-
CEM
LA
BO
RA
TO
RY
XC NHN GI TR S DNG
TNG CC MI TRTRUNG TM QUAN TRC MI TR
CEM
LA
BO
RA
TO
RY
XC NHN GI TR S DNG
CA PH
(Method Validation)
H
CEM
LA
BO
RA
TO
RY
XC NHN GI TR S DNG
TNG CC MI TRNGTRUNG TM QUAN TRC MI TRNG
XC NHN GI TR S DNG
CA PHNG PHP
(Method Validation)
Ni, 30/8/2013
Ngi trnh by: on Vn Onh
-
CEM
LA
BO
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TO
RY
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?
CEM
LA
BO
RA
TO
RY
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
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
tuyn tnh & Khong lm vic
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
1. Xc nhn gi tr s dng (MV)
Ti sao thc hin MV l cn thit?
cn thc hin MV?cn thc hin MV?
2. Thng s c trng khi thc hin MV
(Selectivity)
chch (Bias)
tuyn tnh & Khong lm vic (Linearity & Working Range)
Page 2
tuyn tnh & Khong lm vic (Linearity & Working Range)
(Detection Limit)
(Quantitation Limit)
Yu t nh hng (Ruggedness and Robustness)
3. c lng khng m bo o
-
CEM
LA
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TO
RY
MV l g?
Khng nh bng kim tra vrng cc yu cu xc nhph hp vi mc ch v iuthc
hin.
CEM
LA
BO
RA
TO
RY
thc hin.
chm
chn lc
chch
nhy
tuyn tnh & Khong lm vic
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
tuyn tnh & Khong lm vic
Gii hn pht hin
Gii hn nh lng
Cc yu t nh hng
v cung cp bng chng khch quancho vic s dng phng php l
iu kin phn tch c th c
tuyn tnh & Khong lm vic
Page 3
tuyn tnh & Khong lm vic
hng
-
CEM
LA
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TO
RY
Ti sao cn thit phi thc hin
Tnh khoa hc: Nguyn lnh
Tnh thc tin: Phm vi ca
CEM
LA
BO
RA
TO
RY
Tnh thc tin: Phm vi cahin; s an ton; nhu cul; kh nng so snh v
ph
Cung cp cc kt qu tin quyt nh no a ra da
p ng yu cu ca ISO/IEC 17025
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
p ng yu cu ca ISO/IEC 17025
Ti sao cn thit phi thc hin MV?
l khoa hc; tch chnh xc; tnh n
ca phng php; ngun lc thcca phng php; ngun lc thcca khch hng/chp
nhn ca php
ph bin; thi gian v gi thnh
tin cy m bo rng bt k mtda trn kt qu l ng tin cy
ISO/IEC 17025
Page 4
ISO/IEC 17025
-
CEM
LA
BO
RA
TO
RY
Khi no cn thc hin MV
Phng php mi i vi mt vn
Chnh sa phng php
- Ci tin
CEM
LA
BO
RA
TO
RY
- Ci tin
- Mi rng phm v
Khi QC ch ra rng phng php
Khi phng php thit lp
- Trong mt PTN khc
- Vi ngi phn tch khc.
- Thit b khc
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
- Thit b khc
chng t s tng
cn thc hin MV?
vi mt vn c th
Chnh sa phng php c
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.
Page 5
ng gia 2 phng php
-
CEM
LA
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TO
RY
Tin trnh thc hin
Xc nh cc yu cu phn tch
La chn ph
CEM
LA
BO
RA
TO
RY
Gii php
1. Thm nh li phng php2. nh gi li phng php3. Xc nhn gi tr s
dng
sang phng php khc4. MV mt phng php mi
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
4. MV mt phng php mi
K hoch thc hin MV
cc yu cu phn tch
La chn phng php p dng
php thc hin MV
php MV trc php khi c bt k mt s thay i no
dng khi thay i t phng php nykhc
mi
Page 6
mi
K hoch thc hin MV
-
CEM
LA
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TO
RY
Tin hnh
1. nh gi thng tin phcc yu cu)
Tin trnh thc hin (tip)C
EM
LA
BO
RA
TO
RY
cc yu cu)2. Nu cn thit, ci tin3. Khi cc yu cu c
phng php v kt qu
Xc nh th tc kim sot cht l
S dng phng php tin cy,
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
S dng phng php tin cy,cht lng ph hp v thamcung cp bng chng tin
cy
hnh thc hin MV
ph duyt (so snh cc s liu to ra vi
Tin trnh thc hin (tip)
phng php p ng yu cuc p ng cn phi vn bn hoqu ph duyt phng php
th tc kim sot cht lng
cy, cng vi cc quy trnh kim sot
Page 7
cy, cng vi cc quy trnh kim sottham gia th nghim thnh tho cy lin
tc.
