L Nht Tm UIH- IBF

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PHNG PHP PH UV-VIS
Cc ni dung chnh
+ Vng ph UV-Vis v ngun gc ca s hp th
+ Ph UV-Vis
+ S chuyn dch in t ca cc hp cht hu c
+ Nhm chc
+ Cc yu t nh hng n bc sng hp th
+ H thng thit b quang ph UV-Vis
+ Cc loi my quang ph
+ ng dng ph UV-Vis trong phn tch thc phm
Vng ph UV-Vis l vng nm cn UV cho n cn IR.
c xc nh t khong 180-1100nm.
y l vng ph c nghin cu nhiu v c p dng
nhiu v mt nh lng.
Qu trnh nh lng c tin hnh bng cch o mt vi
bc sng hp thu ca hp cht, sau p dng nh lut
Lambert-Beer tnh ton.
Nhiu th h thit b ra i da trn phng php ny, v ngy
cng ti u ha qu trnh.
Php ph UV-Vis cn c p dng cng vi cc phng php
khc nh Phng Php Sc k trong qu trnh nghin cu.
PHNG PHP PH UV-VIS
PH UV-VIS V NGUN GC CA S
HP TH
Vng ph ny thng c chia lm 3 vng ch yu: cn UV
(185400 nm), kh kin (400700 nm) v cn hng ngoi
(7001100 nm).
Ngun gc ca s hp th trong vng ny ch yu l s tng
tc ca cc photon ca bc x vi cc ion hay phn t ca
mu.
S hp th ch xy ra khi c s tng ng gia nng lng
photon v nng lng cc in t ngoi cng (ca ion hay
phn t) hp th.
Kt qu ca s hp th l c s bin i nng lng in t
ca phn t. Chnh v vp ph UV-Vis c gi l ph in t

PH UV-VIS V NGUN GC CA S
HP TH
PH UV-VIS
S hp th nng lng in t trong vng sng nh
sng t ngoi gn (190-400nm) v kh kin (400-
780nm) ca cc cht gy ra s chuyn dch ca cc
in t t trng thi c bn sang trng thi kch
thch.
Biu bin din s tng quan gia cng hp
thu theo bc sng ca mt cht c gi l ph
UV-Vis ca cht y trong iu kin xc nh
Cc quang ph k UV-Vis o truyn quang T hay
hp th A ca bc x khi truyn qua mu lng
truyn quang T c tnh:

Hay :

hp th A c xc nh :
PH UV-VIS
Ty vo trng thi ca mu o m ph thu c c nhng
ng nt khc nhau:
PH UV-VIS
S CHUYN DCH IN T CA CC HP
CHT HU C
Phn ln cc hp cht hu
c c nghin cu trong
vng ph UV-Vis.
Qu trnh chuyn tip bao
gm cc in t , or
hay in t n nm trn cc
orbital ca cc nguy t
nh nh H, C, N, O
CHUYN MC *
S chuyn v ca e trong lin kt ca cc hp cht hu
c t orbital lin kt ln phn lin kt *.

S chuyn v ny i hi mt nng lng kh ln, v vy
qu trnh chuyn v nm trong vng t ngoi xa ( UV).
S CHUYN DCH IN T CA CC HP
CHT HU C
CHUYN MC n *
S chuyn v ca cc in t t obital n ln cc orbital
* trong cc nguyn t nh O, N, S .
Xy ra vng ph t ngoi gn c cng khng
ln. S dch chuyn ny dao ng 180nm cho alcol,
dn xut halogen ca n l 190nm. i vi cc amin l
220nm
V d : Ete c max= 190nm ( =2000)
Metanol c max= 183nm ( =50)
Etylamin c max= 210 nm ( =800)
S CHUYN DCH IN T CA CC HP
CHT HU C
CHUYN MC n *
y l qu trnh thng xy ra trong phn t c mt
nguyn t cha in t khng lin kt nh nhng
phn t cha nhm chc cacbonyl (C=O) v bc sng
hp thu t 270nm- 295nm. C cng hp thu thp
Bn cht ca cc dung mi c nh hng n bc sng
hp thu v n tc ng n lin kt trong phn t.
S CHUYN DCH IN T CA CC HP
CHT HU C
CHUYN MC *
Cc hp cht ng phn vi etylen cha lin kt i
trong phn t c kh nng hp thu mnh trong khong
bc sng 170nm
V tr hp thu ph thuc vo s hin din ca nhm th
v d etylen c max= 165nm ( =16000)
Nhng hp cht khng mu thng c ph hp thu
trong vng cn t ngoi. Khi chng hp thu bc x th
chng s chuyn t orbital cho in t s chuyn ln
orbital nhn in t c mc nng lng cao hn.
S CHUYN DCH IN T CA CC HP
CHT HU C
TM TC CC BC CHUYN
S CHUYN DCH IN T CA CC HP
CHT HU C
CHUYN MC d d
S chuyn mc xy ra cc orbital d, nht l cc
kim loi vng chuyn tip
Cc phi t c cp in t t do tham gia lai ha
vi nhng orbital ny chuyn in t vo cc
orbital ny gy ra s chuyn mc.
Mu to ra ca cc phc lm cho phc c kh nng
hp thu nhng bc sng vng kh kin
S CHUYN DCH IN T CA CC HP
CHT HU C
CC NHM CHC
Nhm chc l nhng nhm nh c to thnh t
nhiu nguyn t, quyt nh tnh cht ca hp cht
hu c.
Ti v tr cc nhm chc trong phn t cc dch
chuyn in t xy ra, nn ph ca phn t hu c
lin quan ti cc nhm chc trong phn t.
Ngi ta tm c cc bc sng hp thu cc i
cho tng nhm chc.T c th d on s tn ti
ca cc nhm chc thng qua hp thu ca n.


