QUANG PH UV-VIS

Ths.Dng Th Trc Ly
i tng: DK27 & DK38

1
MUC TIEU
1. Trnh bay c c s ly thuyet ve
phng phap quang pho UV-VIS.
2. ng dung c phng phap quang pho UV-VIS
trong phn tch.

2
- Bc xa vung UV- Vis chia thanh 3 vung nho:
* * ()m()n* ( - )n( - )n
UV xa
UV (chan khong) 200 UV 400 Vis 800
(nm)

- Vung t ngoai chn khng (UV xa) = 50-200nm it c s dung vi:

+ Co nng lng kha ln, khi va cham gy v lin kt / phn t
+ Bi hp thu manh bi hu ht dung mi va oxy cua khng khi
+ Bi hp thu bi thach anh (dung lam cc o)

- Vung t ngoai gn (UV gn): = 200 - 375 nm

- Vung kha kin (Vis) : = 375 - 800 nm

3
C phi hp cht no cng cho ph UV-VIS?
iu kin no c ph UV-VIS?
V du : xet phan t formaldehyd (HCHO):
ien t tao day noi n
ien t tao day noi oi
ien t n khong tao day noi

4
S kich thich in t t orbital phn t ang chim ng (orbital lin
kt) hoc khng lin kt (n) sang orbital phn lin kt

5
 E *>E n * > E * > E n *

- in t :
+ Lin kt n trong phn t
+ * cn E ln ( <150 nm: UV chn khng).
+ Hidrocarbon no: dung lam dung mi
- in t :
+ Lin kt tham gia lin kt i va ba,
+ T * cn E nho hn ( UV-Vis)
+ Phn t co ni i, ba, ni i lin hp
- in t t do n (phi lin kt):
+ in t n co cac di t O, S, N, X.
+ n * cn nng lng gia UV gn v xa
+ n * cn E thp hn *
Cac phn t co cu truc lin hp co nhiu in t va n
tham gia n*, * 6
- Cac hp chat co he thong noi oi lien hp se hap thu anh
sang vung UV-Vis, so day noi lien hp cang tang th hp
chat hap thu photon anh sang vung co bc song dai hn.
Do vay co the noi pho UV - VIS cho ta cac thong tin ch yu
ve day noi oi.

- Cac hp chat no khong cho hap thu UV v the ta s

dung chung lam dung moi hoa tan cac hp chat khac khi
o pho UV-VIS.

7
Ten goi Chuyen Cau truc max va
cua dai dch (nm) max
-C=O; thay oi
R n* -NO2 ;-C=S ; <100
radikalartig -N=N-..
K polyen,
Konjugierto
* styren, 104
acetophenon
benzaldehyd

B * nhan thm, 100 < <104

Benzenoid 230< max<280
d vong thm

E * Hp chat khong 2.103<

bao hoa, vong <14.103
Ethylenic kn lien hp

8
9
 O HAP THU CUA VAI CAU TRUC THEO KIEU DAI R
Cau truc Cong thc Dai R yeu
:
bc song
Cac aldehyd va ceton R C =O, -CHO gan 280 nm
mach thang
Cac hp chat nitro (- NO2) khoang 270 nm

Thioceton C=S dai hn 460 nm

Cac hp chat azometan CH3 N = N CH3 gan 347 nm

10
 HAP THU CUA POLIEN TANG THEO SO NOI OI

Cau truc polien n max max

1 174 24000

2 227 24000

CH3 (CH = CH)n CH3, 3 275 30000

4 310 76500

5 342 122000

11
3. CAC AC TRNG E MO TA PHO UV-VIS
V tr (1), cng (2), hnh dng 3): vn ph
Abs
334
0.9
0.8
0.7
Mu chun
0.6
0.5
242
Mu th
0.4
0.3
0.2
0.1
0.0
nm
200 250 300 350 400 450

12
4.1. MAU SC
- Co tm quan trong ring bit cho 1 cht.
- Mau cua 1 cht lin quan vi s hp thu va phan xa cua 1 cht.
- Mt ngi nhin thy mau b tr cho mau hp thu.

