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KY THUT PHN TICH SINH HOA

CU 1. NHNG CHT LAM KT TUA PROTEIN VA THU C KT TUA Phng php ta c s dng tng i rng ri thu nhn cc phn t sinh hc m nht l protein. ta, ngi ta c th dng nhiu cch khc nhau: ta bng mui, ta bng cc dung mi hu c hoc thay i pH ca dung dch c cha protein. 1. Ta bng mui y l cch ph bin ta protein. Kh nng ha tan ca protein ty thuc vo nhiu yu t: c tnh l ha t nhin ca protein, pH, nhit , nng ca mui nng mui thp, tnh tan ca protein tng nh (salting in). Tuy nhin, nng mui cao, tnh tan ca protein gim mnh (salting out). u tin, gi thuyt salting in nng mui thp c gii thch bi Debye - Huckel. Trong dung dch, protein c bao bc xung quanh bi cc ion mui mang in tch tri du. Chnh c tnh ny gia tng hot tnh ca cc dung mi, lm gim cc phn t protein khng mang in tch, do lm tng tnh tan ca protein trong dung mi. Gi thuyt ca Debye Huckel cho rng: tnh tan ca protein c biu din bng mt hm logarit, gi tr ca hm t l vi cn bc hai ca cng ion trong dung dch. Thut ng salting out trong mi trng c nng mui cao c gii thch bi Kirkwood. S gia tng lng ion mui trong dung dch lm gim qu trnh Solvate ha, gim tnh ha tan ca protein dn n s ta. nng mui cao, ha tan tun theo cng thc sau ca Cohn: log S = B - KI Trong S: ha tan ca protein B: hng s (ty thuc vo chc nng ca protein, pH v nhit ) K: hng s salting out (tu thuc vo pH, hn hp v lng mui c trong dung dch) I: cng ion ca mui. th hm log biu din ha tan ca protein bin thin theo nng mui: Hnh 1. tan theo nng mui (http://sosnick.uchicago.edu/precpsalt.html) Nhng ng biu din khc nhau ty theo tng protein khc nhau. V th, khi mun tch 1 protein t mt hn hp gm nhiu protein khc nhau, ta c th la chn nng mui thch hp sao cho ph hp nht vi protein mc tiu.

dc ca ng salting out m t trn ph thuc vo chc nng ca protein v nng mui, khng ph thuc vo nhit v pH. Ngoi ra, nu trng lng phn t ca protein tng th lng mui cn cho phng php ta gim xung. Hiu qu ta protein ca cc anion mui khc nhau th khc nhau, c th xp theo th t gim dn nh sau: citrate > phosphate > sulphate > acetate/chloride > nitrate > thiocyanate. 2. Ta protein bng cc dung mi hu c Khi thm dung mi hu c vo mi trng, hng s in mi ca mi trng gim xung. Cng thc biu din mi quan h gia ha tan v hng s in mi: ln (S/Sw) = (A/RT) (1/Dw - 1/D) Trong S: ha tan ca protein trong dung mi hu c Sw: ha tan ca protein trong nc D: hng s in mi ca mi trng sau khi b sung dung mi hu c vo Dw: hng s in mi ca nc R: hng s kh T: nhit tuyt i A: hng s kh Cng thc trn cho thy: khi thm dung mi hu c vo mi trng, hng s in mi tng ln, kh nng ha tan ca protein gim, v th to s kt ta. Tuy nhin, cc dung mi hu c li c i lc vi cc b mt k nc ca phn t protein. Kt qu l chng lm bin tnh protein trong sut qu trnh ta. Do , khi ta, ch nn s dng cc dung mi hu c nng thp (mc d mt s dung mi nh: 2 methyl 2,4 pentanediol (MPD), dimethyl Sulfoxide (DMSO) v Ethanol c th c s dng nng cao.