-
CEM
LA
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TO
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Thng s c trng thc hin MV i vi cc php phn tch
CEM
LA
BO
RA
TO
RY
Principles of Method Validation Part 1
CEM
LA
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RA
TO
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Thng s c trng thc hin MV i vi cc
Page 8
-
CEM
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Mu th nghim
Mu thc hin MV phi p ng
C lng mu ph hp
c dn nhn ph hp
i din cho loi mu ang c
CEM
LA
BO
RA
TO
RY
i din cho loi mu ang c
c vn chuyn v lu ginguyn vn ca mu
c phn tch trong khong
thi im ly mu
Vt liu/Ha cht c thmnhn din y
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
nhn din y
Vt liu c s dng cho thit
Ngi phn tch cn c mtm o lin quan trong qubin i c th c trong
qu
ng cc yu cu sau:
c th nghimc th nghim
gi di cc iu kin c th duy tr tnh
khong thi gian ph hp tnh t
thm vo n nh mu cn phi c
Page 9
thit b ly mu cn phi c quy nh
mt s hiu bit, nhn thc v khngtrnh ly mu v n nh hng n strnh phn
tch
-
CEM
LA
BO
RA
TO
RY
chm (Precision)
Mc gn nhau gia cc kt qu th nghim c trong cng iu kin quy
chm thng c biu din bi cc thut ng sau
CEM
LA
BO
RA
TO
RY
lch chun (s)
lch chun tng i
lch chun trung bnh (SDM) ca cc th nghim lp
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
lch chun trung bnh (SDM) ca cc th nghim lp bit nh sai s chun ca
cc gi tr trung bnh
chm (Precision)
gn nhau gia cc kt qu th nghim c lp thu iu kin quy nh
biu din bi cc thut ng sau
(RSD) hoc h s bin thin (CV)
lch chun trung bnh (SDM) ca cc th nghim lp c
Page 10
lch chun trung bnh (SDM) ca cc th nghim lp csai s chun ca cc gi
tr trung bnh
-
CEM
LA
BO
RA
TO
RY
chm/ lp li (Repeatability)
lp l php o s bin i ca cc kt qu o tronggian ngn
lp li l chm ca cc kt qu c o di iu
CEM
LA
BO
RA
TO
RY
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
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
- Trong khong thi gian ngn
chm/ lp li (Repeatability)
bin i ca cc kt qu o trong thi
lp li l chm ca cc kt qu c o di iu lp li l chm ca cc kt qu c o di
iu
Cng vt liu th nghim
Cng phng th nghim
Trong khong thi gian ngn
Page 11
Trong khong thi gian ngn
-
CEM
LA
BO
RA
TO
RY
ti lp l php o s bin i ca cc kt qu o trongthi gian di
chm/ ti lp (Reproducibility)C
EM
LA
BO
RA
TO
RY
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
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
- Khc ngi thc hin
- Khc thit b o
- Trong thi gian di
bin i ca cc kt qu o trong
chm/ ti lp (Reproducibility)
ti lp l chm ca cc kt qu c o di iu
Cng vt liu th nghim
Khc phng th nghim
Page 12
-
CEM
LA
BO
RA
TO
RY chm trung gian l php o s
trong thi gian di trong mt phng th nghim
chm trung gian l chm ca cc kt qu c o di
chm/ chm trung gian (Intermediate)C
EM
LA
BO
RA
TO
RY
chm trung gian l chm ca cc kt qu c o di iu kin:
- Cng mt PTN
- Cng phng php
- Cng vt liu th nghim
- C s thay i ln v
Khc ngi thc hin
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
Khc ngy thc hin
Khc thit b o
- Trong khong thi gian di (1
l php o s bin i ca cc kt qu o di trong mt phng th nghim
chm trung gian l chm ca cc kt qu c o di
chm/ chm trung gian (Intermediate)
chm trung gian l chm ca cc kt qu c o di
Cng vt liu th nghim
iu kin o
thc hin
Page 13
Khc ngy thc hin
Trong khong thi gian di (1-3 thng)
-
CEM
LA
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TO
RY
chm - nh hng ca cc iu kin khc nhauC
EM
LA
BO
RA
TO
RY
Principles of Method Validation Part 1
CEM
LA
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RA
TO
RY
nh hng ca cc iu kin khc nhau
Page 14
-
CEM
LA
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TO
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c tnh chm
S lng cc php phn tch ln, cng ln
CEM
LA
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RA
TO
RY
7 - 15 php phn tch c
Yu cu ti thiu trong phn tch n
chm ph thuc vo nn mu v nng phn tnh, c tnh trn ton di nng lm
vic.
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
cc php phn tch ln, tin cy c tnh chm
c lp cn c xc nh
Yu cu ti thiu trong phn tch nc 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.
