Ch nht,5/12/2010

Nguyn Vn Tng
MSSV:0814251
MSTT:CN-E2

XC NH ST TRONG NC BNG 1, 10
PHENANTROLIN
Phng php dy chun
(Bui sng-mu kim tra:Q1B)
I.
-

II.

Nguyn tc:
S dng phng php trc quang xc nh hm lng Fe2+ c trong mt
mu cht no bng cch thit lp ng chun ph thuc ca mt quang
vo nng Fe2+. T ng chun v s liu o quang ca mu cht xc nh
c nng ca Fe2+.
Ion Fe2+ to phc mu cam vi 3 phn t 1, 10 phenantrolin gi tn l
Ferroin.

Phc tn ti dng cation khong pH rng 2.0 9.0, c hp thu cc di

508 nm v h s hp thu phn t () ti bng 1.1*104 L.mol-1.cm-1. Phc rt
bn, c cng mu khng thay i trong nhiu thng. Khong tun theo
nh lut Beer l 0.13 5 g/mL.
Phn ng mu chn lc gia 1,10 phenantrolin v Fe2+ nn c th xc nh
ring lng Fe2+ ngay c khi c mt Fe3+.
C th xc nh hm lng ca Fe3+ bng cch sau: dng tc nhn kh mnh
nh hydroxylamine kh Fe3+ thnh Fe2+, sau xc nh hm lng tng
ca ion st c trong dung dch da trn lng Fe2+ (Fe2+ c sn trong dung dch
v sau khi kh Fe3+). Hm lng Fe3+ c tnh bng hm lng tng ca ion
st tr cho hm lng ca ion Fe2+ c sn trong dung dch (c xc nh khi
khng tin hnh kh Fe3+).

Ha cht, dng c:

Ch nht,5/12/2010

Dung dch Fe2+ chun 10 ppm.

Dung dch m pH = 5.
Dung dch hydroxylamine 10%.
Dung dch 1, 10 phenantrolin 0.5%
My so mu, bnh mc 50 ml, becher, pipet cc loi.

III.
-

Thc hnh:

1) Pha dy chun:
Pha dy chun vi fiol 50ml vi thnh phn nh sau:
S th t
Fe (chun), ml

1
0

2
0.50

3
1.00

4
2.00

5
4.00

6
6.00

m pH 5 , mL

5.0

NH2OH 10%, mL

1.0

Phenantrolin 0.5%, mL

1.0

H2 O

Thm n vch mc (50ml)

7
8.00

8
10.00

Sau khi thm cc thuc th, n nh trong vng 10ph ri tin hnh o
quang.
Cc mu 2, 3, 4 th tch chun c ly bng pipet 2ml.
Mu 5 th tch mu c ly bng pipet 5ml.
Mu 6, 7, 8 th tch mu c ly bng pipet 10ml.
2) Chun b mu cn xc nh:
Mu cn xc nh c nh mc 100 ml.
S dng 6 bnh nh mc 50ml tin hnh pha cc mu xc nh nng .
Trong c 3 bnh c thm hydroxylamine (a), 3 bnh khng c thm
hydroxylamine (b).
Trc khi pha 6 bnh pha trc 2 mu nhp vi 1 mu c hydroxylamine
v 1 mu khng c hydroxylamine xc nh lng th tch mu cn ly sao
cho khi ln mu, dung dch c mt quang trong khong 1/3 n 2/3 ng
chun.
Trng hp ny xc nh c l rt 4ml mu em i ln mu, xc nh
nng
Bnh
Fe (chun), ml

a
4.00

a
4.00

m pH 5 , mL
NH2OH 10%, mL

a
4.00

b
4.00

b
4.00

5.0
1.0

Phenantrolin 0.5%, mL

1.0

H2 O

Thm n vch mc (50ml)

b
4.00

Ch nht,5/12/2010

Th tch mu c ly bng pipet 5 ml.