3. Ta protein bng phng php im ng in Khi thay i pH ca mi trng, mc ta ca protein cng thay i. pH thp, protein tch in dng v nhm amide b proton ha (thu nhn proton). gi tr pH cao, protein tch in m v cc nhm carbocyl trong phn t protein b mt i proton (mt H+). Ti gi tr pI (Isoelectrics point - im ng in), protein khng tch in. iu ny lm gim tnh tan ca protein v protein khng cn kh nng tng tc vi mi trng, khi , cc phn t protein s tch ra khi mi trng. Hin tng ny c gii thch bng phng trnh Cohn. th biu din ha tan ca 2 protein khc nhau thay i theo pH

Hnh 2. T l protein ta theo pH (http://sosnick.uchicago.edu/precpph.html ) Phng php ny thng dng cho cc protein u nnh (c im ng in khong 4.6) 4. Ta protein bng cc non ionic polymer Phng php ny c thc hin bng cch cho thm non ionic vo trong dung dch protein. Khi thm vo, s lng cc phn t nc c th tng tc c vi protein gim xung. Ngi ta thng s dng dextrans v polyethylen glycols. i = i0 + RT (ln mi + fii mi + fij + mj) Trong i0: in th ha hc chun ca phn t i i: in th ha hc ca phn t i R: hng s kh T: nhit tuyt i mi, mj: nng molar ca phn t i v j fii: h s tng tc ca phn t i fij: h s tng tc gia phn t i v j Khi protein tn ti nng cao, hay pH mi trng vt ra ngoi im ng in mi xy ra qu trnh tng tc gia cc cu t trong protein.

Mum thm loi non ionic polymer no vo trong dung dch protein ty thuc vo trng lng phn t ca protein . Cng tng t nh ta protein bng phng php im ng in. Ti 1 gi tr pH no (sau khi thm non ionic polymer) th kh nng ha tan ca protein cng gim xung. Cc ion non ionic polymer thng s dng l cc dung mi hu c nh Ethanol hay Acetone. Cc dung mi khc t s dng v kh thao tc v c th gy bin tnh protein. Ch : - Nhit thp lm tng hiu qu ta v gim s bin tnh protein. - Nng ion: 0.05 0.2. - i vi cht tan c trng lng phn t cng cao th lng dung mi cn cho qu trnh ta cng t. Theo Scopes (1982), khi bt u ta protein bng Acetone, nn tun theo cng thc sau: [(v/v)%] = 1.8 - 0.12 ln [MW] Trong (v/v)% l th tch dung mi cn cho qu trnh ta tnh theo % MW: trng lng phn t ca cht tan Nu trong hn hp c s hn din ca 2 protein, tnh tan ca protein ny s gim v s hin din ca protein kia. Thun li ca phng php ta bng cch s dng non ionic polymers l protein sn phm bn, v c th s dng nhit phng. 5. Ta bng ion kim loi Trong phng php ta ny, ion kim loi s gn vi mt phn ca phn t protein. Thun li ca phng php ny l c th ta c protein trong mt dung dch rt long. Cc ion kim loi c th chia lm ba nhm. Cc ion nh Mn2+, Fe2+, Co2+, Ni2+, Zn2+ v Cd2+ s gn cht vo cc acid carboxylic v cc hp cht c cha nitrogen. Cc ion nh Ca 2+, Ba2+, Mg2+ v Pb2+ s gn vi nhm chc acid carboxylic. Cc ion nh Ag2+, Hg2+, v Pb2+ s gn cht vi cc nhm c cha lu hunh. MT S PROTOCOL THAM KHO Ta protein bng acetone 1. Lm lnh acetone xung 200C. 2. Chuyn mu protein vo trong cc tube eppendorff thch hp. 3. Thm 4 ln th tch acetone lnh vo tube. 4. Vortex, - 200C trong 60 pht. 5. Ly tm 13.000 15.000 x g trong vng 10 pht. 6. Cn thn loi b dch ni. Trnh chm ti phn cn protein. (Nu mun loi b hon ton cc cht tp nhim, c th lp li cc bc t 2 6) trc khi tin hnh bc 7) 7. cho acetone bay hi t do nhit phng trong vng khong 30 pht. Ta bng Acetone/TCA (Trichloroacetic acid) 1. Chun b dung dch TCA 10% trong aceton. Bo qun - 200C cho n khi s dng.

