Phn 1

PHN TCH TRC QUANG

02/09/15

Phn 1: PHN TCH TRC QUANG

nh ngha Nguyn tc
Phn tch trc quang l tn gi chung ca cc phng php phn tch
quang hc da trn s tng tc chn lc gia cht cn xc nh vi
nng lng bc x thuc vng t ngoi, kh kin hoc hng ngoi.
Nguyn tc ca phng php trc quang l da vo lng nh sng
b hp thu bi cht hp thu tnh hm lng ca cht hp thu.

02/09/15

Phn 1: PHN TCH TRC QUANG

c trng nng lng ca min ph

02/09/15

Phn 1: PHN TCH TRC QUANG

c trng nng lng ca min ph

nh sng c bc sng nh hn 200nm, b hp thu bi oxi khng kh, hi
nc v nhiu cht khc, v vy ch c th o quang bc sng nh hn
200 nm bng my chn khng.
nh sng c bc sng t 200 400 nm, c gi l nh sng t ngoi
(UV), trong vng t 200 300 nm c gi l min t ngoi xa, cn
vng t 300 400 nm gn min kh kin c gi l min t ngoi gn.
nh sng c bc sng trong khong t 800 2000 c gi l nh
sng hng ngoi (IR). S hp thu nh sng min ph ny t c s
dng gii quyt trc tip cc nhim v phn tch, nhng c s dng
rng ri nghin cu cu to ca phn t.
02/09/15

Phn 1: PHN TCH TRC QUANG

c trng nng lng ca min ph
nh sng vng UV c bc sng trong khong: 200 400 nm
nh sng vng IR c bc sng trong khong: 800 2000 nm
nh sng vng VIS c bc sng trong khong: 396 760 nm
Trong phng php trc quang phng php hp thu quang
hc, chng ta thng s dng vng ph UV VIS c bc sng
t 200 800 nm

02/09/15

c trng nng lng ca min ph

Da cam

Vng

Lc

739 - 610

610 - 590

590 - 560

560 - 510

Lam

Chm

Tm

510 - 490

490 - 430

430 - 400

02/09/15

Lu
Nhng hp cht mu l nhng hp cht c kh nng hp thu mt hoc mt vi
mu ph ca nh sng t nhin, c th hp thu hon ton hoc mt phn cng
ca mu ph.
Nu ch hp thu duy nht mt mu ph, th mu ca dung dch chnh l mu b
sung (t hp mu ph v mu b sung tr thnh khng mu)
Th t
1
2
3
4
5
6
7
8
9
10

02/09/15

(nm)
400 -430
430 480
480 -490
490 500
500 560
560 580
580 -595
595 650
650 730
730 760

Mu ph

Mu b sung

Tm
Chm
Chm lc
Lc chm
Lc
Vng lc
Vng
Cam

ta

Vng lc
Vng
Cam

ta
Tm
Chm
Chm lc
Lc vng
Lc

Phn 1: PHN TCH TRC QUANG

Phn loi cc phng php trc quang
Phng php hp thu quang: phng php ny da trn
vic o cng dng nh sng b cht mu hp thu chn
lc.
Phng php pht quang: phng php ny da trn vic
o cng dng nh sng pht ra bi cht pht quang khi ta
chiu mt dng nh sng vo cht pht quang.
Phng php o c: phng php o c da trn
vic o cng dng nh sng b hp thu hoc b khuyt tn
bi h keo c iu ch t cht cn phn tch
02/09/15

Phn 1: PHN TCH TRC QUANG

Cc i lng c trng ca nh sng
Bc sng l khong cch gia hai im dao ng ng pha gn
nht, n v o l A0, m, , nm...(1nm=1m=10A0=10-9m).

02/09/15

Tn s sng = c trong tc nh sng trong chn khng bng

3.10 m/gy hoc 3.10 nm/gy, khi v c n v cm thi n v ca l gy-1

10

S sng

17

= 1 l s bc sng trn 1cm chiu di, n v l cm-1.

