BMCNTP HBK TPHCM

NGUYN L

Da trn nhng bin i gy ra bi s tng tc gia
bc x vi vt cht

BMCNTP HBK TPHCM
BC X IN T
Bc x in t: l dng nng lng truyn i trong
khng gian
Bc x in t c bn cht sng v ht
BMCNTP HBK TPHCM
BC X IN T
Bn cht sng: Bc sng, chu k, tn s, s sng, vn
tc truyn sng, s giao thoa

Bn cht ht: c trng bi cc quang t (photon)
mang nng lng

BMCNTP HBK TPHCM
BC X IN T
Cc i lng c trng
Cc i lng Phng trnh n v
Bc sng, nm
S sng, = 1/ cm
-1
Tn s, = c/ Hz hay s
-1
Nng lng, E E = h = h.c/ J hoc Kcal
BMCNTP HBK TPHCM
NNG LNG CA VT CHT
Nng lng ca vt cht:
E = E
t
+ E
d
+ E
q
E
t
: nng lng in t
E
d
: nng lng sinh ra do giao ng tng tc gia cc
nguyn t
E
q
: nng lng do s quay ca phn t xung quanh trng
tm ca n
BMCNTP HBK TPHCM

Phn vng bc x in t:
BMCNTP HBK TPHCM
10Mm
30Hz
Bc sng
Tn s
BC X IN T
Nguyn t hay
phn t mc
nng lng thp,
b kch thch bi
bc x c mc
nng lng ph
hp, c th thu
nng lng,
nhy ln mc
cao hn

Bc x t ngoi v kh kin
c kh nng di chuyn in t t mc
c bn ln mc nng lng cao
Bc x hng ngoi gy nn s dch
chuyn mc nng lng dao dng
ca cc nguyn t

Bc x c mc nng lng thp
hn gy nn s dch chuyn
mc nng lng quay ca phn t
Bc x tng tc ln vt cht
BMCNTP HBK TPHCM
BMCNTP HBK TPHCM
K THUT QUANG PH
Khi bc x tng tc vi vt cht th bin thin nng
lng l 1 i lng c th o c hoc l tn hiu c
th ghi c
BMCNTP HBK TPHCM
Bc x Vt cht
o
KQ
Ghi tn hiu
K THUT QUANG PH
Th hin:
1. S hp thu: vt cht s gi li 1 cch chn lc nng lng ca
mt s bc x

2. Ph hp thu: kt qu ca s hp thu l lm cho nng lng
ca bc x b gim , c s thay i tnh cht sng ca bc x
(xut hin vch ti tng ng vi bc sng b hp thu trn
quang ph vch) hoc th hin ng biu din s gim
nng lng ca bc x

3. S pht x: Vt cht hp thu nng lng, chuyn ln mc
nng lng cao, trang thi kch thch khng bn, c xu
hng quay v trng thi bn, khi n pht ra bc x (s
pht hunh quang, s pht ln quang)
BMCNTP HBK TPHCM
Nguyn l hot ng ca mt s
my phn tch quang ph

- Quang ph T ngoi kh kin (UV-Vis): nhng cu t trong mu c
kh nng hp thu chn lc bc x vng t ngoi v kh kin, to mi
hp thu c trng

- Quang ph pht x hunh quang: khi hp thu nng lng t bc x,
phn t pht x mt phn nng lng hp thu

- Quang ph hng ngoi (IR): Phn t cht hu c hp thu nng lng
bc x trong vng hng ngoi, s hp thu ny xy ra khi lng t c
tn s dao ng bng vi tn s dao ng t nhin trong phn t

- Quang ph hp thu nguyn t (AAS): 1 nhit nht nh, mt m
hi c kh nng pht x bc sng no th cng c kh nng hp thu
bc sng .
BMCNTP HBK TPHCM
K THUT QUANG PH
Cc b phn chnh trong my o quang:
1. Ngun bc x
2. B phn to nh sng n sc
3. Mu
4. B phn nhn kt qu
5. My o (my ghi)
BMCNTP HBK TPHCM
S my o quang

