) e" !ethod de&elo$ed fo a $aticula $oble!

) Established !ethod used in diffeent

laboatoies diffeent e,ui$!ent o diffeentstaff

) Out3of3contol situations "ithin intenal ,ualityassuance

) on3successful $atici$ation in *-s

) De!onstation of e,ui&alence bet"een t"o!ethods e.(. a a$id ne" test a(ainst a standad

!ethod


9/599

'alidation State(y

Chaacteiation

of the test !ethod

Co!$aison

of !ethod

chacacteistics

"ith

e,uie!ents

'eification

to (i&e

$oof that the

e,uie!ents ae

fulfilled

9 C 7

S$ecific test e,uie!ents defined by custo!e o !aet

A


10/5910

7escription of test *ethod !clause 5):):'

Identification / Sco$e *aa!etes in "hich !ati#

ea(ents and !ateials C6sE,ui$!ent and en&ion!ental

conditions

CalibationSa!$lin( and

sa!$le $e$aation

-est $efo!anceincludin( safety instuctions

uality assuance

Docu!entation e$otin(

e&aluation citeia

8ncetainty o $ocedue fo

its esti!ation


11/5911

-ools

7eter*ination of *ethod characteristics

Chaacteistics

of calibation

Chaacteistics

of the an(e

Chaacteistics

of accuacy

Selecti&ity o

S$ecifity

obustness /u((edness


12/5912

-ools


Chaacteistics

of calibationLinearity2ensiti/ity

Perfor* *ore

point calibrationCoelation

coefficient9: ; 0.


13/5913

Commission Decision !""!#$%'C


14/5914

Calibration

) Calibation &alues should be a$$lied "ithin the "oin( an(e.

) +o"est $oint should be the li!it of ,uantification.

) =o calibation cetified standad solutions shall be used.

) In addition also standad solutions ha&e to be added to blan

!ateials analysed o&e all $ocess ste$s.

) =o linea calibation functions at least fi&e concentations ae necessay.

) -est calibation functions fo lineaity and hi(hli(ht failin(

lineaity.

'alidation of testin( !ethods


15/5915

Sensiti(ity )resolution*


16/5916

Linearity

Definition based on sa!$les "ith &ayin( concentations

and the calculation of the e(ession of esults.

Si(nal and concentation do not ha&e to be fully elated.

=i&e standads suffice to $oduce a calibation cu&e if

lineaity is o..

6oe standads ae necessay once the lineaity is

unsatisfactoy.

E#a!ine e$eat sa!$les and standads o&e a $aticula

"oin( an(e to e#$eience "hethe a eliable line can be

da"n bet"een $oof and detection li!it.



17/5917

Ho*o;eneity of /ariances

Chaacteistic &alues fo the co!$aison

of standad &aiations fo diffein(

concentation.

In case of (eat diffeences bet"een

standad de&iations

in3ho!o(eneity of &aiances

the (or>in; ran;e needs to be split)


18/5918

-ools


Chaacteistics

of calibation

Chaacteistics

of the an(e

Li*it of detection

Li*it of 1uantification

Chaacteistics

of accuacy

Selecti&ity o

S$ecifity



19/5919

in; ran;e

=indin( out the a$$o$iate "oin( an(e usin(

) diffeent !atices

) &ayin( concentations.

Concentation an(e "ithin the achie&e!ent of

acce$table

) accuacy and

) $ecision is $ossible.

+n ;eneral the (or>in; ran;e is broader than the linear ran;e)

alidation of testin; *ethods


20/5920

+rere,uisites for calibration

Standads !ust be faultless.

*ecision has to be si!ila thou(hout the entie "oin( an(e.

-he !odel function is a$$licable9 eithe linea o cu&ed.

Eos only !ay occu ando!ly "ithin si(nals.

Eos ha&e to follo" the no!al distibution.


+OD


21/5921

+OD

Characteristics of the ran;e

Li*it of detection

+OD #+OD> ?)s+/ b @lan &alue !ethod

#+OD> 4 )s#o Calibation function !ethod

#+d> +i!it of detection

s+> Standad de&iation of the blans

S#o> Standad de&iation of the calibation functionb > Slo$e of calibation function

'aious

con&entionsA+OD> 6ean of blans B ? # S+

bycalculation

+O


22/5922

+O


Li*it of 1uantificationby

calculation

#+O> 11 )s#o Calibation function !ethod

#+O> +i!it of ,uantification

s+> Standad de&iation of the blans

S#o> Standad de&iation of the calibation functionb > Slo$e of calibation function

