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VN PHNG CNG NHN CHT LNG

Bureau of Accreditation (BoA)


H THNG CNG NHN PHNG TH NGHIM VIT NAM
Vietnam Laboratory of Accreditation Scheme (VILAS)

HNG DN CC V D C LNG KHNG M BO O


TRONG PHN TCH HO HC NH LNG
M s: AGL 19
Ln ban hnh: 1.04
Ngy ban hnh:

H tn

Bin son

Xem xt

Ph duyt

Hong Thanh Dng

V Xun Thy

TS.H Tt Thng

K tn
THEO DI SA I TI LIU
TT

V tr

Ni dung sa i

Ngy sa i

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Li m u: Hng dn ny c xy dng da trn nguyn tc ca cc ti liu sau:


1. EURACHEM: Quantifying Uncertainty in Analytical Measurement, Laboratory of the
Government chemist, London, UK, 1995. ISBN 0-948926-08-02.
2. Guide to the Expression of Uncertainty in Measurement, ISO, Geneva, Switzerland 1993.
ISBN 92-67-10188-9.
3. Protocol for uncertainty evaluation from validation data, Valid Analytical Measurement,
report number LGC/VAM/1998/088, January 2000.
4. ISO 5725:86: Accuracy (trueness and precision) of measurement methods and results Part
1-6.
5. SAC-SINGLAS Technical Guide 2. A guide on measurement uncertainty in chemical
analysis, First edition, April 2000

GII THIU CHUNG


Hng dn ny a ra cc v d c th v c lng khng m bo cho cc ch tiu th
nghim. Trong hng dn s a ra cc v d c th cho cc ch tiu phn tch ho hc nh
lng. T v d n gin n v d c lng KB cho cc ch tiu phn tch phc tp,
nhiu bc trong qu trnh th nghim. Mc ch ca hng dn l h tr, cung cp cho
cc phng th nghim ha hc c mt bc tranh tng qut v cc bc c lng KB
cho cc ch tiu c th v c th p dng c lng KB cho cc ch tiu c th ca
PTN.
Ni dung hng dn gm 3 phn chnh nh sau:
I.

c lng khng m bo thnh phn t nhng d liu c sn

II.

c lng khng m bo cho mt s bc trong qu trnh phn tch

III.

c lng khng m bo cho mt s ch tiu th nghim c th

AGL 18

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Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

I.

C LNG KB THNH PHN T NHNG D LIU C SN

1.

Qui nh k thut ca nh sn xut cung cp cho bnh nh mc 100mL cp A l


0.08mL. Tnh khng m bo chun ca th tch bnh?
Qui v Phn b dng hnh ch nht - khng m bo chun l:
0.08/ 3 = 0.046 mL

2.

Qui nh k thut ca nh sn xut cho pipet 2mL l 0,01mL. Tnh khng m bo


chun ca th tch cht lng chuyn qua pipet?
Qui v Phn b dng hnh ch nht - khng m bo chun l:
0.01/ 3 = 0.0058 mL

3.

Chng ch hiu chun cho cn 4 s cho bit khng m bo o l 0,0004g vi mc


tin cy khng t hn 95%. Tnh khng m bo chun?
Mc tin cy 95% nn qui v phn b chun v KB chun bng KB m rng
chia 2
0.0004/2 = 0.0002 g

4.

tinh khit ca mt hp cht ho hc c nh cung cp a ra l (99,9 0,1) %.


Xc nh khng m bo chun ca tinh khit ca hp cht?
Qui v Phn b dng hnh ch nht - khng m bo chun l:
0.1/ 3 = 0.058 %

5.

Mt qu cn hiu chun c chng nhn l 10.00000g 0.04mg vi mc tin cy t


nht l 95%. Tnh khng m bo chun ca qu cn?
Mc tin cy 95% nn qui v phn b chun v KB chun bng KB m rng
chia 2
0.04/2 = 0.02 mg

6.

lch chun ca cc ln cn lp li ca qu cn 0,3g l 0,00021g. Tnh khng m


bo chun?
lch chun chnh l khng m bo chun nn KB chun l: 0.00021 g

7.

Chng ch hiu chun ca pipet 25mL cp A c ghi khng m bo l 0.03mL.


khng m bo ny c da vo khng m bo chun nhn vi h s ph k=2,
cho mt mc tin cy xp x l 95%. Tnh khng m bo chun ca th tch cht
lng chuyn qua pipet?
C h s ph k = 2 nn qui v phn b chun v KB chun l:
0.03/2 = 0.015mL

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8.

khng m bo ca th tch dung dch trong bnh nh mc


dung mi hu c ho tan vo bnh nh mc 100ml n vch nh mc. Tnh
khng m chun ca th tch cht lng trong bnh nh mc?
Cho cc d liu sau:
- Kt qu ca 10 ln thc hin (cn v) vo bnh nh mc cp A 100mL c lch
chun l 0.01732mL
- Qui nh k thut ca nh sn xut cho bnh l 0.08mL. H s n ca th tch ca
dung mi hu c ho tan l 1x10-3 0C-1
- S khc nhau gia nhit phng th nghim v nhit hiu chun bnh nh mc
c c lng l 3 0C vi mc tin cy l 95%.
Tnh

khng m bo chun do s khc nhau gia nhng ln ong v cn chnh l


lch chun: 0.01732mL

T qui nh k thut ca nh sn xut qui v phn b hnh ch nht v tnh ra


khng m bo chun ca th tch bnh l 0.08/ 3 = 0.046mL

khng m bo do s khc nhau gia nhit phng th nghim v nhit hiu


chun bnh c c lng l Vx3x1x10 -3mL vi V l th tch ca bnh, 3 l s
bin thin nhit c th v 1x10 -3 l h s n th tch ca dung dch cht hu c. V
th tch gin n ca cht lng ln hn nhiu th tch gin n ca bnh nn ch quan
tm n th tch gin n ca dung dch. S khc nhau v th tch do nh hng nhit
c tnh (da vo mc tin cy l 95%) l:

100 x 3 x 1 x 10-3 = 0.3mL


V c chuyn thnh lch chun bng cc chia cho h s ph k=2
khng m bo do nh hng ca nhit l 0.3/2=0.15mL
Tng hp 3 thnh phn khng m bo trn s c khng m bo ca th tch
dung dch trong bnh nh mc l:

uv = 0.01732 + 0.0462 + 0.1502 = 0.16 mL


9.

khng m bo ca th tch cht lng chuyn qua pipet


Mt pipet 2mL cp A c s dng pha dung mi hu c. Tnh khng m bo
chun th tch cht lng c chuyn qua pipet?
Cho cc d liu sau:
- Cc th tch t 10 ln lp li vic chuyn cht lng t pipet 2mL cp A c lch
chun l 0.0016mL
- Qui nh k thut ca nh sn xut pipet l 0.01mL. H s n th tch ca dung
mi hu c l 1x10-3 0C-1
- S khc nhau gia nhit phng th nghim v nhit hiu chun bnh nh mc
c c lng l 30C vi mc tin cy l 95%.

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Tnh

khng m bo chun do vic ong lp li dung mi qua pipet chnh l lch


chun: 0.0016mL

khng m bo do hiu chun pipt t nh sn xut c qui theo phn b dng


hnh ch nht. Vy khng m bo do hiu chun pipet l:
0.01/ 3 = 0.0058mL

khng m bo do s khc nhau gia nhit phng th nghim v nhit hiu


chun pipet c lng l V x 3 x 1 x 10 -3 mL vi V l th tch ca pipet, 3 l s
thay i nhit v 1x10-3 l h s gin n th tch ca dung mi hu c. V th tch
gin n ca cht lng ln hn th tch gin n ca pipet nn ch cn tnh gin n
ca dung mi. S bin thin v th tch do nh hng ca nhit c tnh l:

2 x 3 x 1 x 10-3 = 0.006 mL
Sau chuyn thnh lch chun bng cch chia cho 2 nn khng m bo do nh
hng ca nhit l:
0.006/2 = 0.0030 mL
Tng hp 3 thnh phn khng m bo trn s cho khng m bo ca th tch
dung mi chuyn qua pipet l:

uv = 0.0016 + 0.0058 + 0.0030 = 0.0067


2

10.

mL

khng m bo cn
Mt phng php yu cu cn mt chun ni b 100mg trn cn 4 s. Tnh khng
m bo chun ca vic cn?
- Chng ch hiu chun ca cn c nu khng m bo o l 0.0004g vi mc
tin cy khng di 95%
- Cn lp li qu cn 100mg trn cn 4 s c lch chun l 0.000041g
Tnh
khng m bo t vic hiu chun cn c tnh ton t chng ch hiu chun.
khng m bo c trch dn l 0.0004g vi mc tin cy l 95%. Bin i
thnh lch chun bng cch chia khng m bo cho 2. khng m bo
hiu chun l:
0.0004/2= 0.0002g = 0.200mg

khng m bo do s bin thin ca cc ln c cn l lch chun ca cc


php cn lp li:
0.000041g = 0.041mg
Tng hp khng m bo trn s cho khng m bo ca khi lng vt liu
l:

u w = 0 .200 2 + 0 .041 2 = 0.208 mg


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11.

khng m bo ca nng mt dung dch


Dung dch chun ni b c chun b bng ho tan khong 100mg vt liu (cn bng
cn 4 s) trong mt dung mi hu c ho tan v y vo bnh nh mc ti vch
100mL . Tnh nng ca dung dch theo mg/L. Tnh khng m bo chun v
khng m bo m rng ca nng dung dch ?
D liu:
- 100.5mg vt liu c cn. khng m bo chun lin quan ti vic cn ny c
tnh v d 10 trn.
- tinh khit ca vt liu c trch dn t nh sn xut l (99.9 0.1)%.
- khng m bo chun ca th tch cht lng trong bnh nh mc 100mL c
tnh trong v d 9 trn.
Tnh
Cc ngun khng m bo gp phn vo ton b khng m bo ca nng dung
dch l:

Vic cn vt liu chun b dung dch;

tinh khit ca vt liu;


Th tch cui cng ca dung dch.

Cc khng m bo lin quan l:


Cn vt liu (uw): 0.208mg
tinh khit ca vt liu (up): 0.1/ 3 = 0.00058%
Th tch cui cng: 0.16 mL

Nng ca dung dch l C (mg/L) c tnh theo cng thc sau:


C (mg / L) =

W P
1000
V

Trong :
W:
P:

khi lng ca vt liu s dng (mg)


tinh sch ca vt liu s dng (% tinh sch chia 100)

V:

Th tch cui cng ca dung dch (mL)

Nng ca dung dch l:

100 ,5 99 ,9
1000 = 1004 , 0 mg / L
100

V cch tnh nng ch gm php nhn v chia, cc thnh phn khng m bo c


tng hp nh cc lch chun tng i (relative standard deviations)
T chng ta c th tnh khng m bo tng hp ca nng dung dch chun ni
b l:
u
uc
= w
C
W
2

u p uv
+ +
P V
2

0,208 0,00058 0,16


u c = 1004
+
+
= 2,69 mg/L
100,5 0,999 100
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Gi tr ca khng m bo chun ca nng dung dch chun ni b l 2.69mg/L


khng m bo m rng l 5.38mg/L c tnh ton bng cch s dng h s ph
k=2
Nng ca dung dch c th c cng b trong bo co l: 1004 5 mg/L v ghi ch
l khng m bo c tnh da vo khng m bo chun nhn vi h s ph
k=2 v cho mt mc tin cy l xp x 95%.

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II.

C LNG KB CHO MT S BC TRONG QU TRNH PHN TCH

1.

CN

1.1

Mc ch

1.1.1 Cn 500mg Cu bng cc phng php cn khc nhau


1.1.2 Cc bo co khi lng
Wt bnh ng + Cu, g
Wt bnh ng, g

32,5829
32,0822

Wt Cu, g

0,5007

Ch thch: Wt l Khi lng


1.2

Xc nh ngun khng m bo ca php phn tch

1.2.1 th nguyn nhn v kt qu/nh hng


PCu tinh khit
Khi lng WCu
lp li
Khi lng m (b)
Khi lng m (tng)
Tuyn tnh
nhy
Hiu chun
1.3

Tuyn tnh
nhy

lp li
Khi lng m

Hiu chun

nh lng cc khng m bo thnh phn

1.3.1 S tinh tinh khit ca kim loi ng


Cc nh cung cp a ra thng bo v tinh khit ca Cu trong chng ch phn tch
khi lng Cu l 99,99 0,01% m khng cp n tin cy ca n.
V khng ai a ra gii hn tin cy ca tinh sch ny, chng ti a ra 1 php tnh
v khng m bo theo phn b dng ch nht nn khng m bo chun u
(PCu) l

0 , 0001 / 3 = 0 , 000058
1.3.2 Qui trnh cn
1.3.2.1. Hiu chun tuyn tnh
Hiu chun bn ngoi ca cn c s dng tuyn b rng s khc nhau gia trng
lng tht trn a cn v s (trng lng) c trn thc chia trong khong
0,05mg vi tin cy 95%.
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Vi s phn b bnh thng, tin cy 95% cho mt h s l 1,96. Bi vy khng


m bo lin quan din t nh lch chun l
0,05/2 = 0,025mg
Ch : khng m bo ca thnh phn ny tng ln gp i bi lin quan n 2
ln cn mt ln l trc khi thm kim loi Cu v 1 ln l sau khi thm Cu
1.3.2.2. lp li
Lp li 10 ln php o c b v trng lng tng s c 1 lch chun ca cc sai
khc gia cc ln cn l 0,06mg vi khong trng lng trong khong t 20mg n
100mg
Ch : Chng ta tnh lp li ch duy nht 1 ln bi v n c tnh v s khc
nhau ca trng lng a n mt lch chun ca cc ln cn khc nhau
1.3.2.3. nhy
nhy ca cn c th khng c quan tm v nhng trng lng khc nhau c
o trn cng 1 cn phm vi rt hp
1.3.2.4. Tnh khng m bo chun tng hp trong quy trnh cn
u (mCu) =
1.4

Tng kt cc gi tr ca khng m bo
M t

1.5

2(0,025) + 0,062 = 0,07mg

Gi tr x

u(x)

u (x)/x

tinh khit ca kim loi Cu, P

0,9999

0,000058

0,000058

Wt ca kim loi Cu (mg)

500,7

0,07

0,00014

Tnh khng m bo tng hp v khng m bo m rng


Bi v khng m bo tng hp u (WCu) / WCu

0 , 000058

+ 0 , 00014

= 0,00015 vy u (wCu) = 0,00015 . 500,7 = 0,07

khng m bo m rng vi h s ph k = 2 l:
U (WCu) = 0.07 x 2 = 0.14
khi lng ng 500.7mg bo co khng m bo l:
500,7 0,14 mg vi h s ph k=2 [mc tin cy xp x 95%]
Ghi ch phn b khng m bo ca tinh khit ca ng l rt nh c th
khng tnh n.

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2.

S CHUN B TH TCH

2.1

Mc ch

2.1.1 chun b mt axt ng ho dung dch chun nitrat Cu t 500.7mg Cu pha trong bnh
nh mc 500mL
2.1.2 Cc bc tin hnh l:
a)
b)

Cn 500mg Cu tinh khit trong cc cn


Dng 5mL axit nitrit c ho tan Cu

c) Khi phn ng ngng v Cu ho tan hon ton trong dung dch axit. Chuyn
dung dch ny vo bnh nh mc 500mL. Thm nc ct n vch nh mc.
2.2

Xc nh ngun khng m bo

2.2.1 S nguyn nhn v nh hng


U (W)/W

500mL dung dch


Hiu chun th tch V
lp li
nh hng T

2.3

Xc nh khng m bo thnh phn

2.3.1 S khng m bo khi cn l yu t hnh thnh u tin khi cn 500,7mg 0,14mg vi


h s ph l 2
2.3.2 Hiu chun th tch ca nh sn xut
Tuyn b ca nh sn xut l bnh nh mc 500mL c sai s 0,15mL nhit 20 0C.
Khng c 1 tuyn b v tin cy no. Bi vy chng ta cho rng c mt phn b dng
tam gic v th tch tht dao ng gn tm hn l khong gii hn xa. Do vy khng
m bo trong hiu chun l 0,15/ 6 = 0,06mL.
2.3.3 Lp li cc phn tch th tch
Lp li 10 ln y v cn bnh nh mc 500mL cho mt khng m bo chun
di dng lch chun l 0,04mL s ny c dng tnh trc tip kt qu cui
cng.
2.3.4 nh hng ca nhit
Theo nh sn xut, bnh nh mc c hiu chun 200C, bi th trong phng th
nghim gii hn thay i trong khong 40C. khng m bo sinh ra t nh hng
ny c th c tnh t vic c lng khong nhit v h s n th tch. V n

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th tch ca cht lng ln hn n th tch bnh nh mc nn n ca cht lng


cn thit c coi trng, h s n ca nc l 0,00021 0C-1
Do th tch n l:
500mL x 4 0C x 0, 000210C-1 = 0,420mL
Tnh khng m bo chun i vi s thay i nhit bng s dng phn b
theo dng ch nht: 0,420/ 3 = 0,25mL
2.3.5 Tnh khng m bo chun tng hp cho php o th tch u (V)

0,062 + 0,042 + 0,252 = 0,26

u (V) =
2.4

2.5

Tm tt cc gi tr ca nhng khng m bo trong vic chun b th tch


M t

Gi tr X

U (x)

U (x)/x

Trng lng Cu, mg

500,7

0,07

0,00014

Th tch V, mg

500

0,26

0,0005

Tnh khng m bo tng hp v m rng


khng m bo tng hp ca vic chun b 500,7mg Cu trong 500ml dung dch xem
nh l khng m bo ca vic cn v ong th tch l:
u (Conc) / Conc =

[u (W ) / W ]2 + [u (V ) / V ]2

[0,075 / 500,7]2 + [0,26 / 500]2

= 0,00052
Cng nh nng ca dung dch Cu l 500,7mg/500mL = 1001,4mg/L
U(conc) = 0,00052 x1.001,4mg/L = 0,52mg/L
khng m bo m rng ca vic chun b 500,7 mg Cu trong 500ml dung dch hay
nng 1001,4 mg/L l 0,52 x 2 = 1,04mg/L vi h s ph k=2
Bi th, nng ca dung dch Cu l 1001,4 1,0mg/L vi h s ph k=2
2.6

Nhn xt

2.6.1 Vi nhng iu trnh by trn, phn b ca khng m bo khi cn nh hn nhiu so


vi vic chun b th tch

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3.

