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BO CO V MY UV Vis 1800 I.

. Khi qut v phng php ph t ngoi - kh bin Khi phn t hp th bc x t ngoi hoc kh kin th nhng electron ho tr ca n b kch thch v chuyn t trng thi c bn ln trng thi kch thch. V th ph hp th c gi l ph t ngoi - kh kin (Ultraviolet and Visible Spectra, vit tc l UV-Vis) v cng oc gi l ph hp th electron. 1.1. Cc kiu chuyn mc electron Khi chng ta ni v chuyn mc electron th cn bit n mi quan h gia cc mc nng lng.

Hnh : Biu th hin mi lin quan gia cc mc nng lng S chuyn t trng thi c bn ln trang thi kch thch t phng din nng lng. l s chuyn mc nng lng thp ln mt mc nng lng cao hn. Trong phn t cc electron cc obitan khc nhau (, , khng lin kt, phn lin kt) ng vi mc nng lng khc nhau. Tu thuc vo mi phn t c th, c th c cc obitan 1, 2,, 1, 2,, na, nb, vi cc mc nng lng khc nhau. Mi trng thi electron u c m t bng nhng ng cong th nng ring. Nh vy c th xy ra nhiu kiu chuyn mc electron t trng thi c bn ln trng thi kch thch cao hn.

Hnh : S phn b cc mc nng lng ca cc obitan phn t Cc kiu chuyn mc hay gp:
Chuyn mc N V: l s chuyn electron t trng thi lin kt ln trng

thi phn lin kt c nng lng cao hn. Chuyn mc N V i vi electron gi l chyn mc * v n ng vi gi tr E ln nht nn thng th hin vng t ngoi xa, i vi electron gi l chuyn mc * v n ng vi gi tr E nh nht nn thng th hin vng t ngoi gn hoc vng kh kin khi c nhiu electron lin kt vi nhau.

Hnh: S chuyn mc *

Chuyn mc N Q: l s chuyn electron t trng thi khng lin kt ln

trng thi phn lin kt c nng lng cao hn. l gp cc phn t c cha cc nguyn t vi cc cp electron cha tham gia lin kt (nh nguyn t halogen: oxygen, nitrogen,). C hai loi chuyn mc N Q: chuyn mc n * v

chuyn mc n *. C hai chuyn mc u c trng bi cng thp (gi tr nh). Chuyn mc n * thng th hin vng t ngoi, chuyn mc n * thng th hin vng t ngoi gn hoc vng kh kin.

Hnh: Chuyn mc nng lng n *

Chuyn mc N R: l s chuyn electron t trng thi c bn ln trng

thi nng lng rt cao theo hng ion ho phn t. Chuyn mc ny i hi nng lng rt ln nn th hin hin vng t ngoi xa. Ph thu c trong trng hp ny thng dng xc nh nng lng ion ho phn t. Chuyn mc km theo chuyn dch in tch: l nhng chuyn mc m trong electon chuyn t mt nguyn t hoc mt nhm nguyn t ny n mt nguyn t hoc mt nhm nguyn t khc. S hp th km theo chuyn dch in tch thng gp cc hp cht v c v phc cht. 1.2. Phn bit cc kiu chuyn mc electron
Chuyn mc n * c nhng c im sau: 1. Chuyn mc n * c h s hp th mole nh, thng t khi vt qu

103.
2.

Bc sng cc i (max) ca vn n * chuyn dch v pha bc sng

ngn (chuyn dch xanh) khi chuyn t dung mi khng phn cc sang cc dung mi phn cc mnh hoc c kh nng to lin kt hydrogen. Thng th ng dng dung mi ny vo khong 5 20 nm. Dung mi phn cc c mt kh nng lm gim max (chuyn dch xanh) i vi chuyn mc n * l do h thp nng lng trng thi c bn v lm tng nng lng trng thi kch thch. Cc dung mi to lin kt hydrogen vi cht tan thng gy ra s chuyn dch mnh vn hp th n * v pha sng ngn do dung mi to lin kt hydrogen vi chnh i electron n nhn trch nhim hp th bc x trong chuyn mc ang xt. Mt khc trng thi kch thch ch cn mt electron n nn lin kt hydrogen b yu i, n khng th lm gim nng lng ca trng thi kch thch nh lm i vi trng thi c bn. Trng hp phn t c