13
Bc song Nang lng Mau sac Mau sac thay , nm
cua (kj/mol) anh sang c
vach hap hap thu cua chat Violet 400-420
thu, (nm)
Indigo 420-440
380- 420 299- 274 Tm luc vang
420- 440 274- 249 lam vang Blue 440-490
440- 490 249- 244 lam-luc nhat da cam Green 490-570
490- 500 244- 238 luc-lam nhat o
500- 560 238- 214 luc o ta Yellow 570-585
560- 580 214- 206 luc-vang tm
Orange 585-620
580- 600 206- 200 vang lam
600- 650 200- 198 da cam lam luc nhat Red 620-780
650- 750 198- 149 o luc-lam nhat

Mau hp thu va mau b tr

14
15
4. 2. NHOM MANG MAU (CHROMOPHORE)
La nhom chc cha no, lin kt ng hoa tri trong phn
t gy ra s hp thu bc xa trong vung UV-VIS ( > 200nm)
- CHROMOPHORE chuyn dich n * thng co
300nm.
- CHROMOPHORE chuyn dich * thng co
ranh gii UV gan va xa va co ln .

16
 Mt s nhom mang mau va max tng ng:

Nhom mang mau Cong thc Th du max (nm)

Carbonyl (ceton) RRC = O Aceton 271
Carbonyl RHC = O Acetaldehyd 293
(aldehyd)
Carboxyl RCOOH Acid acetic 204
Amid RCONH2 Acetamid 208
Etylen RCH = CHR Etylen 193
Acetylen C=C Acetylen 173
Nitril C=N Acetonitril 160
Nitro RNO2 Nitromethan 271

17
 Trong h lin hp, tinh bt i x cua lin kt cang tng thi cac
chromophore cang d hp thu
o hap thu cua phan t tuy thuoc v tr cua 2 nhom chromophore
- Ngan cach bi C A = tong o hap thu cua moi chromophore
va chuyen dch ve pha bc song dai
- ke nhau A tang len (hyperchromic) va cc ai hap thu chuyen
ve bc song dai hn

- S hin din cua dai hp thu tai mt c bit l mt ch thi tt cho

nhm mang mu.
- hp thu cc ai khng c inh ma con tuy vao iu kin: pH, dung
mi, nhit ,...

18
4.3. NHOM TR MAU (auxochrome)

- La nhng nhom th no cha cc d t gn vao h thng hp

thu lam thay i ca bc song ln cng cua dai hp thu

cc ai.

- Thng lam chuyn dch max v phia dai hn.

Vi du: -OH, -NH2, -Cl, -SH...

- Lam tinh hp thu tng do lam giam nng lng cn hp thu.

19
20
4.4. CAC HIU NG VA S CHUYN DICH
- S chuyn dch sang o (bathocromic) = red shift
- S chuyn dch sang xanh (hypsocromic) = blue shift
- Hiu ng tng cng (hypercromic effect)
- Hiu ng giam cng (hypocromic effect)

21
22
 5. CAC YU T ANH HNG N HP THU
5.1. CU TRUC PHN T

5.2. MI TRNG
Mt s yu t nh:
- Dung mi
- Nng
- pH
- Nhit
Cac thng s nay phai c kim soat am bao chinh
xac cao nht va khi so sanh cac ph c o di nhng
iu kin khac nhau

23
5.2.1. DUNG MI

24
hyperchromic

bathochcromic

25
5.2.1. DUNG MI

- Dung mi co th hp thu bc xa UV-VIS. Khi khao sat phai chu thich dm

c dung hoa tan mu.
- phn cc cua dm co th lam bin i mi trng in t cua nhom hp
thu mang mau v ln cua s chuyn dch co th lin quan vi phn
cc cua dm.
Vi du: hp thu cua aceton co th thay i t 259nm - 279nm tuy vao dm
c s dung.
- phn tich so sanh, nn s dung mt dm duy nht cho tt ca cac ln o.
- Tng tac lng cc (dipole-dipole): thng xut hin khi cht tan va dm
u la cht phn cc. Tac dung nay se co th gy s hiu ng
bathochromic hoc gy hiu ng hypsochromic.