3) o mu, kt qu:
Ly mu trng (mu 1) v mu bnh s 5 tin hnh qut sng, xc nh max.
S dng 2 cuvet tin hnh qut sng: 1 cuvet ng mu trng, 1 cuvet ng
mu s 5.
Cho mu trng vo trc, rezero i cho mt quan hin my v 0 ri ly
mu trng ra, cho mu 6 vo o v ghi nhn gi tr mt quang.
Tin hnh nh trn vi cc gi tr bc sng tng t 400nm n 600nm.
(nm)
A
(nm)
A
400
0.052
506
0.154
410
0.073
508
0.154
420
0.083
510
0.154
430
0.096
512
0.155
440
0.105
514
0.154
450
0.114
516
0.151
460
0.122
518
0.148
470
0.133
520
0.145
480
0.142
530
0.116
490
0.145
540
0.077
492
0.145
550
0.042
494
0.146
560
0.023
496
0.146
570
0.011
498
0.147
580
0.005
500
0.150
590
0.002
502
0.151
600
0.001
504
0.152

Ch nht,5/12/2010
0.18

TH BiU DiN MT QUANG

THEO BC SNG

0.16

MT QUANG A

0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
350

400

450

500 SNG550
BC
(nm)

600

650

700

Chnh bc sng ca my v gi tr max thu c v tin hnh o quang dy

chun. max = 5126 nm.
o dy chun t dung dch c nng thp n dung dch c nng cao.
hp thu quang ca dy chun bc sng max= 506 nm:

ng chun:

STT

C(ppm)

0.00

0.000

0.10

0.015

0.20

0.038

0.40

0.075

0.80

0.154

1.20

0.236

1.60

0.320

2.00

0.401

Ch nht,5/12/2010

4) Tnh ton:
T dng ca ng chun thit lp c phng trnh tuyn tnh bc nht:
A = a + bC

)
(

(
)
Tnh cc h s a, b:

( )

( )

Tnh phng sai d:

)
(

Ch nht,5/12/2010

lch chun cho cc h s hi quy a v b:

Khong tin cy :

Vy phng trnh ng hi quy c thit lp:

A = (0.20264 0.00120)C (0.0053773 0.00135)

5) Xc nh nng Fe2+ v Fe3+ c trong mumu kim

tra:Q1B-bui sng)
Sau khi tin hnh o quang xc nh dy chun, tin hnh o quang cc
mu.
-Rt 6ml mu cho vo 2 fiol 50ml
Trong a:mu c cha NH2OH
-b:mu khng cha NH2OH

Kt qu cc mu:
a
a
0.405
0.409
0.407

Mu
A
Atb
6)
a) Tnh tng nng ion st:

a
0.407

( )
ng

b
0.336

b
0.336
0..336

b
0.338

Ch nht,5/12/2010

(
-

Sai s:
(
( )
[

)
(

(
(

)
(

) [
)

) ]

) ]
(

0.0905
-

Nng tng ion st ca mu:

Ctng mu = 16.9585 0.0905 ppm

b) Tnh nng ion Fe2+:

(

( )

Sai s:
(
( )
[

(
)

) ]
)
(

) [
(

) ]

Ch nht,5/12/2010

Nng tng ion Fe2+ ca mu:

CFe2+ = 14.038 0.0896(ppm)
3+
c) Tnh nng Fe :
-

(
(
-

Nng tng ion Fe3+ ca mu:

CFe3+ = 2.9205 0.127 ppm

IV.
-

Nhn xt, lu :

Qua th ng chun cho thy dng th lch so vi nh lut Lambert

Beer. th dng y = ax + b ch khng phi y = ax (ng chun khng qua
gc ta ). Tuy nhin chnh xc ca ng chun chp nhn c vi R2
= 0.999.
My ph UV c s dng l my UV mt chm tia. V th trc khi s dng
phi cho my khi ng khong 20ph cho ngun sng t n n nh v
khng tin hnh o lu qu, khi n s qu nng s nh hng n ngun
sng pht ra, lm kt qu o b sai lch.
Tin hnh qut li bc sng ca phc cht xc nh c bc sng cho
hp thu quang cc i ng vi iu kin th nghim.
V phc kh bn nn c th tin hnh o quang ca phc sau (do thiu my
phi ch i).
So snh vi vic dng phng php trc quang xc nh hm lng nitrit c
trong mu, ta thy gii hn pht hin ca vic xc nh Fe2+ bng phng php
quang cao hn. Hm lng nitrit c th xc nh nng thp hn so vi
vic xc nh hm lng Fe2+ (nh hn 10 ln).
C th thy y, th t cho cc thuc th l quan trng. mu, nu ta cho
1,10 phenantroline trc ri mi cho hydroxylamine th qu trnh kh Fe3+
thnh Fe2+ khng c thc hin hiu qu do Fe3+ to phc khng mu vi cho
1,10 phenantroline lm hn ch phn ng kh st vic xc nh tng nng
cc ion st s sai lch.
Cuvet c s dng trong bi thc tp khng tt lm. Cuvet c mu c th
nh hng n kt qu o quang.
C th thy y sai s b nh hng rt ln bi sai s mu, sai s dng c nh
hng khng nhiu lm. (sai s mu do qu trnh pha, thm thuc th, thi gia
i i o quang).