TNH TRNG LNG PHN T CA DUNG DCH

3.1

Mc ch
chun b 1 mol hoc dung dch thng, trng lng phn t ca dung dch cn bit
v khng m bo trong nh gi khi lng phn t. VD, chng ti yu cu tnh
khng m bo bng cch tnh khi lng phn t KMnO4

3.2

Hi ng IUPAC v khi lng nguyn t v s d tha ng v


IUPAC xy dng mt danh sch nhng nguyn t vi khi lng nguyn t ring r v
khng m bo lin kt trong bi Pure Appl.chem, vol 69,pp.2471-2473 (1997).
Bng danh mc y ca tt c cc nguyn t v khng m bo ca chng c th
tm thy trong trang web sau: http://www.chem.qmw.ac.uk/iupac/AtWt/

3.3 Tnh khi lng phn t ca KMnO4 v khng m bo


3.3.1 Khi lng nguyn t v khng m bo c lit k (t IUPAC) i vi tng
(thnh phn) nguyn t cu to ca KMnO4
khng m
bo u (e)

khng m bo
chun u (e)/3

Nguyn t

Khi lng nguyn t


AW (e)

39,0983

0,0001

0,000058

Mn

54,938049

0,000009

0,0000052

15,9994

0,0003

0,00017

3 c s dng t bng IUPAC sau khi tnh ton khng m bo trong


nh gi do s to nn cc lin kt bng phn b dng ch nht.

Ch :

3.3.2 Khi lng phn t KMnO4 l:


NWKMnO4 = 39,0983 + 54,938049 + 4x15,9994
= 158,0339 g.mol-1
u (NW KMnO4) = 0,000058 + 0,0000052 + (4 x 0,00017)
= 0,0007 g.mol-1
2

3.4

S phn b ca cc nguyn t KMnO4 l s tp hp n gin nhng s phn b ca


nguyn t n. Bi vy, khng m bo tng hp c tnh l bnh phng ca tng
bnh phng ca phn b ca tng nguyn t.

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Danh mc mt s nguyn t thng thng

Nguyn t
H2
C
N2
O2
F
Na
Mg
Al
P
S
Cl2
K
Ca
Cr
Mn
Fe
Co
Ni
Cu
Zn
as
Br
Ag
Cd
Ti
Sb
I
Ba
Hg
Pb

AGL 18

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Khi lng nguyn t


1.00794
12.0107
14.00674
15.9994
18.9984032
22.989770
24.3050
26.981538
30.973761
32.066
35.4527
39.0983
40.078
51.9961
54.938049
55.845
58.933200
58.6934
63.546
65.39
74.92160
79.904
107.8682
112.411
118.710
121.760
126.90447
137.327
200.59
207.2

KB lin quan
0.00007
0.0008
0.00007
0.0003
0.0000005
0.000002
0.0006
0.000002
0.000002
0.006
0.0009
0.0001
0.004
0.0006
0.000009
0.002
0.000009
0.0002
0.003
0.02
0.00002
0.001
0.0002
0.008
0.007
0.001
0.00003
0.007
0.02
0.1

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4.

NG CONG HIU CHUN

4.1

Quan h tuyn tnh


Mt phng php hoc mt dng c phn tch thng c hiu chun bng kho st
nhng xu hng, y, tm nhng gi tr ca x. Trong phn ln cc trng hp quan
h ny l mi quan h tuyn tnh v d:
y = a + bx vi a,b bit th v c mt ng cong hiu chun. Trong trng hp
ny nng xobs bit t phn tch 1 mu m quan st c s tr li yobs t c
cng thc: xobs = (yobs - a)/b
Trong vi trng hp, phng php phn tch yu cu quan h tuyn tnh i t 0 VD:
a = 0, trong trng hp ny quan h tuyn tnh l y=bx v xobs = yobs/b
Phng php thng thng ca quan h tuyn tnh da trn tng cp hiu chun (xi,
yi) c s dng phng php hiu chun bnh phng tuyn tnh nh nht (cng
hoc khng cng nh hng, tc ng = 0). .

4.2

Ngun khng m bo
C 4 ngun khng m bo chnh ng quan tm khi nh gi khng m bo ca
xobs
a. S thay i ngu nhin ca php o y (gm yi v yobs)
b. nh hng ngu nhin qui cho nhng gi tr ca xi
c. Hng s khng bit gia xi v yi
d. Gi nh khng c gi tr tuyn tnh
Trong 4 ngun trn th mt ngun c ngha nht l (a). Phng php nh gi (a)
thng qua s thay i cn li S, S c th tnh bng cng thc:
S2 = (yi - yc)2/(n-2)
Trong :
yi: l s c im hiu chun th i
yc: l s c tnh t y = a+bx
n: s im hiu chun
v u (xobs,y)= var(x) vi var (x)=S2/b2

4.3

V d
Trong v d ny c 3 chun s dng hiu chun
tp trung, xi

Thu c, yi

125

50

1,197

200

4,754

y = a + bx thch hp vi hiu chun trn a v b c tnh nh sau


AGL 18

Ln ban hnh : 1.04

Trang : 13

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

xi+yi+ - nx y
b=

a = y - bx
xi - nx
2

Trong v d ny
x

x2

xy

125

625

25

50

1197

59850

2500

200

4754

950800

40000

Tng

255

6076

1011275

42525

Trung bnh

85

2025,333

Trong :
xiyi+ - nxy

1011275 - 3 x 85 x 2025,333

b=

= 23,732

xi2 - nx2

42525 - 3 x 852

a = y - bx = 2025,333 23,7321343 x 85 = 8,102


V th
y = a + bx = 8,102 + 23,732x
Trong v d ny c th tnh ton thu c yc t gi tr bit ca x v bnh phng ca
hiu (y - yc)2
x

5
50
200

125
1197
4754

yc c tnh
126,76259
1194,7086
4754,5288

(y - yc)
3,10672
5,25036
0,27961

V th
S2 = (yi - yc)2/(n-2)
= (3,10672 + 5,25036 + 0,27961)/(3 - 2)
= 8,63669
var(x)

= S2/b 2
= 8,63669 / 23,73213432
= 0,0153346

u(xobs,y) =

AGL 18

var(x) =

0.0153346= 0,124

Ln ban hnh : 1.04

Trang : 14

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

5.

NG DNG CA GC-MS SC K KHI PH

5.1

Mc ch
V d di ch ra khng m bo o trong k kh khi ph GC- MS
(gas chromatographic - mass spectrometric).

5.2

Th t cc bc nh gi khng m bo lin quan


Bc 1: Cc yu cu k thut
Ho cht s dng cho k thut GC-MS phn tch nng biphenyl tinh khit trong
benzen. Cht chun s dng hiu chun l 50ug/ml dung dch chun v dung dch
trng (vd 0ug/ml).
Nng (ug/mL) ca biphenyl trong benzen c th c tnh ton s dng phng
php hiu chun hai im (Phng php Bracketing/ng hng)
Cspl = Aspl x C50/ (A50 - A0)
Trong :
Aspl :
Vng tr li/phn ng ca GC-MS cho biphenyl mu phn tch
A50 : Vng tr li/phn ng ca GC-MS cho biphenyl ca chun
A0 : Vng tr li/phn ng ca GC-MS cho biphenyl mu trng
C50 : Nng biphenyl dung dch chun c nng 50ug/ml biphenyl
Bc 2: Xc nh ngun khng m bo
chm
C50
Tuyn tnh

Tinh khit

Ho tan

Cn

chm

chm
Cspl

A0
chm

A50
chm

Aspl
chm

Bc 3: Xc nh khng m bo tng hp
C50:
- Dung dch chun c chun b t cht rn biphenyl, u tin cn v sau ho tan
v pha long trong benzen.
Vi cc ln cn khc nhau tnh c khng m bo chun l 0.000206g i vi
0,052g biphenyl

AGL 18

Ln ban hnh : 1.04

Trang : 15

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

- tinh khit ca biphenyl c nh cung cp tuyn b l ln hn 99.0% . Do s


tinh khit ca nguyn liu c tnh ton l 99.5% vi khng m bo lin quan
( 100% - 99%)/2 3 = 0,289%
- Cht rn byphenyl sau c ho tan v pha long thnh 1,000mL vo bnh nh
mc. Chi tit k thut ca bnh nh mc 1L s dng tuyn b cp chnh xc l
1000.22 0.20mL. Trong trng hp ny phn b hnh ch nht c s dng nh
mt phn ca kim tra vic kim sot cht lng (QC) ch ra rng trung tm phn
b ph hp hn cc vng gn phm vi. khng m bo chun t chng ch hiu
chun ca bnh gm thu tinh l 0,20/ 6 = 0,0816mL
Lp li vic v cn ch ra c khng m bo chun l 0,15mL.
V nh hng ca nhit khc nhau i vi khi lng benzen s dng ho tan
v pha long khng c d liu ca h s n ca benzen chng ta cho rng h s n
ca n l gp i nh nc ti nhit thng (m c h s n l 2,1x10-4mL/0C).
T kinh nghim chng ta bit rng c lng ny c th y . Nhit phng dao
ng trong khong 40C vi mc tin cy xp x 95%
khng m bo chun pht sinh t s thay i nhit
1000,22x (4,0/2) x 4,2 x 10-4 = 0,840mL
khng m bo chun v ho tan v pha long l
(0,816 + 0,15 + 0,840 ) = 0,857mL
2

Bi vy, C50 v khng m bo chun ca n c tnh ton nh sau:


m

Gi tr

0.052

99.5%

1000.22

KB

0.000206

0.289%

0.857

0.052

0.052206

0.052

0.052

99.5%

99.5%

99.789%

99.5%

1000.22

1000.22

1000.22

1001.077

C50

51.72862

51.9335

51.8789

51.6843

0.20492

0.15025

-0.04428

0.04199

0.02257

0.00196

0.066529
u (C50)

AGL 18

0.257933

Ln ban hnh : 1.04

Trang : 16

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

A0 , A50 , Aspl
o lp li a ra kt qu sau:

1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
Trung bnh
lch chun
lch chun trung bnh

Vng phn ng ca GC - MS
A0
A50
2
390
0
397
0
395
1
394
0
398
2
396
2
391
1
392
0
396
1
395
0.9
394.4
0.876
2.633
0.277
0.833

Aspl
265
260
269
266
263
268
265
262
267
265
265
2.749
0.869

Cc lch chun ca cc gi tr trung bnh bng trn c s dng trc tip nh


khng m bo chun lin quan v cc gi tr trung bnh m s c s dng trong tnh
ton cui cng
Tuyn tnh: Hiu chun hai im p dng tuyn tnh trong phm vi tp trung xc
nh. Tuy nhin cc nghin cu ch ra rng bng cch phn tch dung dch biphenyl ti
cc mc tp trung khc nhau, lch ln nht t kt qu thc l 1.0ug/mL. Phn
b dng hnh ch nht c p dng v khng m bo chun thay v tuyn tnh l
1.0/ 3 = 0.577.
Bc 4: Tnh ton khng m bo tng
Nh s tuyn tnh l kt qu cui cng th s bao gm
u tin cc khng m bo chun thay v C50, A0, A50 v Aspl l phi hp/t hp
bi phng php chia bng nh trnh by bng di a ra tp trung trong mu
l 34.836ug/mL vi khng m bo chun 0.266ug/mL.
Nh vy tng khng m bo chun l:

AGL 18

(0 , 222

+ 0 , 577

) = 0 , 618 ug

/ mL

Gi tr
KB

C50
51.72862
0.257933

A0
0.9
0.277

A50
394.4
0.833

Aspl
265
0.869

C50
A0
A50
Aspl

51.72862
0.9
394.4
265

51.98655
0.9
394.4
265

51.72862
1.177
394.4
264

51.72862
0.9
395.233
265

51.72862
0.9
394.4
265.869

Cspl

34.8363

34.861
0.0245
0.0006

34.7627
-0.07359
0.00542

34.9505
0.11424
0.01305

u(Cspl)

0.04924
0.2219

35.01
0.173703
0.030173

Ln ban hnh : 1.04

Trang : 17

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Tnh khng m bo m rng vi mc tin cy 95% h s ph k = 2.26 c s


dng nh ch xc nh 10 ln (bc t do =9). khng m bo m rng l
U (Cspl) = 0.618 x 2.26 = 1.397 ug/mL
Bi vy kt qu l:
34.8 1.4 (ug/mL)*

khng m bo c bo co l khng m bo m rng tnh ton s dng


h s ph k=2.26 vi bc t do l 9 m a ra mc tin cy tng ng l 95%.
5.3

Cch la chn cc khng m bo chun tng hp


C50 = 1000000 x mP/V
V th,
Cspl = Aspl x C50 / (A50 - A0) = 1000000 x mPAspl/ [V (A50 - A0)]:
m

A0

A50

Aspl

Gi tr

0.052

99.5%

1000.22

0.9

394.4

265

KB

0.000206 0.289%

0.857

0.277

0.833

0.869

0.052

0.052206 0.052

0.052

0.052

0.052

0.052

99.5%

99.5%

99.789%

99.5%

99.5%

99.5%

99.5%

1000.22

1000.22

1000.22

1001.077 1000.22

1000.22

1000.22

A0

0.9

0.9

0.9

0.9

1.177

0.9

0.9

A50

394.4

394.4

394.4

394.4

394.4

395.233

394.4

Aspl

265

265

265

265

265

265

265.869

Cspl

34.8363

34.9743

34.9375

34.8065

34.8608

34.7627

34.9505

0.13801

0.10118

-0.0298

0.02454

-0.0736

0.11424

0.01905

0.01024

0.00089

0.0006

0.00542

0.01305

0.04924
u(Cspl)

0.221902
Bng ny a ra kt qu tng t nh bng kt qu trc

AGL 18

Ln ban hnh : 1.04

Trang : 18

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

6.

SAI S RING/C TRNG CA CC PHNG PHP C BIT

6.1

Gii thiu

6.1.1 khng m bo ca c tnh cht liu ca vt cht nh trng lng v khi lng
ca dung dch chun b c mi lin h n gin v khng phc tp xc nh. Cc
c tnh phn tch ca vt cht m gi tr thc v bn cht ca thnh phn c xc
nh l phc tp bi v trong tin trnh phn tch chng ta tin hnh khng ch php
phn tch cui cng nhng hu ht cc trng hp mt th tc tin phn tch m c th
t bn thn n l ngun gy sai khc ln hn l s sai khc c tm thy trong bn
thn php phn tch.
6.1.2 Cc th tc phn tch thng da vo cc c tnh ho hc ca vt cht v d dng ca
hp cht cho php phn tch ngay hay tch ra qua tnh cht cht liu c s dng
tch v d tnh tan, tnh bay hi/d bay hi...Cc tnh cht s dng cho cc th tc
phn tch khng l cc tng chng l cc ch s mt hn ch nh hng n
chnh xc v cp chnh xc ca cc kt qu phn tch bi nguyn nhn sai s c th
ca khng m bo a ra trong phng php. V d: cc kt ta khng tan thng
tan trong mt chng mc no , qui trnh khng tch c th lin quan nh s hon
ho v ...v.v.
6.1.3 Khng th tho lun tt c cc nguyn nhn c th m c th ch dn cc sai s trong
cc qui trnh phn tch ring v nh phn tch phi bit trc cc sai s h thng v
hiu chnh chng khi khng m bo phn tch c tnh n. Hn na bt c
phng php phn tch v bt c s thay i no trong phng php l i tng ca
cc sai s c th ca bn thn phng php . y chng ta phi cp duy nht
cc ngun chung ca sai s c th m tnh cht quan trng ca sai s c th c din
t bng s.
6.2

Sai s trong vic ho tan kt ta

6.2.1 Ngun c th ca sai s trong php phn tch trng lng qua s kt ta l ho tan ca
cc kt ta.
S cn bng
ABn A + Bn
M c thit lp trong s khng ho tan ca mt tnh cht c th ho tan ca ABn. l
cc c im bi sn phm ho tan
Ksp = [A][B] n
tp trung [A] v [B] s dng trong sn phm pha long phi thng c din t
nh nng phn t gam. Cc gi tr Ksp ca mt vi vt cht quan trng c a ra
bng di:

AGL 18

Ln ban hnh : 1.04

Trang : 19

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Cc sn phm c th tan ca mt vi hp cht v c khng ho tan


ti nhit (C) trong ngoc ( )
Hp cht
Hp cht halogen
AgCl
AgBr
Agl
Hp cht hydro
Fe (OH)3
Al (OH)3
Zn (OH)2
Hp cht sunphat
CaSO4
BaSO4
PbSO4
Hp cht cacbon
CaCO3
MgCO3
BaCO3

Ksp
1.56 x 10 -10 (25)
7.7 x 10-13 (25)
1.5 x 10-16 (25)
1.1 x 10-36 (18)
1.1 x 10-15 (18)
1.6 x 10-14 (18)
2.45 x 10-5 (25)
1.08 x 10-10 (25)
1.5 x 10 -8 (18)
0.87 x 10-8 (25)
2.6 x 10 -5 (12)
7.0 x 10 -9 (16)

6.2.2 V d 1
tnh mi quan h sai s, nguyn nhn do ho tan ca phn ca cht kt ta trong qu
trnh ra bng cch gn/cht: 0.1g ca AgCl c ra vi 250ml nc ti 250C
y
Ksp = [Ag+][Cl-] = 1.56 x 10-10 mol2.L-2
Nu mt mol ca AgCl ho tan trong mt lt nc th: [Ag+] = [Cl -] = a
T vic ho tan ca mt mol AgCl, mt mol ion Ag v mt mol ion Cl c sinh ra
Thay th vo s din t cho Ksp chng ta c: Ksp = a2 = 1.56 x 10-10 mol2.L-2
v do : a = 1.25 x 10-5 mol.L-1
Nng ca AgCl g/L l a ra bi cng thc nhn trng lng gram ca AgCl
1.25 x 10 -5 mol.L-1 x 143.3g.mol-1 = 1.8 x 10-3 g/L
Bi vy ch 4.5 x 10 -4 g ca AgCl s ho tan trong 250ml nc ra s dng
Sai s i km l:
4.5 x 10 -4 x 100
% = 0.45%
0.1
Ch thch: Sai s ny khng nh nn khng th b qua c. Sai s ny phi c hiu
chnh trong vic c lng khng m bo.
6.2.3 V d 2
tnh nguyn nhn sai s trong phn tch trng lng ca 0.1g AgCl bng ra cht kt
ta vi 250ml dung dch HCl 0.01M
Trong trng hp ny, [Cl-] = 0.01M hoc 10-2mol.L-1

