BMCNTP HBK TPHCM

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

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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 Bc sng, S sng, Tn s, Nng lng, E Phng trnh = 1/ = c/ E = h = h.c/ n v nm cm-1 Hz hay s-1 J hoc Kcal

BMCNTP HBK TPHCM

NNG LNG CA VT CHT

Nng lng ca vt cht:

E = Et + Ed + Eq Et: nng lng in t Ed: nng lng sinh ra do giao ng tng tc gia cc nguyn t Eq: nng lng do s quay ca phn t xung quanh trng tm ca n

BMCNTP HBK TPHCM

BC X IN T
Phn vng bc x in t:
Bc sng
10Mm

30Hz

Tn s

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Bc x tng tc ln vt cht

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
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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

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

o Bc x Vt cht KQ Ghi tn hiu

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K THUT QUANG PH
Th hin: S hp thu: vt cht s gi li 1 cch chn lc nng lng ca mt s bc x
1.

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 phn t pht x mt phn nng lng hp thu

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

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CC B PHN CHNH CA MY O QUANG

Ngun bc x:
Ngun bc x lin tc: Pht ra quang ph lin tc

+ n t ngoi: Hydrogen, deuterium cha kh H2, 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.

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CU TO V HOT NG CA MY QUANG PH LNG KNH

C J F L S1 L1 S2 L2

F
Quang ph ca ngun J

ng chun trc

Lng knh

BMCNTP HBK TPHCM

QUANG PH VCH PHT X

C
J
Na H2

L1

L2

Quang ph vch pht x

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QUANG PH VCH HP THU

Quang ph vch pht x

C
J S

L1

L2

n hi H2 Na

Quang ph lin tc

Hin Quang ph tng o vch hp sc th

BMCNTP HBK TPHCM

QUANG PH LIN TC
C J
S

L1

L2

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 T ngoi xa trung bnh Vng kh kin gn

100 286

200 300 145 95

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400 72

800 nm 36

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)

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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

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QUANG PH T NGOI V KH KIN (UV-Vis)

IR IA

I0

IT

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QUANG PH T NGOI V KH KIN (UV-Vis)

Cng hp thu:

+ truyn sut :

T=IT/I0 hay T(%) = IT/I0.100

+ hp thu: A = lg (1/T) = lg (I0/IT)

BMCNTP HBK TPHCM

nh lut Lambert Beer

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
BMCNTP HBK TPHCM

Tnh cht ca nh lut Lambert Beer

Tnh cng tnh: Thay i chiu dy cuvet tng hoc gim hp ph quang

A = e lC = el1C +el2C + + elnC = eCni=1 li

A = e lC = elC1 +elC2 + + elCn = elni=1 Ci

Thay i nng cht mu (thm cht mu) hoc pha long dung dch Gii bi ton phn tch nhiu cht mu trong dung dch

A = e lC = e1lC1 +e2lC2 + + enlCn = l.ni=1 ei Ci

BMCNTP HBK TPHCM

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: 5-40oC 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)
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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 = 8,2.103M-1cm-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= 0,58/8,2.103 = 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 Amu = Cm.l. + Thm 1 lng cht chun xc nh vo dd mu. hp thu ca dd l: Amu+chun = (Cm+Cc).l. Khi :

Cc Am Cm Ac m Am

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 Cx: 1 x1, 2 x2 Cu t Y, nng Cy: 1 y1, 2 y2 Tng ng, c 2 cc i ti 2 bc sng A1 = x1 lCx + y1 lCy A 2 = x2 lCx + y2 lCy
BMCNTP HBK TPHCM

A C Cx
x

A1 y 2 y1 x 2 x1 y 2
2 y1

A 2 y1 A1 y 2

l ( y1 x 2 x1 y 2 ) A1 x 2 A 2 y1

Cy

l ( y1 x 2 x1 y 2 )

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
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ng dng phng php quang ph UV-Vis

BMCNTP HBK TPHCM

My UV- Vis Shimadzu

BMCNTP HBK TPHCM