XNGTSD Cua PP (Sept11) - 3

La chn v
xc nhn gi tr s dng
ca phng php th
Kha o to phng th nghim VILAS, 2011
nbt.11
La chn v xc nhn GTSD phng php

_____________________________________________
Ni dung :
1. Tng quan v XNGTSD phng php th
2. Qu trnh XNGTSD
3. Kim sot phng php trong hot ng thng nht v
s dng s liu XNGT thu c
4. Bi hc c kt
nbt.11
La chn v xc nhn GTSD phng php

_____________________________________________
Mc tiu:
Gip hc vin
Hiu ng v XNGTSD phng php v cc yu cu
Bit p dng k thut thng k cn thit trong qu trnh
XNGTSD
Bit la chn v s dng loi/ kiu XNGTSD thch hp
C th nh gi s ph hp ca phng php th vi
mc ch s dng
Nhn thc mi lin kt gia XNGTSD v KB
nbt.11
La chn v xc nhn GTSD phng php th
Phn 1.
Tng quan v XNGTSD mt phng php
nbt.11
1.

Tng quan v XNGTSD phng php

_________________________________________
Xc nhn gi tr s dng (GTSD) phng php l g ?

Ti sao cn XNGTSD phng php ?
Khi no v lm th no PTN xc nhn GTSD mt
phng php ?
Cc thng s c trng
nh gi s ph hp vi mc ch s dng ca mt
phng php nh th no ?
nbt.11
1.

_________________________________________
Mt kt qu phn tch tt ch c c khi:

Phng th nghim c vn hnh theo mt h thng

m bo cht lng (ISO 17025)

S dng phng php th thch hp, c gi tr ,
S dng thuc th, cht chun/mu chun ph hp,
Tham gia cc chng trnh th nghim thnh tho/ lin
phng
nbt.11
1.

_________________________________________
Xc nhn gi tr s
dng phng php l g ?
S khng nh bng cch kim tra v cung cp cc
bng chng khch quan cho thy cc yu cu c th i
vi mt mc ch s dng nht nh c p ng
(ISO/IEC 17025)
Trong :
Mc ch s dng nht nh= yu cu th nghim
Bng chng khch quan= s liu thc nghm
(cc thng s c trng ca phng php)
S khng nh (t vic so snh yu cu vi bng chng c c)
nbt.11
1.

_________________________________________
Ti sao cn xc nhn gi tr s

dng phng php?
V o c:
- Cn m bo s ph hp vi mc ch s dng ca
khch hng (ngi s dng kt qu)
- Cn m bo phng php s dng l tt (tnh khoa hc)
V kha cnh thng mi:
ch tin cy/ cht lng ca sn phm a ra
V tnh php l:
Qui nh ti cc vn bn php qui
Phm vi p dng ca phng php
Tnh so snh c gia cc th nghim vin/ phng th nghim/
cc quc gia
nbt.11
1.

_________________________________________
Khi no cn xc nhn gi tr s
dng m t phng
php?
Trong qu trnh xy dng, trin khai mt phng php th
Trc khi p dng bt k mt phng php no vo th

nghim trn mu:
- Kim tra kh nng ca chnh PTN c p ng s liu
cng b
- Kim tra tnh ph hp so vi yu cu th nghim
Khi c thay i phm vi p dng ca phng php/ mi
trng th nghim/ th nghim vin
Sau mt khong thi gian khng s dng phng php
nbt.11
1.

_________________________________________
Ai thc hin xc nhn gi tr s
dng phng php?
Th nghim vin/ ngi phn tch:
- xy dng mt phng php th ni b v xc nhn GTSD cc

phng php mi
- kim tra thc nghim cc thng s ca mt phng php
c xc nhn trc
Phng th nghim:
Trin khai phng php, quyt nh hnh thc xc nhn GTSD v
phn cng thc hin v ph duyt
Cc t chc tiu chun ha/ ngh nghip/ ban chuyn :
Xc nhn GTSD ca cc phng php thng qua nghin cu lin
phng
nbt.11
10
1.

_________________________________________
Xc nhn gi tr s
dng phng php nh th no?
Pht trin / nhn
dng phng php
d kin
Xc nh yu
cu phn tch
Lp k hoch xc
nhn thc nghim
.
Bc quan trng
trc khi bt u XNGT
Cng b gi
tr s dng
Khng
Yu cu
c p
ng ?
nbt.11
Tin hnh thc

nghim
S dng s liu
nh gi s ph hp
vi mc ch
11
1. Tng quan v XNGTSD phng php

_________________________________________
ISO 17025 ni g?
Mc 5.4.2

PTN phi s dng cc phng php, k c phng php ly mu,

p ng yu cu ca khch hng v thch hp vi php th (ch
tiu/ thng s cn th)
u tin s dng phng php tiu chun quc t, quc gia hoc
khu vc
Phng TN phi m bo s dng bn tiu chun mi nht, tr khi
n khng ph hp vi mc ch hoc khng c kh nng thc hin
Phng TN cng c th s dng phng php th ni b, phng
php th khng tiu chun nu thch hp vi mc ch s dng v
phi c xc nhn gi tr.
nbt.11
12

_________________________________________
Th no l
mt phng php th
thch hp ?
nbt.11
13

_________________________________________
C bng chng cho thy phng php cho kt qu

ph hp vi mc ch s dng, th hin qua cc c
trng k thut ch yu:

chm (Precision): bao gm lp li (repeatability),

ti lp (reproducibility)
chch, hoc thu hi (Bias/ recovery)
nbt.11
14

_________________________________________
Phm vi o (working range, applicability)

(Gii hn pht hin-LOD (limit of detection), gii hn
nh lng (LOQ); tuyn tnh)
Tnh chn lc (selectivity)/ c hiu (specificity)/ nhy
(sensitivity)
n nh (Ruggedness)
nbt.11
15

_________________________________________
c trng k thut/ hiu nng ca mt phng php
(Performance characteristics/ parameters)
Mt phng php m gi tr ca cc thng s k trn

c thit lp thng qua nghin cu lin phng v cng
b r rng th c coi l phng php xc nhn gi
tr y (fully validated)
Phn ln cc phng php tiu chun/ chnh thc gn
y u thuc loi ny.
nbt.11
16