Ch nht,5/12/2010

Chn pipet rt cc dung dch sao cho ph hp vi th tch cn rt.

Do lng st trong nc my kh ln, nn phi s dng nc ct tin hnh
ton b qu trnh. Trng dng c bng nc ct trc khi s dng.
C th xc nh hm lng st trong mu bng vic o quang phc ca st vi
SCN-, hay vic to phc vi XO. Tuy nhin trong xc nh hm lng st bng
to phc vi XO, mu trng c mu nn c th xy ra hin tng overlap gia
hp thu ca nn v mu vic xc nh mu kh khn.
Do phng php xc nh da trn mu ca phc nn pH ca dung dch phi
c n nh mt khong pH nht nh s dng dung dch m.

XC NH HM LNG NITRIT
PHNG PHP DY CHUN (chiu, mu
kim tra Q2B)
1) Nguyn tc:
-

Xc nh Nitrit da trn s tng hp phm mu azo. Hm lng nitrit t l vi

cng mu ca hp cht diazo.
Xc nh cng mu ca dung dch, ta xc nh c hm lng NO2- c
trong mu.
HO3S

NH2 + NaNO2

HCl

HO3S

N + NaCl + H2O

NH2
H2N
HO3S

N N

SO3H

Hp cht diazo to thnh c max= 520 nm.

Vic tin hnh th nghim thng qua 2 bc tng hp diazo:
o Bc 1: s to thnh mui diazonium t HNO2 v mui ca amin thm
bc nht.
o Bc 2 : s ghp cp mui diazonium l tc nhn thn in t vi hp
cht thm c tnh thn hch tng i cao nh cc amin, phenol to
hp cht azo c mu.
Vic xc nh nng ca NO2- theo hp thu quang c thc hin theo
nh lut Lamber-Beer :

Ch nht,5/12/2010

Vic xc nh lng NO2- theo phng php dy chun tc l kho st s thay

i hp thu quang theo nng C, chiu dy cuvet l= const.

2) Ha cht dng c:
-

Dung dch NO2- 2ppm 0.0010 ca phng th nghim.

Dung dch acid sulfanilic 1%.

Dung dch -naphtylamin.

My o UV.
Fiol 50ml.
Pipet cc loi.

3) Thc hnh:
-

a) Pha dy chun:
Pha dy chun vi fiol 50ml vi thnh phn nh sau:
S th t
-

NO2 (1 ppm), mL

1
0

2
0.50

3
1.00

4
2.00

5
4.00

6
6.00

Acid sulfanilic 1%, mL

1.0

-naphtylamin 0.6%, mL

1.0

H2 O

Thm n vch mc (50ml)

7
8.00

8
10.00

Sau khi thm acid sulfanilic, i 3 4 pht ri mi cho -naphtylamin vo.

Tin hnh qut sng xc nh li max ca hp cht diazo.
Tin hnh o hp thu quang ca cc dung dch sau phn ng.
b) Chun b mu cn xc nh:
Mu cn xc nh c nh mc 100 ml.
Tin hnh pha mt lng mu th v so snh hp thu quang vi cc dung
dch trong dy chun.
T hp thu quang ca mu nhp, xc nh lng mu cn ly sao cho
hp thu quang ca mu sau nh mc 50ml c hp thu quang t 1/3 n 2/3
dy chun.
Tin hnh pha mu v o 3 ln, ly kt qu trung bnh 3 hp thu quang.
c) Tnh ton:
hp thu quang ca dy chun bc sng max= 520 nm:
STT

C(ppm)