Quang thng l nng lng nh sng bc x theo mi phng ca

ngun im trong mt n v thi gian.
Cng nh sng I l dng sng pht ra t ngun im trong mt n
v gc khi l strian: I =
4
hc
Nng lng bc x in t: E=

Khi hp thu nh sng ni nng ca phn t tng t mc c bn E 0 n

mc E1 cao hn. Phn nng lng hp thu l nng lng ca photon, n t
l vi tn s nh sng
02/09/15

E = E1 - E 0 = h =

hc

10

C s l thuyt ca phng php

Nu

dung dch hp thu bc x vng t ngoi, nh

sng trng truyn sut hon ton n mt, dung dch
khng mu.
Dung dch c mu khi cha cu t c kh nng hp
thu bc x vng thy c, do khi nh lng
bng phng php quang ph hp thu thy c cn
c gi l phng php so mu hay o mu.
Dung dch mu c nng cng cao, kh nng hp
thu ca mu cng mnh, cng nh sng n mt
cng yu, dung dch c mu cng sm.
02/09/15

11

nh lut Bouguer Lambert Beer

Chiu bc x n sc c bc sng I c
cng I0 qua dung dch cha cu t kho
st c nng C. B dy dung dch l l. Ti
b mt cuvet o, mt phn bc x b phn x
c cng IR, mt phn bc x b hp thu
c cng IA. Bc x ra khi dung dch c
cng I.
02/09/15

12

nh lut Bouguer Lambert Beer

IR
I0

IA

Do : I0 = IR + IA + I
Chn cuvet o c b mt nhn, truyn sut IR = 0
I0 = IA + I

02/09/15

13

nh lut Bouguer Lambert Beer

I0
log =A= .l.C
I
Trong : l mt hng s t l c tn hp thu
phn t biu th hp thu ca dung dch c
nng cht tan l 1M c ng trong bnh
dy 1cm v c n v l l.mol-1cm-1.
By gi ta c th p dng d dng nh lut Beer
vo vic xc nh nng cc cht tan bng cch
o hp thu A ca chng.
02/09/15

14

Cng hp thu bc x ca cu t c
xc nh bng 2 i lng

truyn sut T (Transmittance)

I
I
T = hay T% = 100
I0
I0

hp thu A (Absorbance) hay mt

quang OD (optical density)
I0
1
100
A = log = log = log
= 2 - logT%
I
T
T%
02/09/15

15

 Nu o hp thu quang ca mt lot

dung dch bng mt dng sng n sc
(ti mt gi tr ) th A = f(l,C) l hm
bc nht, ng biu din l mt
ng thng, cn ng T = f(C) l
mt ng cong.
V vy trong phn tch trc quang ch
dng ng A = f(C) m khng dng T
= F(C).
02/09/15

16

A = lC

02/09/15

17

Bng tm tt tnh cht cc i lng trc quang

i lng

T
A (hay D)

02/09/15

Cng thc

n v

I
T=
I0
I0
A=lg
I

A
=
Cl

cm2/mol

Yu t ph
thuc

Yu t
khng
ph
thuc

Ghi ch

Khng c
tnh cng
tnh

,C,l

I0

,C,l

I0

C tnh cng
tnh

, bn cht
cht mu,
bn cht
dung mi, t0

I0,C,l

c trng
cho nhy
v phn ng
mu

18

ng dng tnh cht cng tnh ca A

Tnh cng ca mt quang hay hp thu A
A = AA + AB = 1lC1 + 2lC2

Mt quang o c khi cht tan ho tan trong mt dung mi l mt

quang tng cng ca dung dch .
A = AX + Adm
A phn nh ng AX th Adm rt nh ( 0). tho mn iu kin
ny, ta nn chn dung mi c ph hp thu rt xa ph hp thu ca cht
tan.

02/09/15

19

Dung dch mu tun theo nh lut hp thu

c bn nu tho mn cc iu kin sau:

C s trng kht cc ng ph - i vi cc dung dch c nng

khc nhau.
th biu din s ph thuc A C khi l = const l mt ng thng i
qua gc to .
Khi pha hai dung dch 1 v 2 sao cho C1l1 = C2l2 th cng t ta s c
A1 = 1lC1 = A2 = 2lC2
Cc ng ph A - vi nng Cn khc nhau u c cng max
th biu din mi quan h gia truyn qua T v lgC c im un
nm gi tr T = 0.368
02/09/15