_________________________________________

Mt s phng php khc c xy dng, ph bin, tuy

nhin mi ch c xc nhn mt phn GTSD hoc
Cha xc nhn
(gi tr cc thng s/ c trng k trn cha c hoc
cha c nghin cu y ).
Phng th nghim s phi xem xt, tin hnh nh
gi, xc nhn cc gi tr ca phng php (Validation).
nbt.11
17

_________________________________________
Ti sao phi s dng v nh gi/ xc nhn cc phng
php ny ?
Phng php tiu chun tng ng cha sn c

K thut/ cng ngh thng pht trin nhanh hn qu
trnh xy dng tiu chun
nbt.11
18

_________________________________________
nh gi Ph hp vi mc ch s
dng nh th no?

Phn tch d liu t cc thng s k thut c/thu c

Cc gi tr mc tiu c t c ? (da trn chun mc
qui nh)
- C = phng php ph hp vi mc ch
- KHNG, cn tm kim phng php khc hoc ci tin, xc nhn
thm cc gi tr
Vic xc nhn gi tr ca phng php c coi l hon

tt khi tuyn b ph hp vi mc ch s dng !
nbt.11
19

_________________________________________
Lu
Mt s t chc quc t v Tiu chun ha ,
Hip hi ngh nghip,
C quan cng nhn
ban hnh cc ti liu Hng dn/ lin quan n
hot ng xc nhn gi tr s dng phng php,
V d:
ISO, AOAC, IAEA, IUPAC, Codex, NATA,
nbt.11
20
La chn v xc nhn GTSD phng php th

_________________________________________
Phn 2
QU TRNH XC NHN GI TR S DNG
CA PHNG PHP
nbt.11
21
2. Qu trnh xc nhn GTSD ca phng php (I)

_________________________________________
Vic xc nhn gi tr s dng c th tin hnh cc mc

(loi/ kiu) khc nhau, ty theo hin trng s dng
mt phng php c th ti PTN,
Bao gm :
Thm nh nng lc thc hin ca PTN i vi mt
phng php (Verification)
v
Xc nhn gi tr s dng (y ) ca mt phng php
(Validation)
nbt.11
22
2. Qu trnh xc nhn GTSD ca phng php

_________________________________________
Cc thut ng trn c dng m t chnh xc nhng

g phng th nghim phi thc hin trc khi a mt
phng php phn tch no vo p dng chnh thc
trong hot ng phn tch/ th nghim hng ngy
Vy khi no p dng
Thm nh nng lc thc hin mt phng php
(Verification) ?
hay
Xc nhn gi tr s dng ca phng php (Validation)?
nbt.11
23

_________________________________________
nh ngha 1 (trong phm vi bi ny):

Thm nh nng lc thc hin mt phng php
(Verification):
L hot ng t kim tra nng lc thc hin ca mt

phng th nghim i vi mt phng php m cc
thng s ca n c xc nhn gi tr v cng b
trc (fully validated method).
PTN ch cn chng minh nhn vin ca mnh c th p
dng phng php mt cch ng n v phng php
c th p dng cho mc ch nh, vi cc gi tr
hp l cng b.
nbt.11
24

_________________________________________
nh ngha 2 (trong phm vi bi ny):

Xc nhn gi tr s dng ca phng php
(Validation):
L hot ng kim tra v xc nh gi tr y cc c trng k
thut/ hiu nng ca mt phng php th hin qua cc thng s
(nh nu trn).
Xc nhn GTSD c th c thc hin bi mt PTN (single
laboratory/ internal validation), hoc qua nghin cu lin phng
(collaborative study)
L mt qu trnh cn thc hin chng minh rng phng php
c chn c th o/ kim ci mnh cn
,
nbt.11
25

_________________________________________
Trong thc t,
Phn ln cc PTN p dng hon ton qui trnh t cc
phng php th tiu chun, xc nhn (standard
method/ validated method),
Mt s PTN khc v nhng l do ring, c th mun sa

i, hoc thay th cc phng php ny bng phng
php th n gin hn, phng php nhanh / cc b Kit
bn sn
nbt.11
26

_________________________________________
ISO 17025 yu cu
Phng TN phi khng nh c th p
dng mt cch ng n phng php
tiu chun trc khi bt u th nghim
nbt.11
27

_________________________________________
Thm nh nng lc thc hin mt phng php tiu chun
Performance verification of a standardised method
(I) Eurachem Guide:

Xem phng php c th p dng ngay cho PTN?
Chng minh nng lc nhn vin p ng cc gi tr cng b ca
phng php, v d : LOD, lp li,
(II) IUPAC (Harmonized Guidelines for Single Laboratory Validation of Method of

Analysis):
PTN s dng mt phng php tiu chun/c nghin cu lin phng ch
cn chng minh nng lc c th t cc c trng k thut cng b ca
phng php.
PTN cn tin hnh cc thc nghim v chm, chch (trn cc nn
mu khc nhau)
nbt.11
28

_________________________________________
Tin hnh thm nh (verification) khi no?

Trc khi a mt phng php tiu chun mi,

cng b gi tr vo p dng = Method verification
Khi mt phng php tiu chun p dng trong PTN
nhng nay mun p dng cho mt hoc vi i tng
mu khc= Matrix verification
Khi a mt sn phm thng mi mi, c cng b cc
gi tr vo p dng
nbt.11
29

_________________________________________
Xc nhn gi tr s dng ca phng php (Validation),

Khi no?
Khi PTN p dng phng php t xy dng/ ni b
Khi p dng mt phng php tham kho, mt sn
phm thng mi (kit th) cha xc nh / cng b gi
tr s dng
Khi PTN mun sa i mt vi ni dung ca phng
php tiu chun:
cn phi chng minh c phng php vn cho cc
gi tr bng hoc tt hn so vi phng php gc / tham
chiu khi c s thay i y
nbt.11
30

_________________________________________
Ty hin trng phng php tham chiu d kin s dng Phng th
nghim s quyt nh mc nh gi, xc nhn thch hp,
cn c trn :
Nguyn tc chung: mt phng php khi p dng phi c h tr

bi cc bin php kim sot cht lng (vi cc thng s c bn:
chm, chch, n nh)
Vic xc nhn phi t ti mc cn thit c th so vi phm vi p

dng ca php th/ yu cu phn tch
Vic xc nhn phi c xem xt li, khi cn
nbt.11
31
Bng1: m t cc mc thm nh khc nhau