AGL 18

Ln ban hnh : 1.04

Trang : 20

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Khng ch vic so snh nng ion Cl m c th l nguyn nhn bi s ho tan


AgCl, chng ta thu c:
[Ag+] = Ksp /[Cl-] = (1.56 x 10 -10mol2.L-2) / 1 x 10-2mol.L-1
= 1.56 x 10-8 mol.L-1
Tng ng vi:
1.56 x 10 -8 mol.L-1 x 143.3g.mol-1 = 2.24 x 10 -6 g/L cho s tan ca AgCl
Bi vy 5.6 x 10-5 ca AgCl tan trong 250 ml ca axit s dng
Trong trng hp ny sai s l:
5.6 x 10 -5 x 100
% = 0.056%
0.1
Ch thch: Gi tr ny c th khng tnh ng trong cc phn tch chnh xc. Hn na
n c th cn nhc trong thc hin, tnh v t l khng hon ton c thit lp gia
dung dch ra v cht kt ta nn sai s thc t l nh hn nhiu so vi mt tnh ton.
6.3

Sai s trong chun cht kt ta

6.3.1 Sn phm tan c cng c s dng tnh ton trong chun cht kt ta m
im cn bng c th c ch ra bi s to thnh ca ho tan t t cht kt ta qu
mc chun v trong s phc tp chun m mt cht kt ta c ho tan li.
6.3.2 V d 3
Trong chun Mohr dung dch nitrat bc vo dung dch NaCl m thm cromat kali.
AgCl kt ta hon ton trong khi chun nhng rt nhanh chng nng Cl t mt
gi tr hin ti, dng clo bc nu ch ra mc cn bng hoc im kt thc. Tng cc
ion Cl khng c chun tng ng ti nng ion bc dng clo bc.
Tng s khng chun ny c th c xc nh t sn phm
K(Ag2CrO4) = [Ag+]2[CrO42-] = 2 x10-12mol3.L-3
Bng tnh nng ion trong dung dch v thay th gi tr ny vo cng thc ton.
Nu chng ta cn nhc trng hp m 1mL ca 5% w/v dung dch clo kali c thm
vo ho 25mL ca 0.1M dung dch Clo v 0.1 nitrat bc, nng ion Clo l:
0.05
1000
[CrO4 ] = ---------- x -------- mol.L-1 = 5.15 x 10-3 mol.L-1
194.2 50
2-

Khi nng ion Clo khng c chun l tng ng vi nng ion bc c


a ra l:
[Ag+] =

2 x10 12
1
mol.L = 2 x 10 -5mol.L-1
3
5.15 x10

Tng clo khng c chun l:

AGL 18

Ln ban hnh : 1.04

Trang : 21

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

50
--------- x 2 x 10 -5 mol = 10-6 mol
1000
Nh tng tnh l
25
-------- x 0.1mol
1000

V th sai s km theo l:

10 -6 x 100
--------------- %, hoc 0.04%
2.5 x 10-3

C th thy t tnh ton trn trong trng hp ny sai s chun ph thuc vo nng
ca cht ch th thm vo; iu ny c din t s cn nhc rng cht ch th
vn chuyn vai tr ca cht lm kt ta. Nu cho qu nhiu clo vo dung dch, clo bc
c nh dng trc khi t c im kt thc; mt khc nu nng clo l qu nh,
thm nitrat bc thm vo dung dch vt qu v bi vy m mu clo bc s kh nhn
thy hn. V th sai s c th lm nh i bi vic gi iu kin chun (nh tng
cht ch th nng v dung tch ca dung dch...) gn nh ging nhau qua s tiu
chun ho ca chun bng chun ca clorua natri tinh khit s dng sau khi
chun trong xc nh mu.
6.4

Sai s ca phng php chit

6.4.1 Trong m hnh phn tch ho hc cc phng php chit l thng c s dng
phn chia cc cht. Qui tc ca cc phng php l s phn b mt cht trong hai dung
mi. Th tc thc hin bao gm vic lc dung dch, thng mt dung dch m vic
lc khng ho trn c m cht c chit d ho tan hn theo s phn chia
c hc ca hai gia on.
6.4.2 Trong phn tch ho hc hu c dung dch - s chit dung dch c s dng cho thi
gian di; trong phn tch ho hc v c, hp cht phc tp (thnh thong mu) ca cc
kim loi khc nhau c chit trong dung mi phn cc thp. S phc tp ca mu c
th o c ngay lp tc bi my o mu. Hoc dung mi c th c bay hi sau khi
chia, v thnh phn chit c a tr li dung dch nc v xc nh c im chia.
6.4.3 Kh nng trng hp khc trong vch v d thc hin chit li vo dung dch nc ca
mt thnh phn khc (thng pH khc nhau) t dung dch/ho tan khi s chit bc
u kt thc. Trong cc phn tch vt s chit l thng bao gm quang ph hc hoc
trc quang s ht v pht ra nguyn t.
6.4.4 Nng ca cht trong dung dch gim i bi s chit t nng nguyn C0 ti nng
C1 l:
VA
C1 = C0 ---------------DVB + VA
Nu D l h s phn b v VA, VB l dung tch ca giai on A v B s dng trong chit.
Trong hot ng chit tip theo nng ca cht c chit gim so vi nng ban u l
AGL 18

Ln ban hnh : 1.04

Trang : 22

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

VA
------------------DVB + VA

C2 = C0

v sau n ln thc hin


VA
Cn = C0 ------------------DVB + VA

6.4.5 T nhng mi lin quan ny c tnh theo:


a. Th tch dung mi s dng c th ln;
b. Nhng chit hon ton s t c bng trung bnh ca chit lp li vi th tch dung
mi nh hn vi chit mt ln vi th tch ln;
c. Tuy nhin i lng chit hon ton c th khng bao gi t c bi vy chng ta
phi tho mn vi s chit hon ton thc nghim.
d. Vai tr quan trng c vn hnh bng gi tr ca i lng phn b, D nu cha
ln, vic chit hon ton c th t c vi s chit n; tuy nhin h s ny thp
vic chit phi c lp li mt vi ln.
6.4.6 V d 4
Khi chit 0.20g iot t 100mL ca dung dch nc vi 50mL phn CCl4 (D=85) phn
cn li sau ln chit u tin l
C1 = 0.2

100
------------------85 x 50 + 100

= 0.0046 g

v khng m bo chun gy ra bi s chit khng hon ton ln u tin l:


0.0046 x 100
=
1
------------------- = 2.3 % (mt sai s ng k)
0.2
Sau ln chit th hai cn li l:
=

C2 0.2

100
------------------85 x 50 + 100

2
= 0.0001057 g

v gy ra khng m bo l:
2

0.0001057 x 100
------------------0.2

= 0.05 % (khng c tnh n)

Do khi thc hin chit CCl2 l chit hon ton iot t dung dch nc.

AGL 18

Ln ban hnh : 1.04

Trang : 23

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

7.

CHUN B CHUN HIU CHUN

Gii thiu: V d ny tho lun v vic chun b mt dung dch chun cho thit b quang ph
hp ph nguyn t (AAS), t kim loi c tinh khit cao (trong v d ny l 1000mg/L Cd
pha trong HNO3). D v d ny khng i din cho mt php phn tch nhng vic s dng
chun hiu chun l mt phn ca hu ht cc php phn tch v php phn tch thng thng
l cc php o m c lin quan ti cc chun cung cp vic lin kt chun ti h n v SI.
Bc 1: Yu cu k thut
Mc tiu ca bc 1 l vit ra tuyn b r rng l i lng no c o. Yu cu k thut
v d ny l m t vic chun b chun hiu chun v php ton th hin mi lin quan ca i
lng o v cc thng s m i lng o ph thuc.
Qui trnh/th tc
Chun b chun hiu chun bao gm cc bc sau:
Chun b chun cadmi (Cd)
Lm sch b mt kim
loi

Cn kim loi

Ho tan v pha
long

Kt qu
Cc bc tin hnh l:
B mt ca kim loi c tinh khit cao c x l vi hn hp axit loi b cc oxit
kim loi. Phng php lm sch c nh cung cp kim loi a ra v cn c th thu
c kim loi tinh khit nh cng b trong chng ch.

Bnh nh mc (100mL) c cn theo 2 cch l khng c kim loi tinh khit v c kim
loi tinh khit bn trong. Cn s dng c phn gii 0.01mg.

1mL axit nitric (HNO3) (65%m/m) v 3mL nc kh ion c thm vo bnh nh


mc ho tan Cd (tng ng 100mg, c cn chnh xc). Sau bnh nh mc
c y nc kh ion n vch nh mc v lc bnh trn u t nht 30 ln.

Tnh ton
i lng o trong v d ny l nng ca dung dch hiu chun v ph thuc vo khi
lng tinh khit ca kim loi c tinh khit cao (Cd), Dung tch ca dung dch c pha
long. Nng ca dung dch c th hin theo cng thc:

AGL 18

Ln ban hnh : 1.04

Trang : 24

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

C Cd =
Trong :
CCd:

1000.m.P
mg / L
V

nng ca chun hiu chun [mg/L]

1000:
m :

thng s chuyn i t [mL] sang [L]


khi lng kim loi tinh khit [mg]

P
V

tinh khit ca ca kim loi


dung tch ca dung dch chun hiu chun [mL]

:
:

Bc 2: Xc nh cc ngun KB trong phn tch


Mc tiu ca bc th 2 l lit k tt c cc ngun gy ra KB ca tng thng s m nh
hng ti gi tr ca i lng o.
tinh khit:
tinh khit ca kim loi (Cd) c nh cung cp a ra trong chng ch l 99.99
0.01 %. do P l 0.9999 0.0001. Cc gi tr ph thuc vo hiu qu ca vic lm
sch b mt kim loi tinh khit. Theo th tc ca nh cung cp th khng c KB
trong vic nhim b mt kim loi do cc oxit kim loi v do khng c KB no
c ghi trong trong chng ch. Tuy nhin c th kim tra lp li v thy rng phn b
KB trong phn ny c th b qua.
Khi lng (m)
Giai on th hai ca vic chun b bao gm vic cn kim loi tinh khit.
Khi lng ca Cd c xc nh bng vic cn c b thu c m = 0.10028g
Ti liu ca nh cung cp xc nh l c 3 ngun KB trong vic cn c b: lp li;
kh nng c ( phn gii s/ digital resolution) ca cn v phn b KB trong
chc nng hiu chun ca cn. V chc nng hiu chun c 2 ngun KB c xc
nh l nhy v tnh tuyn tnh. nhy c th c b qua v cc khi lng khc
nhau c cn trn cng mt cn trong khong hp.
Th tch (V)
Th tch ca dung dch cha trong bnh nh mc c 3 ngun gy ra KB:
- KB ca chng ch ca bnh nh mc
- Phng sai ca cc ln cn v nh mc ti vch
- Bnh v dung dch trong bnh trong mt iu kin v nhit khc vi nhit hiu
chun bnh v dung tch.
Cc nh hng khc c th hin trong s xng c di y:

AGL 18

Ln ban hnh : 1.04

Trang : 25

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

tinh khit (P)

V
Nhit
Hiu chun
lp li

c(Cd)
phn gii

phn gii

m(b)
Tuyn tnh
nhy

m(tng)
Tuyn tnh
lp li

lp li

Hiu chun

nhy

Hiu chun
m

Bc 3: nh lng cc thnh phn KB


Trong bc 3 l nh lng cc ngun gy ra KB cho i lng o, s dng cc kt qu
thc nghim hoc t cc phn tch ton hc c lng.
tinh khit
tinh khit ca Cd trong chng ch ca Cd l 0.9999g 0.0001g. V khng c thm mt
thng tin no khc v KB nn gi thit l phn b hnh ch nht. c c KB
u(P) l chia 0.0001 cho

3
u ( P) =

0.0001
3

= 0.000058

Khi lng m
KB lin quan vi khi lng ca Cd c c lng s dng d liu t chng ch hiu
chun cn v khuyn co ca nh sn xut v vic c lng KB l 0.05mg. Vic c
lng ny l t 3 ngun KB c nu trong bc 2.
Th tch V
Chun b th tch c 3 ngun nh hng chnh: hiu chun, lp li v nhit .
- Hiu chun: nh sn xut cung cp s liu cho bnh nh mc l 100mL 0.1mL ti iu
kin 20 0C. Gi tr KB c cung cp khng c mc tin cy hoc thng tin v phn
b, do d on phn b theo hnh tam gic v KB c tnh theo cng thc:
0.1mL
6

= 0.04mL

- lp li: KB qua c lng thc nghim t vic cn v y 10 ln ti 100mL


cho lch chun l 0.02mL v c s dng trc tip thnh KB chun.
- Nhit : theo nh cung cp bnh nh mc th bnh c hiu chun 20 0C do nhit
PTN dao ng trong khong 4 0C. KB do nh hng ny c th c tnh t khong
AGL 18

Ln ban hnh : 1.04

Trang : 26

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

dao ng nhit v h s n ca dung tch. H s n dung tch ca dung dch ln hn ca


bnh nn ch cn nhc n h s n ca dung dch. T h s n ca nc l 2.1x10-40C-1,
th tch thay i l:
(100 x 4 x 2.1 x 10-4) = 0.084 mL
Gi thit phn b hnh ch nht nn KB chun l:
0.084mL
3

= 0.05mL

Ba phn b KB trn c tng hp thnh KB chun u(V):


u (V ) = 0.04 2 + 0.02 2 + 0.05 2 = 0.07mL
Bc 4: Tnh ton KB tng hp

C Cd =

Cng thc tnh CCd

1000.m.P
mg / L
V

S dng cc gi tr thu c th hin trong bng trn tnh c nng ca chun hiu chun
CCd nh sau:
C Cd =

1000 100.28 0.9999


= 1002.7mg / L
100.00

Ngun KB, gi tr v gi tr KB lin quan v KB tng i c th hin qua


bng sau:
M t

Gi tr x

u(x)

u(x)/x

tinh khit ca kim loi P

0.9999

0.000058

0.000058

Khi lng ca kim loi m

100.28

0.05 mg

0.0005

Dung tch ca bnh nh mc V (mL)

100.00

0.07 mL

0.0007

Da vo cng thc tnh KB tng hp vi trng hp cng thc tnh i lng o l php
nhn v chia ta thu c KB tng hp:
2

u c (Cd )
u (V )
u( P)
u (m)
2
2
2
=
+
+
= 0.000058 + 0.0005 + 0.0007 = 0.0009
C Cd
P
m
V
u(cCd) = CCd x 0.0009 = 1002.7mg/L x 0.0009 = 0.9 mg/L
KB m rng U(cCd) c tnh bng cch nhn KB tng hp vi h s ph k = 2:
U(cCd) = 2 x 0.9mg/L = 1.8 mg/L
Qua phn tnh KB tng hp chng ta nhn thy KB chun do nh hng ca vic
chun b dung tch bnh nh mc l ln nht sau n nh hng do cn kim loi. nh
hng do tinh khit ca kim loi gy ra KB rt nh, khng ng k.

AGL 18

Ln ban hnh : 1.04

Trang : 27

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

* Vic tnh ton KB tng hp c th c tnh theo cch s dng bng tnh di
y:
A

Gi tr

0.9999

100.28

100.00

KB

0.000058

0.05

0.07

4
5

0.9999

0.999958

0.9999

0.9999

100.28

100.28

100.33

100.28

100.00

100.00

100.00

100.07

c(Cd)

1002.69972

1002.75788

1003.19966

1001.99832

10

u(y,xi)

0.05816

0.49995

-0.7014

0.00338

0.24995

0.49196

11

u(y) ,u(y,xi)

0.74529

12
13

u(c(Cd))

0.9

Ch thch: Cc gi tr ca cc tham s c in t C2 n E2. KB tng ng c


in vo dng di t C3 - E3. Copy cc gi tr t C2 E2 vo ct th 2 t B5 B7. Kt qu
tnh nng C(Cd) tnh c t cc gi tr trn v a ra trong B9. C5 ch gi tr ca P t C2
cng thm KB C3. Kt qu tnh C(Cd) s dng cc gi tr t C5 - C7 th hin trong C9.
Ct D v E tnh C(Cd) cng theo cch trn. Cc gi tr ch ra dng 10 (C10 E10) tnh
bng cch ly cc gi tr t dng (C9 E9) tr i gi tr B9. Dng 11 (C11 E11) l cc gi
tr ca dng 10 (C10 E10) c bnh phng ln v tng cc gi tr ny c th hin trong
B11. C13 a ra gi tr KB chun tng hp tnh bng cch ly cn ca B11.

AGL 18

Ln ban hnh : 1.04

Trang : 28

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

III. C LNG KB CHO MT S CH TIU TH NGHIM C TH


CHUN AXIT/ BAZ: XC NH NNG CA DUNG DCH

1.

AXIT CLOHIRIC (HCL)


1.1

Phng php.
Xc nh dung dch NaOH 0,1 M bng chun trn c s dung dch KHP 0,1 M

(potassium hydrogen phthalate);


Sau xc nh nng ca 0,1M HCl bng chun trn c s dung dch 0,1M
NaOH.

1.2

1.3

Th tc.
-

Bc 1: Cn 5 g KHP
Bc 2: Ho tan KHP trong nc v nh mc ti 250 ml th tch

Bc 3: Tnh ton nng phn t gam ca KHP


Bc 4: Ho tan 2 g NaOH ht trong nc v nh mc ti 500 ml th tch

Bc 5: a 25 ml dung dch NaOH vo 1 bnh tam gic


Bc 6: Chun dung dch NaOH bng KHP t mt Burette 50 ml

Bc 7: Tnh ton nng ca dung dch NaOH


Bc 8: a 25 ml dung dch NaOH vo bnh tam gic

Bc 9: Chun dung dch NaOH c chun ho bng dung dch HCl t mt


Burette 50 ml

Bc 10: Tnh ton nng ca dung dch HCl


S nguyn nhn v kt qu xc nh nng dung dch NaOH

tinh khit (KHP)

Khi lng (KHP)


Nng (NaOH)

Th tch (KHP)

1.4

Trng lng phn t (MW - KHP)

nh gi cc thnh phn khng m bo

Bc 1: Cn KHP
Cng vic
Hp ng + KHP

33,5895 g

Hp rng

28,5130 g

Khi lng KHP

AGL 18

Ln ban hnh : 1.04

5,0765 g

Trang : 29

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Cc ngun khng m bo
Khi lng (KHP)
Hiu chun
Hiu chun
nhy
tinh khit (KHP)

nhy

lp li

tuyn tnh

lp li

tuyn tnh

Khi lng (th - c b)

Khi lng (b)


c (NaOH)
Th tch (KHP)

Trng lng phn t (KHP)

a) Kt hp vi vic hiu chun cn c s dng.