20

Cc nguyn nhn gy sai lch khi nh lut Beer

Mc n sc ca nh sng ti. nh sng khng n sc
thng dn n lch m. Cht mu hp thu cc i max v
ch max mi c s tuyn tnh gia Aimax Ci v th Aimax Ci
l mt ng thng, khi mt quang l cc i. Mc
n sc cng ln, kh nng tun theo nh lut Lambert Beer
cng ln.
Nng ln ca dung dch kho st: Nng ca dung dch
ln s xy ra tng tc in, i lng thay i, thng thng
khi tng nng dung dch, gi tr gim. S sai lch khi
nh lut Lambert Beer thng l sai s m.
02/09/15

21

 S trng hp hoc kh trng hp phn t, s

solvat ho hay hydrat ho xy ra khi thay i
nng cht hp thu; s to thnh cc hp cht
trung gian, phc ph, cc hp cht ng phn,
to h keo hay s c mt ca cc cht in ly
mnh, pH u c kh nng lm thay i hp
thu ca dung dch, lm sai lch khi nh lut
Beer.

02/09/15

22

Ph hp thu
ng biu din s ph thuc ca hp thu A hoc vo
di sng (hay tn s sng ) gi l ph hp thu ca cht kho st.
Ph hp thu ca phn t l ph m gm mt hoc mt s m
hp thu, mi m u c dng ng phn b xc sut chun v
khc nhau bi cng hp thu v bc sng cc i max ca
m. Trong trng hp n gin phn t ch c mt tm mang mu
th ph A = f() ch c mt gii ph c dng i xng hnh chung.

02/09/15

23

02/09/15

max

24

Phn tch nh lng bng phng php trc quang

Nguyn tc v c s nh lng ca phng php
Phng php ng chun
Phng php thm chun
Phng php vi sai
Phng php chun trc quang
Phng php so snh
02/09/15

25

Nguyn tc v c s nh lng ca phng php

Nguyn tc chung ca phng php phn tch trc
quang
-

Chuyn cu t thnh hp cht c kh nng hp th

nh sng.

o s hp th nh sng ca hp cht to thnh v suy

ra hm lng cht cn xc nh X.

Nguyn tc chung ca cc phng php phn tch

nh lng
-

o quang ca dung dch mu.

So snh cng mu (hoc hp th quang) ca

dung dch nghin cu vi dung dch chun.
02/09/15

26

C s nh lng
nh lut Bougher-Lampere-Beer: khi
chiu mt chm photon n sc qua dung
dch th mc hp th ca dung dch t l
thun vi cng sut chm photon v nng
cc phn t hp th
Cng thc: A = .l.C

02/09/15

27

Phng php ng chun

QUI TRNH
- Pha mt lot dung dch chun c Ctc tng dn mt cch u n.
(thng 5 8 Ctc) (Cc dung dch chun phi c cng iu kin nh
dung dch xc nh)
- Tin hnh o A hoc T ca dy chun chn.
- Dng th AX = f(Cx). Vit PTHQ tuyn tnh ca ng chun.
- Tin hnh pha ch dung dch xc nh.
- Do A hoc T ca mu.
- Cn c vo PTHQ tuyn tnh ca dy chun v AX m xc nh
nng ca
cht X trong mu
02/09/15

28

Phng php ng chun

- th A = f(Ctc) tu theo cch o ta thu c 2 dng ng
chun:
+ Dng 1: i qua gc ta
+ Dng 2: khng i qua gc ta
- Khi chn vng nng xy dng ng chun phi ch :
+ Vng nng ca dy chun phi bao gm c CX
+ Vi vng nng chn dung dch phi tun theo nh lut
Beer
+ Cc gi tr Atc ng vi nng chn phi sao cho khi o trn
my c lp li cao v bo m s tuyn tnh A = f(C)
02/09/15

29

Phng php ng chun

U V NHC IM CA PHNG PHP

U IM
- Vi mt ng chun cho php phn tch hng lot mu.
- Dung dch cng khng i hi phi tun theo nh lut Beer
mt cch nghim ngt.
NHC IM
- chnh xc ca phng php khng cao.
- Khng loi c nh hng ca nn mu
02/09/15