(ngun: IUPAC)
1.Phng php th
Thm nh ng v chm
xc nhn y
2. PP xc nhn y nhng s
dng cho mt nn mu mi/ mt thit
b mi
Thm nh - nh trn v c th + LOD
3. PP c xy dng hon chnh,

nhng cha XNGT y
-nh trn + xc nh S R
4. PP xut bn trong cc n phm

khoa hc v cng b cc thng s
quan trng
-nh trn-
5. PP xut bn trong cc n phm

khoa hc nhng khng c cc thng
s quan trng
Xc nhn gi tr y ti PTN
6. PP do phng th nghim t xy
dng
Xc nhn gi tr y ti PTN
nbt.11
32
2. Qu trnh xc nhn GTSD ca phng php (II)

_________________________________________
Trc khi thc hin thm nh

PTN cn bin son d tho Qui trnh ni b (SOPs)
bng vn bn, da trn phng php tham chiu, m t
ngn gn, chnh xc cc yu cu :ha cht, thit b, iu
kin mi trng v cc bc tin hnh

Lp k hoch thm nh
Phn cng thc hin
nbt.11
33

_________________________________________
Khi lp k hoch
PTN cn xem xt, tnh n cc im sau:
Nhu cu c th ca khch hng, hoc
Yu cu t cc c quan qun l lin quan
Ci g thc t c th phn tch c?
Cc iu kin thc t ca phng th nghim
- iu kin k thut c th trong php th ?
- Thit b ?
- Cc ngun lc khc ?
nbt.11
34

_________________________________________
Khi lp k hoch

Nn nghin cu, t ra mt s cu hi chi tit c th ,

nhm:
Thit k ni dung thm nh st vi yu cu nht
Kt qu thm nh c ngha, s dng c
Tit kim thi gian v ti chnh
nbt.11
35

_________________________________________
Lu trong thm nh/ xc nhn gi tr

Trc tin, hy chn cc phng php quc t hoc
quc gia c th ph hp vi mc ch s dng
Nu khng sn c, hoc khng th p dng c, hy
tin hnh xc nhn gi tr ti PTN (single laboratory
validation) theo mt qui trnh c chp nhn rng ri
V d: - CAC/ GL 49: Hng dn thm nh/xc nhn ni b PTN i
vi phng php phn tch,

- Quyt nh 2002/657/EC: lin quan n xc nh mt vi d cht
ha hc trong ng vt v sn phm ng vt)
nbt.11
36

___________________________________________
Thc hin xc nhn

Nhn vin thc hin phi l ngi c kin thc v
kinh nghim thch hp
Xc nh cc c trng ca phng php mc cn
thit
Kh nng p dng c th m rng (nhiu loi sn phm,
i tng mu)
Tm kim thm gi tr chnh xc t cc nghin cu
lin phng
B sung/ xem xt li khi thch hp
nbt.11
37
2. Qu trnh xc nhn GTSD ca phng php (III)

Cc thng s k thut
___________________________________________
Cc thng s k thut
chnh xc (Accuracy)
l thc o cht lng ca kt qu th, ngha l
kt qu hu ch cho khch hng hay nhng
ngi quan tm nh th no
bao hm hai thnh phn l:
chm v ng
nbt.11
38

Cc thng s k thut
___________________________________________
Cc thng s k thut
1. chm (Precision)
p dng cho php th nh lng/ bn nh lng

L thc o v tc ng ca cc sai s ngu nhin

l mc gn nhau ca cc kt qu phn tch c lp
thu c trong cc iu kin nh
chm thng c cng b dng lch chun
(s) hay lch chun tng i (RSD)
C 2 thc o chm, th hin bng lp li (Sr)
v ti lp (SR)
nbt.11
39

Cc thng s k thut
___________________________________________
Cc thng s k thut
Xc nh chm
(p dng cho php th nh lng/ bn nh lng)

c th cng b chm phn nh trung thc nht

hiu nng ca mt phng php trong iu kin bnh
thng ca n, PTN cn xc nh r cc iu kin ny,
Vt liu/mu th c chn cn phi thuc loi in
hnh ca cc mu thng ngy phi th
Mu th c chn phi bn, n nh trong qu trnh
xc nh
nbt.11
40

Cc thng s k thut
___________________________________________
Xc nh chm
(p dng cho php th nh lng/ bn nh lng)
lch chun lp li Sr (Repeatability standard deviation):

c tnh ton t
Cc kt qu thu c vi cng mt phng php th
trn cc mu cng loi, thc hin ti cng mt phng th
nghim bi mt kim nghim vin, s dng cng thit b
v trong cc qung cch thi gian ngn
nbt.11
41

Cc thng s k thut
___________________________________________
Xc nh chm
lch chun ti lp SR (Reproducibility standard deviation)
c tnh ton t
Cc kt qu thu c vi cng mt phng php th

trn cc mu cng loi, thc hin ti cc phng th
nghim khc nhau, bi cc kim nghim vin khc nhau,
s dng thit b khc
nbt.11
42

Cc thng s k thut
___________________________________________
Xc nh chm
ti lp ni b PTN/ chm trung gian
(Intermediate precision or Infra-/within-laboratory reproducibility SDw)
c tnh t
Cc kt qu thu c vi cng mt phng php th trn cc mu
cng loi, thc hin ti cng mt phng th nghim bi cc kim
nghim vin khc, s dng cng thit b nhng trong cc khong
thi gian khc nhau
ti lp ni b PTN/ chm trung gian l thng s rt quan trng
trong xc nhn GTSD phng php v c tnh KB
nbt.11
43