Page 15
-
CEM
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chm Vs Nng cht phn tch
RS
D (
Rep
rod
uci
bil
ity)
%30
20
CEM
LA
BO
RA
TO
RY
RS
D (
Rep
rod
uci
bil
ity)
%
10
0
-10
-20
-30
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
-30
10
pp
b
10
0 p
pb
1 p
pm
Nng cht phn tch
chm Vs Nng cht phn tch
Page 16
10
pp
m
10
0 p
pm
0.1
%
1 %
10
%
10
0 %
Nng cht phn tch
-
CEM
LA
BO
RA
TO
RY
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
CEM
LA
BO
RA
TO
RY
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
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
chch c th m hoc dng
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 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 phng th nghim
chch phng php
chch c th m hoc dng
Page 17
chch c th m hoc dng
-
CEM
LA
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TO
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chch (tip)
Nguyn nhn sai s
Mu phn tch
Phng php
CEM
LA
BO
RA
TO
RY
Thit b o
thu hi khng hon ton
Phng th nghim hoc ng
Sai s tng th c xc nh
Phn tch cc cht chun nn ph hp
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
nn ph hp
Th nghim xc nh
So snh lin phng
Phng php i chng
thu hi khng hon ton
Phng th nghim hoc ngi phn tch
nh t
Phn tch cc cht chun bit trc nng (CRM) c
Page 18
thu hi mu thm chun
chng c lp
-
CEM
LA
BO
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TO
RY
c lng sai s t vic thm chun C
EM
LA
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RA
TO
RY
R4 lng cht thm chun
Sai s = R3 - R4V d: Xc nh Atrazine trong nc
(R4)
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
V d: Xc nh Atrazine trong nc
Sai s = (45.9 ng/L 32.4 ng/L)
Sai s = 13.5 ng/L 15.0 ng/L
Sai s = -1.5 ng/L
sai s t vic thm chun
R2-R1
lng cht thm chun
V d: Xc nh Atrazine trong nc
Page 19
V d: Xc nh Atrazine trong nc
32.4 ng/L) 15.0 ng/L
15.0 ng/L
-
CEM
LA
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TO
RY
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 tn ti ca chng
CEM
LA
BO
RA
TO
RY
Hiu chnh php o hng ngy
Bo co gi tr o v sai s ring r
khng m bo o (MU) kt hp vi hiu chnh sai s c th s c tnh n
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY Khi c mt sai s ln m khng th loi b
s dng s liu phn tch cn phi c cnh bo s
o hng ngy
o v sai s ring r
o (MU) kt hp vi hiu chnh sai s
Page 20
-
CEM
LA
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TO
RY
ng ( chch) Vs chmC
EM
LA
BO
RA
TO
RY
Inaccurate &imprecise
Khng ng v Khng Chm
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
Inaccurate butprecise
Accurate butng v Khng Chm v Khng ng
ng ( chch) Vs chm
Page 21
Accurate butimprecise
ng v Chm
ng v Khng Chm
-
CEM
LA
BO
RA
TO
RY c trng ca mt phng
pht hin v xc nh rtrong mt hn hp, mthnh phn khc
chn lc (Selectivity)C
EM
LA
BO
RA
TO
RY
thnh phn khc
Ph hp th nguyn t, PhPh khi
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
time
phng php: Mc m n c thrng mt cht phn tch nht nhkhng c s nh hng t
cc
chn lc (Selectivity)
vs.
pht x nguyn t, Ph hunh quang,
Page 22
vs.
time
-
CEM
LA
BO
RA
TO
RY
Khong tuyn tnh v khong lm vic(Linearity and Working range)
Khong tuyn tnh: khongphng php phn tch chonng cht phn tch.
CEM
LA
BO
RA
TO
RY
nng cht phn tch.
Khong lm vic: khongthit b cho tn hiu p ng
Tnh ton tuyn tnh v
Phn tch cc mu chun
Ti thiu phn tch ti 6
V th t cc kt qu
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
V th t cc kt qu
- nh gi tuyn tnh
- Xc nh khong tuyndi ca khong lm
Khong tuyn tnh v khong lm vic(Linearity and Working range)
khong nng ca cht phn tch mcho kt qu phn tch t l thun vi
khong nng ca cht phn tch mng tt.
v khong lm vic
chun bit nng
6 cp nng
qu thu c
Page 23
qu thu c
tnh bng h s hi quy r2 > 0,99
tuyn tnh tng i, cn trn v cnlm vic.