26
 Dung moi Hang so ien ly Max (nm)
Nc cat 78,5 < 195
Hexan 1,9 199
Ethanol (tuyet oi) 24,3 207
Methanol 32,6 210
Cyclohexan 2,0 211
Chloroform 4,8 246
Dimethylsulfoxid Khong 270
Aceton 20,7 331
Pyridin 305
Benzen 280
Carbon tetraclorid 265
BuOH 210
Isopropanol 210
Dicloromethan 235
27
 KHOANG BC SONG HAP THU CUA MOT SO DUNG MOI

o dai song Cac dung moi hap thu

180-195nm Acid sulphuric (96%), nc, acetonitril, cyclohexan, isooctan

200-210nm Cyclopentan, n-hexan, glycerol, methanol, ethanol

210-220nm N-butyl alcohol, isopropyl alcohol, cyclohexan, ethyl ether, 1,4-

dioxan

245-260nm Chloroform, ethyl acetat, methyl format

265-275nm Carbon tetrachlorid, dimethyl sulphoxid/formamid, acetic acid

280-290nm Benzen, toluen, m-xylen

300-400nm Pyridine, aceton, carbon disulfit

28
5.2.2. NNG
Thng thi nng ch anh hng n cng cua dai hp thu.

nng cao, tng tac phn t (nh la dimer hoa) co th gy nn s

thay i v dang va vi tri cua dai hp thu lam khac i tinh tuyn tinh

cua hp thu theo nng va dn n kt qua inh lng khng

chinh xac.

Ph UV-VIS ca benzaldehyd
thay i theo dung mi va nng 29
5.2.3. pH
- Khi thay i pH thi cu truc phn t cua mt cht c th thay i se
anh hng rt ln ph hp thu chu yu l dich chuyn gia hai dang
cn bng khac nhau.

Vd: cc gi tri pH khc nhau th cu trc cua ch thi pH c th thay

i nn se hp thu cc ai bc sng khc nhau.

- Nu ang khao sat mt cht ma pH anh hng n ph cua mt mu

o thi nn dung h m lam n inh pH mi trng ri mi khao
sat hp thu cua cht nay.

30
 (Helianthin)

Chu y: hu ht cac h m hp thu co y nghia va co th anh hng n

dai song cc ai i vi cac phep o. Do vy, phai lam song song mu
trng.

31
 Phenol Phenolat
* max = 210 =6200 max = 235 =9400
n* max = 270 =1450 max = 287 =2600
- Phenol/ mi trng trung tinh: phenol th hin dai E va dai B nh benzen
- Phenol/ kim, phenol phenolat: OH (tr mau) bathochromic dai B
(gop chung cac electron lam tng tng tac n*)
ng dung:
- Nghin cu s co mt cua phenol.
- o pKa cua acid yu (pK=10).

32
- Mi trng trung tinh: ph anilin tng t vi ph phenol.

- Mi trng acid: nhom amin t proton hoa tao ion anilinium. Khng con
electron t do trn nit nn giam tng tac n* va k o xay ra hiu
ng hypso/ hypochromic.

33
nh hng cua pH n ph UV-VIS

34
5.2.4. NHIT
- S trng n n gian cua dm co th lam thay i hp thu biu kin va
do o cung co th lam anh hng n ung cua kt qua.

- S cn bng vt ly hay hoa hoc: in hinh la khi nhit tng se pha huy
cu truc cua mt s hp cht.

Nh vy:

- Nu khi o ph, nhit co anh hng n mu thi phai s dung cc o

n nhit khng lam thay i hp thu biu kin.

35
 6. MAY QUANG PH T NGOAI
Spectrophotometer

Light Source Monochromator

(filter,
wavelength selector)
Detector
Sample

Data Processing

36
6.1. CU TAO

S may UV-VIS
1: en ngun 2: cach t hay lng kinh
3: cc cha dung dich o 4: t bao quang in
5: b phn khuch ai 6: may ghi tin hiu
37
6.2. Phn loai

Quang ph k 1 chum tia Quang ph k 1 chum tia (dai diod)

Quang ph k 1 chum tia (dai diod) 38

 39
Quang ph k 2 chum tia
 6.3. Cc b phn cu to Ngun sng

en Deuterium : s dung cho vung UV en Tungsten: s dung cho vung

VIS vi co cng cao / vung
vi vung VIS bi nhiu nhiu.
nay. Half-life 10.000 gi
Half -life 1.000 gi