Cc thng s k thut
___________________________________________
Xc nh chm - Cc iu kin:
a. Chung cho cc trng hp :
- Ti thiu xc nh trn 7 mu lp (hoc mu kp)/ mi nn mu
- Nu c th, nn chn cc mu ti mc / c cha nng thch hp
ca cht cn km tra (theo yu cu ca ngi s dng kt qu/ vn
bn php qui)
- Tt nht, PTN nn tin hnh xc nh lp li, trn cc phn mu
ring r, c chun b t mt mu th, hoc t mu chun c
chng nhn (CRMs), mu chun khc (RM/ QC sampes) di iu
kin thao tc bnh thng, trong mt khong thi gian di hn (t
nht trong khong 2 tun l)
- Cng c th s dng biu kim sot lin tc da vo cc mu
nh trn, hoc mu c gy nhim/ thm chun hoc bt k loi
mu no kim thng xuyn trong PTN
nbt.11
44

Cc thng s k thut
___________________________________________
a. Chung cho cc trng hp :
- Thng thng, hot ng ny lin quan n ti lp ni b PTN
(nh cp).
- Tnh gi tr trung bnh, lch chun v lch chun tng i
(RSD)/ h s bin thin (CV %)
theo cng thc :
S =
(x
RSD% = CV% =
SD
100
X
n 1
Trong : x = xi / n
n= s kt qu ring l thu c
nbt.11
45

Cc thng s k thut
___________________________________________
b. i vi php th
ha hay sinh hc:
chm c th thay i ty theo nng cht cn phn
tch / mt vi sinh vt c mt trong mu
cn xc nh b sung chm khi nng cht/ vi sinh
phi phn tch d kin dao ng hn 50% so vi mt gi
tri trung bnh thng thy
Trong mt vi trng hp, cng c th ch cn xc nh
chm ti mt n hai gi tr c ngha c bit vi
ngi s dng kt qu, v d: gii hn cnh bo hoc ti
mc qui nh (hoc c th mc 20% v 80% ca
ng chun)
nbt.11
46
V d:
chm (trong phn tch sng lc)
lp li

3 nng :
1x; 1,5x; 2x MRPL
0,5; 1x; 1,5x MRL
6 mu lp/mc nng
3 ln xc nh
Trong cng iu kin th
ti lp ni b PTN

nbt.11
3 nng :
1x; 1,5x; 2x MRPL
0,5; 1x; 1,5x MRL
6 mu lp/ nng
3 ln xc nh
Trong cc iu kin khc
nhau (th nghm vin, iu
kin mi trng, lot ha
cht pha ch)
47

Cc thng s k thut
________________________________________________
2. ng (trueness) / chch (bias)

ng ca mt phng php m t mt kt qu th gn vi mt
gi tr chun/ tham chiu mc chp nhn c.
Vic thiu ng (khng ng) chng t li h thng
chch l cch din t mang tnh nh lng ca ng.
ng ca mt kt qu c ci thin khi chch gim (nh)
thu hi trong phn tch l mt chch thng gn lin vi qu
trnh x l mu, chit tch cht cn phn tch ra khi mu v cc qui
trnh phn tch khc trc khi xc nh chnh thc.
nbt.11
48

Cc thng s k thut
________________________________________________
2. ng (trueness) / chch (bias)

Cch tt nht l phn tch mt nn mu c chng nhn (CRM) c

cha mt nng n nh (hoc mt lng) ca cht cn phn tch
thu hi c bo co theo %, vi gi tr xc nh c/ nng
cng b
Nu khng sn c mt CRM thch hp, thu hi c th xc nh
bng cch phn tch mt mu trc v sau khi thm mt lng
bit cht cn phn tch (mu m/ mu thm chun). Tnh chnh lch
gia cc kt qu thu c
nbt.11
49

Cc thng s k thut
________________________________________________
Xc nh chch (I)
Ti thiu phi c 7 kt qu lp
Mu chun (CRM) cn thuc nn mu tng t (v nng cht
phn tch cng nn ph hp vi mu th thng thng)
Trng hp khng sn c mu chun,
C th s dng mt mu c chun b trong PTN, c hm lng
cht cn phn tch bit.
Hm lng cht ny phi c kim tra k lng bng hai (hoc
nhiu hn) phng php c nguyn tc vt l/ ha hc khc nhau
v tt nht nn th ti nhiu hn mt PTN
Khi c th, mu chun ni b nn c hiu chun da trn mt
CRM
nbt.11
50

Cc thng s k thut
________________________________________________
Xc nh chch (II)
Cng c th so snh kt qu thu c vi kt qu t
mt phng php chun ha.
S dng kt qu t chng trnh nghin cu lin phng/
th nghim thnh tho
La chn b sung: thu hi
thu hi (din t l R) c s dng cho cc
phng php c cc bc x l mu, tch chit cht
cn phn tch,
nbt.11
51

Cc thng s k thut
________________________________________________
Xc nh chch (II)
Tnh thu hi da trn mu c hm lng bit.

R = Q tm c
Q gc
Tnh thu hi da trn mu c thm chun:
R = Q tm c Q mu ban u
Q thm
nbt.11
52

Cc thng s k thut
________________________________________________
Xc nh chch (II)
C th s dng Control charts:
X-charts : kim sot chch (t CRMs).
C th tnh c lch chun ti lp ni b t biu
ny
R-charts : kim sot chm ca thu hi
C th tnh c lch chun lp li hoc
lch chun ti lp ni b t biu
nbt.11
53

Cc thng s k thut
________________________________________________
3. n nh (Ruggedness)
L mn cm ca mt phng php phn tch i vi cc thay i
nh trong iu kin thc nghim,
v d:

Thnh phn mu th,

Cc m ha cht/mi trng khc nhau
pH, thi gian chit mu,
iu kin bo qun
iu kin mi trng (nhit , p sut)
iu kin x l mu
nbt.11
54

Cc thng s k thut
________________________________________________
3. n nh (Ruggedness)
Th n nh nh th no ?
(ch trong trng hp xy dng phng php)
Nhn dng cc yu t c th gy nh hng n kt qu
(cht cn phn tch, dung mi, pH, nhit , tc nng nhit..)
Thay i nh tng yu t
Nu mt yu t c tm thy c nh hng n kt qu ca phn
t i din th tin hnh thm cc thc nghim
xc nh gii hn chp nhn i vi yu t ny (ghi li)
Khuyn ngh ca CRL (chu u): phn tch 10 mu trng v 10 mu thm

chun cng nng , thay i nh cc yu t pht hin nh hng
nbt.11
55

Cc thng s k thut
_____________________________________________
4. Tnh bn vng (stability)
Khng cn th nu d liu v tnh bn vng c
(t phng th nghim khc hoc trong cc n phm, ti
liu km)
ghi s liu ny vo bo co thm nh
Th tnh bn vng:
- Cht phn tch trong dung dch
- Cht phn tch trong nn mu
Cc phn ly t dung dch mi pha hoc t mu th
c bo qun trong cc iu kin khc nhau (nhit /
thi gian)
nbt.11
56