Giy chng nhn hiu chun ch ra rng ti tin cy 95%, mt ln cn t c bng
s khc nhau trong cng mt phm vi l 0,1 mg ca gi tr c hin th. Thnh
phn khng m bo c th c biu din nh lch chun bng cch chia
0,1/ 2 ta c 0,05 mg.
b) lch chun ca nhng ln cn lp li (1- 50 g) theo bo co QA ca PTN ch ra rng
mt lch chun 0,09 mg
Nhng khng m bo phi hp khi cn u (Wk)
u (Wk) =

(0,05 2 + 0,09 2 ) = 0,103 mg

Bc 2: Chun b dung dch KPH chun


Cng vic: Ho tan 5 g KPH v nh mc ti 250 ml trong bnh nh mc.
Cc ngun khng m bo
Khi lng (KHP)
nhy

Hiu chun

Hiu chun
lp li

tinh khit (KHP lp li

nhy
Tuyn tnh

Tuyn tnh
Cn (tng)
Cn (b)
c (NaOH)

lp li
Hiu chun
Nhit

Dung tch (KHP)

AGL 18

chch

lp li

im kt thc

Ln ban hnh : 1.04

Khi lng phn t (KHP)

Trang : 30

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

a.

khng m bo ca th tch i vi bnh nh mc c s dng: Catalog ca nh


sn xut ni rng bnh nh mc 250 ml c khng m bo 0,15 ml khng cp
ti tin cy. Do , mt phn b hnh ch nht ca sai s c chp nhn vi 3.
lch chun ca dung tch khi l 0,15/ 3 = 0,087 mL.

b.

khng m bo o trong vic nh mc n th tch thit k: lch chun i vi


nhng th tch khc nhau ca bnh nh mc c tnh ton sau mt lot cc ln nh
mc lp li (10 ln) v cn nc n vch nh mc c a ra l 0,014 mL. Gi tr
s c s dng trong ln tnh ton cui cng v khng m bo o trong php o
th tch.

c.

S khc nhau gia nhit dung dch v nhit hiu chun ca bnh nh mc.
Coi nh ch h s gin n th tch ca dung dch bi v n l con s ng k ln hn
dung tch gin n ca bnh nh mc thu tinh, cho mc ch thc hin.
Ly h s gin n i vi nc l 2.1 * 10-4 trn mt C v nu s khc nhau v nhit
gia dung dch v nhit hiu chun l 5 .. i vi dung tch 250 ml c s
dng s a ra mt khong tin cy 95% ca:
250 x 5 x 2.1 x 10-4 mL = 0.263 mL
Do s khc nhau lch chun ca nhit l:
0.263/ 2 = 0.132 ml
khng m bo o phi hp ca th tch chun KHP u (Vk) l:
u (Vk) =

(0,087 + 0,014 + 0,132 ) = 0,16 mL


2

Bc 3: Tnh ton nng phn t gam KHP


Cng vic: Nng ca dung dich KHP, Mk c tnh ton theo cng thc:
Mk = (Wk x P x 1000)/ (Vk x MW)
Trong :
Wk : Khi lng ca KHP c s dng (5,0765 g)
P: tinh khit ca KHP (99,8 0,2%)
Vk : Th tch ca dung dch c nh mc (250 mL)
MW: Khi lng phn t ca KHP theo cng thc C8H5O4K
B sung cho khng m bo ca Wk v Vk c kim tra sm hn, c hai khng m
bo c xc nh,:
a.
khng m bo i vi tinh khit ca KHP
tinh khit ca KHP do nh cung cp a ra l 99.8% 2%, c ngha P l 0.998
0.002. Khi tin cy khng c cng b cho khng m bo, Chng ta phi
a ra mt phn b hnh ch nht v sai s ng vi
u (P) = 0,002/
b.

3 , v ta c.

3 = 0,0012

khng m bo ca khi lng phn t KHP

AGL 18

Ln ban hnh : 1.04

Trang : 31

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Cng thc phn t ca KHP l C8 H5O4K . Xem bng trng lng nguyn t ca cc nguyn
t, C, H, O, K bao gm cc c lng khng m bo c ban hnh bi IUPAC tp ch
Ho hc tinh khit v ng dng, s 66.
Nguyn t

KB

Khi lng nguyn t

KB chun

12.011

0.001

0.00058

1.00794

0.00007

0.00004

15.9994

0.0003

0.00017

39.0983

0.0001

0.000058

Ch : Cc khng m bo chun c tnh bng cch chia khng m bo cho

S tp hp khng m bo ca mi nguyn t i vi khi lng phn t ca KHP c


tnh bng cch tng cng tng khng m bo chun bng s lng cc nguyn t ca
mi nguyn t trong cng thc phn t, v kt qu c lp bng nh di y:
S nguyn t
trong cng thc

Trng lng c
tnh ton

Kt qu c
tnh

KB c
tp hp

C8

8 x 12.011

96.088

0.0046

H5

5 x 1.00794

5.0397

0.00020

O4

4 x 15.9994

63.9976

0.00068

1 x 39.0983

39.0983

0.000058

204.2236
Do khi lng phn t ca KHP l 204,2236 v khng m bo tng hp ca u(MW)
l cn bc 2 ca tng bnh phng ca cc khng m bo ring bit. V d
u(MW) =

(0,0046 + 0,0002 + 0,00068 + 0,000058 ) = 0,0047


2

Tt c nhng tp hp khng m bo o, chng ta c th tm tt chng nh di y:


Nhn t KB

Gi tr c s
dng, V

KB

lch chun lin


quan

Wk

5.0765 g

0.000104

2.05 * 10 -5

0.998

0.0012

1.202 * 10-3

Vk

250 ml

0.16

6,4 * 10-4

MW

204.2236

0.0047

2.3 * 10-5

Do khng m bo chun ca nng (Mk) trong dung dch chun KHP c biu
din nh sau:
u (Mk)/ Mk =

(0,0000205 + 0,001202 + 0,00064 + 0,000023 ) = 0,0014 mol L-1


2

Nng dung dch KHP, Mk, c tnh theo cng thc (1) nh sau:
Mk = (5,0765 * 0,998 * 1000) / (250 * 204,2236) = 0,0992 mol L-1
AGL 18

Ln ban hnh : 1.04

Trang : 32

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Do ta c khng m bo chun u (Mk) trong dung dch KHP l:


u (MK) = 0,0014 * 0,0992 = 0,00014
T ta c nng dung dch KHP l 0.0992 mol L-1 vi khng m bo chun 0,00014
mol L-1.
Bc 4: Chun b dung dch NaOH
Dung dch NaOH c chun b c chun ho bng phn tch ho trc tip bi dung
dch chun KHP, khng m bo lin quan ti vic chun b dung dch NaOH khng c
lu mc d tinh khit ca dung dch NaOH v th tch dung dch c chun b c
khng m bo.
Bc 5: Cho 25 ml dung dch NaOH (bng Pipet) vo bnh nh mc
Nh trong bc 2, cc phn di y c lu khi chuyn 25 mL NaOH chun :
a.
khng m bo trong th tch nh mc pipet c s dng
Nh sn xut pipet tuyn b rng pipet c s dng c khng m bo 0,03 ml. Gn
ng vi phn b hnh ch nht bi v tin cy khng c bit, lch chun ca th tch
pipet c tnh l 0,03/ 3 hoc 0,017 mL.
b.

khng m bo trong nh mc pipet n 25 mL

Cc php o cn lp li ca th tch 25 mL vi pipet a ra mt lch chun 0,001 mL,


N c s dng trc tip trong vic tnh ton cui cng ca khng m bo chun.
c.

khng m bo trong thay i th tch do tc ng ca nhit .

(Nhit ca php phn tch Vs nhit hiu chun ca pipet)


ly s thay i nhit c th ca 5 0C v h s gin n th tch ca thu tinh l 2.1 x 10-4, do
vi tin cy ca php phn tch th tch l 95% th nhn t nhit l:
25 x 5 x 2,1 x 10-4 = 0,0263 mL
T , lch chun i vi s thay i nhit l 0,0263/ 2 = 0,013mL.
Phi hp tt c 3 ngun khng m bo trn, chng ta c khng m bo u (Vs) trong vic
chuyn i dung dch NaOH nh cn bc 2 tng bnh phng 3 lch chun trn, c kt
qu sau:
u (Vs) =

(0,017 + 0,001 + 0,013 ) = 0,021


2

Bc 6: Chun dung dch NaOH trn c s dung dch KHP chun (Va)
25 dung dch NaOH c chun trn c s dung dch chun KHP t mt burette 50 mL.
Mt khc, chng ta cn lu cc ngun khng m bo t quan im ca cc 3 yu t tng
ng c tho lun trn:
a.

khng m bo trong th tch c cng bo ca Buretter l 50 mL

AGL 18

Ln ban hnh : 1.04

Trang : 33

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Nh sn xut cng b rng buret c s dng c khng m bo 0.05 mL. Gn ng


vi phn b hnh ch nht bi v tin cy khng c bit, lch chun th tch ca pipet
c tnh l 0,05/ 3 = 0,029 mL
b.

khng m bo trong th tch dung dch chun KHP c s dng chun .

Khi mong mun s dng khong 25 mL dung dch chun trong vic chun , vic cn v c
lp li ca th tch 25 mL t buret c kim tra v a ra mt lch chun 0,012 mL.
Chng ta phi s dng con s ny nh khng m bo chun ca th tch c s dng.
c.

khng m bo ca nh hng nhit gia nhit chun ti nhit phng


v nhit hiu chun ca buret .

Ly s thay i nhit c th 5 0C nh trc v h s gin n th tch ca thu tinh l 2,1


x 10-4 0C, Vi tin cy 95% ca php o th tch th yu t nhit l:
25 x 5 x 2,1 x 10-4 ml = 0,0263 mL
Do , lch chun i vi s thay i nhit l 0,0263/2 = 0.013 ml.
Trong vic chun ny, 25,20 ml dung dch KHP c dng s dng trong vic t
c im cui vi dung dch NaOH. T y, s dng nhng s c c trn, khng
m bo chun phi hp u (Va) c tnh ton nh sau:
u (Va) =

(0.0292 + 0.0122 + 0.0132 )

hoc 0.034 ml

Bc 7: Tnh ton nng dung dch NaOH.


Cng thc c s dng trong vic tnh ton ca dung dch NaOH l:
Ms = (Mk x Va)/Vs
Ms: Nng dung dch NaOH
Mk: Nng dung dch KHP chun
Va: Th tch dung dch KHP chun c s dng
Vs: Th tch dung dch NaOH c chun
Lu ton b nhng tp hp khng m bo trong b 3 n bc 6, chng ta c th tm
tt chng nh di y:
Nhn t khng
m bo

Gi tr c s
dng, V

khng m
bo

lch chun
lin quan

Mk

0.0992 mol l-1

0.00014

1.41 x 10 -3

Vk

25.2 ml

0.034

1.35 x 10 -3

Va

25.0 ml

0.021

8,40 x 10 -4

khng m bo chun trong nng dung dch NaOH (Ms) c tnh nh sau:
u (Ms)/ Ms =
AGL 18

(0.00141 + 0.00135 + 0.00084 ) = 0.0021 mol L-1

Ln ban hnh : 1.04

Trang : 34

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Nng ca dung dch NaOH, Ms, c tnh theo cng thc (2) nh sau:
Ms = (25.20 x 0.992)/ 25.0 = 0.100 mol L-1
T , khng m bo chun u (Ms) trong nng dung dch NaOH l:
u (Ms) = 0.0021 x 0.1000 = 0.00021 mol L-1
Bc 8: Ly 25 ml dung dch NaOH chun HCL (Vb)
25 ml th tch dung dch NaOH chun c s dng cho chun , nhng lu tng t c
th c p dng nh trong bc 5, 25 ml dung dch vi khng m bo chun l u (Vb)
0.021 ml.
Bc 9: Chun NaOH trn c dung dch HCL (Vc)
Nh trong bc 6, dung dch HCL chun 25 ml NaOH t buret 50 ml. Do khng
m bo chun tng hp, u(Vc) l 0.034ml.
Bc 10: Tnh ton nng dung dch HCL (Mh)
Cng thc c s dng trong vic tnh ton dung dch NaOH l:
Mh = (Ms x Vb)/Vc
Mh: Nng dung dch HCL
Ms: Nng dung dch NaOH chun
Vb: Th tch dung dch NaOH chun c s dng
Vc: Th tch dung dch HCL c chun
Lu ton b nhng tp hp khng m bo trong bc 3 n bc 9, chng ta c th
tm tt chng nh di y:
Nhn t khng
m bo

Gi tr c s
dng, V

khng m
bo

lch chun
lin quan

Ms

0.100 mol l-1

0.00021

2.13 x 10 -3

Vb

25.00 ml

0.021

8.4 x 10-4

Va

25.10 ml

0.034

1,35 x 10 -3

khng m bo o chun lin quan trong nng dung dch HCL (Mh) c tnh nh
sau:
u( M h )
Mh

(0,002132 + 0,001352 + 0,000842 ) = 0,0027 mol L-1

Nng ca dung dch HCL, Mh, c tnh theo cng thc (3) nh sau:
Ms = (25,00 * 0,100)/ 25,30 = 0,0988 mol L-1
T , khng m bo tng hp chun u (Mh) v nng dung dch HCL l:
u (Mh) = 0,0027 x 0,0988 = 0,00027 mol L-1

AGL 18

Ln ban hnh : 1.04

Trang : 35

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bc 11: Tnh ton khng m bo m rng


khng m bo m rng U (Mh) c tnh bng cch nhn khng m bo tng hp
chun vi h s ph k, k = 2.
U(Mh) = 0,00027 x 2 = 0,00054 mol L-1
T ,nng dung dch HCL c phn tch l:
0,0984 mol l-1 0,00054 mol L-1
1.5

Nhng lu :

Trong chun axit/bazo ny, cc vn di y l nhng ngun c th gy ra KB v


chng c a ra xem xt trong v d ny. Nu cc ra KB b sung c nghi th cng
nn lu .
Cn khi lng Hiu chun cn v lp li;

Cn trng lng - nh hng bn ngoi ca khng kh trong PTN, c th khi mt cn


vi lng c s dng;

nh hng nhit s khc nhau gia nhit phng v nhit hiu chun;
tinh khit ca ho cht c s dng trong chun ho;

khng m bo ca khi lng phn t ca ho cht c s dng;


khng tinh khit c th , v d cht kim khc trong cc vin xt NaOH;

Sai s h thng trong bnh thu tinh dung tch;


Thay i trong vic tm kim im kt thc, v d vic nh gi c nhn;

Cc phn ng cnh tranh, nh s dnh bm ca carbon dioxide t khng kh.

AGL 18

Ln ban hnh : 1.04

Trang : 36

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

2.

XC NH AXIT LINOLEIC TRONG SA BO. PHN CHIT T SA BT


BI GC-FID

2.1

Chi tit k thut

a.
b.

Cn chnh xc 10g mu sa bt
Mu c chit di s c mt ca NH4OH vi ru cn, ete etilic v ete du.

c.

Cc este metyl axit bo ca vic chit cht bo c chun b bng s dng NaOCH3 v
este ho BF3CH3OH

d.
e.

Hn hp dung dch thu c sau lm bay hi ti gn kh di s bc hi ca nit.


Phn cn li sau c ho tan vo heptan v dung dch thu c chuyn vo bnh

f.

nh mc 25ml v thm heptan (C4H10) cho ti vch trn cng ca bnh.


Methyl linoleic c chia v xc nh s lng bi GS ch s ion ho vch hng.
ng cong hiu chun tuyn tnh s dng xc nh s lng c xy dng da
vo 4 mc tp trung ca methyl linoleic vi s y n im 0 (s dng im trng/
mu trng).