nhiu hn mt electron n th s to lin kt hydrogen i vi i electron ny s gy hiu ng cm ng i vi i electron kia v do cng dn ti chuyn dch xanh.
3. Vn n * thng b trit tiu trong mi trng acid mnh. l s

proton ho hoc s to thnh sn phm cng lin kt mt i electron n.


4. Vic gn cc nhm y electron vo nhm mang mu cha electron n

cng thng lm cho vn n * chuyn dch v pha sng ngn. Chuyn mc * thng c cng ln, gi tr thng t 103 n 105. Ngc vi chuyn mc n *, khi chuyn sang dung mi phn cc mnh, hoc khi a cc nhm y electron vo phn t th vn ph ng vi chuyn mc * s dch chuyn v pha sng di (chuyn dch ). Khi phn bit chuy mc * vi n * khng nn ch da vo gi tr m phi kt hp vi hiu ng dung mi v nh hng ca nhm th. Chuyn mc d - d v chuyn mc km theo chuyn dch in tch thng xut hin vng t ngoi gn hoc vng kh kin. Chuyn mc d - d c trng bi h s hp th mole rt nh (thng khng qu 102) cn chuyn mc km chuyn in tch li c ln (c 104). Ngi ta nhn thy rng v tr ca vn hp th ng vi chuyn mc km chuyn in tch thay i theo kh nng solvate ho ca in dung : Trong nhng dung mi solvate ho tt cc i hp th chuyn dch v pha sng ngn. 1.3. Quy tc chn lc ph electron Cng hp th ng vi mi chuyn mc c biu din bi cng thc sau : = 0,87.1020.P.a Trong : P l xc sut chuyn (nhn gi tr t 0 n 1), a l din tch hng bc x ca h hp th. H hp th y c hiu l phn phn t nhn trch nhim hp th bc x tc l nhm mang mu (chromophore). i vi ph dao ng cc quy tc chn lc l tng i n gin. i vi cc chuyn mc electron, cc quy tc chn lc l phc tp hn nhiu v chng l hm ca tnh i xng v bi ca c trng thi c bn v trng thi kch thch. Di y l tm tt ni dung:
Tt c cc hm sng trong phn t u c phn thnh chn (k hiu l g)

hoc l (k hiu l u). i vi cc phn t i xng, cc chuyn mc g u hoc u

g c php, cn cc chuyn mc g g v u u l b cm. Quy tc ny gi l quy tc chn lc theo chn l.


Chuyn mc gia cc trng thi c bi khc nhau l b cm. Chng hn

chuyn mc singlet triplet l b cm do bi . Chuyn mc b cm do bi c khng vt qu 1. Chuyn mc cc phn t khng c tm i xng th ph thuc vo tnh i xng ca trng thi u v trng thi cui. Cc vn ph vi nh hn 103 l kt qu ca cc chuyn mc b cm theo cc m hnh n gin nhng li xy ra c do phn t thc l khc vi m hnh n gin m ta xy dng cho n. 1.4. Bc x v s hp th bc x 1.4.1. Bc x in t Bc x in t c c trng bi bc sng (qung ng m n i c sau mi dao ng y ), hoc tn s v (s dao ng trong mt giy). Chng lin h vi nhau bi biu thc sau:

Vi:

C l tc nh sng (trong chn khng c = 2,99.1010 cm/s)

Nng lng (E) ca lng t vi tn s v c tnh nh sau:

Hay:

Vi: h l hng s Plank (M .Planck), h =6,626.10-34 J.s. Biu thc cho thy nng lng ca bc x t l thun vi tn s v s sng, t l nghch vi bc sng. Ton b dy sng c chia thnh cc vng ph khc nhau (hnh 1). Mt ngi ch cm nhn mt phn ph in t rt hp, l vng kh kin (vng nhn thy c) bao gm cc bc x c bc sng 396 760 nm.