Cc thng s k thut
____________________________________________
5. Gii hn pht hin (LOD)/ Gii hn nh lng (LOQ)

Khng phi lun lun cn xc nh LOD, tuy nhin i vi cc php

th c hc hoc kim sot mi trng th n c bit cn thit
Cn lu y khng phi l LOD ca thit b nn mu s dng xc
nh LOD phi c x l trong ton b qui trnh theo phng
php.
L tng nht nn s dng mt mu khng cha cht ch (mu
trng); nu khng, hy tm mt mu c hm lng cht ny thp ti
mc c th.
Cn thu thp s liu phn tch t t nht 7 ln trong khong thi gian
t nht l 3 tun l.
nbt.11
57

Cc thng s k thut
____________________________________________
5. Gii hn pht hin (LOD)/ Gii hn nh lng (LOQ)
c 2 cch xc nh LOD:
Tnh SD t cc mu t nhin hoc mu chun vi hm lng rt
thp ca cht cn phn tch, hoc t ng chun.
Tnh SD t cc mu t nhin hoc mu trng khng cha cht cn
phn tch. Nu khng c tn hiu t mu trng, nhiu nn (S/N) thit
b c th c khuych i
LOD = b + 3.So (b= gi tr trung bnh ca mu trng; So= SD ti
zero)
LOQ (thng c bit) = 3 LOD
nbt.11
58
2. Qu trnh xc nhn GTSD ca phng php (IV)

Thm nh/ xc nhn gi tr ban u
____________________________________________
6. Thm nh/xc nhn ban u

-
Hng dn ca 2002/657/EC thch hp i vi

cc php phn tch phc v kim tra/ thanh tra
c bit lin quan n xut nhp khu
D dng trong quyt nh chp nhn/ bc b mt l hng
khi kt qu bo co km theo KB (hng dn
Eurochem)
nbt.11
59

____________________________________________
Thm nh/xc nhn phng php sng lc (nhanh)
(screening method)
Phng php sng lc c s dng pht hin s c mt ca
mt cht hoc mt nhm cht mc cn quan tm kim sot, (v
d: ELISA, plate test, biosensor, sc k, quang ph)
Yu cu ti thiu s dng mt phng php/ k thut

sng lc l:
Cn phi c thm nh/ xc nhn (tnh truy xut)
Phi c t l chp nhn sai nh hn 5% ( -erro) mc
hm lng cn kim sot
nbt.11
60

____________________________________________
Thm nh/xc nhn phng php sng lc/nhanh
(screening method)
Cn xc nh r phm vi ca phng php (scope):

Cht hay hp cht cn phn tch

Di hm lng quan tm
Danh sch cc nn mu (matrix)
nbt.11
61

____________________________________________
Thm nh/xc nhn phng php sng lc (nhanh)
(screening method)
Cc k thut, kt th thng c s dng trong PTN ?

R-Biopharm, Ridascreen
Euro-Diagnostica
Transia GmbH
CER, laboratoire dhormology,
Randox ??
nbt.11
62
Performance characteristics for method

validation (screening)
(theo 2002/657/EC)
Qualitative method: identifies a substance on basis of its chemical, biological or
physical propriety (binary response: +/-, absence/presence)
Quantitative method: determines the amount or mass fraction of a substance
(response: numerical value of appropriate unit)
Detection
Capability
CC
Decision
Limit
CC
Trueness/
Recovery
Precision
Selectivity/
Specificity
Applicability/
Ruggedness/
Stability
Qualitative
methods
Quantitative
methods
+ = determination is mandatory
nbt.11
63

____________________________________________
Thm nh/ xc nhn ln u
(initial validation)
Thm nh ln u vi cc nn mu thng kim tra nht:
Kh nng pht hin (CC)
Tnh chn lc/ c hiu
Kh nng p dng/ tnh n nh/ bn vng
chm (ch trong php th bn nh lng/nh lng)
Nu c th, s dng cc ngun mu trng khc nhau, cc KNV khc nhau,

cc ngy khc nhau vi cng loi mu c gy nhim/ thm chun
nbt.11
64

____________________________________________
Kh nng pht hin (CC):
L hm lng nh nht ca cht cn kim tra c th
pht hin, nhn dng v nh lng c trong mu th
vi mt xc sut sai li l

Vi MRPL: CC= hm lng nh nht m ti phng php c th pht

hin ng cc mu b nhim vi m bo thng k l 1-
Vi MRL: CC= hm lng ti phng php c th pht hin c mc
nng ti a cho php (MRL) vi m bo thng k l 1-
nbt.11
65

____________________________________________
Kh nng pht hin (CC): xc nh bng 2 cch
Trong trng hp khng c mc gii hn cho php
1. Phn tch 20 mu trng CC = 3 x S/N
Phn tch 20 mu trng c thm chun mc CC
CC = CC + 1.64 x SDRW
2. Dng ng chun:
Phn tch cc mu trng c thm chun mc: 0; 0.5; 1 ; 1.5 v 2 MRLP
Dng kt qu phn tch ln th (trc y) tng ng vi nng (trc x)
CC = y-intercept (trng) + 2.33 x SDRW
nbt.11
66

____________________________________________

Trng hp khng c mc gii hn cho php,
Nu khng c kt qu nh lng:

Phn tch mu trng c thm chun tai mc v trn mc CC

(n 20 / mc nng )
CC = mc nng ti ch c 5 % cc kt qu ph hp sai
nbt.11
67
Kh nng pht hin

Detection capability (CC)
Blank
CC
+2.33xSDblank
CC
+1.64xSDRW
=1%
=5%
Signal or
Concentration
nbt.11
68

