CEM LABORATORY

TNG CC MI TRNG
TR
TRUNG TM QUAN TRC MI TRNG
TR

XC NHN GI TR S DNG
CA PHNG
PH
PHP
(Method Validation)

Ngi trnh by: on Vn On

H Ni, 30/8/2013

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Ni dung

1. Xc nhn gi tr s dng (MV)

MV l g?
Ti sao thc hin MV l cn thit?
Khi no th cn thc hin MV?
2. Thng s c trng khi thc hin MV
chm (Precision)
chn lc (Selectivity)
chch (Bias)
nhy (Sensitivity)
tuyn tnh & Khong lm vic (Linearity & Working Rang
Gii hn pht hin (Detection Limit)
Gii hn nh lng (Quantitation Limit)
Yu t nh hng (Ruggedness and Robustness)
3. c lng khng m bo o
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MV l g?

Khng nh bng kim tra v cung cp bng chng khch quan

rng cc yu cu xc nh cho vic s dng phng php l
ph hp vi mc ch v iu kin phn tch c th c
thc hin.

chm
chn lc
chch
nhy
tuyn tnh & Khong lm vic
Gii hn pht hin
Gii hn nh lng
Cc yu t nh hng

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Ti sao cn thit phi thc hin MV?

Tnh khoa hc: Nguyn l khoa hc; tch chnh xc; tnh n
nh

Tnh thc tin: Phm vi ca phng php; ngun lc thc

hin; s an ton; nhu cu ca khch hng/chp nhn ca php
l; kh nng so snh v ph bin; thi gian v gi thnh
Cung cp cc kt qu tin cy m bo rng bt k mt
quyt nh no a ra da trn kt qu l ng tin cy
p ng yu cu ca ISO/IEC 17025

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Khi no cn thc hin MV?

MV

Phng php mi i vi mt vn c th
Chnh sa phng php c
- Ci tin
- Mi rng phm v
Khi QC ch ra rng phng php thay i theo thi gian
Khi phng php thit lp c a vo s dng
- Trong mt PTN khc
- Vi ngi phn tch khc.
- Thit b khc
chng t s tng ng
gia 2 phng php

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Tin trnh thc hin

Xc nh cc yu cu phn tch
La chn phng
ph
php p dng
Gii php thc hin MV

1. Thm nh li phng php MV trc

2. nh gi li phng php khi c bt k mt s thay i no
3. Xc nhn gi tr s dng khi thay i t phng php ny
sang phng php khc
4. MV mt phng php mi

K hoch thc hin MV

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Tin trnh thc hin (tip)

Tin hnh thc hin MV

1. nh gi thng tin ph duyt (so snh cc s liu to ra v
cc yu cu)
2. Nu cn thit, ci tin phng php p ng yu cu
3. Khi cc yu cu c p ng cn phi vn bn ho
phng php v kt qu ph duyt phng php
Xc nh th tc kim sot cht lng
l

S dng phng php tin cy, cng vi cc quy trnh kim so

cht lng ph hp v tham gia th nghim thnh tho
cung cp bng chng tin cy lin tc.

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Thng s c trng thc hin MV i vi cc

php phn tch

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Mu th nghim
Mu thc hin MV phi p ng cc yu cu sau:
C lng mu ph hp

c dn nhn ph hp
i din cho loi mu ang c th nghim
c vn chuyn v lu gi di cc iu kin c th duy tr tnh
nguyn vn ca mu
c phn tch trong khong thi gian ph hp tnh t
thi im ly mu
Vt liu/Ha cht c thm vo n nh mu cn phi
nhn din y
Vt liu c s dng cho thit b ly mu cn phi c quy nh
Ngi phn tch cn c mt s hiu bit, nhn thc v khng
m o lin quan trong qu trnh ly mu v n nh hng n s
bin i c th c trong qu trnh phn tch
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chm (Precision)
Mc gn nhau gia cc kt qu th nghim c lp thu
c trong cng iu
iu kin quy nh
chm thng c biu din bi cc thut ng sau
lch chun (s)
lch chun tng i (RSD) hoc h s bin thin (CV)

lch chun trung bnh (SDM) ca cc th nghim lp c

bit nh sai s chun ca cc gi tr trung bnh

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chm/ lp li (Repeatability)
lp l php o s bin i ca cc kt qu o trong thi
gian ngn

lp li l chm ca cc kt qu c o di iu
kin:
- Cng phng php
- Cng vt liu th nghim
- Cng phng th nghim
- Cng ngi thc hin
- Trong khong thi gian ngn