1.4.2. S hp thu bc x v mu sc ca cc cht nh sng nhn thy bao gm dy bc x c bc sng t 760 396 nm vn c gi l nh sng trng. Khi cho nh sng trng chiu qua lng knh, n s b phn tch bng mt s tia mu (, da cam, vng, lc, lam, chm, tm). Mi tia mu ng vi mt bc sng hp (bng ?????). Nu nh sng chiu vo mt cht no m b khuych tn hon ton hoc i qua hon ton th i vi mt cht c mu trng hoc khng mu. Thu tinh thng hp th cc bc x vi bc sng nh hn 360 nm, n trong sut vi cc bc x kh kin. Thu tinh thch anh hp th bc x vi bc sng nh hn 160 nm, n trong sut i vi bc x kh kin v c bc x t ngoi gn. Nu mt cht hp th hon ton tt c cc tia ca nh sng trng th cht c mu en. Nu s hp th xy ra mt khong no ca vng kh kin th cc bc x khong cn li khi n mt ta s gy cho ta cm gic v mt mu no . Hai tia ph nhau khi trn vo nhau s to ra nh sng trng. Quan h gia mu ca tia b hp th v mu ca cht hp th (cc cht mu ph ca nhau). Bng ????: Quan h gia tia b hp th v mu ca cht hp th

Tia hp th Mu Tm Xanh Lc xanh Lc Lc vng vng Da cam , nm (wavelength) 400 430 430 490 490 510 510 530 530 560 560 590 590 610 610 - 730

Mu ca cht hp th (mu ca tia cn li) Vng lc Vng da cam ta Tm Xanh Xanh lc Lc

mt hp cht c mu, khng nht thit max ca n phi nm vng kh kin m ch cn cng hp th ca n vng kh kin ln.

II. Gii thiu chung my UV Vis My UV-Vis l mt thit b c s dng ph bin trong phng phn tch. Cc my ph hin thng c ni vi my vi tnh, do vic ghi ph ht sc thun li nh c nhng chng trnh o t ng theo cc ch khc nhau. Ngoi ra, cn c th lu gi ph i chiu v so snh khi cn thit. Nh s dng my vi tnh, b t ghi cn th ghi ra nhng s liu cn thit nh gi tr bc sng (), hp ph ( Abs) v ta cng c th xc nh h s hp th mol () nh vo nh lut Bouguer Lamber Beer:
A=
lg P0 =.l.c P

Trong : A l hp th. c l nng cht tan(mol/L)

l l b dy ca cell cha mu (cm). l h s hp th mol (Lmol-1cm-1). ngha ca h s hp th mol : H s hp th mol c trng cho cng hp th ca cht nghin cu bc sng cho. H s hp th Mol khng ph thuc vo nng v b dy ca lp cht hp th . ch ph thuc vo bn cht cht hp th v bc sng ca bc x hp th Do c trng cho cng hp th bc x ca cht c kho st.Khi ln ta ni cht hp th mnh(cng hp th ln), ngc li khi nh- cht hp th yu(cng hp th nh). vng t ngoi - kh kin nh lut Bouguer-Lamber-Beer lun tun theo v vy gi tr thng lun c xc nh v c lp li tt. Gi tr ca cc vn hp th ng vi cc chuyn mc electron khc nhau thay i trong nhng khong rt rng. iu ny rt quan trng khi nghin cu cc hp cht thin nhin vi lng thu c nh. Dung mi dng o UV-Vis khng c hp th vng ph cn o.Ngi ta thng dng cc loi dung mi nh: methanol, ethanol, ncNgoi ra ngi ta cn s dng cc loi dung mi khng mu nh chloroform, dioxane, benzen Trong khi o UV-Vis th dung mi ng vai tr quan trng nn dung mi phi c tinh ch mt cch cn thn. Nu dung mi c ln tp cht ch mt lng nh th cng lm sai lch kt qu o. o c chnh xc trong mt s trng hp ta phi chy Baseline li nh: - o cc mu vi cc dung mi khc nhau. V d nh mu tan trong dung mi l methanol nhng mu khc li dng dung mi l ethanol. - mt thi gian lu ta khng s dng th ta phi chy Baseline li. Khi ta tin hnh o UV-Vis th ta s thu c hp th mol v cng hp th cua cht.