'aious

con&entionsA+O > 5 o o 10 # S+

+O

C i i D i i !""!#$%'C


23/59

23

Commission Decision !""!#$%'C

+OD


24/59

24

+OD


Li*it of detection by

si;nal to

noise ratio

Fro* 5 ? " to " ? "

Li*it of 1uantification

Fro* # ? " to 5 ? "


25/59

25

Effect of peak shape on LO ! LO"

- l


26/59

26

-ools


Chaacteistics

of calibation

Chaacteistics

of the an(e

Chaacteistics

of accuacy

2electi/ity or

2pecifity

obustness ou((edness

S l ti it / S ifit


27/59

27

Selecti&ity / S$ecifity

2electi/ity2pecifity

8se of

antibodies

E#tent to "hich $aticula analytes

can be dete!ined in co!$le# !i#tues. !ethod "hich is selecti&e fo an analyte

is said to be s$ecific.

Selecti&e detectos

in instu!ental

analysis

Selecti&e !edia in

!icobiolo(y

Cross-reactions =

Th diff b t l ti it d ifi it


28/59

28

Selecti(ity

) selecti&e !ethod (i&es

coect esults fo all

inteestin( analytes "heeas a

s$ecific test !ethod (i&escoect esults fo the

inteestin( analyte "heeas

othe analytes !i(ht intefee

each othe.

The difference bet(een selecti/ity and specificity

S-ecifity

) Outlines the analytical

e#tent to "hich an

analytical substance o

substance (ou$ can be

dete!ined "ithout

intefeence fo! sa!$le

elated co!$onents.

@o proble* usually both (ords are *i.ed 6

lid ti f t ti th d


29/59

29

Selecti(ity

) Outlines the e#tent to "hich an analytical substance

can be dete!ined "ithout intefeence fo! othe

co!$onents.

) 6ethod selecti&e to one analytical substance

s$ecific.

) ssess selecti&ity "hile a$$lication to $ue solutionu$ to co!$le# !atices.

) Docu!ent distubances and estictions of the

!ethod.


-ools


30/59

30

-ools


Chaacteistics

of calibation

Chaacteistics

of the an(e

Chaacteistics

of accuacy

Selecti&ity o

S$ecifity




31/59

31

Robustness

) *at of the outine.

) +ean fo! othe analytical institutes as the !ethod3establishin( laboatoy "ill ha&e $o&ed

obustness befoe $ublishin( a ne" !ethod.

7e;ree of proneness of a *ethod toconscious and unconscious alterations

of a (or>in; instruction)


Robustness


32/59

32

Robustness

2ensiti/ity de;ree of the test *ethod a;ainsts*all de/iations in e.peri*ental conditions)

E#a!$les9

) -i!es "ithin $ocess ste$s

) En&ion!ental conditions e.(. te!$eatues

) 6ino $ocess chan(es e.(. $ flo" ates in *+C

Ru;;edness accordin; to U2P


33/59

33

Ru;;edness accordin; to U2P

Ru;;edness is the de;ree of reproducibility obtainedunder a /ariety of conditions e.pressed as relati/e 27

e);)?

) diffeent analysts

) diffeent e,ui$!ent

) diffeent tade !as of ea(ents

) etc.

+nternal reproducibility

-ools


34/59

34

-ools


Chaacteistics

of calibation

Chaacteistics

of the an(e

Chaacteistics

of accuacy9

Correctness

Uncertainty

Precision

!Repeatability$Reco/ery'

Reproducibility

Selecti&ity o

S$ecifity

obustness o

u((edness

ccuacy > *ecision and coectness I


35/59

35

ccuacy > *ecision and coectness I

Accuracy

E.actness of an

analytical *ethod

Precision

7e;ree of

repeatability of an

analytical *ethod

UncertaintyReproducibility

Reco/ery $

9ias

Repeatability

Correctness $

Trueness

2yste*atic error $

Correct /alue


36/59

36

p#ecise

andco##ect

not p#ecise

$%tco##ect

p#ecise

$%t

on'

not p#ecise

and

on'

(andom and systematic de)iations

ccuacy > *ecision and coectness II
http://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPThttp://../!!!VORTR%C3%A4GE/SKC/POWERPOINT_PR%C3%A4SENTATION/POWER_POINT_HOCHFORMAT_1.PPT


37/59

37

Characteristics of accuracy

Coect3

ness

*ecision

6easue of coectness co&ein( syste!atic and

non3syste!atic !istaes use of C6 co!$aison

to a "ell chaacteied !ethod

6easue of the de(ee of e$eatability

co&ein( syste!atic !istaes

e$eatability9 one laboatoyF

@ette9 intenal e$oducibility

e$oducibility9 !oe than one laboatoy

ccuacy *ecision and coectness II



38/59

38

Repeatability

6easue!ent of efeence !ateials.

