HP CHT THIN NHIN

NI DUNG
Chng 1: M U Chng 2 : PHENOL-AXIT PHENOL V DN XUT Chng 3 : FLAVONOIT Chng 4: TINH DU-HP CHT LACTONTECPENOIT Chmg 5: STEROIT Chng 6: SAPONIN Chng 7:GLYCOZIT TIM V GLYCOZIT XIANOGEN Chng 8: ANKALOIT V CAROTENOIT Chng 9: SASCARIT- AMINOAXIT- PROTEIN -AXIT NUCLEIC

Chng 1:

M U

1.1.KHI NIM V PHN LOI HP CHT THIN NHIN

1.1.1 I TNG V LCH S NGHIN CU

H p ch t thin nhin l cc sn phm hu c ca cc

qu trnh trao i cht trong c th sng. Ngnh ha hc nghin cu tnh cht v cu trc ca cc hp cht thin nhin c gi l ha hc cc hp cht thin nhin L ch s nghin c u Thi xa xa Th k 19: Mt trong nhng cng trnh c gi tr l qui tc isopren v cu to ca tecpenoit (Wallch, 1887) Th k 20:Xc nh cu trc hp cht thin nhin c s tin b vt bc nh cc k thut hin i, cc phng php ph

1.1.2 PHN LOI CC HP CHT THIN NHIN

1.1.2.1 Da vo tnh thit yu i ng thc vt: ngi ta chia thnh 2 nhm

Ch t trao i s c p : L nhng cht thin nhin cn thit cho s sng gm cacbonhidrat, protein, axit nucleic, cc lipit v dn xut ca chng. Cc hp cht ny c sn sinh t cc c th sng, khng ph thuc vo loi. Qu trnh trong cc cht trao i s cp c to thnh c gi l qu trnh trao i s cp Ch t trao i th c p: L nhng hp cht thin nhin khng hn khng cn thit cho s sng ca ng thc vt. Cc cht trao i th cp thng ph thuc nhiu vo loi. Cc hp cht th cp bao gm : tecpenoit, steroit, flavonoit, ankaloit.Chng l sn phm ca qu trnh trao i th cp.

 Trong hp cht thin nhin thng c cc nhm chc c bn: + Hp cht hidrocacbon cha no + Ancol- phenol ete +Andehit- xeton + Axit hu c v dn xut + Amin + D vng + Hp cht tp chc

1.1.2.2 Da vo b khung cacbon, cc nhm chc v theo tnh ph bin ca hp cht:

Cc hp cht thin nhin thng c phn loi thnh: + Cht bo- lipit + Hidratcacbon- Gluxit ( monosacarit, oligosacarit, polisacarit) + Axit amin- Protit + Tecpenoit (monotecpen,sesquitecpen, ditecpen, tritecpen) + Steroit + Coumarin + Flavonoit + Ankaloit + Tanin + Cht khng sinh + Vitamin

1.2 PHNG PHP CHIT XUT

1.2.1 NGUYN LIU 1.2.2 TNH PHN CC CC HP CHT THIN NHIN 1.2.3 DUNG MI 1.2.4 CC PHNG PHP CHIT XUT

1.2.1.NGUYN LIU
Trong iu kin cho php nn dng nguyn liu ti. Nguyn liu thu hi xong nn n nh bng cch nhng vo cn hay nc un si trong vi pht, sau ro nc hay lm kh t nhin trong khng kh. Ht sc trnh dng nhit cao lm kh nguyn liu Phi s b x l nguyn liu ban u nh: vt b nguyn liu c su bnh, Phi xc nh ng tn khoa hc ca cy. Do thng khi thng bo khoa hc phi ghi a ch ngi xc nh, c quan ngi c c th lin h tham kho khi cn thit

1.2.2 TNH PHN CC CC HP CHT THIN NHIN Trong cy, cc hp cht hu c tn ti dng ha tan trong nc, trong du bo hoc tinh du - Cc hp cht ha tan trong nc (dch t bo) l cc hydratcacbon c phn t lng thp (monosaccarit, mt s oligosacarit nh pectin, gm); cc glycozit, mui ankaloit ca cc axit hu c; cc aminoaxit, mui ca aminoaxit; cc hp cht phenol ha tan di dng glycozit. Ni chung cc cht tan trong nc l cc cht phn cc. Cc hp cht c nguyn t H lin kt trc tip vi nguyn t m in nh O, N, F, Cll nhng nhm phn cc, cng nhiu nhm phn cc trong phn t th tnh phn cc cng ln