Khi bc x chiu vo cc phn t, n c th b khuch tn hoc b hp th bi cc phn t. S khuch tn khng lm thay i tn s ca bc x l s khuch tn thng, cn khuch tn lm thay i tn s ca bc x c gi l khuch tn t hp. Chng ta quan tm n s hp th bc x bi v vic ghi ph chnh l ghi li s hp th bc x bi phn t. nhit khng tuyt i, lp v electron ca phn t khng b kch thch, s quay ca phn t cng khng xy ra, nhng phn t c mt nng lng dao ng no gi l nng lng dao ng im khng. Khi nng lng tng dn d tr nng lng nhit ca phn t n gi tr 0,03- 0,3 kcal/ mol, phn t chuyn sang nhng trng thi quay b kch thch nhng trng thi dao ng v trng thi electron vn khng i. Khi nng lng chuyn ng nhit tng ln ti 0,3- 12 kcal/mol, trng thi kch thch ca phn t cng cha b kch thch nhng trng dao ng b kch thch. Mun kch thch electron cn phi c nng lng ln hn nhiu, vo khong hng chc n hng trm kcal/mol. Nng lng ng vi cc bc x thuc vng kh kin hay t ngoi. Nu phn t hp th cc bc x c nng lng ln hn nh bc x kh kin hoc t ngoi th nng lng electron ca chng b thay i. Nu ch c trng thi electron thay i th vch hp th tng ng s c tn s: Vel = E/hc. S thay i trng thi electron ng thi c c s thay i trng thi dao ng v trng quay nn ta khng thu c cc vch vi tn s v el = vdd + vqy. Ph thu c trong trng hp ny c gi l ph hp th electron. Hay gn hn l ph electron. V ph electron th hin vng t ngoi kh kin (UV-Vis) nn cng c gi l ph t ngoi kh kin. phm l: Khi nim m mu, nht mu ca dung dch trong thc t lin quan ti gi tr ca cht hp th v c nng ca n. T ph UV-Vis v h s hp th mol, ta c th suy ra tnh cht ca sn

- Hiu ng m mu ca dung dch c o bng hp th. S chuyn dch cc i hp th v bc sng di (hiu ng Batocrom) th tng. - Hiu ng nht mu l hiu ng lm gim cng hp th l gim . Da vo ph ta bit c bc sng, hp th, h s hp th mol t ta c th so snh vi mu chun xc nh sn phm c t c nhng yu cu hay khng (v d nh l v nhm chc, v tinh khit ca sn phm) Da vo hp th ca mu, ta c th xc nh nhm chc c trng ca sn phm. V d nh nhng hp cht hp th trong vng 220-800 nm thng l cc hydrocacbon hoc l dn xut n gin ca chng nh(CH3Cl, C2H5Cl). Hoc nu nh l hydrocacbon loi anken, ankin th ni i, ni ba trong phn t v tr c lp. 1. Cu to chung ca my UV/VIS spectrophotometer Nhng b phn ch yu ca my ph UV Vis l: ngun pht bc x, b to n sc phn chia chm sng, b phn o so snh cng nh sng ri chuyn thnh tn hiu (detector).

Hnh : S cu to chung ca my UV Vis

Hnh : Cu to ca my UV Vis Chc nng ca tng b phn nh sau:


Curve: vt liu chnh dng lm curve o vng t ngoi - kh kin l

thch anh hay thu tinh thch anh, silica nung chy v thu tinh v cc vt liu tr bc sng t 800 400 nm.
Ngun sng: pht c bc x t ngoi ngi ta dng n Deuterium

Arc (teri) vi phm vi bc sng 190 420 nm, cn vi n Tungsten (vonfram) vi phm vi bc sng 350 2,500 nm c th pht c c bc x t ngoi v kh kin hoc cng c th s dng n Xenon (n kh tr) c phm vi bc sng l 190 800 nm.