2. Thm 10 th tch TCA 10% trong acetone vo mu. Vortex. - 200C t nht 3h (tt nht nn qua m). 3. Ly tm 15 000 x g trong 10 pht. Loi b dch ni. 4. Thm 1 th tch acetone lnh ( - 200C). Vortex. - 200C t nht 10 pht. 5. Ly tm 15000 x g trong 5 pht. Loi b dch ni. 6. cn kh t nhin trong khng kh. Ta bng TCA/DOC (Deoxycholate) 1. Chun b dung dch DOC 2%. 2. Trn mu vi DOC 2% theo t l 1/1000. trn 30 pht. 3. Thm TCA 100% vo mu sao cho nng cui cng ca TCA l 15%. Vortex ngay ( trnh cc khi ta ln hnh thnh c th manh theo cc phn t tp nhim). 4. yn t nht 1h. Tt nht l qua m. 5. Ly tm 15 000 x g trong 10 pht. Loi b dch ni c cha TCA v cc cht tp nhim 6. Thm Ethanol lnh hay Acetone lnh vo cn ra. 7. Vortex k cho cn tan hon ton. nhit phng trong 5 pht. 8. Ly tm 15 000 x g trong 10 pht. Cn thn loi b dch ni. 9. Lp li bc ra, ly tm, loi dch ni. 10. Lm kh cn. Ta bng Chloroform/Methanol 1. Thm 4 th tch Methanol vo mu. Vortex k. 2. Thm 1 th tch Chloroform. Vortex. 3. Thm 3 th tch nc. Vortex. 4. Ly tm 15 000 x g trong 2 pht. (Lc ny, protein c th cn trn phn dch ni). Loi i phn nc lp trn. 5. Thm 4 th tch Methanol. Vortex. 6. Ly tm 15 000 x g trong 2 pht. Cn thn loi b hon ton dch ni. Trnh chm n phn cn. 7. Lm kh mu bng bung ht chn khng hay bng khng kh.

CU 2 TRANG THAI VT LY HOA TAN CUA PROTEN Khi ho tan protein thnh dung dch keo th n khng i qua mng bn thm. Hai yu t m bo bn ca dung dch keo:

S tch in cng du ca cc protein. Lp v hidrat bao quanh phn t protein.

C 2 dng kt ta: kt ta thun nghch va khng thun nghch:

Kt ta thun nghch: sau khi chng ta loi b cc yu t gy kt ta th protein vn c th tr li trng thi dung dch keo bn nh ban u.

Kt ta khng thun nghch: l sau khi chng ta loi b cc yu t gy kt ta th protein khng tr v trng thi dung dch keo bn vng nh trc na

CU 3.TRANG THAI KEO LA GI?????????? Sol, cn gi l dung dch keo, l mt h phn tn cc ht rn k dung mi c kch thc t 1 n 1000 nanmt trong mt cht lng, c th c to thnh t mt huyn ph hay bng cch ha c. Cc k thut huyn ph bao gm c vic nghin nt cht rn trong mt my nghin. Ha c hay cc phng php kt ta dng cht kt ta (mui) hay thay i nhit cc ht keo chuyn t trng thi dung dch sang trng thi h keo. Ngi ta dng cc cht to huyn ph tng bn cho sol. a). Khi nim v phn loi h keo Cc cht tn ti di dng cc ht c kch thc khc nhau. Mt s ht nh ht ct c th nhn thy bng mt thng; mt s khc nh cc t bo vi khun khng th nhn thy bng mt thng nhng c th quan st thy nh knh hin vi quang hc; vi cc knh hin vi chuyn dng hin i c th pht hin c c nguyn t. Ngi ta nh ngha Cc ht c kch thc hn phn t v ion nhng khng ln c th quan st c bng knh hin vi quang hc c gi l cc ht keo. Ht keo l mt h phc tp to nn bi mt s lng ln khong t c khi lng khong nguyn t, n 10-7 cm.