Nh phng php th bao gm cc qu trnh chit v dn xut bi vy xc nh chnh xc ca


mt phn tch l rt c lp trong hiu qu ca cc qu trnh ny. Theo tng bc nh gi s
hiu qu, nghin cu thu hi c tin hnh song song vi cc php phn tch bnh thng.
Vi t l thu hi R nng ca axit linoleic trong mu sa bt (Cspl) c th c tnh nh
sau:
Cspl = (Cml x V x Fla) / (R x Fml x Wspl)
Trong :
Cspl:
nng axit linoleic trong mu sa bt (mg/g)
Cml:
nng thu c t ng cong hiu chun methyl linoleic (mg/mL)
V: dung tch cn li ca dung dch trc khi phun 25mL/ injection
Fla: khi lng axit linoleic (g/mol)
Fml:
khi lng methyl linoleic (g/mol)
Wspl:
2.2

khi lng mu sa bt (g)

Xc nh ngun khng m bo
Nguyn nhn v kt qu c th c xy dng nh sau;

Fml

Fla

Cml
Cspl

Thu hi(R) chnh xc(r)

AGL 18

Ln ban hnh : 1.04

Wspl

Trang : 37

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

2.3

Xc nh khng m bo thnh phn

Cml kt qu thu c t ng cong hiu chun ca mu l 7.15mg/mL


cc phn tch ny 4 mc ca chun hiu chun vi nng l 1mg/mL, 2mg/mL, 4mg/mL
v 10mg/mL tng ng c chun b t 1000 2mg/mL methyl linoleic trong chun
chnh heptan. 4 chun hiu chun v mu trng ca
sau:
Nng
Phm vi phn
(mg/mL), xi
ng
0
2
1
135
2
280
4
560
10
1194

chng c o v thu c kt qu
Phm vi phn
ng thc
0
133
278
558
1192

Mi quan h ca im 0 bnh phng thch hp l y = bx a n ng cong hiu chun l:


xiyi
b=
x2
Nng xobs ca phn tch mt mu vi cc kt qu quan st thu c phn ng yobs sau
a ra bi xobs/b. Khng m bo u (xobs,y) trong gi tr d on xobs vi s thay i ca y
c th c lng t gi tr khc nhau ca phn cn li S nh v d 4 trn

Tng

xy

x2

133

133

278

556

558

2232

16

10

1192

11920

100

17

2161

14841

121

Do
xiyi
= 14841/121 = 122.6528926 y = 122.6528926x. V th:

b=
x

AGL 18

(y - yc)2

Tnh yc

133

122.652893

107.06263

278

245.305785

1068.9117

558

490.61157

4541.2005

10

1192

1226.52893

1192.2467

Ln ban hnh : 1.04

Trang : 38

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Do ,
S2 = (yi - yc)2/(n-2) = (107.06263 + 1068.9117 + 4541.2005 +1192.2467)/ (5-2)
= 2303.14
var(x)= S2/b2 = 2303.14/122.65289262 = 0.1531
u(xobs,y) = var(x) = 0.1531 = 0.391
Kt qu l u(Cml) = 0.391mg/mL
V: Bnh dung tch s dng y dung dch cui cng n 25mL c chng ch gi tr
25.040 0.015 mL ti 200C thu c t ti liu k thut ca nh cung cp. Trong nghin cu
ny phn b dng hnh ch nht c la chn. Bi vy khng m bo hiu chun l
0.0153 = 0.00866mL
Bnh thu tinh c s dng trong mi trng m nhit thay i 4.00C (mc tin cy
95%, nu khng nu mc tin cy th phn b hnh ch nht c tha nhn)
H s n ca heptan theo s thay i nhit l khng c bit n, c tha nhn n
khong gp hai nh h s n ca nc 2.1x10-4 mL/0C. Trong trng hp ny h s n ca
heptan c tha nhn l 4.2x10 -4 mL/0C. T kinh nghim ca chng ta chng ta bit rng s
tha nhn l trong c ln hn phm vi nhit ca PTN ni m tin hnh phn tch.
khng m bo theo s thay i nhit cho mt dung tch 25.04mL l t ti im ln nht
ca 25.04x(4.0/2)x4.2x10-4 = 0.0210mL
Bi vy, u (V) =

(0.008662 + 0.02102 ) = 0.0227

Fla: nh v d trc trng lng phn t (NW) ca axit linoleic (C18H32O2) v khng m
bo ca n c tnh nh sau:

Gi tr

12.0107

1.00794

15.9994

KB

0.00046

0.00004

0.00017

12.0107

12.01116

12.0107

12.0107

1.00794

1.00794

1.00798

1.00794

15.9994

15.9994

15.9994

15.99957

Fla

280.44548

280.454

280.447

280.446

0.00832

0.00128

0.00035

6.9E-05

1.6E-06

1.2E-07

7.091E-05
u (Fla)

AGL 18

0.008421

Ln ban hnh : 1.04

Trang : 39

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Fml: Tng t, NW ca mehtyl linoleic (C19H34O2) v khng m bo l:


C

Gi tr

12.0107

1.00794

15.9994

KB

0.00046

0.00004

0.00017

12.0107

12.01116

12.0107

12.0107

1.00794

1.00794

1.00798

1.00794

15.9994

15.9994

15.9994

15.99957

Fml

294.47206

294.481

294.473

294.472

0.00878

0.00136

0.00035

7.7E-05

1.8E-06

1.2E-07

7.902E-05
u (Fml)

0.0088895

Wspl: khi lng ca mu l 10.0232g vi nhiu ln cn khc nhau. Bo co hiu chun ch


ra lch ln nht l 0.4mg t gi tr nu ca cn chun a ra l 0.4/ 3 = 0.231mg ca
khng m bo chun cho mi ln cn. Nh cn ca mu bao gm c trng lng ca mu
v bao b khng m bo tnh t lch c nhn hai:
u (Wspl) =

(0.231 + 0.231 ) = 0.327 mg 0.000327g


2

Thu hi (R): theo cc bc nghin cu trc trn mt mu tng t th thu hi lp c


th hin trn mt mu n v tm thy c trung bnh thu hi l 91.3% vi lch chun l
5.4%. V th lch chun lin quan l 5.4%/91.3% = 0.0591. i vi mu hin ti s thu hi
l 0.950 (95%). V th khng m bo chun theo thu hi l 95%x0.0591 = 5.61%.
chnh xc(r): trc, mt mu c th lp li. Qua m nghin cu tnh chnh xc
nng axit linoleic c tm thy trung bnh l 5.65mg/g vi lch chun l 0.96mg/g.
V th khng m bo chun lin quan l 0.96/5.65 = 0.170
2.4 Tnh ton khng m bo tng
Cc khng m bo chun theo Cml, V, Fla, R, Fml v Wspl c tng hp u tin bi
phng php chia bng sau:

AGL 18

Ln ban hnh : 1.04

Trang : 40

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Cml

Fla

Fml

Wspl

Gi tr

7.15

25.04

280.44548

95.0%

294.47206

10.0232

KB

0.391

0.0227

0.008421

5.61%

0.00889

0.00033

Cml

7.15

7.541

7.15

7.15

7.15

7.15

7.15

25.04

25.04

25.0627

25.04

25.04

25.04

25.04

Fla

280.44548

280.445

280.445

280.4539

280.445

280.44548

280.445

95.0%

95.0%

95.0%

95.0%

100.61%

95.0%

95.0%

Fml

294.47206

294.472

294.472

294.47206

294.472

294.48095

294.472

Wspl

10.0232

10.0232

10.0232

10.0232

10.0232

10.0232

10.0235

Cspl

17.906665

18.8859

17.9229

17.907203

16.9082

17.906125

17.9061

0.97923

0.01623

0.0005376

-0.9985

-0.000541

-0.00058

0.95889

0.00026

2.89E-07

0.99695

2.922E-07

3.4E-07

1.9561078
u(Cspl)

1.3986092

khng m bo chun trn thu c l tng hp m t s chnh xc c a ra cho


tng khng m bo thnh phn. S chnh xc thu c trn l lch chun lin quan, v
th
u(Cspl)/Cspl =

[(1.3986/17.9067) + 0.170 ] = 0.187


2

V th, tng khng m bo thnh phn l


u(Cspl) = 0.187xCspl = 0.187 x 17.90667 = 3.35mg/g
khng m bo m rng U(Cspl) vi mc tin cy 95% thu c t khng m
bo tng hp vi h s ph k=2 l:
U(Cspl) = 2x3.35 = 6.70mg/g
Nng ca axit linoleic trong th nghim sa bt tnh c l:
17.9 6.7 mg/g*
Bo co khng m bo l khng m bo m rng c tnh bng cch s dng h s
ph k=2 vi mc tin cy tng ng l 95%
2.5 Nhn xt
nh gi cc thnh phn quan trng
Theo qui lut nhn ln ca sai s s thm vo v bt i lin quan, v d x = x1 + x2 khng
m bo tng hp l cn ca bnh phng ca tng cc khng m bo thnh phn. v d

AGL 18

Ln ban hnh : 1.04

Trang : 41

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

( )

( )]

u(x) = u x1 + u x2 . V th s tp trung ca tng thnh phn ti khng m bo tng


hp c th so snh trc tip gia u(x1) v u(x2).
Tuy nhin trong trng hp khi mi lin quan l nhn v/hoc chia. v d x= x1/x2 khng
m bo chun tng hp c tnh l:
2

u(x)/x =

{[u(x )/ x ] +[u(x )/ x ] }
2

Bi vy trnh t so snh phn b ca mi thnh phn ti khng m bo tng mt l


so snh mi lin quan ca khng m bo chun ca cc thnh phn v d: u(x1)/x1 v
u(x2)/x2
i vi v d tho lun trn mi lin quan l nhn v chia. V th s so snh phn b mi
thnh phn , mi thnh phn khng m bo phi c th hin mi lin h vi khng
m chun.
Gi tr

KB chun

KB lin quan

Cml

7.15mg/g

0.391 mg/g

0.0547

25.04mL

0.0227 mL

0.0009

Fla

280.44548g/mol

0.00842 g/mol

0.00003

Fml

294.47206g/mol

0.00889 g/mol

0.00003

Wspl

10.0232

0.000327 g

0.00003

95.0 %

5.61 %

0.0591

r
Cspl

0.170
17.90667mg/g

3.35 mg/g

0.192

T bng trn r rng s gp phn chnh l t Cml, R v r. Thc t mt c th trnh by


khng m bo tng hp ch t ba ngun bng cch l i khc ngun nh khc. Trong trng
hp ny,
u(Cml)/Cml =

(0.05472 + 0.05912 + 0.1702 ) = 0.188. V th u(Cml) = 0.188x17.90667 =

3.37mg/g. V vy r rng l khng m bo ch c tnh t cc ngun chnh l khng c


ngha khc nhau t s thu c sau khi cn nhc tt c cc ngun.

AGL 18

Ln ban hnh : 1.04

Trang : 42

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

3.

XC NH HM LNG AXITS TRONG DU

3.1 Phng php


c s ca mu du c ho tan trong s trung ho IPA v chun da vo cht chun l
dung dch KOH 0.1mol/L vi cht ch th phenolphthalein.
3.2

S nguyn nhn v kt qu
CKOH

W (du)

MW (KOH)

Gi tr ca axit
Chun (V)

3.3 Xc nh cc khng m bo
3.3.1 Dung dch chun KOH 0.1M vi HCl 0.5M
A

Chun b HCl 0.5M


V phng din thng mi chun b dung dch HCl cha 18.230 g HCL (mHCl) c
s dng chun b Cst HCl 0.5M dung tch1000mL
Bnh nh mc s dng chun b dung dch c dung tch l 1000mL 0.4mL ti
200C. lch chun thch hp ca s hiu chun dung tch s dng phn b hnh ch
nht l 0.4/ 3 hoc 0.23mL
T nhit thc v nhit hiu chun bnh nh mc l -3 0C vi mc tin cy 95% h
s n dung tch ca nc l 2.1 x 10-4/0C, s bin i th tch c th l 1000 x 3 x 2.1 x
10-4 hoc 0.63mL
lch chun tng ng l 0.63/1.96 hoc 0.32mL
Tng hp hai phn b khng m bo u(V), chng ta c:
u(V)/V =

(0 .23

+ 0 . 32

)/ 1000

= 0.00039

Nng ca HCL l mHCL/ MHCL. V ni MHCL l khi lng phn t ca HCL.


Nh sn xut dung dch HCL ch dn mt lch c th ca chun 0.02% cho mi
0

C. Ly mt s khc nhau c th trong PTN ca nh sn xut - 20 C (vi tin cy


95%), khng m bo chun ca m HCL l:
u(mHCL) = 18.230 x 0.02 x 2 / (100 x 1.96) = 0.004 g
u(mHCL)/ m HCL = 0.00022

AGL 18

Ln ban hnh : 1.04

Trang : 43

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

khng m bo chun ca trng lng phn t HCL, theo khi lng nguyn t
IUPAC v phn b hnh ch nht, l:
u(mHCL) = 0.000043
Cn lu rng u(MHCL)/ MHCL l khng ng k khi so snh vi u(V)/V v
u(mHCL)/ mHCL , khng m bo chun lin quan l:
u(Cst)/ Cst =
B.

2
2
(0.00039 + 0.00022 ) = 0.00045.

Xc nh CKOH
Nng chnh xc ca dung dch KOH c thit lp trc khi s dng bng chun
trn c s dung dch HCL chun.
Do , CKOH = Cst * Vst/ VKOH
Ni, Vst l th tch (ml) dung dch HCL chun c s dng cho chun th tch
VKOH (ml) ca dung dch KOH.
Nh c trnh by trn, u(Cst)/ Cst = 0.00045
i vi vic chuyn dung dch KOH n bnh tam gic, mt pipet thu tinh dung tch 5
0.01 ml c s dng. Ly s bin i nhit c th l 30C vi tin cy 95%,
v lp li ca pipet ( lch chun) 0.0033 mL, n c th tnh u(VKOH)/VKOH =
0.0015
Vic chun c hon thnh s dng mt buret vi lng 5 ml c vch chia 0.01
ml ( chnh xc hiu chun ca nh sn xut l 0.01 ml)
S sai khc nhit c th l tng t nh nu trn, lch chun ca vic ong l
0.0033 mL v lch chun ca im ch cui cng tng ng vi c ca buret
(0.017mL) l 0.0098mL
V th gi tr ln nht ca u(Vst)/Vst = 0.013 nu CKOH = 0.1 mol/L v tng ng Vst
= 1 mL
khng m bo u(Cst)/Cst v u(VKOH)/VKOH l khng ng k trong so snh ti
u(Vst)/Vst
V vy u(CKOH)/CKOH = u(Vst)/Vst = 0.013

3.3.2 Xc nh gi tr ca axit
Gi tr ca axit l:
AV = MKOHVKOHCKOH / m
Lu phng php th s dng v khi lng phn t KOH l 56.1 thay th hon ton
gi tr MKOH = 56.10564; bi th trong trng hp ny
u(MKOH)/MKOH = 0.00564/(56.1 x 3 ) = 0.00006
chit axit bo t do bng dung dch KOH chng ta s dng buret 5mL ni trc
u(Vst)/Vst = u(Cst)/Cst = u(Vst)/Vst = 0.013
AGL 18

Ln ban hnh : 1.04

Trang : 44

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

khng m bo ca mu du trng lng 2.5g l u(m)/m = 0.0023


Tht r rng khng m bo ca trng lng phn t KOH v ca mu du l khng ng
k, v th

u( Av) / Av =

[u(VKOH ) /VKOH ] + [u(CKOH ) / CKOH ]2 = 0.018

khng m bo m rng vi h s ph k 2 l
U(AV)/AV = 2 x 0.018 hoc 0.04
Ghi ch: s ch ca im kt thc ca chun l ngun nh hng khng m bo ln. Nu
buret v d c c l 0.043mL khng m bo m rng c th tng n 0.07
Hn na mu ca du v s thay i c th trong cht ch th chy ti gn im kt thc trong
dung dch du ho tan l khng c tnh ti. Tng t mi lin quan ti nh hng ca CO2
khng kh trong CKOH

AGL 18

Ln ban hnh : 1.04

Trang : 45

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

4.

C LNG KB T D LIU PH DUYT PHNG PHP.


XC NH THUC TR SU PHOSPHO HU C TRONG BNH M

4.1

Tm tt

4.1.1 Mc ch
Xc nh d lng thuc tr su phospho hu c trong bnh m dng phng php chit v
sc k kh (GC)
4.1.2 Qu trnh o
Qu trnh xc nh d lng thuc tr su phospho hu c trong bnh m c th hin
hnh 4.a
Hnh 4.a. Phn tch thuc tr su phospho hu c
ng nht

Tch chit

Lm sch

C c

Chun b chun
hiu chun

Xc nh GC

Hiu chun GC

Kt qu

4.1.3 i lng o
D lng thuc tr su phospho hu c trong bnh m c tnh theo cng thc sau:
Pop =

I op .cref .Vop
I ref . Re c.msample

.Fhom .10 6 mg.kg-1

Trong :
Pop: hm lng thuc tr su c trong mu (mg.kg-1)
Iop : Cng ti a ca phn chit ca mu
Cref : Nng khi lng ca cht chun (g.ml-1)
Vop : Th tch cui cng ca dch chit
106 : chuyn t g.g-1 sang mg.kg-1
Iref : Cng ti a ca cht chun

AGL 18

Ln ban hnh : 1.04

Trang : 46

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Rec : phn thu hi/ thu hi


msample : khi lng mu
Fhom : h s hiu chnh cho mu khng ng nht
4.1.4 Xc nh cc ngun gy ra khng m bo o
Cc ngun gc ca khng m bo o c cc nguyn nhn c th hin hnh 4.b
Hnh 4.b Phn tch ngun gc ca khng m bo o
ti lp

c(ref) tinh
khit (ref)

V(op)

m(ref)

I(op)
I(ref)
m(ref)
V(ref)
pha long

tuyn
tnh

hiu chun

I (op)

nhit

hiu chun

nhit

V(ref)
hiu chun

hiu chun

V(op)

hiu chun

Pha long

m (sample)

P(op)
m(gross)

m(tare)

tuyn
tnh

hiu chun

tuyn
tnh

hiu chun
hiu chun

F(hom)

I(ref)

thu hi

m mu

4.1.5 nh lng cc thnh phn ca khng m bo o


Da trn d liu ph duyt phng php th ni b, c 3 yu t to nn KB o. Cc yu
t ny c lit k trong bng 4.c v th hin trn s 4.d (cc gi tr c suy ra t bng
4.e)
Bng 4.c : khng m bo o trong phn tch thuc tr su
Gi tr

KB
chun u(x)

j KB chun
tng i u(x)/x

lp li

1.0

0.27

0.27

Da trn 2 ln th ca 2
mu khc nhau

chch

0.9

0.043

0.048

mu thm

Cc ngun gc khc
( ng nht...)

1.0

0.2

0.2

nh gi trn c s
phng php s dng

u(Pop)/Pop

--

--

0.34

KB chun tng i

M t cc ngun

AGL 18

Ln ban hnh : 1.04

Ghi ch

Trang : 47

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Hnh 4.d khng m bo o trong phn tch thuc tr su


u(y,xi) (mg/kg)
ng nht
chch
lp li
P(op)
0

0.1

0.2

0.3

0.4

Cc gi tr ca u(y, xi) = (y/xi).u(xi) ly t bng 4.k


4.2

Hng dn chi tit

4.2.1 Gii thiu


V d ny minh ha mt mt phng php tnh KB bng cch s dng cc d liu ph
duyt phng php th ni b. Mc ch ca php o l xc nh tng d lng thuc tr su
phospho hu c trong bnh m. Qui trnh ph duyt v cc lin kt thit lp nghin cu thc
nghim bng cch tin hnh trn cc mu thm. Gi nh KB do s khc nhau trong kt
qu o ti s thm v phn tch trong mu l nh so vi KB o tng.
4.2.2 Bc 1. c im k thut
c trng ca php o i vi cc phng php phn tch c m rng s tt hn nu c
m t cc bc khc nhau ca qu trnh phn tch v m t cng thc ton hc tnh kt qu
o/th.
Qui trnh:
Qu trnh o c m t trong s hnh 4.a Qu trnh ny c chia thnh cc giai on
sau:
i)

ng nht: Mu c chia thnh nhng phn nh (khong 2cm), ly ngu nhin 15


phn trong s ny v ng ho chng. Nu c phn no nghi ng khng c
ng ho tt phi c ly li trc khi trn u.

ii)
iii)

Cn trng lng ca khi mu ny c khi lng l msamble


Chit : lng dch chit ca mu phn tch vi dung mi hu c, em gn v sy qua

iv)

ct Na2SO4 v c c dch chit bng cch s dng thit b Kuderna-Danish.