40
en xenon co th la ngun cua vung UV-VIS
nhng nhiu nhiu hn en deuterium va tungsten.
en xenon chi c s dung o s khuch tan phan xa,
chu yu la cng cao.
41
B PHN PHT HIN (DETECTOR)

DETECTOR DIOD ARRAY 42

CC O

43
3 method c s dung trn may quang ph UV-VIS 2 chm tia:
- Photometry (o im): o hp thu cua 1 mu 1 (2,3,...)
bc song bt ky. Vi du o A cua dung dch kalipermanganat
bc song 450nm.
- Wavelength scan (quet ph): xac nh ph cua 1 cht trong 1
vung bc song nao o. Vi du: quet ph dung dch
kalipermanganat vung 400-650nm.
- Timescan : theo di s thay i hp thu cua 1 cht theo thi
gian. Vd: theo di thi gian kt thuc phan ng cua ion canxi vi
EDTA tao phc Ca-EDTA.
Cuvet (cc o): 1 cp ng nht (1 cai cho mu trng, 1 cai cho mu cn o).
44
Mt may quang ph chun la may phai c hiu chun v:
7.1. ung bc song (phai c chun hoa v )
Thng thng chun bng:
- Cac kinh holmium peroxid (quet ph so sanh vi ph chun)
hay didymium.
- Cac cht a bit trc cc ai hp thu nh: vitamin B12 co cc
ai 361nm, cobalt clorid co cc ai 510nm.

45
46
47
7.2. Phai cho s chi ung v hp thu
Vi dd kali cromat 0,040g/1000ml KOH 0,05M, phai cho hp
thu cac bc song ung vi hp thu chun (xem bang)-
hoc dd kali cromat 60mg/l H2SO4 0,005M.
Tra kt qua (Dc in My) hp thu cac bc song ung
vi hp thu chun.

48
Gia tr hp thu chun cua dd K2Cr2O7 0,040g/lit /KOH
0,05M 250C (cc o 10mm)

Bc Bc Bc Bc
o o o o
song song song song
hap thu hapthu hap thu hap thu
(nm) (nm) (nm) (nm)
220 0,4559 290 0,4295 360 0,8297 430 0,0841
230 0,1675 300 0,1518 370 0,9914 440 0,0535
240 0,2933 310 0,0458 380 0,9281 450 0,325
250 0,4962 320 0,0620 390 0,6841 460 0,173
260 0,6345 330 0,1457 400 0,3872 470 0,0083
270 0,7447 340 0,3143 410 0,1972 480 0,0035
280 0,7235 350 0,5528 420 0,1261 490 0,0009
500 0,0000

49
 phng ng nn

50
Ph ao ham bc 2

51
 phn giai

52
Cc o: kim tra s khac nhau truyn quang gia 2 cc o

53
 8. NG DUNG
8.1. INH TINH
-Ph hp thu cua mt cht la ng biu din hp thu cua cht o
theo bc song cua anh sang chiu ti: A = f ().
- T ng biu din nay co th xac nh c cac hp thu cc ai.
CC AI HP THU: bc song ma o cht o co hp thu ln
nht.
- Ph UV -VIS thng thng chi th hin mt s it dai rng nn cung
cp thng tin it hn quang ph IR (ph nay co nhiu dai hep).
- S hp thu nhiu nht cua cac cht hu c la do s co mt cua cac
ni (khng bao ha).

54
8.1.1. nh tinh trong trng hp khng co cht chun

- So sanh ph cua cht khao sat vi ph co trong tai liu, trong

cac b ngn hng ph.

Phai thc hin ung theo cac iu kin a ghi (tai liu) v dung
mi, nng , loai may.

- Trng hp n gian co th so sanh max hay max & A11

cung u.
A11
Vd: cc ai hp thu cua dexamathason l 241nm, cua
dexamethason bc sng 241nm l 297.

55
8.1.2. nh tinh trong trng hp co cht chun

So sanh ph cua mu o vi ph cua cht chun:

- Cung nng , hai ph phai cho max va max ging ht nhau.

- Tin hanh: ghi ph cua mu khao sat (cn nh tnh) va mu chun trong
cung dung mi, may, nhit , ri so sanh ph.