____________________________________________
Trng hp c mc gii hn cho php (MRL):
1. Phn tch 20 mu trng c thm chun mc MRL
CC = MRL + 1.64 x SDRW
Phn tch 20 mu trng c thm chun mc CC
nbt.11
69
Kh nng pht hin (Detection capability)

CC
MRL
+1.64xSDMRL
CC
CC
+1.64xSDRW
=5%
=5%
Signal or
Concentration
nbt.11
70

____________________________________________
Trng hp c mc gii hn cho php (MRL))
(tip)
2. Dng ng chun:
Phn tch cc mu trng c thm chun mc: 0; 0.5; 1 ; 1.5 v
2 MRL
Dng kt qu phn tch ln th (trc y) tng ng vi nng
(trc x)
CC = MRL + 1.64 x SDRW
nbt.11
71

___________________________________________
Tnh chn lc/ c hiu

c hiu: l kh nng mt phng php c th phn
bit c cht cn phn tch vi cc hp cht khc
Vn lin quan n nhiu (do k thut o, nhm hp cht, nn mu)
nbt.11
72

_________________________________________
Tnh chn lc/ c hiu

Th c hiu trong phng php sng lc nh tnh nh th no ?
Phn tch 20 mu trng v 20 mu dng tnh ( lu li hoc thm chun),
thc hin trong cng mt ngy/ KNV hoc vo cc ngy khc nhau/ KNV
(NA= pht hin m tnh; N- = m tnh tht)
c hiu = 100 * NA/ N nhy =
100 * PA/ N+
( PA= pht hin dng tnh; N+ = (+) tht)
T l dng tnh gi (FP) = 100* FP/ (N- + N+ )
T l m tnh gi (FN)
= 100* FN/ (N- + N+ )
chnh xc= 100* (PA + NA)/ (N- + N+ )
nbt.11
73

__________________________________________
Tnh chn lc/ c hiu
Th c hiu trong phng php sng lc bn nh
lng/ nh lng nh th no ?
- La chn cc cht tim n gy nhiu (VD: cc cht
chuyn ha, cc dn xut)
- Phn tch mt lng mu trng thch hp (n 20)
- Phn tch cc mu c thm cht gy nhiu mt
nng thch hp
nbt.11
74

____________________________________________
Tnh chn lc/ c hiu
c tnh nh hng ca cht gy nhiu :

- nh danh gi/ nhn dng sai ?
- nh hng trong nh lng ?
- Vic nh danh cht ch b cn tr ?
nbt.11
75

____________________________________________
Kh nng p dng
Phm vi p dng ca phng php cn phi xc
nh r:

Loi nn mu (cht lng/ rn; loi m: c, gan, c cht

bo/ khng bo, c ng/ khng ng)
Chng loi ng vt, thc vt
Di nng c th nh lng chc chn
nbt.11
76

____________________________________________
Kh nng p dng
p dng phng php cho mt nn mu mi:

Phn tch t nht 10 mu trng khc nhau c b sung cht cn

phn tch nng quan tm (cc) + th kh nng nhiu
Nu 10 mu dng tnh phng php c th p dng cho nn
mu mi ny
Nu c 1 mu m tnh th thm 10 mu na
Nu vn c 1 mu m tnh cn tnh ton li cc cho nn mu mi
ny
nbt.11
77

____________________________________________
Tip cn mi: Hng dn ca CRL 20/ 1/ 2010 i vi
thm nh phng php sng lc d lng (thuc th y)

Vi cc php phn tch c xc nhn

Cc cht c MRL qui nh (hoc MRPL)
Nng sng lc ch s = 1/ 2 MRL (hoc MRPL)
Nu kt qu c 1 hoc 0 mu m tnh gi trong s 20
mu th Screen positive l chng minh rng CC
nh hn mc MRL v nh hn hoc bng mc MRL
Mc chp nhn: CC 5 % m tnh gi
nbt.11
78

____________________________________________
Tip cn mi: Hng dn ca CRL 20/ 1/ 2010
Trong phn tch hng ngy:
Chn K1= CC ( MRPL), v d Clenbuterol = 0,5 g/kg
K2 = 2* CC
= 1,0 g/kg
Tin hnh th nh qui trnh, + mu trng
Ghi nhn gi tr cc hp th,
Tnh c hp th gii hn (Abs limit): Abs limit = neg 2. SD
Abs. blank > Abs limit
Abs K1 v K2 < Abs limit
Cc mu c Abs. < Abs limit s c phn tch li
nbt.11
79

____________________________________________
Tip cn mi: Hng dn ca CRL 20/ 1/ 2010

Trong phn tch hng ngy:
Mu QC 1= ( MRL), v d Trifluralin = 0,5 g/kg
Mu QC 2 = MRL
= 1,0 g/kg
Tin hnh th + mu trng
Cc mu c Abs < abs K1 phn tch li/ khng nh
nbt.11
80

____________________________________________
Khuyn ngh:
i vi phng php xc nhn/ kt th thng mi:
Theo di lch s nh gi cc tnh nng
Thu thp s liu t nh cung cp v vic xc nhn
C phng n thc nghim nh gi k nng ca KNV
S dng cc mu QC trong phn tch hng ngy
Tham gia vo cc chng trnh th nghim thnh tho
PTs)
nbt.11
81

____________________________________________
Php th vi sinh - Method verification
thng thng yu cu tt c hoc mt vi yu t di y:

Ti thiu 5 mu th (bao gm kt qu (+) v (-),

Cc mu t nhin, hoc mu ch ng gy nhim u c th s
dng (nu gy nhim, mt chn phi tng t nh d kin c
trong mu t nhin, hoc cc di t 0-100; 100-1000, 10 000 ty
mc ch)
Mu trng: phi c thanh trng, hoc c bng chng xc nhn
thc nghim l khng cha/ hoc ch cha rt t VSV ch v
thm VSV nn khng gy nhiu ng k n vic c kt qu
nbt.11
82

____________________________________________
Php th vi sinh - Method verification

Cc chng chun v chng hoang d nn cng c

s dng trong qu trnh kim tra kim chng bt k
sai khc c th xy ra
Cc mu chia nh c giao cho mt s th nghim
vin thc hin m bo kim sot cht lng ni b
Thc hin phn tch mu trng, mu th v mu kim
sot theo ng qui trnh ca phng php.
nbt.11
83