Hnh : n Tungsten
B to n sc: thng dng lng knh thch anh hoc cch t c nhim v

tch ring tng dy sng hp (n sc). Ngun sng nhiu mu sc (polychromatic) s qua khe vo (Entrance slit) v n thit b tn sc, di tc dng ca thit b tn sc s to ra nh sng n sc khi qua khe ra (Exit slit) v i ra ngoi.

Khe h vo
nh sng trng

nh sng n sc

Khe h ra

Hnh : B to n sc

Thit b tn sc: c ba loi thit b tn sc. o

Th nht l filter (lc): c tc dng lc vn hp th v lc nh

sng so snh.

a)

b)

Hnh

: a) Cu to ca thit b lc b) ng dng trong bnh lc chuyn ng trn u (Filter Wheels)

Lng knh (prism): l thit b tn sc c bit n nhiu nht c tc

dng lm tn sc chm sng khi i qua n.

Hnh : Chm nh sng i qua lng knh b tn sc


o

Grating (cch t): ging nh lng knh n c tc dng tn sc chm tia i

qua n. C hai loi: cch t nhiu x phng v cch t nhiu x lm.

a)

b)

Hnh : a) Cch t nhiu x lm. b) Cch t nhiu x phng.


Detector: l b phn phn tch c nhim v phn tch cng chm nh

sng i qua dung dch v i qua dung mi. Detector c dng cho vng UV Vis l detector ph hp th quang phn t UV Vis. V nguyn tc cu to ca detector gm nhng phn nh sau:
o

Ngun sng : l n D2 (vng ph UV), hay n W Halid (vng ph

Vis). o o Bung mu v mi trng hp th. B n sc thu chm sng, phn ly v chn tia sng cn o.

o o

B phn in t thu nhn v khuych i tn hiu o. B phn ch th kt qu o.

2. Nguyn tc hot ng ca my UV/VIS spectrophotometer 2.1. Nguyn tc hot ng ca my UV Vis mt chm tia

Hai ngun sng pht ra bc x t ngoi kh kin t n Deutrium v n Tungsten n gng lm di phn x ca gng lm s to ra mt tia sng trng i qua thu knh chun trc (collimating) v qua khe vo (entrance slit) ti gng lm. Di s pht x ca gng lm ngun nh sng trng s i n grating nhm to ra tia sng n sc v pht x li gng lm. Cng di s pht x ca gng lm, tia sng n sc s i n gng phng b phn x v chiu qua khe ra (exit slit) n filter wheel (bnh lc chuyn ng trn u). T filter wheel tia sng i n mt gng phng v tip tc xuyn qua mu. Pha sau mu c mt mng trp (shutter) c th hay khng th cho ngun sng i qua to ra s ngt qung. Kt qu s c detector do tm v sau s c c kt qu trn my tnh.

Hnh : Nguyn tc hot ng ca my UV Vis mt chm tia

2.2.

Nguyn tc hot ng ca my hai chm tia

Hai ngun sng pht ra bc x t ngoi kh kin t n Deutrium v n Tungsten n gng lm di phn x ca gng lm s to ra mt tia sng trng i qua thu knh chun trc (collimating) v qua entrance slit ti gng lm. Di s

pht x ca gng lm ngun nh sng trng s i n grating nhm to ra tia sng n sc v pht x li gng lm. Cng di s pht x ca gng lm, tia sng n sc s i n gng phng b phn x v chiu qua exit slit n filter wheel (bnh lc chuyn ng trn u). T filter wheel tia sng i n mt gng phng. Gng phng pht ra tia phn x i n mt b phn chia chm sng s hng chm sng n sc lun phin i ti curve ng dung dch mu v dung dch dung mi. B phn phn tch detector s so snh cng chm sng i qua dung dch v i qua dung mi v s pht tn hiu cho ra kt qu o.