NHNG YU T NH HNG TI S
CHUYN MC
nh hng ca dung mi
nh hng ca s lin hp
nh hng ca pH
NH HNG CA DUNG MI
Bc sng hp thu v cng hp thu ca cc hp
cht chu nh hng ca dung mi.
S tc ng ca nhng dung mi khc nhau ln cc
phn t lm thay i mc nng lng gia cc trng
thi kch thch v c bn.
S tc ng ca dung mi ln phn t lm sinh ra
:chuyn dch xanh v chuyn dch
NHNG YU T NH HNG TI S
CHUYN MC
CHUYN DCH XANH
L hin tng hp thu bc x ca cc hp cht hu c
c bc sng ngn hn trong nhng dung mi c tnh
phn cc cao
Hin tng tm thy qu trnh chuyn dch n * ca
nhm cacbonyl.
Nguyn nhn l do s lm bn trng thi n ca dung
mi.
NHNG YU T NH HNG TI S
CHUYN MC
NHNG YU T NH HNG TI S
CHUYN MC
CHUYN DCH
L hin tng cc hp cht hu c c xu hng hp thu
nhng bc x c bc sng di hn trong nhng dung
mi c phn cc cao hn
Hin tng c tm thy cc phn t hu c m trong
cu trc phn t ca n c s lin hp.

NHNG YU T NH HNG TI S
CHUYN MC
CHUYN DCH
Nguyn nhn ca hin tng ny l:
+ Do khi mch C cng di th hiu ng lin hp cng tng,
dn ti lch nng lng gia hai trng thi gim.
+ Trong phn t hu c c hiu ng lin hp cng di th
bc sng hp thu cng ln

NHNG YU T NH HNG TI S
CHUYN MC
CHUYN DCH
NHNG YU T NH HNG TI S
CHUYN MC
TM TC CHUYN DCH XANH V
CHUYN DCH
NHNG YU T NH HNG TI S
CHUYN MC
NH HNG CA pH
nh hng bn ca phc
nh hng n s to phc
nh hng dng tn ti
NHNG YU T NH HNG TI S
CHUYN MC
nh hng ca s lin
hp
S lin hp p- hay -
u lm cho trng thi kch
thch ca in t
*


bn hn
c nng lng thp hn u
ny dn ti bc sng hp
thu di hn
NHNG YU T NH HNG TI S
CHUYN MC
THIT B QUANG PH HP THU UV-VIS








Ngun sng c nhim v cung cp bc x tng thch vi
qu trnh o. Bc x c cung cp bi ngun sng
thng l chm bc x a sc, n bao trm mt khong
rng ca ph.
NGUN SNG
+ n Tungsten Halogen , l mt ngun sng ph bin
dng trong my quang ph. n ny cha mt si dy
mnh tungsteng c t trong thy tinh. Khong bc x
m n cung cp l t 330 n 900 nm, c dng trong
vng visible.
+Thi gian s dng n ny khong 1200h.
+ Vi U= 6v v cng rt ln dy
tungten b nung a bu khi tr
( neon, Argon)ln trng thi kch
thch v pht bc x



TUNGSTEN LAMP
n hydrogen or deuterium cung cp bc x trong vng
Ultraviolet tng ng vi di bc x t 200 n 450
nm.
Trong hai n th n Deuterium n nh hn v c thi
gian s dng khong 500h. y l n cho ph lin tc




Hydrogen / Deuterium Lamps
Hydrogen / Deuterium Lamps
Thu nhn chm bc x a sc pht ra t n, v cho
bc x n sc i ra..
C hai loi thit b ph bin gm lng knh c cch t


THIT B TO BC X N SC
Lng knh (Prism)
+ Nhng bc x c bc sng khc
nhau s b b gy nhng
gc khc nhau khi i ra khi lng .
+ Lng knh c th c lm t thy
tinh hay thch anh.
+ Ty thuc vo vt liu lm lng lnh
m n c th tch nhng bc x trong
vng no (Lng kinh thy tinh ph hp
vi cc bc x trong vng visible
nhng lng knh thch anh th bao ph
c hai vng Ultraviolet v Visible) .