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chm/ ti lp (Reproducibility)
ti lp l php o s bin i ca cc kt qu o trong
thi gian di

ti lp l chm ca cc kt qu c o di iu
kin:
- Cng phng php
- Cng vt liu th nghim
- Khc phng th nghim
- Khc ngi thc hin
- Khc thit b o
- Trong thi gian di

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chm/ chm trung gian (Intermediate)

chm trung gian l php o s bin i ca cc kt qu o

trong thi gian di trong mt phng th nghim

chm trung gian l chm ca cc kt qu c o d

iu kin:
- Cng mt PTN
- Cng phng php
- Cng vt liu th nghim
- C s thay i ln v iu
kin o
Khc ngi thc hin
Khc ngy thc hin
Khc thit b o
- Trong khong thi gian di (1-3
(1 thng)

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chm - nh hng ca cc iu kin khc nhau

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c tnh chm

S lng cc php phn tch ln, tin cy c tnh chm

cng ln

7 - 15 php phn tch c lp cn c xc nh

Yu cu ti thiu trong phn tch nc
n
l 10 bc t do (n-1)
chm ph thuc vo nn mu v nng phn tnh, c
tnh trn ton di nng lm vic.

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30

RSD (Reproducibility) %

20
10
0
-10
-20

100 %

10 %

1%

0.1 %

100 ppm

10 ppm

1 ppm

100 ppb

-30
10 ppb

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chm Vs Nng cht phn tch

Nng cht phn tch

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chch (Bias)
Mc sai khc gia k vng ca cc kt qu th nghim v
gi tr qui chiu c chp nhn
Nghin

cu chch l nghin cu tng sai s h thng. S

sai khc h thng so vi gi tr quy chiu c chp nhn
cng ln th chch cng ln

chch gm:
chch phng th nghim
chch phng php
chch c th m hoc dng

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chch (tip)
Nguyn nhn sai s
Mu phn tch
Phng php
Thit b o
thu hi khng hon ton
Phng th nghim hoc ngi
ng
phn tch
Sai s tng th c xc nh t
Phn tch cc cht chun bit trc nng (CRM) c
nn ph hp
Th nghim xc nh thu hi mu thm chun
So snh lin phng
Phng php i chng c lp
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c lng sai s t vic thm chun

R2-R1

(R4)

R4 lng cht thm chun

Sai s = R3 - R4
V d: Xc nh Atrazine trong nc
Sai s = (45.9 ng/L 32.4 ng/L) 15.0 ng/L
Sai s = 13.5 ng/L 15.0 ng/L
Sai s = -1.5 ng/L
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X l vi cc sai s
Khi c mt sai s ln m khng th loi b
Ngi s dng s liu phn tch cn phi c cnh bo s
tn ti ca chng
Hiu chnh php o
o hng ngy
Bo co gi tr o
o v sai s ring r
khng m bo o
o (MU) kt hp vi hiu chnh sai s
c th s c tnh n

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ng ( chch) Vs chm

Inaccurate &
Khng
ng v
imprecise
Khng Chm

Inaccurate but

Chm v
Khng
precise
ng

Accurate but
ng
v Khng
imprecise
Chm

Principles of Method Validation Part 1

ng v Chm

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chn lc (Selectivity)
c trng ca mt phng php: Mc m n c th
pht hin v xc nh r rng mt cht phn tch nht nh
trong mt hn hp, m khng c s nh hng t cc
thnh phn khc
Ph hp th nguyn t, Ph pht x nguyn t, Ph hunh quang,
Ph khi

vs.

time
Principles of Method Validation Part 1

time
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Khong tuyn tnh v khong lm vic