Ho lng

v)

Axeton nitril/hexane lng, ra axeton nitril bng hexane, sy hexane qua ct natri
sulphate

vi)

C c dch ra bng ga vi ngn la nh cho n khi dch chit gn nh kh hon


ton

AGL 18

Ln ban hnh : 1.04

Trang : 48

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

vii) Pha long n th tch tiu chun Vop (khong 2ml) trong ng nghim chia vch 10ml
viii) o : Tim 5l dch chit v o GC c c cng mi (peak) Iop
Chun b dung dch chun 5l .ml-1 (nng khi lng Cref)
Hiu chun GC s dng 5l dung dch chun, tim v o GC c cng mi

ix)
x)

chun Iref
Hnh 4.a. Phn tch thuc tr su phospho hu c
ng nht

Tch chit

Lm sch

Bulk up

Chun b chun
hiu chun

Xc nh GC

Hiu chun GC

Kt qu

Tnh ton kt qu
Nng khi lng Cop ca mu cui cng c tnh nh sau
C op = c ref .

I op
I ref

g.ml-1

v nh gi Pop ca hm lng thuc tr su trong mu c (mg.kg-1) c tnh bi:


Pop =

cop .Vop
Re c.msample

. 106

mg.kg-1

Hoc, thay Cop vo cng thc c


Pop =

I op .c ref .Vop
I ref . Re c.m sample

.10 6 mg.kg-1

Trong :
Pop: Mc thuc tr su trong mu (mg.kg-1)
Iop: Cng mi ca mu chit
Cref: Nng khi lng ca mu chun (g.ml-1)

AGL 18

Ln ban hnh : 1.04

Trang : 49

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Vop: Th tch cui cng ca dch chit (ml)


106: i t (g.g-1) ra (mg.kg-1)
Iref : Cng pick ca mu chun
Rec: thu hi
msample: Khi lng ca chia [g]
Phm vi:
Phng php phn tch ny c kh nng p dng cho cc thuc tr su c bn cht ho hc
tng t vi nng t 0.01 n 2 mg.kg-1 vi cc loi mu bnh m khc nhau.
4.2.3 Bc 2: Xc nh cc ngun KB o
Xc nh v phn tch cc nguyn nhn gy ra khng m bo o l mt qu trnh phn
tch tng hp, cch tt nht l phn tch theo s nguyn nhn v kt qu. Cc thng s
trong biu thc ton hc ca php th c th hin bi cc nhnh chnh trn s . Cc
thng s mi c thm vo s theo tng bc trong qu trnh phn tch (hnh 4.b) cho
n khi cc thng s c lit k y .
S khng ng nht ca mu khng phi l mt thng s trong cng thc ton hc tnh kt
qu ca php th nhng n li nh hng ng k n qu trnh phn tch. V th mt nhnh
chnh mi cho thng s F(hom), i din cho s khng ng nht ca mu c thm vo
trong s nguyn nhn v kt qu (hnh 4.e)
Hnh 4.e:
S phn tch cc ngun gc KB o trong trng hp mu khng ng nht

hiu chun

c(ref) tinh
khit (ref)

tuyn tnh

I (op)

V(op)

m(ref)

chm

nhit

hiu chun chm

nhit

V(ref)

hiu chun
hiu chun

hiu

chm

chm

Pha long

P(op)
m(khng b)
m(b)

chm
tuyn
tnh

hiu chun

nhy

nhy
F(hom)

AGL 18

thu hi

Ln ban hnh : 1.04

I(ref)

hiu chun

tuyn
tnh

chm
hiu chun
m mu

Trang : 50

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Tm li s th hin cc nguyn nhn dn n khng m bo o th s khng ng nht


ca mu bao gm trong s tnh ton ca php th. ch ra cc kt qu ca khng m
bo o mt cch r rng, tt nht nn vit cng thc:
I op .cref .Vop

Pop =

I ref . Re c.msample

.Fhom .10 6 mg.kg-1

Trong F(hom) l mt s hiu chnh gi thit cho tnh thng nht trong php tnh gc. iu
ch r rng khng m bo o trong h s hiu chnh phi c bao gm trong s nh
gi khng m bo o tng. Cng thc trn cng ch ra khng m bo o s c p
dng.
Ch thch: Cc h s hiu chnh: cch tip cn ny kh l khi qut v c th rt c gi tr. V mt
nguyn l, mi php th u phi c cc h s hiu chnh thng nht. V d: khng m bo o Cop
c th c din t bng khng m bo o chun i vi Cop hoc khng m bo o chun
i din cho khng m bo o trong cho h s hiu chnh. Trong trng hp sau, gi tr xc nh
khng m bo o Cop c din t bi lch tiu chun tng i.

4.2.4 Bc 3 : nh lng cc thnh phn ca khng m bo o


nh lng cc thnh phn khc nhau ca KB o t d liu nghin cu phng php
th ni b v nghin cu ph duyt phng php:
- S c lng tt nht ca phng sai tng ca ton b qu trnh phn tch qua nhiu
qu trnh th nghim (th nghim lp li, ti lp)
S c lng tt nht ca tng chch (Rec)v khng m o ca n

S nh lng ca bt c khng m bo o no lin quan n kt qu tnh ton


khng y i vi ton b qu trnh.

Mt vi s sp xp li cc nguyn nhn v biu hiu qu rt c gi tr to thnh mi lin


h v a ra nhng thng tin r rng hn. (hnh 4.f)
Hnh 4.f: S nguyn nhan v kt qu sau khi phn b li c cc d liu nghin cu
hiu chun(2)

ti lp(1)

I (op)

V(op)

c(ref) tinh
khit (ref)

V(op)

m(ref)

I(op)
I(ref)
m(ref)
V(ref)
pha long

tuyn
tnh

nhit (2) hiu chun(2)

nhit (2)

V(ref)
hiu chun(2)

hiu

hiu chun(2)

pha long

m (sample)

P(op)
m(c mu v b)
m(b)

tuyn
tnh
hiu chun(3)

tuyn
tnh

hiu chun(2)
hiu chun(2)

F(hom) (3)

AGL 18

thu hi (2)

Ln ban hnh : 1.04

I(ref)

m mu

Trang : 51

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Ch thch: Thng thng mu c chy tng l mu nh, mi l mu chy u phi hiu chun thit
b, mu kim tra thu hi kim sot chch v mu p ngu nhin kim tra chm. S phi
thc hin hnh ng khc phc nu trong trng hp cc ln th ny c du hiu cho cc kt qu lch
nhau ng k. Kim sot cht lng c bn ny p ng c cc yu cu chnh ca vic nh gi
KB t d liu ph duyt phng php.

Vic chn thm mt yu t nh hng lp li trong s cho ta cng thc tnh Pop nh
sau:
Pop =

I op .c ref .Vop
I ref . Re c.m sample

.Fhom .10 6 .Fref

mg.kg-1

cng thc *

iu ch ra lp li ging nh ng nht. Cch tnh ton theo dng ny thun tin hn


v c trnh by di y.
Cc nh gi ca cc nh hng khc nhau s c cn nhc
Nghin cu chm
Ton b phng sai ca qu trnh phn tch qua nhiu ln phn tch c thc hin i vi
cc mu th p (cng mt mu ng nht, tch chit v qu trnh xc nh) i vi d lng
thuc tr su phospho hu c c tm thy trong cc mu bnh m khc nhau. Kt qu c
trnh by trong bng 4.g
Bng 4.g: Kt qu nghin cu th nghim lp thuc tr su
D1
[mg.kg-1]

D2
[mg.kg-1]

Malathion

1.30

1.30

Malathion

1.30

Malathion

Trung bnh
[mg.kg-1]

D1 D2

D1-D2/
trung bnh

1.30

0.00

0.000

0.09

1.10

0.40

0.364

0.57

0.53

0.55

0.04

0.073

Malathion

0.16

0.26

0.21

-0.10

-0.476

Malathion

0.65

0.58

0.62

0.07

0.114

Pirimiphos Methyl

0.04

0.04

0.04

0.00

0.000

Chlorpyrifos Methyl

0.08

0.09

0.085

-0.01

-0.118

Pirimiphos Methyl

0.02

0.02

0.02

0.00

0.000

Chlorpyrifos Methyl

0.01

0.02

0.015

-0.01

-0.667

Pirimiphos Methyl

0.02

0.01

0.015

0.01

0.667

Chlorpyrifos Methyl

0.03

0.02

0.025

0.01

0.400

Chlorpyrifos Methyl

0.04

0.06

0.05

-0.02

-0.400

Pirimiphos Methyl

0.07

0.08

0.75

-0.10

-0.133

Chlorpyrifos Methyl

0.01

0.01

0.10

0.00

0.000

Pirimiphos Methyl

0.06

0.03

0.045

0.03

0.667

D lng

AGL 18

Ln ban hnh : 1.04

Trang : 52

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Cc d liu thu c khc nhau (khc so vi trung bnh) cung cp mt kt qu cho phng sai
tng ca qu trnh th nghim lp li. a ra s c lng KB chun tng i cho
tng th nghim n l, lch chun phi chia cho 2 thnh lch chun cho KB
chun cho cc gi tri n. T suy ra gi tr ca KB chun t phng sai ca qu trnh
th nghim lp li tun theo ton b cc bc trong qu trnh phn tch nhng loi tr nh
hng ng nht 0.382/ 2 = 0.27.
Ch thch: Ban u c th cc php th p ny khng a ra bc t do. Nhng n khng c mc
ch l a ra s lng chnh xc i v chm ca qu trnh phn tch i vi mt loi thuc tr
su c bit trong mt loi bnh m c bit no . iu quan trng hn trong qu trnh phn tch
php th l nghin cu th nghim c trn nhiu loi mu th v mc nng d lng trong
mu a ra s la chn i din ca cc loi thuc tr su phospho hu c. Cch thc c hiu qu
nht l th nghim lp li trn cc loi mu bnh khc nhau nghin cu bc t do cho mi loi
mu nghin cu.

Nghin cu v chch
chch ca phng php phn tch c nghin cu ni b trong phng th nghim bng
cch nghin cu cc mu thm (ng nht mu v chia mu thnh cc mu nh v thm mt
hm lng vo mi mu nh) bng 4.h a ra cc kt qu ca mt qu trnh nghin cu cc
mu thm trong mt thi gian di trn cc dng khc nhau.
Bng 4.h. Kt qu nghin cu thu hi thuc tr su
Loi d
lng
PCB

Nng
[mg.kg-1]
10.0

OC

Thc n gia sc tng hp 1

s2) (%)

Trung bnh
2)
(%)
84

0.65

33

109

12

OC

0.325

100

90

Du thc vt v ng vt

OC

0.33

34

102

24

Brassicas 1987 (ci bp)

OC

0.32

32

104

18

Bnh m

OP

0.13

42

90

28

Bnh biscot

OP

0.325

30

84

27

thc phm ch bin t


xng v tht

OC

0.325

95

12

Gluten ng

OC

0.325

92

ci du

OC

0.325

11

89

13

La m

OC

0.325

25

88

tng

OC

0.325

13

85

19

La mch

OC

0.325

84

22

C cht
Du thi

N1)

(1): s nghin cu thc nghim


(2): Trung bnh v lch chun mu s tnh % thu hi
AGL 18

Ln ban hnh : 1.04

Trang : 53

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

ng tuyn tnh c suy ra khi lm th nghim trn cc mu bnh m (c


nh du bng mu ghi) ch ra rng H s thu hi trung bnh t 42 mu l 90% vi
lch chun (s) l 28%. KB chun c tnh t lch chun ca trung bnh
u (Re c) = 0.28 / 42 = 0.0432
Mt ngha ca php th c s dng xc nh cho h s thu hi trung bnh t so snh s
khc nhau ng k so vi 1.0. Theo th nghim thng k t c tnh ton theo cng thc sau:
t=

1 Re c
u (Re c)

(1 0.9)
= 2.315
0.0432

Gi tr ny c so snh vi gi tr ti hn l 2, i vi bc t do n-1 v tin cy l 95%


(trong n l s lng kt qu dng c lng Re c ). Nu t >=t
k so vi 1. t = 2.31 tcrit,41 = 2.021.

crit

th Rec l khc ng

Trong v d ny, h s hiu chnh (1/ Re c ) c s dng v do vy r rng trong cc kt qu


tnh ton Re c c th hin.
Cc ngun gc khc ca khng m bo o.
Trong s 4.i ch ra cc ngun gc khc ca khng m bo o bao gm cc d liu v
chm (1), d liu v h s thu hi (2) hoc cc ngun gc khc (3) c kim tra v
tnh ton trong khng m bo o.
Tt c cc cn v dng c o th thch quan trng phi c kim sot nghim ngt. chm
v h s thu hi phi c tnh n trong s nh hng ca vic hiu chun cc dng c dng
o th tch bi v trong sut qu trnh th nghim u phi dng cc dng c o th tch v
pippet. Nghin cu cc phng sai m rng trong gn na nm cho thy c nh hng ca
nhi mi trng ln kt qu. Mt s cc ngun ng gp khc vo KB l tinh khit
ca mu chun, s khng tuyn tnh ca thit b GC khi tr li kt qu (i din bi vic hiu
chun cho cc thng s Iref v Iop) v vic mu ng nht cng l mt thnh phn thm vo
trong yu cu ca vic nghin cu.
Hnh 4.i nh gi cc ngun gc khc ca khng m bo o
hiu chun(2)

lp li (1)

tuyn tnh

I (op)

V(op)

m(ref)

I(op)
I(ref)

nhit (2) hiu chun(2)

m(ref)
V(ref)
pha long
V(op)

c(ref) tinh
khit (ref)

nhit (2)

V(ref)
hiu chun(2)

hiu chun(3)

hiu chun(2)

pha long

m (sample)

P(op)
m(gross)

m(tare)

tuyn tnh
hiu
chun(3)
F(hom) (3)

AGL 18

thu hi (2)

Ln ban hnh : 1.04

hiu chun(2)

tuyn
tnh
hiu chun(2)

I(ref)

m mu

Trang : 54

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

(1) ti lp c tnh n trong qu trnh nghin cu phng sai ca th tc phn tch


(2) tnh n trong qu trnh nghin cu chch ca th tc nghin cu
(3) c tnh n trong s nh gi cc ngun gc khc ca khng m bo o
tinh khit ca chun m nh sn xut cung cp l 99.53% 0.06%. tinh khit cng l
mt ngun gc ca KB o vi KB o chun l 0.0006/ 3 = 0.00035 (phn b hnh
ch nht). Nhng gi tr ny qu nh so vi nghin cu KB t nghin cu chm nn
c th b qua.
tuyn tnh ca kt qu thuc tr su photpho hu c trong phm vi gii hn nng c
thit lp trong sut qu trnh nghin cu. Hn na vi cc ch s trong bng 4.g v 4.h
khng tuyn tnh cng ng gp vo phn b chm.
ng nht ca mu bnh m l nguyn nhn cui cng gy ra khng m bo o.
Khng c d liu v phn b d lng thnh phn hu c trong cc sn phm bnh m. ( l
mt iu ngc nhin v phn tch i vi a s thc phm ng nht him hn i vi cc
mu khng ng nht). ng thi thc t khng thc hin cc php o trc tip ng nht.
Bi vy, s nh gi khng m bo o da trn c s ca phng php ly mu.
Mt s phn b d lng thuc tr su c cng b v mt vi php thng k n gin
c s dng tnh ton KB o chun cho ton th mu phn tch v vic tnh ton
lin quan n khng m bo o tiu chun trong ca thuc tr su c trong mu cui
cng l :
D lng phn b pha trn cng b mt ch c: 0.58
D lng phn b ph u trn b mt ch c: 0.20
D lng phn b u trong mu nhng b gim nng do bay hi hoc bi phn hu t
bn trong ln b mt : 0.05 0.10 (ph thuc vo dy b mt).
Trng hp (a) l c bit cung cp vi t l ly mu hoc ng nht hon ton : n s xy ra
trong trng hp thm cht ph gia ln b mt. Trng hp (b) bi vy c coi nh l
trng hp xu nht. Trng hp (c) c xem nh c kh nng nht nhng khng th phn
bit d dng vi (b), trn c s gi tr 0.20 c chn.
4.2.5. Bc 4 : Tnh ton khng m bo o tng hp
Trong qu trnh nghin cu phng php ni b ca thao tc phn tch, ti lp, chch v
tt c cc ngun gc khc ca khng m bo o c nghin cu k lng. Cc gi tr
ny c ghi trong bng 4.c
Cc gi tr tng i c kt hp bi cng thc:
u c(Pop) / Pop = (0.27 2 + 0.048 2 + 0.2 2 = 0.34
uc (Pop) = 0.34 x Pop
Bng tnh i vi trng hp ny c ch ra trong bng 4.k Ch rng KB o tuyt i
(0.373) i vi kt qu hiu chnh danh ngha l 1.1111, t c gi tr 0.373 / 1.11 = 0.34

AGL 18

Ln ban hnh : 1.04

Trang : 55

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 4.c : khng m bo o trong phn tch thuc tr su


Gi tr

KB
chun u(x)

j KB chun
tng i u(x)/x

lp li

1.0

0.27

0.27

Da trn 2 ln th ca 2
mu khc nhau

chch

0.9

0.043

0.048

Mu thm

Cc ngun gc khc
( ng nht...)

1.0

0.2

0.2

nh gi trn c s
phng php s dng

u(Pop)/Pop

--

--

0.34

KB chun tng i

M t cc ngun

Ghi ch

Kch thc tng i ca 3 thnh phn ny c th c so snh bng cch s dng s


Hnh 4.d ch ra cc gi tr [u(y, xi)] c suy ra t bng 4.k.
Hnh 4.d khng m bo o trong phn tch thuc tr su

u(y,xi) (mg/kg)
ng nht
chch
lp li
P(op)
0

0.1

0.2

0.3

0.4

Cc gi tr ca u(y, xi) = (y/xi).u(xi) ly t bng 4.k


lp li gp phn ln nht trong phn b KB o. Thnh phn ny c ngun gc t tng
phng sai ca phng php nn cc nghin cu thm cn phi tin hnh ngy cng ci
tin cho phng php v gim KB o. V d KB o c th gim ng k nu ng
nht mu c tnh n v thc hin trong ton b cng on trc khi thc hin ly mt mu
phn tch.
khng m bo o m rng U(Pop) c tnh ton bng cch nhn KB tng hp vi
h s ph k = 2. Ta c:
U(Pop) = 0.34 x Pop x 2 = 0.68 x Pop

AGL 18

Ln ban hnh : 1.04

Trang : 56

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 4.k. khng m bo o trong php phn tch thuc tr su


A
1
2
3
4
5
6
7
8
9
10
11
12
13

B
Gi tr
KB o

C
lp li
1.0
0.27

D
chch
0.9
0.043

E
ng nht
1.0
0.2

ti lp
chch
ng nht

1.0
0.9
1.0

1.27
0.9
1.0

1.0
0.943
1.0

1.0
0.9
1.2

Pop
u(y, xi)
u(y)2, u(y, xi)2

1.1111

1.4111
0.30
0.09

1.0604
- 0.0507
0.00257

1.333
0.222
0.04938

u(Pop)

0.377

0.1420

(0,377/1.111 = 0.34 tng ng vi


khng m bo o tng i)

Cc gi tr ca cc thng s c a vo dng th 2 t C2 n E2. KB chun ca


chng c th hin dng di t C3 n E3. a cc gi tr t C2 n E2 vo ct B5 n
B7. Kt qu ca cc gi tr c th hin B9 (=B5 x B7/B6, da trn tnh ton cng
thc *). C5 a ra gi tr lp li t C2 tr i KB ca n C3. Kt qu tnh ton s
dng cc gi tr C5:C7 c C9. Ct D v E theo cc bc tng t. Gi tr a ra dng 10
(C10 n E10) l gi tr dng t C9 n E9 tr i gi tr B9. Ti dng 11 (C11 n E11).
l nhng gi tr ca dng 10 (C10:E 10) l bnh phng v tng ca gi tr th hin B11.
B13 l gi tr KB tng hp bng cch khai cn bc hai ca gi tr ca B11.