=otification of a blan sa!$le "ith the analytical substance.

Co!$aison to a efeence $ocedue.

*atici$ation in a co3o$eati&e intelaboatoy test in"hich the Gtue contentH is no"n.

8se at least one of the follo"in( $ocedues fo dete!ination9




39/59

39

Precision

Info!ation on accuacy of t"o coincidin( andinde$endent analytical esults.

*ecision de$ends on the concentation of the analyte.

e$eatability is $at of the $ecision "ith es$ect to

e$eated !easue!ents9

sa!e !ateial

sa!e !ethod sa!e analyst

sa!e laboatoy and

shot ti!e3s$an in3bet"een the analyses.




40/59

40

Correctness $ Trueness

8se the one of the follo"in( !ethods to assue coectness9

) nalysis of cetified efeence !ateial.

) *atici$ation in intelaboatoy co!$aisons.

) Co!$aison to a no"n $ocedue.

If not a&ailable9

) Docu!ent any data that $o&e coectness.) 6ae an a$$o#i!ation as a fist a$$oach to coectness.

) *uchase efeence !ateial.

) 8se a si!ila !ethod in $aallel.




41/59

41

Reco/ery

) *efo!ed by addition of the analytical substance to the !ati#.

) !ati# shall be fee of the analytical substance.

) If not the sa!$le "ill be eniched "ith a "ea concentation of

the analytical sa!$le.

) O a si!ulated !ati# "ill be used.

) eco&ey can be dete!ined only "hen the analytical sa!$le is

a&ailable in $ue fo!.

) +ist the eco&ey ate in J and the standad de&iation "hen

eco&ey ates ae constant.

) Othe"ise issue eco&ey ate as a function.




42/59

42

Fra*e conditions for the

deter*ination of the reco/ery rate

-he analytical substance needs to be added in

that fo! in "hich it occus in natue.

-he sa!$les e,uies (ood ho!o(enisation.

-he nati&e content shall be belo" the

dete!ination li!it.




43/59

43

Reproducibility

Statistically

Co!$aati&e standad de&iation e$licate standad de&iation

-hou(h $atici$ation in $oficiency testin( sche!es

o $atici$ation in intelaboatoy co!$aisons.


internally

e.ternally

Pro/e reproducibility

-ools


44/59

44

-ools


Chaacteistics

of calibation+ineaitySensiti&ity

Chaacteistics

of the an(e+i!it of detection

+i!it of ,uantification

Chaacteistics

of accuacyCoectness8ncetainty*ecision

e$eatability/

eco&eye$oducibility

Selecti&ity o

S$ecifity

obustness o

u((edness



45/59

45

Classification of the test method

) nalysis of conta!inants e.(. $esticides hea&y !etals

food additi&es

) nalysis of food constituents salt and !ineals

) 6ethods "hich ae dete!inin( tue &alues

) -est !ethods "hich ha&e to be calibated "ith instu!ents

) *hysical !easue!ent $ &alue conducti&ity

) ualitati&e dete!inations

) 6icobiolo(ical !ethods

7istin;uish bet(een




46/59

46


*a#amete# +nalysis of

t#ace

contaminants

+nalysis of

constit%ents

,on)entional

methods

-ethods not

necessa#y to

cali$#ate

*hysical

methods

"%alitati)e

methods

.acte#iolo'ical

methods

esc#iption of

test method

/ / / / / / /

,ali$#ation / / / /

0o#kin' #an'e / / / / / / /

etection limit / / / /

ete#mination

limit

/ /

(eco)e#y / / /(epeata$ility / / / / / /

,o##ectness / / / /

1pecificity / / / / /

(ep#od%ci$i lity if possi$le if possi$le if possi$le if possi$le if possi$le if possi$le if possi$le

Para*eters for /alidation



47/59

47

Common (alidation needs of

known methods

Method .ccuracy +recision S-ecificity#Selecti(ity

/inearity Detection# Determinationlimits

obustness

++12 +E12 3,*2 (4 / / 5/6 / / /

*ola#o'#aphy! Voltamet#y / / / / /

7it#ation / / /

0ate# assay / / / /

,h#omato'#aphy! ,apilla#yElect#opho#esis

/ / / / /

1pect#oscopy / / / / /

(elease / / / / /

7O, / / / /

En8yme 3mm%no +ssays / / / / / /


*ocedues fo !ethod &alidation


48/59

48

0he nalysis of Standard eference Materials

) Keneally acce$ted !ethod fo &alidation.