Hnh : nguyn tc hot ng ca my UV Vis hai chm tia


3. ng dng ca my UV/VIS Spectrophotometer Phn tch cc ho cht v c hay hu c. Phn tch s an ton ca thc phm. Phn tch mu huyt. Phn tch DNA/RNA. Phn tch hm lng nng dc d. Phn tch lng chlorine.

III. Gii thiu v my UV/Vis 1800 My UV/Vis 1800 l mt trong nhng thit b c s dng ph bin trong vic phn tch phn t trong vng t ngoi v kh kin. Vic nghin cu cc c tnh ca cc phn t ha hc nh cc ph hp th vng t ngoi v kh kin c s dng rng ri trong tt c cc lnh vc ca vic nghin cu khoa hc v thc t sn xut.

Hnh : My UV Vis 1800

1. Cu to ca my UV/Vis 1800 Cu to v chc nng ca my UV/Vis 1800 cng ging nh cu to v chc nng ca my UV Vis Spectrophotometer gm c cc b phn: ngun pht bc x (1), b to n sc (2), b phn chia chm sng (3), dung dch cht nghin cu (4), dung mi (5), detector (6) l b phn o v so snh cng nh sng ri chuyn thnh tn hiu in, v b phn ghi ph (7). y vt liu c s dng lm curve l thu tinh thch anh.

Hnh : S cu to ca my UV Vis 1800

2. Nguyn tc hot ng ca my UV/Vis 1800 Nguyn tc hot ng ca my UV/Vis 1800 cng da trn nguyn tc hot ng ca my UV Vis Spectrophotometer hai chm tia.

Ngun sng bc x t ngoi kh kin c pht ra t n Deutrium v n Tungsten n b to n sc (dng grating) v ti y tia sng s c tch ra tng dy sng hp (n sc) v c truyn n b phn chia chm sng s hng chm tia n sc lun phin i ti curve ng dung dch mu v ng dung mi. Sau khi i qua curve ng dung mi v curve ng mu th tia sng s i n b phn phn tch (detector), ti detector s so snh cng chm sng i qua dung dch (I) v i qua dung mi (Io). Tn hiu quang c chuyn thnh tn hiu in. Sau khi c phng i tn hiu s chuyn sang b t ghi v ng cong s ph thuc ca lg Io/I.

Hnh 53: S nguyn l hot ng ca my UV/Vis 1800

3. ng dng ca my UV/Vis 1800 My c bc x trong vng t ngoi v kh kin nn c xem l mt php phn tch nh tnh cng nh php phn tch nh lng, c bit phng php ny cng gp phn xc nh cu trc phn t hp cht hu c, v c v phc cht. My UV/Vis 1800 c nhng ng dng ging nh my UV/VIS Spectrophotometer. Nhng cng ty thng c s dng o cc mu lng theo di phn ng hu c, phn tch nh tnh v nh lng.

4. Hng dn s dng phn mm UV Probe Mt s lu : y l thit b quang ph nn cn: trnh cho my khng b bi ( s dng vi che bi khi khng s dng my), khng t my ni c rung hay m cao, in p cp cho my phi n nh. Bt in v cho my n nh t nht 15 pht trc khi bc vo php phn tch.

Thot khi phn mm UV Probe trc khi tt my UV. Sau khi tt my phi ch ti thiu 10 pht mi m my tr li. Bc 1: Vo chng trnh. m bo my UV c bt in. Bt my tnh. Nhp p chut vo biu tng UV Probe trn mn hnh vo chng trnh. Sau nhp User name v password nu my c hi mt khu. Bc 2: Chn ch o

( bng cch kch chut vo biu tng tng ng trn thanh cng c) C 3 ch o: Spectrum, Photometric, Kineties Bc 3: Kt ni vi my UV.

o trong ch Spectrum

( o qut nhm xc nh cc cc i hp th ca mt mu cht cha bit) Chn thng s cho php o: Kch chut vo nt lnh M s m ca s sau:

Hnh 4

Kch chut vo mc Intrument Parameters s m tip ca s sau ta chon ch ( Transmittance Truyn qua khi mu dng bn mng, Absorbance Hp th - Thng dng vi hu ht cc mu, Energy Nng lng Thng chn ch ny kim tra nng lng n, Reflectance Phn x - Nu chn ch ny cn lp thm b ph kin o phn x).