-
CEM
LA
BO
RA
TO
RY
Khong tuyn tnh v khong lm vicC
EM
LA
BO
RA
TO
RY
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
Khong tuyn tnh v khong lm vic
Page 24
-
CEM
LA
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TO
RY
Gii hn pht hin (LoD) v
Gii hn pht hin (LoD)c th pht hin c nhnglng trong iu kin th
CEM
LA
BO
RA
TO
RY
Gii hn nh lng (LoQtch trong mu c th nhng chp nhn c trong
Gii hn pht hin (LoD(IDL) v Gii hn pht hin
Gii hn nh lng (LoQ(IQL) v Gii hn nh lng
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
(IQL) v Gii hn nh lng
hn pht hin (LoD) v Gii hn nh lng (LoQ)
(LoD) l nng cht phn tch nh nhtnhng khng nht thit xc nh hm
nghim.
LoQ) l nng thp nht ca cht phnnh lng c vi chm v
trong iu kin tin hnh phn tch
(LoD): Gii hn pht hin ca thithin ca phng php (MDL)
LoQ): Gii hn nh lng ca thitlng ca phng php (MQL)
Page 25
lng ca phng php (MQL)
-
CEM
LA
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TO
RY
Gii hn pht hin (LoD) v
LoD = 3 x S/N LoQ = 3 LoD = 9 S/N(Da vo lch chun ca nhiu)
CEM
LA
BO
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TO
RY
Principles of Method Validation Part 1
CEM
LA
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TO
RY
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)
Page 26
lch chun ca nhiu SD
-
CEM
LA
BO
RA
TO
RY EPA 40 CFR Part 136. APPENDEX B, revision
1. D on MDL ca cht phn
2. Chun b 2 mu thm chunMDL d on
Tnh ton gii hn pht hin ca phng php (MDL)C
EM
LA
BO
RA
TO
RY
3. Phn tch 2 mu ny theo quytch
4. Nu hiu sut thu hi t yukhng t yu cu, quay tr
5. Chun b t nht 7 mu thmtch theo quy trnh phn tch
6. Ghi nhn kt qu v tnh ton
Principles of Method Validation Part 1
CEM
LA
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TO
RY
Kim tra li gi tr MDL thu c
1. MDL < Spike < 10xMDL
2. S/N c nm trong khong yu
3. Hiu sut thu hi c nm trong
. APPENDEX B, revision 1.11
phn tch
chun (Spike) c nng bng 5 ln gi tr
Tnh ton gii hn pht hin ca phng php (MDL)
quy trnh phn tch v ghi nhn kt qu phn
yu cu, chuyn sang bc tip theo. Nuli bc (1) v chn MDL cao hn.
thm chun c nng nh bc (2) v phntch
ton MDL
Page 27
c
yu cu khng? 2,5 10?
trong khong yu cu khng? 70 110?
-
CEM
LA
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TO
RY
Phn tch Toluen trong nc bng GC/MS
MDL c tnh l 0,040 ug/L
V d tnh ton MDL C
EM
LA
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RA
TO
RY
MDL c tnh l 0,040 ug/L
Nng Spike 0,21 ng/L vo nn nc ct 2 ln
MDL < Spike < 10x MDL
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
0,084 < 0,21 < 0,84
S/N = 0,20/0,029 = 7,0
AVE H = 96,9%
Phn tch Toluen trong nc
MDL c tnh l 0,040 ug/L
V d tnh ton MDL
MuKt qu
ug/LH%
Mu 1 0,23 110MDL c tnh l 0,040 ug/L
Nng Spike 0,21 ng/L vo
Mu 1 0,23 110
Mu 2 0,21 100
Mu 3 0,24 114
Mu 4 0,19 90
Mu 5 0,18 86
Mu 6 0,22 105
Mu 7 0,17 81
Mu 8 0,16 76MDL < Spike < 10x MDL
Page 28
Mu 8 0,16 76
Mu 9 0,23 110
AVE 0,20 96,9
SD
MDL
S/N
0,029
0,084
7,0
-
CEM
LA
BO
RA
TO
RY
Yu t nh hng (Robustness and Ruggedness)
Xem xt nhy ca ccphp i vi nhng thayphn tch m khng hoctch.
CEM
LA
BO
RA
TO
RY
tch.
Ch ra mc n nh cadng
Principles of Method Validation Part 1
CEM
LA
BO
RA
TO
RY
Yu t nh hng (Robustness and Ruggedness)
cc thng s c trng ca phngthay nh ca mi trng v iu kinhoc t nh hng
n kt qu phn
ca phng php trong qu trnh s
Page 29
-
CEM
LA
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TO
RY
Ti liu tham kho
EURACHEM Guide 1998
The Fitness for Purpose of Analytical
CEM
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TO
RY
IUPAC Technical Report
Harmonized Guideline forMethods, Pure Appl. Chem
Principles of Method Validation Part 1
CEM
LA
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TO
RY 1998
Analytical Methods
2002
for Single Laboratory Validation ofChem. 2002, 74, 835-855
Page 30
-
CEM
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TO
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CEM
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TO
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XIN TRN TRNG CM N!
Principles of Method Validation Part 1
CEM
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XIN TRN TRNG CM N!
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