Ht keo c th l cht v c hay hu c. Hu nh tt c cc cht u c th tn ti dng keo. Mt h keo lun lun bao gm cc ht keo gi l cht phn tn v mt cht lm mi trng phn tn. Mi trng phn tn quan trng thng gp l nc v khng kh. b). Tnh cht ca h keo Cc h keo c tnh cht tng t nh dung dch. Cc ht keo khng tch ra khi h nh cc ht c kch thc ln khc, v c th xuyn qua c giy lc. Tuy nhin tc khuch tn ca cc ht keo trong h chm hn tc khuch tn ca cc ht trong dung dch nh phn t v ion. S tn x nh sng cng l mt thuc tnh quan trng ca h keo. Khi chiu nh sng i qua mt h keo ta c th quan st ng i ca chm sng t mt bn thng gc vi phng truyn ca chm sng. y l hiu ng Tyndall c dng phn bit h keo vi dung dch. tng nhit si, h nhit ng c, gim p sut hi v p sut thm thu cng ph thuc vo cc ht keo c trong h.

Do kch thc nh nn cc ht keo c t l b mt ln so vi th tch ca ht keo nn cc ht keo c kh nng hot ng b mt ln, chng c kh nng hp th cc phn t v ion c mt trong h. CU 4.TAO KO BIT?????? CU 5.HIN TNG PHN LP????// Ph thuc vo 2 yu + Cht ny khng phi l dung mi ha tan +Khi long ring hai cht lng khng tng ng vi nhau CU 6.TAO KO BIT?????? CU 7.TACH CHIT KEO DNA VA PROTEIN BNG CHT GI LA TT????/ DNA Mt s ho cht thng dng dng trong tch chit axt nucleic Ni t lng: Thng thng ph mng/v t bo chng ta phi dng nit lng, v nit lng c 2 tc dng chnh l: + Lm cho mng b cng gin, d b v khi nghin + V ni t lng gi cho mu trng thi lnh, trng thi m cc enzym ph hu axt nucleic khng hot ng cho ti khi chng b c ch bng ho cht. Mt s qui trnh s dng vt liu ng kh, bi vt liu ng kh d ph v mng hn, mt khc s dng vt liu ng kh khng cn gi mu trng thi lnh, v trong mi trng kh tuyt i cc ezim ph hu axt nucleic cng v tc dng. EDTA: Khi mng b ph v, mt lot cc cht c gii phng ra dung dch, trong s chng c cc loi emzim thu phn axt nucleic thnh nhng mnh vn, bi vy cn phi c ch s hot ng ca enzym ny bng EDTA. Cc ion ho tr hai nh: Mg2+ v Ca2+.v.v. rt cn thit cho s hot ng ca enzym thu phn axt nucleic. Nu trong dung dch c EDTA, cht ny s lin kt vi cc ion ho tr hai ni trn, lin kt ny dn ti s c ch hot ng ca enzym thu phn axt nucleic, do vy m axt nucleic s khng b phn hu. Tris: Axt nucleic cn b nh hng rt ln bi axt v kim ca dung dch s dng trong tch chit, axt c th lm cho axt nucleic b gy, chng hn axt clohydric (HCl) lm cho ADN b t nhng v tr purin, trong khi nu nng kim cao th ADN s b tch thnh si n. loi tr nh hng ca axt v kim ngi ta s dng Tris iu chnh pH ca dung dch. CTAB (Cetryl Ammonium Bromide): L ho cht dng trong chit sut axt nucleic c hiu qu cao v ang c s dng ph bin hin nay. CTAB dung dch l mt dung mi c kh nng ho tan cao axt nucleic, v vy m CTAB c dng vi vai tr l cht chnh trong tch chit axt nucleic. tng hiu qu hot ng ca CTAB, mu c x l vi nhit khong t 55OC n 65OC. nhit nh vy, n cn c tc dng lm rch thm mng t bo v mng nhn gii phng ti a lng axt nucleic ra dung dch v lm bin tnh mt s protein, c bit l enzym thu phn cht t: kia