Kch chut vo mc Attachments m ca s khai bo cho cc b ph kin lp thm. V d: khi ta lp mt khay t mu loi 6 cell th phi khai bo loi khay,

khai s cell trong mc Number of cell, sau kch chut vo Initialize my xc nhn s c mt ca n.

Tin hnh php o:

m bo trong bun mu khng t g. Kch chut vo nt Go To WL d chn bc sng m mu phn tch b hp th cc i. t hai cuvet ng mu trng vo khay kch chut vo nt Baseline v ch my qut chun li ng nn. sau o mu, t cuvet ng mu cn o( Sample) vo khay pha ngoi. Sau kch chut vo nt Start my bt u qut. X l kt qu: ( Ph c th v qut xong hoc m mt ph c sn) V d sau khi qut xong ta thu c ph sau:

Hnh 7

Ch : Ty thuc vo mc ch m ta s dng mt hay nhiu chc nng trn.

Ch : Khi nhp chut phi ln ca s cha ph, kt qu hay thng s u hin ra mt danh sach lnh cho php ta tc ng lnh lm thay i c tnh, cho php n hay hin cc phn nm trong ca s . Nguyn tc ny p dng cho tt cc ch o. V d: Nhp chut phi ln ca s cha ph s hin ra cc danh mc sau: Copy: lnh ny cho php ta copy ph sang mt chng trnh ng dng khc. Auto Scale: t ng i gii hn cc ta ph hin ra d nhn.

Cross Hair: chuyn con tr thnh dng si tc hnh du cng, v khi di chuyn con tr cc ta s hin ra hai du si tc gip ta c c hp th ti mi bc sng. Customize: s m ra ca s cho php ta thay i gii hn ta , mu ng ph ... Ch khc: khi mt s menu, ca s thanh cng c cha cc nt lnh b bin mt ta cn vo menu View v nh du hin li chng. Nn nh du tt c cc mc trong menu View.

Ca s di y cho php ta t chn peak trn ph

Thao tc trn ca s ny tng t nh thao tc trn ca s Peak Pick. tnh din tch mt peak no ta c th tin hnh mt trong hai cch sau:
1.

Di chuyn n tng si tc thng ng trn ph ri bm gi chut v

r si tc bn tri n mp u, si tc bn phi n mp cui vng peak. Kt qu s t ng xut hin bn bng kt qu.


2.

Ln lc nhp bc sng u v cui di cn tnh din tch vo ct

Start v ct End, v sau nhp chut vo bn ct Erea hoc Result ta c kt qu. C th nhp chut vo mc Color chn mu cho tng vng peak.

o trong ch Photometric

( Ch nh lng ti mt hay nhiu bc sng, dng ng cong chun) Chn thng s cho php o: Kch chut vo nt lnh M s m ca s sau:

Cc mc quan trng cn kch chut m ca s chn thng s l: Wavelengths, Measurement Parameters, Calibration, Intrument Parameters. Mc Wavelength: nhp bc sng ta cn o ng vi mu cn phn tch.

Sau kch chut vo nt Add cng n vo danh sch bn di.

Nu trong mc Wavelength Type ta chn l Range th ca s s hin ra nh H13. Ta cn nhp bc sng u v cui di vo mc WL Start v WL End ri kch chut vo nt lnh Add cng vo danh sch. Ch : Ta c th cng vo danh sch hn hp bc sng dng im ( Point ) v di (Range). Kch chut vo mc Measurement Parameters s hin ra ca s di y cho php ta nhp cc thng s v vic o lp li ( s ln lp li Sample Repetitions, khong thi gian ngh gia hai ln lp li lin tip ). Thng thng th ta b qua ca s ny.