) Such standads ae $o&ided "ith (uaantee on

the !aet.

) It !ay be necessay to contact the $e$aation

of a uni,ue sa!$le in $aticula !ati# in ode

to utilise this $ocedue fo !ethod &alidation.

) -he analyst !ust de!onstate that the !ethod

$o&ides accuacyand$ecision.




49/59

49

The participation in PT-sche*es or in alaboratory Collaborati/e 2tudy

-he !ost "idely acce$ted $ocedue fo !ethod

&alidation.

Seious $actical da" bacs9 Costly and ti!e consu!in( Effot in co3odination Shi$$in( of sa!$les and data Statistical analysis and inte$etation of esults.

-his !ethod is aely used fo the fist desci$tion

of a !ethod in the liteatue.




50/59

50

Co*parison (ith a currently accepted *ethod

) 8sually done by one analyst

) ort"o usin( a s$lit3sa!$le.

) usin( esults fo! the cuently acce$ted !ethod

fo &eification.(ee!ent su((ests &alidation.

Disa(ee!ent could also su((est that thecuently acce$ted !ethod is in&alid.

) In such case anothe $ocedue has to bee!$loyed fo the !ethod &alidation.

) -he !oe sa!$les ae analysed and the"ide the an(e of concentation the hi(he thecedibility of the &alidation !ethod



51/59

51

The Bero-9lind Method

) One analyst.

) 8sin( sa!$les at no"nle&els of analyte to de!onstateeco&ey accuacy and $ecision.

) =ast si!$le and usefulbut it leads to subLecti&e esults.

) Suitable fo a fist a$$o#i!ation e,uiin( !ini!al ti!e!an$o"e sa!$les and cost.

) In (eneal a (ood stat fo the o&eall &alidation $ocess.

+f this *ethods fails there is no reason to proceed (ith

further /alidation of the *ethod)

Procedures for *ethod /alidation


52/59

52

The 2in;le-9lind Method

) One analyst at the stat.

) Sa!$les ae (i&en to a 2ndanalyst to "ho! le&els of analyte aeunno"n.

) esults ae co!$iled and co!$aed by the 1stanalyst.

) Co!$aison by the fist analyst !aes obLecti&ity ,uestionable.

) -his !ethod still is biased on behalf of the 1stanalyst.

) Suitable afte the eo3blind!ethod has been successful andbefoe additional analysts o the !ana(e!ent "ill be in&ol&ed.



53/59

53

The 7ouble-9lind Method

) -hee analysts.

) 1stanalyst $e$aes sa!$les at no"n le&els.

) 2ndanalyst does the actual analysis.

) ?danalyst o ad!inistato co!$aes bothdata fo! the fist t"o analysts.

) Only the ?d

analyst has access to these data.

6ost effecti&e a$$oach to obLecti&eness



54/59

54

Clearance of /alidated test procedures

alidation

results

Custo*er

re1uire*ents

uality 6ana(e

(



55/59

55

(

7ecision criteria for the e.tent of a /alidation

-echnical feasibility and staff dis$osition%

nalytical &ie"$oints 6easuin( $inci$le !ethod co!$le#ity%

Occuence of the sa!$le "ide3s$ead%

is $otential fo client/ co!$any%

*olitical decisions ctual ai! of the &alidation%

Consideation of no!ati&e

and othe e,uie!ents%

facts

s%pe#:o#dinated c#ite#ia



56/59

56

E.tent of /alidation in analytical sciences

*u$ose

) Suitability of the !ethod

) M6aetabilityG of the !ethod

nalytical sensibility

Costs

e,uie!ents fo! the outside

;


57/59

57

alidation is al(ays a balance bet(een

costs ris>s and technical possibilities)

The /alidation shall be co*pleted

by a state*ent by the laboratory that the*ethod is fit for the intended use)

+23$+EC "4#5 states?


58/59

58

2elf de/eloped soft(are or e.cel

sheets ha/e to be /alidated)

Records for that*ust be a/ailable)

7o not for;et?

If'+ 1orksho-2 +roficiency 0esting 3 4ackground and general as-ects


59/59

1hy -roficiency testing5

* schemes a#e ;%st one impo#tant tool in

validation general

Documents