____________________________________________
Php th vi sinh - Matrix verification
p dung nguyn phng php,thay i nn mu:
Nn mu d kin p dng phi tng t/ gn v nhm
hoc c lin quan vi nn mu xc nhn trong
phng php gc. (VD: c/ tht ? c/ du c? Tm/ mt ong?)
im quan trng cn chng minh kh nng thu hi mt

s lng ng k vi sinh vt c mt trong mu,
Lu k thut x l/ pha long thch hp i vi nn

mu, VD: c nhiu mui/ t mui; c cht bo qun/ khng
nbt.11
84

____________________________________________
Php th vi sinh - Matrix verification

Thay i nn mu
Nhim mu vi lng bit, Rec. phi >70 %

S dng c chng chun v chng trong t nhin gy

nhim chng minh kh nng pht hin di biotype
Cc thng s quan tm cn li tng t nh Method
verification
nbt.11
85

____________________________________________
Php th vi sinh Method verification/validation
Cc trng hp sa i phng php c th bao gm:

Thay mi trng nui cy truyn thng

Thay i iu kin , c bit l nhit , thi gian
S dng k thut ELISA, u d a-xit Nucleic, khc
S dng k thut PCR cho kt qu nhanh
nbt.11
86

____________________________________________
Php th vi sinh Method validation
Xc nhn thng qua so snh

Cc phng php thay th/ sa i phi:
Cho cng nhy, chn lc hoc tt hn

Phng php nh lng: quan tm nht l nhy, s lng

VSV.
i vi mi trng khng chn lc: t nht th 50 mu/matrix, nh
gi thu hi.
Nu l mt mi trng chn lc mi : cn th mt di vi sinh/ lot
nn mu khc nhau m bo mi biotype c th thu hi c.
nh gi PP gc/ phng php tham chiu
nbt.11
87

____________________________________________
Php th vi sinh Method validation
Xc nhn thng qua so snh

Cc phng php thay th/ sa i phi:
Cho cng nhy, chn lc hoc tt hn
Phng php nh tnh: t nht th 50 mu dng

tnh/matrix (bao gm cc chng, loi, typ huyt thanh)
m bo tnh a dng
+ 50 mu m tnh tnh t l (+) tnh v (-) gi,
nh gi kt qu thu c vi kt qu c t phng
php gc/ tham chiu
nbt.11
88
Tnh chn lc/ c hiu

Th c hiu/ tnh chn lc trong phng php sng

lc nh tnh nh th no?
Phn tch 50 mu trng khc nhau v 50 mu dng tnh

(mu m ,cng hoc khc ngy/th nghim vin)
Kt qu
dng tnh
Kt qu m
tnh
Dng tnh
thc (N+)
m tnh thc
(N-)
Dng tnh
ng
(PA)
Dng tnh gi
(FP)
m tnh gi
(FN)
m tnh ng
(NA)
Specificity = 100* NA/N( c hiu)
Cc thng s khc:
chnh xc= 100* (PA+NA)/(N- + N+)
nhy = 100* PA/N+
Dng tnh gi = 100* FP/(N- + N+)
m tnh gi = 100* FN/(N- + N+)
nbt.11
89
2. Qu trnh xc nhn GTSD ca phng php (V)
H s
___________________________________________
nh gi kt qu v tp hp h s
H s thm nh/ xc nhn phng php phi lu gi y :

K hoch v s chun b
Ti liu v d liu quan trc gc (nn mu th, cht chun, thuc
th, mu trng, mu QC, s ln TN, x l thng k)
Ti liu v nh gi s liu thm nh
So snh v nh gi kt qu thu c
Chun b bo co
* Lu : thu thp d liu thch hp cho vic tnh ton MU sau ny
nbt.11
90

_____________________________________________________
nh gi kt qu cui cng so vi mc mong i -lu

k hoch t/ chun mc chung
(Nu cn: s dng F-test)
Kt lun v s ph hp vi mc ch s dng
Nu khng p ng, cn thay i phng php v lp

li qu trnh thm nh/ xc nhn
nbt.11
91

_____________________________________________________
Mc mong i/ chun mc chung:

M t trong phng php gc

Mc gii hn ti a qui chun k thut
Mc yu cu trong hp ng ca khch hng
Chun mc chung t tiu chun khung (ISO, Codex,
AOAC, ASTM,trong trng hp thiu d liu chnh
xc ca phng php gc)
nbt.11
92

_____________________________________________________
Bo co thm nh phng php phi bao gm:

K hoch v s chun b
M t chnh xc cc c trng c nh gi xem xt
Cc bc tin hnh
Thi gian/ ngi thc hin
Cc kt qu thu c v cch tnh ton
Vic nh gi kt qu thu c so vi chun mc
Kt lun r rng (khng mp m, gy hiu nhm)
nbt.11
93
2. Qu trnh xc nhn GTSD ca phng php (VI)

Ph duyt/ ban hnh
_____________________________________________________
Sau khi hon tt qu trnh thm nh phng php :

PTN thng qua trong ni b v trnh bo co ln lnh o

Nu khng cn thc hin thm, PTN hon chnh qui trnh th ni b
(SOP)
Format SOP theo tiu chun ISO, nu r gi tr xc nhn ch yu
Lnh o ph duyt, ban hnh v a vo thc hin chnh thc
trong h thng
o to/ nh gi tay ngh KNV trn phng php mi
Thm nh lin tc (continuous validation):
Thc hin v lu s liu mu kim sot nh gi KB,
______________________________________________
nbt.11
94
3. Kim sot phng php trong hot ng hng ngy

______________________________________________
Khi mt phng php c nh gi /xc nhn v a vo p

dng phn tch hng ngy, PTN cn c k hoch v phn cng
trch nhim kim sot m bo rng hiu nng ca phng
php ph hp vi kt qu trong qu trnh thm nh/ xc nhn
gi tr ban u.
Kim sot lin tc/ thng xuyn:
ng v chm ca phng php cn c kim sot thng
xuyn
Kt qu thu c phi so snh vi cc gi tr c
Ch kim sot nh k do PTN cn nhc, quyt nh mt cch
thch hp nht
nbt.11
95
V d minh ha v kim sot phng php

trong p dng hng ngy
+ 3 std dev
Kt qu o
+ 2 std dev
+ 1 std dev
Mean
- 1 std dev
- 2 std dev
- 3 std dev
Ngy/ gi
nbt.11
96