THIT B TO BC X N SC
+ Cch t c cu to vi v
s nhng khe rt nh trn
mt din tch b mt
khong 200 khe trn mt
rng 1cm
+ Ty thuc vo gc ti ca
chm nh sng v b mt
cch t m hng truyn
ca chm bc x khi phn
x trn b mt cch t theo
nhng hng khc nhau..



CCH T (GRATINGS)
THIT B TO BC X N SC
CCH T (GRATINGS)
Phn loi cch t:
+ Cch t truyn sut: c lm bng thy tinh
+ Cch t phn x : Lm bng nhm
THIT B TO BC X N SC
Quan st ph qua cch t
THIT B TO BC X N SC
+ Khoang hp thu l vng ti nm v tr cui cng ca
ng truyn.
+ Tia bc x n sc sau i c tch ra s i n .
Thng c thit k l mt ng nh gi l Cuvettes.
+ Cuvettes c lm bng nha, thy tinh hay thch anh
cha mu o.

B PHN CHA MU(CUVETTES)
+ C tc dng cm nhn bc x in t sau khi b hp th v
chuyn lng bc x ny thnh dng in.
+ Cng dng in thu c l t l thun vi cng
bc x p vo b mt catot.
+ T bo quang in hay ng nhn quang in l nhng thit
b hu dng trong vic o xc nh




THIT B NHN BIT(DETECTORS)
ng nhn quang in c chc nng t hp cc tn hiu
chuyn i qua vi giai on khuych i trong thn ca
ng. Bn cht ca nguyn liu lm cathode l xc nh
nhy ca ph.
NG NHN QUANG IN
THIT B NHN BIT(DETECTORS)
THIT B NHN BIT(DETECTORS)
T bo quang in
CC LOI MY QUANG PH
+ My quang ph chm tia n l c pht minh ra u
tin, v ton b nh sng i qua mu.
+ Loi ny l r hn v n c thit k kh n gin
C hai loi : My mt chm tia v hai chm tia
My mt chm tia
u im ca my mt chm tia : Gi thnh thp, thng lng
bc x i qua cao v nh vy nhy cao.
Nhc im: l c khong lch thi gian khi tin hnh o gia
cc chun cng nh mu xc nh, v vy c th c vn
vi tri. iu ny xy ra vi nhng thit b c, nhng
thit b hin i vi nhng tnh nng cao v n nh khc
phc nhc im ny. V vy my quang ph mt chm tia
vn c nhng ng dng cao trong cc phng th
My mt chm tia
CC LOI MY QUANG PH
MY QUANG PH DIODE ARRAY
CC LOI MY QUANG PH
+ Thit b o hai chm tia nhm mc ch khc phc tri
xy ra trong qu trnh o. Nguyn nhn gy ra tri l cng
nh sng b mt i trn ng truyn, do thay i ng i,
do phn x, do s chuyn ha thnh dng in khi tng tc
vi detector. S mt i ny khng ng nht khi o mu blank
v mu o ring bit.
+ Trong thit b hai chm tia c hai v tr o, tng ng vi
mu v mu i chng. Chm tia sng ti c phn tch thnh
hai chm c cng bng nhau, mt chm i qua mu o v
mt chm i qua mu i chng.. Nh vy I
0
v I c o cng
mt lc.
Double beam
CC LOI MY QUANG PH
Double beam
CC LOI MY QUANG PH
u im ca thit b hai chm tia
Cho chnh xc cao v mu o v mu i chng c o
cng mt lc.
Nhc im l gi thnh cao, nhy thp do cu trc
quang hc phc tp hn, tin cy thp hn.
Double beam
CC LOI MY QUANG PH
NG DNG PH UV-VIS TRONG
THC PHM
Phn tch protein
Phn tch Carbonhydrat
Phn tch hm lng kim loi trong thc phm
Phn tch mt s ch tiu nc

nh lut Lambert- Beer
nh lut Lambert- Beer
nh lut Lambert- Beer
Mt s sch k hiu K

l

HIU CHNH

SAI LCH TRONG L LAMBERT - BEER
nh lut ch ng vi nhng dung dch c nng
l thp
Do ngun sng khng tht l n sc nn c ln
nhng bc x c bc sng ln cn nhim vo. Dn
ti A= lg I/I
0
khng ng