30

Phng php so snh

So snh 1 chun
-Pha mt dung dch chun c Ctc.
-Tin hnh o A hoc T ca dd chun so vi dd so snh (Atc)
Theo nh lut Lambert Beer: Atc = lCtc.
-Pha dung dch mu vi nng cn xc nh CX (cha bit)
-Tin hnh o A hoc T ca dd mu so vi dd so snh (AX)
Theo nh lut Lambert Beer: AX = lCX.
Khi dung dch xc nh v dd chun c cng bn cht, c th xem

AX
C tc
nh nhau, v l = const. C X =
A tc
02/09/15

31

 nh lng Pb trong mu thc phm, ta tin

hnh cn 5,000 g mu, ho tan thnh dung
dch, sau tin hnh to phc vi thuc
th dithizon, dng phc Pb ditizon tan
trong CHCl3. Tin hnh chit bng CHCl3,
dung dch sau khi chit c nh mc thnh
25 mL. Dung dch chun c chun b tng
t nh mu, cha 10 g Pb2+ trong th tch
dung dch em o l 20,00 mL. Mt quang
ca chun v mu = 545 vi l = 1 cm ln
lt l Ac = 0,320 v Am = 0,225.

02/09/15

32

So snh 2 chun

02/09/15

33

 nh lng Pb trong mu thc phm, ta ti n

hnh cn 5,000 g mu, ho tan thnh dung
dch, sau tin hnh to phc vi thuc
th dithizon, dng phc Pb ditizon tan
trong CHCl3. Tin hnh chit bng CHCl3,
dung dch sau khi chit c nh mc
thnh 25 mL. Dung dch chun c chun
b tng t nh mu, vi mt bnh cha
6,25 g Pb2+ trong th tch dung dch em
o l 25,00 mL v mt bnh cha 12,5 g
Pb2+ trong th tch dung dch em o l
25,00 mL. Mt quang ca chun v m u
= 545 vi l = 1 cm ln lt l A 1 = 0,160,
A2 = 0,320 v Am = 0,225.
02/09/15

34

Phng php so snh

- Dung dch cn xc nh v dung dch tiu chun phi
nm trong khong tun theo nh lut Lambert Beer.
- Thun li khi s lng mu t
- So snh vi nhiu mu chun.
- kt qu chnh xc th Ctc1,Ctc2 v Cx phi c nng
gn bng nhau.

02/09/15

35

Phng php thm chun

Theo phng php ny th mt quang ca dd mu
cha cht cn xc nh c so snh vi chnh dung dch
c thm nhng lng xc nh ca cht cn xc nh.
Trong phng php thm chun, ta thm vo dd xc
nh mt lng dd tiu chun.
C 2 cch thc hin:
-Dng mt dung dch chun v p dng cng thc tnh
-Dng th biu din
02/09/15

36

Phng php thm chun

Dng cng thc tnh
Pha mt dung dch c th tch V cha ion cn xc nh c hm
lng rt nh. Sau tin hnh pha 3 dung dch nh sau:
STT

Bnh 1

Bnh 2

Bnh 3

Dung dch mu

VX

Vx

VX

Dung dch chun C

Vc

Thuc th to mu, m
pH

thm

thm

khng

02/09/15

37

Phng php thm chun

02/09/15

38

Ly 20,00 mL dung dch m u c ch a s t

cho to phc vi thuc th thch hp ri
pha long thnh 50,00mL dung d ch o. o
o hp ca dung dch = 510 nm c gi
tr A = 0,225 (s dng cuvt c l = 1cm).
Ly 20,00 mL dung dch mu cha s t khc
thm vo 4 mL dung dch st chu n 10
mgFe/L cho to phc vi thuc th thch
hp ri pha long thnh 50,00mL dung d ch
o. o o hp ca dung dch = 510 nm
c gi tr A = 0,358. Tnh nng ppm
ca dung dch mu st ban u.

02/09/15

39

Phng php thm chun

Phng php th
Pha mt dy dung dch chun cng chnh l dung
dch nghin cu c cho thm nhng lng chnh xc
ai ca cht cn xc nh nng ca dy chun l
CX + Ca1, CX+ Ca2 t nht l 3 dung dch v mt
dung dch so snh.
o mt quang A tng ng. Dng th AX +ai
Ci
02/09/15

40

Phng php thm chun

Phng php th

02/09/15

41

Phng php thm chun

Phm vi ng dng ca phng php thm chun

Phng php thm chun thng c p dng khi

nng cht phn tch rt nh (vi lng)
u im ca phng php thm chun l c th loi
c nh hng ca nn mu.
Tuy nhin, ch p dng i vi nhng dung dch tun
theo nh lut Lambert Beer .
02/09/15

42

My o quang
Cuvet cha
dung dch
phc mu.