(Linearity and Working range)

Khong tuyn tnh: khong nng ca cht phn tch m

phng php phn tch cho kt qu phn tch t l thun vi
nng cht phn tch.
Khong lm vic: khong nng ca cht phn tch m
thit b cho tn hiu p ng tt.
Tnh ton tuyn tnh v khong lm vic
Phn tch cc mu chun bit nng
Ti thiu phn tch ti 6 cp nng
V th t cc kt qu thu c
- nh gi tuyn tnh bng h s hi quy r2 > 0,99
- Xc nh khong tuyn tnh tng i, cn trn v cn
di ca khong lm vic.
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Khong tuyn tnh v khong lm vic

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Gii hn pht hin (LoD) v Gii hn nh lng (LoQ)

Gii hn pht hin (LoD) l nng cht phn tch nh nh

c th pht hin c nhng khng nht thit xc nh h
lng trong iu kin th nghim.
Gii hn nh lng (LoQ
LoQ) l nng thp nht ca cht ph
tch trong mu c th nh lng c vi chm v
ng chp nhn c trong iu kin tin hnh phn tch
Gii hn pht hin (LoD):
(LoD Gii hn pht hin ca thit
(IDL) v Gii hn pht hin ca phng php (MDL)
Gii hn nh lng (LoQ
LoQ): Gii hn nh lng ca thit
(IQL) v Gii hn nh lng ca phng php (MQL)

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Gii hn pht hin (LoD) v Gii hn nh lng (LoQ)

LoD = 3 x S/N
LoQ = 3 LoD = 9 S/N
(Da vo lch chun ca nhiu)

lch chun ca
nhiu SD

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Tnh ton gii hn pht hin ca phng php (MD

EPA 40 CFR Part 136.. APPENDEX B, revision 1.11

1. D on MDL ca cht phn tch
2. Chun b 2 mu thm chun (Spike) c nng bng 5 ln gi t
MDL d on
3. Phn tch 2 mu ny theo quy trnh phn tch v ghi nhn kt qu ph
tch
4. Nu hiu sut thu hi t yu cu, chuyn sang bc tip theo. N
khng t yu cu, quay tr li bc (1) v chn MDL cao hn.
5. Chun b t nht 7 mu thm chun c nng nh bc (2) v ph
tch theo quy trnh phn tch
6. Ghi nhn kt qu v tnh ton MDL
Kim tra li gi tr MDL thu c
1. MDL < Spike < 10xMDL
2. S/N c nm trong khong yu cu khng? 2,5 10?
3. Hiu sut thu hi c nm trong khong yu cu khng? 70 110?
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V d tnh ton MDL

Phn tch Toluen trong nc
bng GC/MS
MDL c tnh l 0,040 ug/L
Nng Spike 0,21 ng/L vo
nn nc ct 2 ln

MDL < Spike < 10x MDL

0,084 < 0,21 < 0,84
S/N = 0,20/0,029 = 7,0
AVE H = 96,9%

Principles of Method Validation Part 1

Mu
Mu 1
Mu 2
Mu 3
Mu 4
Mu 5
Mu 6
Mu 7
Mu 8
Mu 9
AVE
SD
MDL
S/N

Kt qu
ug/L
0,23
0,21
0,24
0,19
0,18
0,22
0,17
0,16
0,23
0,20
0,029
0,084
7,0

H%

110
100
114
90
86
105
81
76
110
96,9

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Yu t nh hng (Robustness and Ruggedness)

Xem xt nhy ca cc thng s c trng ca phng

php i vi nhng thay nh ca mi trng v iu kin
phn tch m khng hoc t nh hng n kt qu phn
tch.
Ch ra mc n nh ca phng php trong qu trnh s
dng

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Ti liu tham kho

EURACHEM Guide 1998
The Fitness for Purpose of Analytical Methods

IUPAC Technical Report 2002

Harmonized Guideline for Single Laboratory Validation of
Methods, Pure Appl. Chem.
Chem 2002, 74, 835-855

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XIN TRN TRNG CM N!

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