4.2.6. Kha cnh c bit: S dng mu khng ng nht tnh khng m bo o ca


php th xc nh hm lng thuc tr su photpho hu c.
Cho rng tt c cc vt liu c trong mu, u c th tch chit c tin hnh phn tch
khng k n trng thi ca chng, trng hp xu nht l trng hp mu khng ng nht,
mi phn ca mu li cha mt c cht khc nhau. Ti thiu l trng hp c 2 loi vt liu
trong cc phn khc nhau ca mu (k hiu l L1 v L2). H qu ca vic khng ng nht
trong trng hp mu ph c th c c lng bng thng k nh thc. Gi tr mong mun
l trung bnh v lch tiu chun ca lng vt liu trong n phn bng nhau c chn
ngu nhin sau khi chia nh mu.
Cc gi tr ny c tnh bi:
= n.(p1.l1 + p2.l2)
= n.p 1.(l1 l2) + n.l2 [1]
2 = n.p1. (1 p1) . (l1 l2)2 [2]

AGL 18

Ln ban hnh : 1.04

Trang : 57

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Trong l1 v l2 l lng c cht trong cc phn bng nhau thu c t mu L1 & L2 , tng
trng lng l X. p1 , p2 l kh nng xy ra ca cc phn chia c chn t mu (n phi
nh khi i chiu vi tng s cc phn chia c chn)
Coi rng kch thc ca mu l 12 x 12 x 24 cm, chia thnh 432 phn bng nhau c kch
thc 2 x 2 x 2cm v ly 15 phn bng nhau ngu nhin v ng nht trong s . Cng thc
trn c tnh nh sau:
Trng hp (a)
Vt liu c xc nh l n v c b mt rng trn nh ca mu. V th L2 = 0 ging nh l2
v L1 = 1. Mi phn chia bao gm mt phn ca b mt s nhn c mt lng l1 ca vt
liu. Vi kch thc ni trn bng 1/6 ca kch thc mu v th, p1 = 1/6 hoc 0.167 v l1 l
X/72 (v c 72 phn chia)
Ta c:
= 15 x 0.167 x l1 = 2.5 l1
2 = 15 x 0.167 x (1-0.17) x l12 = 2.08 l12
= 1.44l1
RSD = / = 0.58
Ch thch : tnh ton lng X trong ton b mu, gi tr trung bnh ca 432/15 v th X s l:

X = 432/15 x 2.5 x l1 = 72 x X/72 = X.


Kt qu ny ca mt dng ly mu ngu nhin; Cc gi tr trung bnh ny l chnh xc v c th i
din cho gi tr trung bnh ca ton b tp hp. i vi ly mu ngu nhin, th phn b ca
khng m bo o ti khng m bo o tng khc vi phng sai ca vic th nghim lp li v
c din t bi hoc RSD

Trng hp (b):
Vt liu c phn phi trn ton b b mt. Lp lun tng t nh trn, v coi trn ton b
b mt cc phn chia cha cng mt lng l1 ca vt liu, l2 = 0 v p1 c kch thc c tnh
bi:
p 1 = ((12 x 12 x 24) (8 x 8 x 20)) / (12 x 12 x 24) = 0.63
p1 l phn chia ca mu ngoi 2 cm. l1 = X / 272
Ch thch: c s thay i gi tr so vi trng hp (a)

Ta c:
=15 x 0.63 x l1 = 9.5 l1
= 15 x 0.63 x (l1-0.63) x l12 = 3.5 l12
= 1.87 l1
RSD = / = 0.2
Trng hp (c)
Lng vt liu gn b mt gim xung gn n 0 do c s bay hi hoc cc tht thot khc.
Trng hp ny c th tnh ton mt cch n gin nht l coi n nh mt trng hp ngc
li vi trng hp (b), vi p 1 = 0.37 v l1 = X/160
AGL 18

Ln ban hnh : 1.04

Trang : 58

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Ta c:
=15 x 0.37 x l1 = 5.6 l1
2 = 15 x 0.37 x (1-0.37) x l12 = 3.5 l12
= 1.87 l1
RSD = / = 0.33
Tuy nhin, s tht thot ny t n mt su nh hn kch thc ca phn chia, mi phn
s cha mt t lng vt liu l1 v l2 v th, c hai s khng bng 0. Ta ly trng hp m
phn ngoi l 50% v phn trung tm mu l 50%.
Ta c:
l1 = 2 x l2 l1 = X / 296
= 15 x 0.37 x (l1 - l2) + 15 x l2
= 20.6l2
2 =15 x 0.37 x (1 - 0.37) x (l1 - l2) 2 = 3.5 l22
RSD = 1.87/20.6 = 0.09
Trong dng ny, tng ng vi su 1cm m vt liu b mt. Kim tra cc mu bnh m c
v dy 1cm hoc nh hn v s dng. Trong cc dng hin nay th tng ng vi su 1cm
ci m vt liu b mt. Kim tra cc dng mu bnh m c v dy 1 cm hoc nh hn v to
ra su cho vt liu v quan tm n s mt (v cng bn thn n l b ngn cn s mt
di chiu su ny). do phng sai thc trong trng hp (c) s a ra gi tr ca ,
khng nh gi tr 0.09 trn
Ch thch: Trong trng hp trn s gim cc ny sinh KB trong vic mu khng ng nht l
thng s nh hn l ng nht. Ni chung s u mnh vic gim phn b KB. Cn thm vo m
hnh nghin cu cc trng hp s lng ln hn l s lng nh ( ging nh s kt hp cht ch ca
cc ht trong cc khi ln) cha ng gi tr khng t l vi vt liu quan tm. Cung cp rng kh
nng tng hp tr nn khng kt hp vo cc phn mu ng nht l ln, phn b KB s
khng vt qu bt c tnh ton no trong cc trng hp trn.

AGL 18

Ln ban hnh : 1.04

Trang : 59

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

5.

XC NH VITAMIN A V E TRONG SA BT

5.1

Gii thiu phng php

Phng php c dng xc nh nhng ng phn ca tocopherol, retinol v caroten


trong mt s loi thc phm. V d ny xem xt vic ph duyt v tnh ton khng m
bo o c bit cho vic xc nh cc dn xut ca all-trans retinol (vitamin A) v tocopherol (vitamin E) trong sa bt. Mu ng nht c thu phn gii phng cc ng
phn ca retinol v tocopherol. Cc sn phm ny sau c chit bng mt hn hp ete v
sau khi chit c c c. Dng nhng h sc k lng hiu nng cao (HPLC) khc nhau vi
nhng hn hp cht chit c t l khc nhau tch v nh lng nhng ng phn theo yu
cu. Mt h pha thng dng xc nh -tocopherol, trong khi mt h pha ngc c s
dng xc nh all-trans retinol. Trong mi trng hp mu u c chun bng mt
dung dch chun pha long t dung dch gc. Nng ca cht phn tch trong mt mu,
Cvit, c tnh theo cng thc:
Cvit =

As V f C STD
ASTD Ws

Trong :
AS : l din tch mi (nh) ca dung dch mu
ASTD : l din tch mi ca dung dch chun
VF : l lng mu bm (ml)
WF : l trng lng mu dng phn tch (g)
CSTD : l nng ca dung dch chun (gml-1)
Mt s dng chy minh ha cc giai on trong phng php c trnh by hnh 5.1
5.2 Nghin cu chm
Mt nghin cu v nhn hiu thc phm ca cc sa bt sn c trn th trng ch ra rng tt
c u c cc thnh phn c th so snh v cht bo, protein, cacbonhyrat v cha nhng
nng vitamin A v vitamin E tng t. Ba mu c chn kim tra bao ph ht khong
nng c th ca vitamin trong mu. Mi mu c phn tch 4 ln, trong 4 ln chit v
chy sc k lng hiu nng cao (HPLC) khc nhau. Vi mi ln chy HPLC u dng ng
hiu chun mi v dung mi pha ng. Thm vo phn tch mt cht chun c chng
nhn (SRM 1846) c lng Rm . Nhng kt qu ny c tng kt trong nghin cu
chm. Kt qu c trnh by bng 5.1

AGL 18

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Trang : 60

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Hnh 5.1 S minh ha cc bc trong phng php xc nh


all-trans retinol v -tocopherol
Cn mu

Butylat hydroxytoluen
*
Lng thch hp ca
mi dung dch chy
HPLC nn la chn
da theo nng ca
mi vitamin gn vi
chun hiu chun
tng ng.

Thm 1g pyrolgalol vo
150ml dung dch etanolic
KOH
X phng ho trong vng
30 pht bng h hi lu

Chuyn ton b dung dch


vo mt phu tch
Thm 500ml hn hp
ete
Chit vitamin bng hn hp ete

Ra dch chit ete vi nc sch n khi dung


dch sch trung tnh vi phenolphatalein

Chuyn dch ete sch sang bnh ct quay

Ct n kh ti 40 C
nh lng alltrans retinol bng
HPLC pha ngc

Thm 2ml dung


dch BHT

Ho tan vi mt lng
thch hp metanol*

Ly mt lng dung dch thch hp bng pipet

Ct n kh bng my ct quay
Thm 2 ml
dung dch
BHT
nh lng
-tocopherol bng
HPLC pha thng

AGL 18

Ln ban hnh : 1.04

Ho tan bng mt lng thch


hp dung dch pha ng

Trang : 61

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 5.1 Tng kt cc kt qu nghin cu chm phng php xc nh


all - trans retinol v -tocopherol
-tocopherol (vitamin E)

all-trans retinol (vitamin A)


Mu

Trung
bnh
(mg kg-1)

lch
chun
(mg kg-1)

lch
chun
tng i

Trung
bnh (mg
kg-1)

lch
chun
(mg kg-1)

lch
chun
tng i

Sa bt A

9.67

0.146

0.0151

64.08

2.411

0.0376

Sa bt B

10.04

0.247

0.0238

69.05

2.156

0.0312

Sa bt C

6.67

0.217

0.0325

171.93

7.959

0.0463

SRM 1846

5.32

0.285

0.0536

278.51

10.663

0.0383

lch chun quan st c vi vitamin A l theo th t ln dn. iu ny ch ra rng trong


di nng nghin cu, lch chun c lp vi nng phn tch. Trong trng hp ny,
khng m bo lin quan vi chm ca phng php, u(P), cho nhng mu c nng
all-trans retinol thay i xp x khong 5mgkg-1 10mgkg-1, c c lng theo lch
chun chung (pool) theo cng thc sau:
(3 0.146 2 ) + (3 0.247 2 ) + (3 0.217 2 ) + (5 0.285 2 )
= 0.238
S pool =
3+ 3+ 3+ 5

KB u(P) cho vitaminA l 0.238mgkg-1


Ch thch: Kt qu ca v d ny c tnh ton khi s dng bng tnh. Nhng gi tr th hin trong
cc bng v phng trnh c lm trn. Tnh ton li cc gi tr c th c kt qu sai khc.

i vi -tocopherol, lch chun tng i quan st c i vi cc mu t l thun vi


nng -tocopherol ca mu, tc l hm lng cng ln th lch chun cng tng theo.
iu ny ch ra rng lch chun l mt t l xp x vi nng phn tch trong di nng
nghin cu. Trong nhng trng hp nh vy c lng u(P) s dng cng thc tnh lch
chun pool theo lch chun tng i, c kt qu nh sau:
(3 0.0376 2 ) + (3 0.0312 2 ) + (3 0.04632 )(5 0.03832 )
= 0.0387
RSD pool =
3+ 3+ 3+ 5

KB tng i u(P)/P cho -tocopherol l 0.0387 tng ng lch chun tng i


5.3

Nghin cu ng

Tnh Rm v u ( Rm)
Trong trng hp s dng mu chun c chng nhn SRM 1846 do vin Quc Gia v tiu
chun v cng ngh ca Nht (NIST) sn xut. CRM l mu sa bt chun c chng nhn
v s lng vitamin, bao gm vitamin A v vitamin E. Su mu CRM c phn tch. Kt
qu, cng vi nhng gi tr c chng nhn ca mu chun c tng kt bng 5.2

AGL 18

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Trang : 62

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 5.2: Kt qu t nghin cu mu chun sa bt SRM 1846

Ch tiu phn
tch

Gi tr c cng nhn

khng m
Nng
khngm
bo chun
CCRM
bo CRM
u(CCRM)
(mgkg-1)
(mgkg-1)2
(mgkg-1)1

Gi tr quan st c
Trung bnh
C obs
(mgkg-1)

lch
chun sobs
(mgkg-1)

VitaminA

5.84

0.68

0.35

5.32

0.285

vitaminE

271

25

12.8

278.51

10.663

khng m bo l khng m bo m rng vi mc tin cy l 95%

khng m bo chun l khng m bo m rng chia cho 1.96

Tnh Rm v u ( Rm)
i vi vitamin A (all-trans retinol): Rm = 5.32/5.84 = 0.911
0.2852 0.35
+
u ( R m ) = 0.911
= 0.0581
2
6 5.32 5.84
2

xc nh phn b ca Rm ti KB tng hp, gi tr c tnh ton trn c so snh


vi 1, t =

1 Rm
u(R m )

1 0.911
= 1.53
0.0581

Trong trng hp ny t oc so snh vi h s ph, k=2. Khi t nh hn 2 khng c bng


chng no cho thy Rm khc 1 ng k. V th Rm c tha nhn ngang bng 1 vi mt
khng dm bo, u( Rm ) l 0.0581
Tnh u(Rs)
Rs c c lng t nghin cu mu thm cho sa bt A s dng trong nghin cu chm.
Nhng khng th to ra mt lng ln mu ng nht nghin cu mu thm m chun b
cc phn mu thm cng nng ring bit nghin cu. Nhng mu ny c chun b
theo nng nu trong bng 5.3. Bn mu c phn tch mt nng v trong nhng ln
tch v chy sc k lng cao p khc nhau. Nhng mu ny c thm dung dch all-trans
retinol (vitamin A) v -tocopherol (vitamin E) vi nng xp x 10g mu trong mt bnh
x phng ho, trc khi thm nhng thuc th khc. Gi tr trung bnh i vi mi mu c
tnh ton da vp cng thc sau:
Rm =

1 n C obs ( i ) C native
C
n i 1
spike( i )

Cnative l gi tr trung bnh cu nhng kt qu thu c t nghin cu chm. Kt qu c


tng kt bng 5.3.

AGL 18

Ln ban hnh : 1.04

Trang : 63

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 5.3 Tng kt nhng kt qu t nghin cu Rs


i vi vic xc nh vitamin A v vitamin E
Vinamin A
Mc ngu
nhin

Nng
tng ng
(mgkg-1)

Trung bnh
thu hi

14.0

2
3

Vitamin E
n

Nng
tng ng
(mgkg-1)

Trung
bnh
thu hi

0.981

84.0

1.028

18.0

0.991

99.0

1.103

22.0

0.996

114.0

1.167

0.00767

u(Rs)

u(Rs)

0.0696

u(Rs) l lch chun tng i trung bnh thu hi ti tng nng .


Tnh R v u(R)
Trong trng hp ny, khi c sn CRM c lng Rm v u ( Rm) cho c vitamin A v
vitamin E th khng cn tnh Rrep. C Rm v Rs c thy l tng ng vi 1. R v th cng
tng ng 1 u(R) c tnh ton cho vitamin A nh sau:
2

u ( R m ) u ( Rs ) u ( Rref )
= 0.05812 + 0.00767 2 = 0.0586
+
+
u ( R) = Rx

R
R
m s ref
2

Mt tnh ton tng t vi vitamin E cng a kt qu c lng u(R) = 0.0863. Ch rng


v trong c hai trng hp R = 1, v th khng m bo l nh nhau i vi cch din t
lch chun v lch chun tng i.
5.4

nh gi cc ngun khc ca khng m bo

Nghin cu sai s th
C mt s tham s ca phng php m khng i din cho s khc nhau qua nghin cu
chm v ng. Nhng nghin cu ny s dng nghin cu sai s th. Cc bc chit
chit v cc giai on chy sc k lng hiu nng cao (HPLC) ca phng php. Ch c
nghin cu sai s th i vi vitamin A c thc hin. Mt nghin cu tng t cng c
yu cu i vi vitamin E hon thnh c tt c cc ngun gy ra khng m bo.
Nhng tham s c nghin cu v la chn c tng kt trong bng 5.4 v bng 5.5 di
y. Nghin cu c thc hin trn CRM.
Giai on chit v ra sch
Nhng kt qu thu c t kim tra khng thun nht ca qu trnh chit i vi vitaminA
c trnh by bng 5.6. S khc nhau i vi mi tham s, t DxA n DxG, c tnh ton
nh miu t phn B mc 3.3.3.1 Ti liu hng dn c lng KB cho phn tch
ha hc nh lng.