BMCNTP HBK TPHCM
Ngun bc x:
Ngun bc x lin tc: Pht ra quang ph lin tc
+ n t ngoi: Hydrogen, deuterium cha kh
H
2
, He, pht ra bc x = 180-375nm
+ n kh kin: t nng dy Wolfram pht ra
bc x vng kh kin = 320-2500nm
+ n hng ngoi: di in th thch hp,
dng in chy qua dy Ni Cr pht ra bc x hng
ngoi > 750nm

BMCNTP HBK TPHCM
CC B PHN CHNH CA
MY O QUANG
Ngun bc x:
Ngun bc x khng lin tc: pht ra bc x c di
sng nht nh
+ n cathode lm: cathode lm bng kim loi tinh
khit (Pb, Cu, Fe), anode bng dy Wolfram, khi
dng in i qua, kim loi cathode b cation kh y
ra to m my nguyn t, pht ra bc x c trng

BMCNTP HBK TPHCM
CC B PHN CHNH CA MY
O QUANG
B phn to bc x n sc:
+ Knh lc
+ Monochromator: thit b quang c b phn tn sc c
kh nng tch sng

BMCNTP HBK TPHCM
CC B PHN CHNH CA
MY O QUANG
B phn cha mu: Cuvet trong sut:
Thu tinh
Nha
Thch anh
. My o v my ghi: Tn hiu nhn c c khuch
i v ghi di dng ng biu din hoc o trn
my o.

BMCNTP HBK TPHCM
CC B PHN CHNH CA
MY O QUANG
S
L
1

J
L
2

F
C
S
1

S
2
L
F
CU TO V HOT NG CA MY QUANG PH
LNG KNH
ng
chun trc
Lng knh
Quang
ph ca
ngun J
BMCNTP HBK TPHCM
C
J
L
L
1

L
2

F
S
QUANG PH VCH PHT X
Quang ph
vch pht x

Na H
2

BMCNTP HBK TPHCM
C
J
L
L
1

L
2

F
S
QUANG PH VCH HP THU
Quang ph
lin tc
Quang ph
vch hp
th
n
hi
Na
Quang ph
vch pht x
n
hi
H2
BMCNTP HBK TPHCM
C
J J
L
L
1

L
2

F
S
QUANG PH LIN TC
Quang ph
lin tc
BMCNTP HBK TPHCM
u im ca phng php quang ph
Phm vi ng dng rng
C nhy cao, c th xc nh c cc cht c hm
lng rt nh (dng vt)
Thao tc tng i n gin, tin hnh nhanh chng
C th tin hnh nh lng 1 cht khi c mt cht
khc bng cch s dng mu i chng v dung dch
chun
BMCNTP HBK TPHCM
Nhc im ca phng php quang ph
Thit b c cu to phc tp, t tin, d hng

H thng quang i hi n nh cao nn khng th
tin hnh phn tch ti hin trng

Cc thit b quang u cn c bo qun iu kin
kh nghim ngt: trnh m, nhit n nh
BMCNTP HBK TPHCM
QUANG PH T NGOI V
KH KIN (UV-Vis)
S dng bc x t min t ngoi xa (185nm) n
min hng ngoi (1100nm)
Cc bc x t ngoi thng c:
= 200 400nm

Vng t ngoi
chn khng xa
T ngoi
trung bnh gn
Vng kh kin

E
100 200
286 145
300 400
95 72
800 nm
36
BMCNTP HBK TPHCM
QUANG PH T NGOI V KH
KIN (UV-Vis)
Thit b phn tch hot ng trn c s xc nh hm
lng cu t theo cng hp thu bc x vng t
ngoi v kh kin:

My quang ph so mu (Colorimeter)

My quang ph hp thu t ngoi kh kin
(UV-vis Spectrophotometer)
BMCNTP HBK TPHCM
QUANG PH T NGOI V
KH KIN (UV-Vis)
Phng php c dng phn tch mu dng:

+ Nguyn t kh

+ Ion hay phn t trong dung dch
BMCNTP HBK TPHCM
QUANG PH T NGOI V KH KIN
(UV-Vis)
BMCNTP HBK TPHCM
I
0
I
R
I
T
I
A
QUANG PH T NGOI V
KH KIN (UV-Vis)
Cng hp thu:
+ truyn sut :

T=I
T
/I
0
hay T(%) = I
T
/I
0
.100

+ hp thu:
A = lg (1/T) = lg (I
0
/I
T
)
BMCNTP HBK TPHCM
hp thu ca dung dch t l thun vi chiu dy l
ca lp dung dch v t l thun vi nng C ca
dung dch
A = k.l.C
Khi: l = 1cm
C = 1 mol/L th A = l h s hp thu phn t
(L.mol
-1
.cm
-1
) cn gi l h s tt phn t. Ti mi
bc sng, mt cht c h s hp thu phn t khc
nhau
c xc nh bng thc nghim
nh lut Lambert Beer
BMCNTP HBK TPHCM
Tnh cht ca nh lut Lambert Beer
BMCNTP HBK TPHCM
A = e lC = el
1
C +el
2
C + + el
n
C = eC
n
i=1
l
i

A = e lC = elC
1
+elC
2
+ + elC
n
= el
n
i=1
C
i
A = e lC = e
1
lC
1
+e
2
lC
2
+ + e
n
lC
n
= l.
n
i=1
e
i Ci
Thay i chiu dy cuvet
tng hoc gim hp ph
quang
Thay i nng cht mu
(thm cht mu) hoc pha
long dung dch
Gii bi ton phn tch
nhiu cht mu trong dung
dch
Tnh cng tnh:
QUANG PH T NGOI V KH KIN (UV-Vis)
Quang ph so mu
Quy lut nhn bit mu sc: Mu sc t n khng phi l
tnh cht ca nh sng, m nhn thc v mu sc xy ra
qua phn ng kt hp ca h cm gic dy thn kinh no
mt ngi.
nh sng trng l tng hp cc bc x vng thy c
(t 400-780nm)
Dung dch hp thu cc bc x vng t ngoi, nh sng
trng truyn sut qua nn dung dch khng c mu
Mu ca vt rn l mu do s phn x ca nh sng trng
sau khi mt i mt s bc x b vt rn hp thu.
Mu ca dung dch l mu ca s truyn sut n mt ca
nh sng trng sau khi 1 s bc x b dung dch hp ph

BMCNTP HBK TPHCM
My UV Vis Spetronic Genesys 10

Vng o c: T ngoi kh kin
Khong bc sng: 190 - 1100nm
Gii hn tin cy ca bc sng: 0,2nm
Khong gi tr hp thu: 0,3-3A
Gii hn tin cy ca hp thu:
0,005A gi tr 1A
Nhit lm vic: 5-40
o
C
My thuc loi 1 chm tia nhng nhiu
cuvet
BMCNTP HBK TPHCM
My UV-Vis Spetronic Genesys 10

Trnh t tin hnh phn tch:
1. Tm bc sng: dng cht chun, chn bc
sng c hp thu cc i (scan bng chng
trnh AURORA SCAN)
2. S dng bc x n sc (bc sng xc nh)
3. Chn cuvet ph hp (cuvet thch anh, thu tinh
hoc nha)
4. Loi nh hng ca dung mi, thuc th bng
mu trng blank (c x l cng ch vi
mu nhng khng cha mu)
BMCNTP HBK TPHCM
ng dng phng php quang ph
UV-Vis
nh lng cc cht di dng ha tan trong dung dch
hay 1 cu t mang mu
Xc nh cu trc phn t ca 1 cht do c cc nhm
chc hp thu ti nhng bc sng xc nh


ng dng phng php quang ph UV-Vis

o hp thu ti 1 bc sng xc nh:
+ nh tnh:
. Pht hin cu t bng cch so snh hp thu
ca mu ti
max
ca cht chun
. Xc nh h s hp thu phn t : ti mi
bcsng 1 cht c h s hp thu phn t khc nhau