H 14

Kch chut tip vo mc Calibration chn cc thng s lin quan n ng chun ( s im chun, o chun bc sng no, n v, loi ng chun bc my).

Tip kch chut vo vo mc Intrument Parameters chn ch o ( hp th, truyn qua) v mc Attachments ging nh ch o qut. Sau khi chn cc thng s cho php o xong ta tin hnh a mu vo o. V d v php o nh lng ng chun 3 im c chn i qua im c chn di qua im zero nh sau: Tin hnh php o: 1. Kim tra xem my UV c bt in v sn sng o cha.
2. Trong bn kt qu Standard Table ta ln lc nhp tn ca cc mu chun

vo mc Sample ID v nng ca chng vo mc Conc (mi mu chun ng vi mt hng). 3. Trong bng Standard Table ta ch cn nhp tn ca cc mu cha bit vo mc Sample ID. 4. Kch chut vo nt lnh Go to WL bn di mng hnh v nhp bc sng cn o ( y chnh l bc sng m ta nhp trong phn t thng s cho php o).
5. t hai cuvet cng ng mu trng vo khay ri kch chut ln nt lnh

Auto Zero bn di.


6. Nhc cuvet bn pha chm tia qua mu ra, t cuvet ng mu chun th

nht vo ri kch chut vo nt lnh Start. My s c v hin ngay hp th ln bng Standard Table, ng thi xut hin mt im trn th ng cong chun.

7. Tip tc nh vy i vi cc mu chun cn li ta s thu c ng cong

chun. Nu ng cong chun thu c c mt s im khng t yu cu ta cn loi chng ra khi ng cong chun bng cch sau: Pha li mu sau kch chut nh du vo mc Ex trong bng kt qu ca chun vo mt hng mi trong Standard Table, t mu vo ri kch chut ln nt lnh Start o li. 8. Kch chut vo v tr bt k trong bng Sample Table lm ni hng nt lnh bn di mn hnh. t ln lt cc mu cha bit vo v kch nt lnh Start o. Chng trnh s da vo ng cong chun v a ra nng trong bng Sample Table. Ch : Sau khi o khong 3 mu ta nn t 2 cuvet ng mu trng vo v tin hnh Auto Zero. X l kt qu T ng chun to c ta c th lu li, sa i ( nh mc 7 bn trn) v m ra s dng sau ny. Tuy nhin, khng nn s dng li ng chun v iu kin o ( sai s do pha mu, iu kin phng) gia cc ngy o rt khc nhau, kt qu s khng chnh xc. thm hay bt mt s ct trong bng Standard Table hay Sample Table ta kch chut phi ln chng chn Properties, chn tip Columns s hin ra ca s sau:

o trong ch Kinetics

Ch ny thng dng o cc mu sinh hc nh enzym, php o tin hnh trong khong thi gian di.

t thng s php o Kch chut vo nt lnh M thanh cng c bn trn s m ra ca s sau:

Nu chn thi gian l Auto ta cn nhp tng thi gian o vo mc Total Time (n v c mc nh l giy), nhp khong thi gian o nhy vo mc Activity Region, nhp bc sng trong mc Wavelengths ( c th chn dng n hay dng kp nh hnh H17). Nhng mc khc c th mc nh. Nu chn thi gian o l Manual ca s s nh sau:

Ta chn n v trong mc Units, vng thi gian cho mi ln c kt qu trong mc Cycle Time, s ln c kt qu trong mc Number of Raedings. Chng trnh s

t ng tnh ra tng thi gian o v hin ra trong mc Total Time. Cn cc mc khc tng t nh ch Auto. Kch chut tip ln mc Intrument Parameter chn ch o trong mc Measuring Mode. Cc mc khc nh Sample Preparation, File Options c th b qua. Nhp OK thot ra v tin hnh php o. Tin hnh php o Thao tc tng t nh o ch Spectrum X l kt qu Di y l mt v d:

V cui cng l to bo co v in ra.