______________________________________________
Khi no cn thm nh li (re-validation)?

Khi c thay i trong cc iu kin mi trng

Khi c thay i trong qui trnh ca phng php
Khi c thit b mi
Khi chuyn PTN ti c s mi
Khi thay i ng k nhn vin th nghim
Khi phng php khng s dng trong mt thi gian di
nbt.11
97

______________________________________________
Vi s liu kim sot ln, thu thp c trong mt

khong thi gian nht nh, PTN c th tn dng
nh gi thng xuyn nng lc cc nhn vin th
nghim
Quan trng hn, PTN c th c tnh khng m bo
o cho php th c th ca mnh mt cch d dng v
hp l nht
PTN nn nh k cng b cc gi tr s dng mi mt
cch thch hp
nbt.11
98
Xc nhn GTSD ca phng php
4. Bi hc c kt
______________________________________________
1. Vic xc nhn gi tr s dng ca phng php i hi
phi a ra c s liu c ngha.
2. C hai loi phng php tiu chun v phng php ni b u
cn thm nh hoc xc nhn gi tr s dng
3. Xc nhn GTSD ca phng php phi l mt hot ng c lp
k hoch (cc thng s cn thay i theo phm vi p dng)
4. Xc nhn GTSD ca phng php khng th coi l hon thin khi
khng c tuyn b g v s ph hp vi mc ch s dng
5. Xc nhn GTSD ca phng php nn c PTN xem xt nh
mt hot ng 3 trong 1 !
nbt.11
99
Xc nhn GTSD ca phng php

____________________________________________
Cu hi hoc chia s kinh nghim?
Chc thnh cng v

Xin cm n !
nbt.11
100
Ti liu tham kho

ISO/IEC 17025, ISO 5725

Cxg 072-2009 v mt s Cxg khc;
IUPAC TR.; AOAC guidelines
JCGM 200:2008, VIM
NATA Technical note 17, 2009
2002/657/EC; CRLs guidelines 20/1/2010;
Bi ca mt s chuyn gia (J.Engman, Liam O.,
F.Ulberth, N. Van Wouwe, K.H. Lund, UNIDO,
labnetwork,) v mt s ngun khc
nbt.11
101
Thng tin trao i

______________
a ch lin h:
Vn phng Cng nhn cht lng, s 8-Hong Quc Vit
Hoc:
Bi Thanh Nh
e-mail: thanhnha@fpt.vn
nbt.11
102

XNGTSD Cua PP (Sept11) - 3

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

XNGTSD Cua PP (Sept11) - 3

Uploaded by

Copyright:

Available Formats

La chn v

Kha o to phng th nghim VILAS, 2011

La chn v xc nhn GTSD phng php

La chn v xc nhn GTSD phng php

La chn v xc nhn GTSD phng php th

Tng quan v XNGTSD phng php

Xc nhn gi tr s dng (GTSD) phng php l g ?

Tng quan v XNGTSD phng php

Mt kt qu phn tch tt ch c c khi:

m bo cht lng (ISO 17025)

Tng quan v XNGTSD phng php

Tng quan v XNGTSD phng php

Ti sao c n xc nhn gi tr s

Tng quan v XNGTSD phng php

Trc khi p dng bt k mt phng php no vo th

Tng quan v XNGTSD phng php

- xy dng mt phng php th ni b v xc nhn GTSD cc

Tng quan v XNGTSD phng php

Tin hnh thc

1. Tng quan v XNGTSD phng php

PTN phi s dng cc phng php, k c phng php ly mu,

1. Tng quan v XNGTSD phng php

1. Tng quan v XNGTSD phng php

C bng chng cho thy phng php cho kt qu

chm (Precision): bao gm lp li (repeatability),

1. Tng quan v XNGTSD phng php

Phm vi o (working range, applicability)

1. Tng quan v XNGTSD phng php

Mt phng php m gi tr ca cc thng s k trn

1. Tng quan v XNGTSD phng php

Mt s phng php khc c xy dng, ph bin, tuy

1. Tng quan v XNGTSD phng php

Phng php tiu chun tng ng cha sn c

1. Tng quan v XNGTSD phng php

Phn tch d liu t cc thng s k thut c/thu c

Vic xc nhn gi tr ca phng php c coi l hon

1. Tng quan v XNGTSD phng php

La chn v xc nhn GTSD phng php th

2. Qu trnh xc nhn GTSD ca phng php (I)

Vic xc nhn gi tr s dng c th tin hnh cc mc

2. Qu trnh xc nhn GTSD ca phng php

Cc thut ng trn c dng m t chnh xc nhng

2. Qu trnh xc nhn GTSD ca phng php

nh ngha 1 (trong phm vi bi ny):

L hot ng t kim tra nng lc thc hin ca mt

2. Qu trnh xc nhn GTSD ca phng php

nh ngha 2 (trong phm vi bi ny):

2. Qu trnh xc nhn GTSD ca phng php

Mt s PTN khc v nhng l do ring, c th mun sa

2. Qu trnh xc nhn GTSD ca phng php

2. Qu trnh xc nhn GTSD ca phng php

(I) Eurachem Guide:

(II) IUPAC (Harmonized Guidelines for Single Laboratory Validation of Method of

2. Qu trnh xc nhn GTSD ca phng php

Tin hnh thm nh (verification) khi no?

Trc khi a mt phng php tiu chun mi,

2. Qu trnh xc nhn GTSD ca phng php

Xc nhn gi tr s dng ca phng php (Validation),

2. Qu trnh xc nhn GTSD ca phng php

Nguyn tc chung: mt phng php khi p dng phi c h tr

Vic xc nhn phi t ti mc cn thit c th so vi phm vi p

Vic xc nhn phi c xem xt li, khi cn

Bng1: m t cc mc thm nh khc nhau

La chn v xc nhn GTSD phng php

La chn v xc nhn GTSD phng php

Ti sao cn xc nhn gi tr s