AGL 18

Ln ban hnh : 1.04

Trang : 64

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Bng 3.4 : Nhng tham s c nh gi trong nghin cu sai s th


ca qu trnh chit vitaminA
Thng s
Trng lng KOH c s dng trong
ethanolic KOH

Gi tr
A

42g

21g

Thi gian x phng ho

20 pht

40 pht

S ln ca 250ml dch chit ete

Thi gian lc cho mi dch chit ete

2 pht

1 pht

n trung tnh

n kh

Thm 5 pht

50oC

Lng nc ra

**

Thi gian ct quay


Nhit ct quay

30 C

* Phng php hin ti ch ra rng dch chit ete nn c ra vi nc n trung tnh.


i vi thng s (e) dch chit ete c ra vi mt lng nc nhiu hn gp 3ln
** Phng php hin ti ch ra rng mu chit nn c ct quay n kh
i vi thng s f, qu trnh bay hi c tip tc thm 5 pht sau khi kh
Bng 5.5 : Nhng thng s iu tra trong nghin cu sai s th
ca qu trnh sc k lng hiu nng cao i vi vic xc nh vitaminA
Thng s

Gi tr

Tui ct

Mi

a'

Tc dng

0.8ml/pht

b'

1.2ml/pht

c'

T l pha ng
(Metanol/nc)

90/10

d'

95/5

Lng bm

15l

e'

25l

Bc sng

320nm

f'

330nm

Nhit ct

30 C

g'

Bng 5.6 Nhng kt qu t nghin cu sai s th


ca dch chit vitamin A t cng thc chun
Kt qu quan st c (mgkg-1)
s

5.25

3.98

4.22

4.43

5.51

4.78

4.35

4.96

s khcnhau c tnh ton


DxA

DxB

DxC

DxD

DxE

DxF

DxG

-0.4329

0.3898

0.2948

-0.1009

0.2387

0.7036

0.03608

chnh xc i vi phng php c c lng trc l 0.238mgkg-1 vi 14 bc t do .


n Dx i
S dng cng thc t =
, nhng gi tr t c c lng i vi mi tham s.
2s
AGL 18

Ln ban hnh : 1.04

Trang : 65

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Nhng gi tr ny sau c so snh vi gi tr ti hn tcrit = 2.145 (2- tailed, v =14, tin


cy 95%). Kt qu c trnh by bng di :
Bng 5.7: Xc nh cc tham s
c nh hng ng k ti vic chit vitamin A t sa bt
Thng s

Gi tr t

Mc tin cy ngha 95%?

Trng lng KOH c s dng trong


2.573
ethanolic KOH

Thi gian x phng ho

2.316

S ln ca 250ml dch chit ete

1.752

Khng

Thi gian lc cho mi dch chit ete

0.599

Khng

Lng nc ra

1.419

Khng

Thi gian ct quay

4.181

Nhit ct quay

0.214

Khng

Trong nhng trng hp khi m nh hng ca mt thng s c tm thy l khng nh


hng ng k th cng thc sau c s dng tnh KB:
u ( y( xi )) =

2 t crit s real

=
n 1.96 test

2 2.145 0.238 real

= 0.184 real
test
test
4 1.96

Lng dch chit ete


Phng php s dng hai dch chit ete. Trong nghin cu sai s th, nh hng ca vic s
dng mt loi dch chit c nghin cu. Kt qu ch ra rng: s dng mt loi dch chit
gim kh nng thu hi vitamin A. Mc d nh hng khng ng k. Nu phng php c
p dng ph hp th khng chc rng real c th ngang bng vi rest. Tuy nhin c kh nng
xy ra mt vi bin i trong nh hng ca nhng dch chit t s phn tch ny n phn
tch kia qua s sai khc trong vic lc. iu nh hng ny s c tnh n bi vic c
lng khng m bo lin quan vi chm. V nhng l do ny m KB thm
khng bao gm cho tham s ny.
Thi gian lc cho dch chit ete
Phng php c ng dng s dng thi gian lc l 2 pht cho mt dch chit. Trong th
nghim sai s th, thi gian gim xung cn 1 pht. test v th ngang bng vi 1 pht. Ngi
ta c lng rng trong sut qu trnh thc hin phng php thi gian lc c th thay i
gia 1.5 v 2.5 pht. real v th cng l 1 pht. v th u(y(xD) l 0.184mgkg-1
Lng nc ra
Phng php hin thi ch ra rng dch chit ete nn ra vi 150 ml nc cho n khi dch
ra trung tnh vi phenolphtalein. Kt qu ca th nghim sai s th ch ra rng thm lng
nc ra ca dch chit c th dn n lm kt qu thp i. Trong nghin cu khng thun
nht nh hng ca vic tng thm 3 ln nc ra c nghin cu (rest=3). Trong khi s
AGL 18

Ln ban hnh : 1.04

Trang : 66

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

dng phng php ny hng ngy vic thm 3 ln nc ra cha chc c thc hin.
Nhng vic tng thm nc ra 1 ln c th xy ra. Nht l trong trng hp s trung tnh ch
va t ln ra trc . real v th c c lng ngang bng vi 1. v th khng m
bo u(y(xE) l 0.0613.
Nhit ct quay
Phng php ny thng dng vi nhit l 40oC. Ngi ta c lng rng nhit c th
dao ng l 2oC trong vic s dng thng thng. Trong th nghim sai s th nhit
c t ti 30oc v 50 oC, v gi tr ca rest l 20 oC v u(y(xG) l 0.0368mgkg-1.
i vi nhng thng s c xc nh th c khi c mt nh hng ng k i vi dch
chit.
Trng lng KOH s dng chun b dung dch etanolic KOH
Di iu kin vn hnh thng thng, s dng 42 0.1g KOH trong vic chun b dung
dch etanolic KOH theo yu cu cho phn ng x phng ho. Trong nghin cu sai s th,
lng KOH s dng gim xung l 21g. Kt qu thu c ny khc nhau, DxA l 0.4329mgkg-1. H s nhy c tnh nh sau:
cA =

0.4329
= 0.0206mgkg 1 g 1
21

khng m bo thng s ny u(xA), t t vic iu kin gii hn trng lng KOH


ch ra trong phng php, gi nh phn b hnh ch nht. V th u(xA) = 0.1/3=0.0577.
khng m bo kt qu cui cng do s bin i thng s A, u(y(xA)) c tnh theo cng
thc sau:
u ( y( x A )) = u ( x A ) c A = 0.0577 0.0206 = 0.00119mgkg 1
Thi gian x phng ho
Phng php ny thng dng vi thi gian ct quay l 30 pht. Ngi ta c lng rng khi
s dng hng ngy c th dao ng l 5 pht . Trong th nghim sai s th cho thng s ny
bin i t 20 - 40 pht. iu ny dn n kt qu c s sai khc, DxB l 0.3898mgkg-1. H s
nhy cB v th l 0.0195mgkg-1. KB kt qu cui cng do s bin i thng s ny,
u(y(xB) l 0.0564mgkg-1.
Thi gian ct quay
Nhng kt qu ca nghin cu sai s th ch ra rng vic ct quay c tip tc sau khi mu
t kh dn n gim lng vitamin A thu c. C kh nng do s phn hu trong khi phn
tch. Trong nghin cu sai s th, qu trnh ct quay c cho php tip tc thm 5 pht sau
khi kh. iu ny dn n mt s sai khc, DxF l 0.7036 mg kg-1. H s nhy v th l
0.141mg/kg/pht. Ngi ta c lng rng khi p dng phng php ny hng ngy thi gian
ct quay c th tip tc t 0 n 5 sau khi mu kh. iu ny ly c t s phn b hnh ch
nht l 2.52.5pht, uF v th l 1.443pht. khng m bo u(y(xF)) v th l 0.203mg/kg.

AGL 18

Ln ban hnh : 1.04

Trang : 67

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

nh hng ca nhng thng s trn c coi nh t l vi nng phn tch. Nhng


khng m bo v th c tnh theo lch chun chia cho 5.32mg/kh, gi tr trung bnh ca
kt qu thu c t nhng phn tch trc ca SRM 1846 di nhng iu kin phng php
thng thng.
Giai on sc k lng hiu nng cao (HPLC)
Nhng kt qu rt nghin cu sai s th ca qu trnh sc k lng hiu nng cao i vi vic
xc nh vitamin A c trnh by bng 5.8
Bng 5.8: Cc kt qu t nghin cu sai s th
trong qu trnh HPLC xc nh vitamin A
Kt qu quan st c (mg/kg-1)
s

5.47

5.33

5.35

5.07

4.59

4.75

4.79

5.09

DxG'
0.06682

Tnh ton s sai khc


Dx A'

DxB'

Dx C'

DxD'

Dx E'

DxF'

0.4997

-0.04021

-0.01202

0.0091

0.2192

-0.000975

Tnh ton nhng khc nhau qua th nghim cho phng sai ng k so vi 0. lch chun
c s dng trong php th da vo cng thc pool gia cc l d liu chm t 3 dung
dch chun vitamin A, mi nng phn tch 4 ln. iu ny dn n mt lch chun
chung l 0.167mg/kh(v=9).Gi tr xp x tcrit vi tin cy 95% l 2.262
i vi nhng thng s c xc nh l khng c nh hng ng k, KB c c
lng l 0.136x(real /test).
Tc dng
Phng php s dng tc dng l 1ml/pht. Mt s thng tin t cc ti liu cho rng s
bin i c trng trong tc dng l .3%. Trong trng hp ny tng ng l
0.003ml/pht. real v th l 0.006ml/pht. Trong nghin cu sai s th tc dng thay i t
0.8-1.2ml/pht. test v th l 0.4ml/pht. khng m bo trong kt qu cui cng do s
bin i trong tc dng u(y(xB)) v th l 0.0024mg/kg
T l pha ng
Phng php ch ra rng mt pha ng t l l metanol: nc(90:10 v/v). T mt s ti liu
ngi ta d on rng bin i trong hn hp pha ng t khi vt qu 1% trong c hai
thnh phn. Trong nghin cu sai s th t l thay i l 95:5 (v/v) metanol: nc, c th
chp nhn mt t l thay i cao hn trong khi p dng hng ngy. Ln c lng u tin
ngi ta gi nh rng khi s dng thng thng, nh hng ca kt qu cui cng nh c s
bin thin trong hn hp pha ng bng 1/10 kt qu quan st c trong th nghim sai s
th.

AGL 18

Ln ban hnh : 1.04

Trang : 68

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

Lng bm
Thng tin t cc ti liu cho rng 1,5% l h s c trng cho bin ca lng bm i vi
mt mu bm t ng. Phng php s dng lng bm l 20l. test v th l 0,6l. Trong
nghin cu sai s th lng bm bin i 10l. khng m bo u(y(xE)) l 0.00816mg/kg
D bc sng
Trong nghin cu thun nht th d bc sng c thay i l 10nm. Da trn mt s
thng tin th chnh xc mt bc sng l 2nm. khng m bo u(y(xF)) v th l
0.00186mg/kg
Nhit ct
Nh phn nghin cu sai s th, iu tra nh hng ca ct khng c n nhit. Khi
nghin cu th nhit phng trung bnh l 22,2oC vi mt % Cv l 1%. Ct thng c
duy tr ti 30oC. Ngi ta c lng rng, n c m bo trong 1oC. real v test tng
ng l 2oC v 7.8oC. khng m bo u(y(xG)) l 0.0349mg/kg.
nh hng ca cc thng s trn coi nh t l vi nng phn tch. Nhng khng m
bo v th c tnh bng cch chia lch chun tng ng vi 5.32mg/kg
Ch c mt thng s c nh hng c ngha n phng php l tui ct. S dng ct c
cho kt qu thp hn khi s dng ct mi. nghin cu s liu cho thy din tch nh i vi
mu chun tng ct c trong khi din tch nh gim i vi mu, dn n gim nng
c tnh.
Nhng kt qu nghin cu sai s th ch ra rng: khi tui ct thay i th s thc hin phng
php thay i. Khi p dng hng ngy nn iu chnh bng vic s dng chun kim sot cht
lng HPLC. Vic thc hin kim sot thch hp sau khi chnh cc iu kin chy HPLC th
nhng phn chit ca mu s i din cho s phn tch. Do s vn hnh tng i ngn nn
dng nh nghin cu chm c thc hin nh mt phn ca ph duyt phng php s
bao gm phng sai ca phng php do tui ct. Tuy nhin khi dng phng php ny hng
ngy mt mu s c phn tch v d liu thu c s c thu thp trong tng khong thi
gian. Phng sai quan st c trong nhng kt qu ny s bao gm phng sai do s thay
i ct theo thi gian trong nhng gii hn c chp nhn bi chun kim sot cht lng
ca HPLC. Nu c lng chm trong mt thi hn di thu c t mu kim sot khc
c ngha vi c lng chm t nghin cu thc hin trc th n c th c
dng to ra mt c lng khng m bo th cho s liu ln sau. V nhng l do nh
trn m c lng KB do s thay i tui ct s khng bao gm trong KB hin thi.
Cc ngun khng m bo o khc
Nghin cu chm v ng c thit k bao gm cng nhiu KB o cng tt, v
d bng vic phn tch cc khong nng v bng so snh vi cc chun mi v HPLC pha
ng cho tng l th nghim. Cc tham s m khng thch hp s c sai khc th hin qua
nhng th nghim thc nghim nh vic chit v iu kin ca HPLC c nghin cu trong
AGL 18

Ln ban hnh : 1.04

Trang : 69

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

cc th nghim sai s th. Do mt lng nh cc tham s nh hng s khng c


nghin cu trong nghin cu sai s th. Nhng tham s lin quan ti hiu chun dng c thu
tinh v cn s dng chun b chun v mu. V d qua qu trnh nghin cu cng mt cn
c s dng cn khi lng cho tt c cc mu. Mc d chm lin quan ti hot ng
cn ny cng bao gm trong nghin cu chm tng nhng nh hng ca chnh xc ca
cn cng khng c bao gm trong cc ngun KB. Cn c s dng trong nghin cu
c chch 0.0001g. Trong tng lai mt cn khc hoc vn cn c sau khi hiu chun cng
c chch 0.001g. Yu t ny khng bao gm trong cc ngun KB m c th cn nh
gi c lp. Do d cc kinh nghim ca nhng ngi nghin cu cc KB lin quan ti
hiu chun cc cn phn tch v dng c thu tinh l thng nh so vi cc ngun KB
khc nh tng chm v thu hi. Thm vo c lng KB cho cc thng s ny
khng c bao gm trong cc ngun KB.
5.5

Tnh ton KB tng hp v m rng

Bng 5.9 th hin cc KB thnh phn v ln ca tng thnh phn trong phng php
xc nh vitamin A trong sa bt.
Bng 5.9 : cc ngun KB trong phng php xc nh vitamin A trong sa bt
KB chun

Thng s
chm

u(P)

0.238 mgkg-1

thu hi

u(R)

0.0586

Khi lng KOH s dng pha etanolic KOH

u(y(xA))

0.000224

Thi gian x phng ha

u(y(xB))

0.0106

Thi gian lc v chit

u(y(xD))

0.0346

S lng ln ra nc

u(y(xE))

0.0115

Thi gian ct quay

u(y(xF))

0.0382

Nhit ct quay

u(y(xG))

0.00692

Tc dng

u(y(xB'))

0.000383

T l pha ng

u(y(xD'))

0.00256

Lng bm

u(y(xE'))

0.00153

Bc sng

u(y(xF'))

0.0103

Nhit ct

u(y(xG'))

0.00656

Ch thch: cc KB chun a ra khng cng n v l v c din t theo lch


chun tng i
Cc ngun KB c xc nh l t l vi nng phn tch c tng hp c tnh theo
cng thc l cn bc hai ca bnh phng cc ngun
u ( y)
= 0.0809
y

AGL 18

Ln ban hnh : 1.04

Trang : 70

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

y l tng hp KB lin quan vi chm ca phng php c nhn thy l c lp


vi nng phn tch nn KB tng hp l cn bc hai ca tng bnh phng cc KB
thnh phn. V d i vi mu cha hm lng vitamin A l 5mgkg-1 th KB l:
u ( y ' ) = 0.2382 + (5 0.0809) 2
Tnh ton tng t c lng KB cho cc khong nng vitamin A khc nhau:
Bng 5.10 c lng KB chun
cho phng php xc nh vitamin A trong sa bt
Nng (mg kg-1)

KB chun (mg kg-1)

KB tng i

0.47

0.094

7.5

0.65

0.087

10

0.84

0084

Bng 5.11: c lng KB m rng


cho phng php xc nh vitamin A trong sa bt
Nng (mg kg-1)

KB m rng
(mg kg-1)

KB m rng
tng i

0.94

0.19

7.5

1.30

0.17

10

1.69

0.17

Khuyn co v cc ngun khng m bo o: Hnh 5.2 minh ha cc phn b ca cc


KB o cho phng php xc nh vitamin A trong sa bt ti nng 5 mgkg-1. ng
k dc ch ra 1/3 ln so vi ln ca phn b ln nht trong cc thnh phn KB
(trng hp ny l t thu hi). Ch cc tham s m KB ln hn gi tr ny th cn cn
nhc l ngun ng k KB ca phng php. Trong trng hp ny th bn phn b
chnh c xc nh l chm v thu hi v nh hng ca di thi gian lc v chit,
ra nc ct quay v chit cho n kh. Hai ngun sau ca KB thu c khi nh gi
cc d liu qua nghin cu sai s th. Th nghim c thit lp lm r cc tham s c
nh hng ng k ti kt qu thc hin th nghim v cng cung cp c lng ban u
KB ca tng tham s. Nu c lng KB ch ra phn b ng k ca KB th c
ngha l s thay i trong tham s cn c nghin cu thm. Trong trng hp ny th
cc nghin cu ring bit tm ra nh hng ca thi gian lc v iu kin ct quay ln kt
qu phn tch nh th no. S thch hp nu bao gm vic xc nh li c lng cc ngun
KB. Nu KB lin quan vi cc tham s l ln khng chp nhn c th cc tham s
s phi c kim sot mt cch cht ch trong hng dn phng php th v d c th thi
gian lc phi l 2 pht 10 giy. Nu khng thch hp c th kim sot hu hiu v d nu
AGL 18

Ln ban hnh : 1.04

Trang : 71

Hng dn cc v d c lng KB o trong phn tch ho hc nh lng

gii hn kim sot yu cu c th rt cht theo cc phng php gc yu cu th cn c thm


cc nghin cu c th ci tin v pht trin phng php.
Hnh 5.2: Phn b cc KB trong phng php
xc nh vitamin A trong sa bt (khong nng 5mgkg-1)\

Nhit ct
Bc sng
Lng bm
T l pha ng

Tham s

Tc dng
Nhit ct quay
Thi gian ct quay
S lng ln ra nc
Thi gian lc v chit
Thi gian x phng ha
Khi lng KOH
thu hi
chm
0

0.005

0.01

0.015

0.02

0.025

0.03

0.035

0.04

0.045

0.05

0.055

0.06

KB theo dng lch chun tng i

AGL 18

Ln ban hnh : 1.04

Trang : 72

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