0.000

0.02

0.020

Ch nht,5/12/2010

0.04

0.041

0.08

0.079

0.16

0.156

0.24

0.233

0.32

0.309

0.40

0.390

ng chun:
0.45
y = 0.9676x + 0.0013
R = 0.9999

0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0
0

0.1

0.2

0.3

Thit lp c phng trnh tuyn tnh bc nht:

A = a + bC

0.4

0.5

Ch nht,5/12/2010

Tnh cc h s a, b:

Tnh phng sai d:

Khong tin cy :

lch chun cho cc h s hi quy a v b:

( )

( )

Vy phng trnh ng hi quy c thit lp:

A = (0.967 0.000670)C (0.00142 0.000157)
d) Xc nh nng mu:
Bnh mu c nh mc 100ml.
Rt 10.00 ml mu cho vo fiol 50ml, thm cc thuc th, thi gian phn
ng xy ra, tin hnh o quang dung dch.
nh mc 3 bnh nh trn ng thi pha li mu blank.
Kt qu o quang:
Bnh
1
2
3
A
0.148
0.152
0.149
Atb
0.149
Nng ca dung dch pha:

Ch nht,5/12/2010

(
-

Nng ca mu phng th nghim ( dnh mc 100ml):

(

Sai s:
(
[

)
(

(
) [

(
(

) ]
)

) ]

Nng ca mu:
Cmu = 0.775 0.00504 ppm

e) Nhn xt:
My ph c s dng l my UV mt chm tia.
Qu trnh pha cc dung dch chun nn tin hnh ng thi thi gian cc
phn ng xy ra l nh nhau.
Trong phn tch trc quang, th t pha thuc th l rt quan trng. Vic tin
hnh cc bc pha thuc th cn c tin hnh nghim ngt, khng c t
thay i th t cc bc nu khng c kin thc v kh nng lp nn mt
quy trnh mi m khng lm thay i kt qu thc nghim.
Cn phi i cho phn ng gia cht cn phn tch vi thuc th xy ra hon
ton ri mi tin hnh phn ng vi thuc th tip theo hay o quang ng
dng ca trc quang theo i tin trnh phn ng (phn ng xy ra hon tt
th mt quang s n nh ).
Dung dch c chn mang i qut sng thng l dung dch c mu trung
bnh (mu khng qu m cng ko qu nht c xc nh bng mt thng,
y l dung dch s 6). S dng dung dch c mu qu nht s khng thy r
c s thay i ng vi hp thu quang ti cc bc sng gn vi max. S
dng dung dch c mu qu m s nh hng n max.

Ch nht,5/12/2010

Phng php trc quang phn tch c cc cht vi nng nh (ppm).

Trong qu trnh xc nh nng mu, do nng mu cha bit nn cn phi
thc hin mt mu nhp.
Mu nhp l mu c pha t mt lng mu u bt k xem xt hp
thu quang ca mu nhp t mi xem xt pha mu vi nng chnh xc
tin hnh o quang sao cho hp thu quang ca mu nm trong khong t
1/3 n 2/3 ng chun.
Vic phi pha mu sao cho hp thu quang nm t 1/3 n 2/3 ng chun
nhm m bo tnh nh lng ca php phn tch.
Mu trng l mu gm dung mi v cc thuc th, khng c cht cn phn tch
trong . Mu trng c cng dng hiu chnh nn, cho hp thu quang
o c ch l do sn phm c mu ca cht cn phn tch gy ra, khng b nh
hng bi mu ca dung mi cng nh cc thuc th.
Trc khi tin hnh o quang cn phi s dng mu trng hiu chnh nn.
Cc thao tc hc hnh ng vai tr quan trng, nh hng trc tip n kt qu
phn tch:
Cc thao tc rt, nh mc dung dch phi chnh xc.
Khi s dng cuvet khng c cm vo b mt thy tinh nhn.
Gi cho mt ngoi cuvet kh v sch: mt ngoi cuvet ko sch nh
hng n kh nng hp thu nh sng ca dung dch.
Cc lng thuc th lun c dng d so vi cht phn tch cn phn ng,
m bo cho cc cht phn ng l nh nhau trong cng iu kin.
3 bnh mu pha u cho kt qu hp thu quang ging nhau thao tc pha tt.
Do cuvet s dng l cuvet Trung Quc, hay cuvet nha nn ng u ca
2 cuvet cng loi l khng ging nhau khi tin hnh xc nh nng mu,
nn s dng mt cuvet t u n cui.