axt

nucleic. Chloroform: Trong dung dch tch chit, ADN to thnh lin kt vi CTAB, loi b CTAB v ng thi lm bin tnh protein trong dung dch cn s dng chloroform. Sau dung dch tch chit c ly tm, loi b cc tp cht (b ko xung phn pha di ng nghim) cn li axt nucleic ho tan trong lp dung dch nm pha trn v c chuyn sang mt ng nghim mi. Ethanol hoc isopropanol: Ethanol thng c dng trong mi trng dung dch c lc ion, hay nng mui cao, trong iu kin nh vy axt nucleic d dng b kt ta, s kt ta ca axt nucleic s t hiu qu cao hn nu nh c x l trong mi trng lnh. Isopropanol c dng kt ta axt nucleic trong mi trng khng cn c mui, t l lng isopropanol/mu theo th tich l: 1/1. S dng isopropanol c u vit l cc axt nucleic c trng lng phn t thp nh: nhng mnh gy ADN hoc ARN.v.v. s khng b kt ta v chng d dng b loi b cng dung dch. Enzym RNnase v enzym Dnase: Khi kt ta s thu c axt nucleic gm: ADN v ARN: Nu mc tiu l thu ADN, chng cn phi loi b ARN. loi b ARN, ta dng enzym RNase. Enzym RNase s ct ARN thnh nhng mnh vn, do vy sau khi ly tm ADN lng ta v c gi y ng nghim, cn nhng mnh vn ARN vn nm trong dung dch, bi vy khi loi b dung dch th ARN cng b loi b theo. Ngc li nu cn loi b ADN, chng ta dng enzym DNase, cng tng t nh vic loi b ARN, ADN s b ct vn bi DNase v chng cng b loi b cng dung dch Nc ct v TE: Axt nucleic c th c ho tan trong nc ct kh trng hoc dung dch TE. Axt nucleic s c bo qun tt hn, nu n c ho tan trong dung dch TE, v dung dch ny cha Tris v EDTA. PROTEIN

I. Phng php ta bng cc tc nhn gy ta l dung mi hu c (Ethanol 80%, Aceton) ha tan ca protein trong dung dch ph thuc vo nhiu yu t, mt trong s l hng s in mi ca dung dch. Nhn chung, nhng phn t dung mi c hng s in mi ln (nh nc, dimethylsulphoxid) c th n nh tng tc gia chng vi cc phn t protein v to iu kin thun li cho s ha tan ca protein trong dung dch. Ngc li, cc dung mi vi hng s in mi nh (aceton, ethanol) ngn cn s phn tn ca cc phn t protein trong mi trng. Do , ha tan ca nhng phn t protein gim v xy ra kt ta do s lm gim hng s in mi hin hu ca mi trng. iu ny c c bng cch thm mt dung mi ha tan trong nc nh aceton vo dung dch cha protein. S ta bng aceton hoc ethanol 80% c nhiu thun li hn v n tng i r, c sn dng tinh

khit vi t cht tp nhim gy c hay c ch i vi enzym, do nhit bay hi ca dung mi thp nn d tch b dung mi khi ch phm enzym bng phng php sy nh bng qut gi. II. Phng php ta bng mui Ngi ta c th dng mui (NH4)2SO4, NaCl ta protein v cc mui ny va lm trung ha in (do cc ion tc ng tng h vi cc nhm tch in tri du), va loi b lp v hydrat ca phn t protein. Cc protein khc nhau c th b kt ta vi nng mui khc nhau, v vy c th dng mui tch ring cc protein ra khi hn hp ca chng.
CU 8.TINH CHT CUA PHN T Hao nc,a nc. Ki nc.tao ko bit CU 9.TAO KO BIT?????? CU 10.TRICH LY CAM LY VITAMIN A?????????/

CM GO
(Bran rice) I. Gii thiu Trong qui trnh xay xt v ch bin go, sau khi thu c sn phm chnh l go th cn mt sn phm ph c gi tr kh cao l cm go. T lu cm go c cc nh my xay xt thu hi v bn nh l mt sn phm chnh ch sau go. Cm c thu hi di hai dng l cm kh v cm t. Cm kh s c sy thm ln na bo qun c lu hn, cn cm t thng c bn cho cc c s nui c da trn s dng ngay.