- Nu ung 1 hp cht thi 2 ph phai chng khit ln nhau (overlay ph).

56
57
8.1.3. Nhn xt

PP u im Nhc im

UV Vis Co the xac nh chat Phai phoi hp vi nhieu thong so vat

khao sat trong dung ly va hoa hoc, hoa ly khac.
dch rat loang 15 - 40 (iem chay, ch so khuc xa ,
g/ml. SKLM,) mi ket luan.
So dai t, cac phan t ln co cau truc
ch khac nhau t th lai cho pho UV-
VIS giong nhau, t ng dung hn pho
------------------\ IR.
IR
So dai nhieu, a so ----------------------------
cac phan t ln co cau Nong o mau phai am ac hn.
truc khac nhau khong
nhieu th lai cho pho
IR khac nhau.

58
Mt v d v ph IR:

59
8.2. Phat hin tap cht
- Xac nh tap cht trong cac phn t hu c.
-Cac inh la co th xut hin thm do tap cht trong mu, so sanh vi
nguyn liu chun. Cung co th phat hin tap cht bng cach o hp thu
bc song c trng.
Vi du: phat hin tap cht trong paracetamol

60
8.3. Xc nh nhm chc
Pht hin s c mt cua cc nhm chc trong hp cht

61
 8.4. nh lng (nh lut Lambert Beer)
8.4.1 nh lng trc tip
8.4.1.1.nh lng cht khao sat co mt thanh phn
Phng phap o tuyt i (khng s dung trc tip
cht chun)
A
Cx% = 1%
A1cm

iu kin:
-My o phai c chun ha t nht v bc sng v hp
thu.
-Cng iu kin mi trng (dung mi, pH, nhit ).

62
Vd: xc nh lng paracetamol trong trung bnh
1 vin Panadol (mg)?
o hp thu dung dch paracetamol bc sng 257nm
A11
l 0,728. cua paracetamol 257nm l 715, khi lng
bt Panadol a cn l 0,1023g; khi lng trung bnh vin
Panadol l 0,5126g, dung dch em o c pha long 10000 ln
so vi lng bt thuc a cn.

63
nh lng dexamethason phosphat trong thuc nho mt
Neodex bng phng php UV-Vis, bit rng: ht chnh xc 1ml
ch phm (nhn: 1 chai c 5ml cha 5mg dexamethason
phosphat) cho vo bnh nh mc 50ml, thm nc ct va u,
o c hp thu 241nm l 0,658; cho 1%
Acm = 297 (241nm).
Tnh hm lng % dexamethason phosphat trong ch phm?

64
Phng phap s dung h s hp thu mol cua mt cht
Theo cng thc : A = C. l vi (l =1cm)
A
C

( : tham khao tai liu hoc o trn mu chun) co th suy ra Cx
(mol/lit)
Vd: o hp thu cua dd phenol 270nm c A= 0,478;
= 1450 (phenol, =270nm). Tnh nng (g/l) cua dd phenol
trn.

65
Phng phap o tng oi ( a co chuan)
So sanh o hap thu (Ax) cua dung dch th nghiem co nong o (Cx) vi o hap
thu cua dung dch chuan co nong o biet trc (C0)

A x Cx A
Cx x C 0
A 0 C0 A0

Chu y:
- Cx va Co khong c chenh lech qua. Trong thc nghiem, Cx va Co
cang gan nhau ket qua cang chnh xac.

- A0: o hap thu cua dung dch chuan.

66
Phng phap dung ng hi qui chun: (xy dng ng cong chun ).
- Pha cac dd mu chun C1, C2, C3,C4, Cn chinh xac / dm thich hp.
- Ln lt xac nh A1, A2, A3, A4...An max
- Ve th vi truc tung la (A), truc hoanh la (C)
- Xac nh y = ax + b vi R2 = 0,999...
- o Ax cua dd cn khao sat ri cn c vao th tim Cx
(Cx phai nm trong khoang C1 Cn khao sat)

67
8.4.1.2. inh lng hn hp co nhiu thanh phn
Khai nim v s chng ph
Vd: 2 cht trong mt dung dich co 2 dai hp thu