02/09/15

43

Cuvet
trn

02/09/15

44

02/09/15

45

Cu hi trc nghim
1. Trong phng php quang ph hp thu phn t, vng
kh kin (thy c) l vng c t:
a. 100 400 nm
b. 400 800 nm
c. 800 1200 nm
d. 1200 1800 nm
2. Trong phng php quang ph hp thu phn t, vng
t ngoi (UV) l vng c t:
a. 200 400 nm
b. 400 800 nm
c. 800 1200 nm
d. 1200 1800 nm
3. Trong dung mi l nc, aniline hp thu bc sng
280nm vi = 1430 l.mol-1.cm-1. Nu mun pha ch
100mL dung dch aniline c truyn sut 30% i
vi bc x trn th phi cn bao nhiu gam aniline
(C6H5NH2) nguyn cht, (dng cuvet o c l = 1cm).
a. 3,4.10-2 g
b. 3,4.10-3 g c. 3,4 g
d. 34 g
02/09/15

46

4. xc nh hm lng st tng trong mu nc

sng ngi ta tin hnh xy dng ng chun,
o mt quang A v thu c kt qu nh sau:

Phng trnh ng hi quy tuyn tnh l:

a. 0,028 + 0,0532C
b. 0,028C + 0,0532
c. 0,014 + 0,0532C
d. 0,028 + 0,032C
02/09/15

47

5. Trong phng php o quang, gim cng

dng sng sau khi i dung dch c nng
7,9.10-5 M xung 10 ln th chiu dy ca
cuvet cha dung dch l bao nhiu? Bit rng
h s hp th phn t = 6300 l.mol-1.cm-1.
a. 1 cm
b. 2 cm
c. 4 cm
d. 5 cm

02/09/15

48

6. nh lng Fe3+ trong nc bng phng

php trc quang, thuc th KSCN, mi
trng HNO3 (pH = 12). Phc to thnh
c mu , hp thu = 480nm vi =
6300 l.mol-1.cm-1. Tnh nng mol ca
Fe3+ khi phc to thnh c hp thu A =
0,45 dng cuvet o c l = 1cm.
a. 7,14.10-5
b. 71,4.10-2
c. 7,14.10-4
d. 7,14.10-6
02/09/15

49

7. xc nh hm lng st tng trong mu nc sng

ngi ta tin hnh xy dng ng chun nh sau:

Nng ca dy chun ln lt l:
a.2 4 6 8 10 ppm
b.0,02 0,04 0,06 0,08 0,10 ppm
c.0,2 0,4 0,6 0,8 1,0 ppm
d.0,1 0,2 0,3 0,4 0,5 ppm

02/09/15

50

8. Ly 20,00mL dung dch mu c cha st cho to

phc vi thuc th thch hp ri pha long thnh
50,00mL dung dch o. o o hp ca dung dch
= 510nm c gi tr A = 0,225 (s dng cuvt c
l = 1cm).
Ly 20,00mL dung dch mu cha st khc thm
vo 4mL dung dch st chun 10 mgFe/L cho to
phc vi thuc th thch hp ri pha long thnh
50,00mL dung dch o. o o hp ca dung dch
= 510nm c gi tr A = 0,358. Tnh nng
ppm ca dung dch mu st ban u.
a. 3,38b. 1,35
c. 0,80 d. 27
02/09/15

51

9. Trong phng php o quang, khi o

truyn quang mt dung dch trong cuvet
c l=1cm th A = 0,245. Hi %T l bao
nhiu?
a. 68,30%
b. 61,08%
c. 56,88%
d. 57,60%
02/09/15

52

10. Trong phng php o dy chun ca mt

dung dch mu cho kt qu:

Nu mu phn tch c A = 0,672 th nng dung

dch l:
a. 0,2251
b. 0,1390
c. 0,1374
d. 0,1928
02/09/15

53

02/09/15

54