+ Cu to v thnh phn cm go:

Hnh: S lp cm trong ht la Cm go thng c dng bt, mm v mn. Cm go chim khong 10-12% khi lng la cha xay xt. Cm l hn hp ca lp v ngoi ca ht go v lp aloron. Nhng thnh phn c thu hi khi khi xay xt v ch bin go v c gi chung l cm. + Thnh phn dinh dng ca cm go: Thnh phn - Calori - Tng s lipit Khi lng/100g 316 KJ 21g

- Cht bo bo ha - Cht x tiu ha c - Carbohydrat - ng - Protein - Vitamin E - Vitamin B6 - Canxi - Kali

4g 21g 28 g 0,9 g 13,3 g 4,9 mg 4,1 mg 57 mg

Ngun: theo www.Nutritiondata.com Theo bng trn th cm c lng dinh dng rt cao vi lng cht bo cha bo ho cao, vitamin nhm E, nhm B, phylate, km, can xi, kali u rt cao, ngoi ra trong cm cn co chc cht bo omega 3 kh cao. Theo (http://www.ricebranoil.info) th 65% cht dinh dng ca go tp trung cm, thnh thn ca cm c nhiu lai vitamin v cht bo tt, li cn i v c nhiu x d tiu c th xem l rt tt cho c con ngi. Tuy nhin cm thng c cho cc loi gia sc v thy sn n ch khng cho ngi. Nguyn nhn l do cm c mt s enzym (cht men) ni ti hot ng rt mnh s oxy ha cc nhm bo cha no ca cm rt nhanh ch vi gi sau khi ch bin to mi hi kh chu. Thm vo do cng ngh xay xt go cha cao li t c u t theo hng thu cm sch nn cm thng ln rt nhiu loi tp cht (v tru, sn .) Do l do m hu ht lng cm thu c thng c b con nng dn dng nui cc loi gia sc, gia cm. Hng nm trn th gii c khang 40 45 triu tn cm c sn sut v 90% l nm chu (http://www.bannhanong.com/, x l cm go thay th ng cc)

No???? Phng phap.ko tim thy CU 10.QUANG PH HP THU UV_VIS


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 = =.l.c 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 ch ph thuc vo bn cht cht hp th v bc sng ca bc xth . c trng cho cng hp th bc x ca cht c khohp th Do ln ta ni cht hp th mnh(cng hp th ln),st.Khi ngc nh- cht hp th yu(cng hp th nh).li khi vng t ngoi - kh kin nh lut Bouguer-Lamber-Beer lun tun theo thng lun c xc nh v c lp li tt.v vy gi tr ca cc vn hp th ng vi cc chuyn mc electron khc nhauGi tr 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: E/hc.Vel = 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 vel = 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. T ph UV-Vis v h s hp th mol, ta c th suy ra tnh cht ca sn 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. - Hiu ng m mu ca dung dch c o bng hp th. S chuyn dch tng.cc i hp th v bc sng di (hiu ng Batocrom) th .- 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. Ci Labomed UVD3500 l my o quang 2 chm tia, chnh xc th n, nhng m nu lm xc nh hm lng ion kim loi trong nc, hm lng amoni.. th tt qu Nhng bn dc m lm ci ny th hi mt ri, v mi ln c c 01 mu , nu lm vi chc mu chc mt my ngy, hic hic Th na loi my ny thng l c ph quyt thi, m mun xc nh ng hc protein, hay phn r thuc th theo em c Kinetic tin hn nhiu Cng cha bin Bc AnhHB thc mc c th th no, pm em r em t vn ch nhiu ci mun ni m ng ngi qu ngi khng dm

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