BMCNTP HBK TPHCM
ng dng phng php quang ph UV-Vis
+ nh lng 1 cu t:
So snh hp thu ca dung dch mu vi dy dung
dch chun
Tnh nng da vo h s hp thu phn t: o
hp thu ca dung mi, sau o hp thu ca dung
dch mu, tnh nng da vo h s hp thu phn t
Tnh nng theo cht chun: o hp thu ca
dung dch chun v mu cng 1 iu kin
BMCNTP HBK TPHCM
V d
o hp thu ca dung mi, sau o hp thu ca
dung dch mu, tnh nng da vo h s hp thu phn
t
Uraxin c h s hp thu phn t = 8,2.10
3
M
-1
cm
-1
ti
bc sng cc i 260nm. Tnh nng dd uraxin khi o
hp thu dd trong cuvet 1cm l 0,65, hp thu ca
dung mi l 0,07
A= .l.C = (0,65-0,07) =0,58
C= 0,58/8,2.10
3
= 7,1.10
-3
M
ng dng phng php quang
ph UV-Vis
Phng php ni chun: (Thm cht chun vo mu)
+ o hp thu ca mu
A
mu
= C
m
.l.
+ Thm 1 lng cht chun xc nh vo dd mu.
hp thu ca dd l:
A
mu+chun
= (C
m
+C
c
).l.
Khi :
m m c
m c
m
A A
A C
C

ng dng phng php quang ph

UV-Vis
Phng php ngoi chun: Lp ng chun

ng dng phng php quang
ph UV-Vis
nh lng 2 cu
t: trng hp 1
ng dng phng php quang ph
UV-Vis
nh lng 2 cu
t: trng hp 2

ng dng phng php quang ph UV-Vis
+ nh lng 2 cu t trong
trng hp 2: Ti mi bc
sng mi cht c h s hp
thu phn t khc nhau:
Cu t X, nng C
x
:
1

x1
,

2

x2
Cu t Y, nng C
y
:
1

y1
,

2

y2
Tng ng, c 2 cc i ti 2
bc sng
A
1
=
x1
lC
x
+
y1
lC
y

A
2
=
x2
lC
x
+
y2
lC
y


2 1 2 1
2 1 1 2
y x x y
y y
x
A A
C

BMCNTP HBK TPHCM

) (
2 1 2 1
2 1 1 2
y x x y
y y
x
l
A A
C

) (
2 1 2 1
1 2 2 1
y x x y
y x
y
l
A A
C

ng dng phng php quang ph UV-Vis

+ o c ca dung dch: mt long, hp thu
t l vi mt huyn ph (t l vi c)
nh lng cc cht kt ta mn v c hay hu c
nh lng sinh khi vi sinh vt trong qu trnh ln
men
BMCNTP HBK TPHCM

ng dng phng php quang ph UV-Vis
+ Theo di ng hc qu trnh phn ng bng phng
php chun quang:
o hp thu ca dung dch theo thi gian phn ng,
tnh c s bin thin hm lng 1 cht tham gia
phn ng (hoc sn phm to thnh)
BMCNTP HBK TPHCM
ng dng phng php quang ph UV-Vis
o hp thu A = f(): Xc nh cu trc phn t ca
mt cht do c cc nhm chc hp thu ti nhng
bc sng xc nh
BMCNTP HBK TPHCM

ng dng phng php quang ph UV-Vis
trong cng ngh thc phm
So mu: xc nh lng ng kh, mu ca
nguyn liu cng nh bia thnh phm, thnh
phn m amin, hm lng polyphenol, hm
lng cht ng v diacetyl, d lng thuc bo
v thc vt, mu thc phm
o c: nh lng protein, nh lng sinh
khi vi sinh vt trong qu trnh ln men
Scan: xc nh thnh phn cha bit bng ph
UV-Vis, xc nh d lng cht mu bng HP-LC
kt hp UV-Vis
BMCNTP HBK TPHCM
ng dng phng php quang ph UV-Vis
BMCNTP HBK TPHCM
My UV- Vis Shimadzu

BMCNTP HBK TPHCM