Nu 2 dai co A gn nhau va cac Nu 2 dai co cac inh hp thu cac

cc ai tai cac u xa nhau rt gn nhau va A rt khac nhau thi
(20nm) thi se co th thy c 2 dai manh nht se che lp hoan toan
cc ai. dai yu nht.
68
Lut cng tinh mt quang
iu kin: max cua chung cach nhau mt khoang t nht 20nm (ph 2 cht
khng chng ln nhau tng phn tng ng).
Tin hanh:
- Quet ph UV-VIS ring re cua 2 cht X va Y.
- Chon 2 cc ai max1 va max2 c trng cua 2 cht X va Y.
- o ring A tng cht tng bc sng cc ai va tm c trn.
- o A cua dd hn hp cac max1 va max2 cua cac thanh phn vi cc o
co b day 1cm.

69
 -Da trn tinh cht cng tinh cua mt quang, ta co:

mt bc song xac nh thi hp thu cua nhiu hp cht co mt trong

mt hn hp bng tng hp thu cua mi thanh phn.

A1 = 1x X l 1y Y l

A2 = 2x X l 2y Y l
1x , 1y : h s tt mol cua X va Y 1

2x , 2y : h s tt mol cua X va Y 2 (cac h s nay c xac nh khi o

ring bit dung dch cua cac thanh phn).
- Giai h 2 phng trinh 2 n s, tnh c nng tng thnh phn.
- PP nay chi ap dung c cho cac hn hp 2 - 3 thanh phn).
(nu s thanh phn nhiu hn thi rt kho xac nh)

70
Cho KMnO4 (max1=525,5nm; C=4.10-4 M ) v K2Cr2O7
(max2=445,5nm; C=2.10-3 M ); Xac nh nng (mol/l)
tng thanh phn/hn hp Z (K2Cr2O7 & KMnO4)bit:

A KMnO4 K2Cr2O7 HHZ

max1 0,916 0,051 0,31
max2 0,072 0,757 0,538

71
Phng phap quang ph ao ham: 2 cht co max cach xa vai nm, chng
ph

-Cac ph ao ham bc chn (2,4,6,...) thi se co mt min nh v cung

max i vi ph bc 0.
- Cac ph ao ham bc le (1,3,5...) thi A= 0 max cua ph bc 0.

72
 A

A B

Ly ph ao ham cua tt ca cac thanh phn A va B

-Chon bc song nh lng cua cht A (la ni ma ph cua cht B i qua
truc hoanh).
-Chon bc song cua cht B (la ni ma ph cua cht A i qua truc hoanh).
73
Vi du: nh lng paracetamol (A) va propacetamol (B).
Ph hp thu cua 2 cht nay b chng ln nhau. Khi ly ao
ham bc 1, tai bc song 239nm gia tr ph ao ham cua
B bng 0 nn co th xac nh A trong hn hp ma khng b
anh hng bi B. Tai bc song 242nm gia tr ph ao
ham cua A bng 0 nn co th xac nh B trong hn hp
ma khng b anh hng bi A. Nh vy chi cn dng 2
ng chun tai 2 bc song trn.

74
75
8.4.2. INH LNG GIAN TIP

8.4.2.1. Chun o quang ph

Trong phn tich th tich, s thay i mau ng nghia vi kt

thuc chun va thng c quan sat bng mt thng. Qua
trinh nay hay mc sai s. Vic s dung quang ph k phat
hin im kt thuc se giam i iu nay

- Nguyn tc: thay i gia tr hp thu se c dung kim

tra qua trinh chun .

76
Vi du: apotransferrin khng mau. Khi Fe3+ gn vao protein nay mau o
(max = 465 nm): apotransferrin + 2Fe3+ [Fe3+] transferrin.
Tin hanh: chun 2ml dd apotransferin bng nitril triacetat ferric
Kt qua:
- Khi thm Fe3+ vao protein, mau o xut hin va A tng ln.
- Khi protein bao hoa st, se khng co phan ng tao mau nao na va ng
biu din b gay ngay tc khc (off abrupt).
- im kt thuc c ngoai suy t giao im cua hai ng thng.

Abs iem ket thuc

0,400

0,300

0,200

0,100

0 50 100 150 200 250 l Fe(III)

77
8.4.2.2. Tao dn cht hp thu manh / UV-VIS

Nguyn tc:

Thm mt thuc th hu c vao cht khao sat khng hp thu tao thanh

phc cht hp thu manh hn cht ban u ri sau o chit sang mi trng

khac va o trc tip.

Vi ky thut nay thi vic la chon mt TT thich hp se lam gia tng nhay

va tinh chon loc mt cach co y nghia.

ng dung: cac pp phn tich nc va tap cht trong nc.

78
V du:

Dithizon (la diphenylthiocarbazon) tao phc bn vi kim loai, vi

(Pb, Hg) tao phc bn tan c trong dung mi hu c.

Giai thich:

Dithizon khng tan/ nc ma tan/ nhng dm kem phn cc nh

CHCl3. Khi ly dch CHCl3 nay lc vi dd nc (pH thay i) co

cha ion kim loai thich hp (Pb, Hg) thi se tao thanh mt phc

c mau tan /CHCl3 co hp thu vng UV-VIS.

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8.5. NGHIN CU NG HOC PHN NG

Enzyme xc tc s chuyn i 1 hay 1 s c cht thnh 1 hay 1 s san

phm. Ti l cua phan ng xc tc hay hoat tnh cua enzyme c th c xc

nh bng cch o s giam nng c cht hoc s tng nng san

phm. Khi c cht v san phm khc nhau v hp thu, qu trnh phan ng

enzyme c th c theo di bng cch kim sot s thay i hp thu

theo thi gian.

80
8.6. CAC NG DUNG KHAC
8.6.1. Xac inh hng s pKa
Nguyn tc:
Ph UV-VIS cua cac cht hu c co tinh acid hay base thng thay i theo
pH cua mi trng.
Dang va cng cua ph hp thu thay i theo [H+]
Vi du : HB: acid, HB + H2O H3O+ + B-
B- : base lin hp vi dd acid HB.

pKa pH lg
base
pK HAc lg

Ac

lg
( A AB )
acid HAc ( AHB A)

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- Gia tr pH o bng may o pH.
- Gia tr cua ty s may o quang ph. T o tinh c pKa.
Giai thich:
Mt tht acid se o c gia tr hp thu cc ai cua dang HB.
Mt tht kim se o c gia tr hp thu cc ai cua dang B-.
Mt trung gian o c gia tr hp thu cc ai cua ca 2 HB va B-.
hp thu o c se la s t hp tuyn tinh cua 2 dang o.
p dung lut cng tnh mt quang.

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8.6.2.Xac inh s tiu phn hp thu trong dung dich (xac inh im ng
quang)

- Nu 2 cht vi nng bng nhau / dung dch va co nhng dai

hp thu xen phu nhau (ct nhau) thi tn tai mt bc song ma
o h s hp thu mol cua chung bng nhau.

- Nu tng nng cua 2 cht / mt dung dch = hng s thi du ti

s nng cua chung thay i nhng s hp thu tai bc song
noi trn vn khng thay i.

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Thng gp nht la 2 cht nm trong 1 cn bng ma

ng cong hp thu cua chung giao nhau tai mt bc

song nao o. Giao im o khng b thay i khi thay i

cac yu t anh hng n cn bng (pH). Noi mt cach

khac trong cac trng hp trn co tn tai mt bc song

ma o h s hp thu mol cua 2 cht o bng nhau.

im giao nhau tai o goi la im ng quang.

Vd: o methyl trong cc mi trng pH khc nhau

84
- pH = 4,5: o methyl la hp cht mau o/ dd (dang acid HIn)
- pH = 7,1: mau vang c tao thanh/ dd (dang kim In-)
- pH = 4,5 -7,1: dd se cha ca hai dang aid HIn va dang kim In-
(hinh cho thy tt ca ph cua 2 dang 1 [ ] moi pH ct tai mt
im).

Cac ng biu din ph hp thu cua cac dd cha o methyl

85
 iem ang
quang

Ph cua 1 hp cht c ghi nhn nhng pH khac nhau (cng nng )

co cung 1 im co hp thu mt dai song xac nh. im nay c
goi la im ng quang.

Vi du: ph UV-Vis cua methyl da cam cac pH khac nhau.

86