Cc hp cht tan trong du bo hoc tinh du: cc hidrocacbon, monotecpen, sesquitecpen, sterol, carotenoit Cc cht tan trong du bo v tinh du l cc cht t phn cc. Tuy nhin tnh phn cc ca chng cng khc nhau ty thuc vo khi lng nguyn t v nhm chc c trong phn t hp cht. Thng thng cc hp cht c mch cacbon di km phn cc,

1.2.3 DUNG MI
1.2.3.1. Tnh phn cc ca dung mi
C s la chn dung mi chit l phn cc ca cc hp cht cha trong nguyn liu v phn cc ca dung mi. Ngi ta phn bit cc dung mi theo phn cc + Dung mi phn cc mnh: nc, cc ancol thp (metanol, etanol.). + Cc dung mi phn cc yu hoc va: etyl axetat, cloroform, axeton, + Cc dung mi khng phn cc: ete, ete-du ha, benzen, toluen, hexan

1.2.3.2. Cht tan trong nc v dung mi phn cc

Cc cht in ly nh mui v c Cc hp cht hu c ni chung khng ion ha, nhng nu chng c nhng nhm to c lin kt hydro vi nc th tan c trong nc.

 Cng nhiu nhm phn cc th phn t y cng d tan trong nc, nu mch cacbon cng di th ha tan cng gim.Cc hp cht tan c trong nc khi: + Nu 1 nhm phn cc trong phn t c kh nng to thnh lin kt hidro vi nc nu phn t ca cht c mch cacbon khng qu 5 hoc khng qu 6 nu hp cht c mch nhnh + Nu phn t c nhiu nhm phn cc (t 2 tr ln) th t l ny gim xung: mt nhm phn cc cho 3 hoc 4 nguyn t cacbon trong mch th phn t y tan c trong nc.

1.2.3.3. Cht tan trong ete v cc dung mi khng phn cc

Ni chung cc cht khng phn cc u tan trong ete v cc dung mi khng phn cc,ngc li khng tan trong nc v cc dung mi phn cc khc. Cc phn t c mt nhm phn cc trong phn t c th tan c trong ete. Hu ht cc cht hu c tan trong nc th khng tan trong ete. Nu mt cht va tan trong nc va tan trong ete th cht phi l cht khng ion ha, c s cacbon khng qu 5, c mt nhm phn cc to lin kt hidro nhng khng phi l phn cc mnh.

1.2.4 CC PHNG PHP CHIT XUT

Ni chung, khng th c mt phng php chung no c th p dng c cho tt c nguyn liu. Trong phn ny ch nu nhng phng php chit xut nhm nghin cu s b khi cha bit r thnh phn ha hc ca nguyn liu.

1.2.4.1 Phng php chung

Dng mt dy cc dung mi t khng phn cc n phn cc mnh chit phn on cc cht ra khi nguyn liu. v d dy ete- du ha, ete, cloroform, cn v cui cng l nc. Cch chit thng dng nht l chit nng lin tc trn my soxhlet hoc chit hi lu. Sau mi ln chit vi mt loi dung mi, cn lm kh nguyn liu ri mi tip tc chit vi dung mi tip theo. Mi phn on chit, thu hi dung mi v tin hnh phn tch ring.

Chit nng bng soxhlet v hi lu

Da vo tnh phn cc ca dung mi v c th d on s c mt ca cc cht c mt trong cc dch chit. Trong phn on ete, ete du ha s c hidrocacbon bo hoc thm, cc thnh phn ca tinh du nh monotecpen, cc cht khng phn cc nh cc cht bo caroten, cc sterol, cc cht mu thc vt, clorofyl

 Trong dch chit cloroform c sesquitecpen, ditecpen, coumarin, quinon cc aglycon do cc glycozit thy phn to ra, mt s ankaloit bazo yu Trong dch chit cn s c mt glycozit, ankaloit, flavonoit, cc hp cht phenol khc, nha, axit hu c, tanin Trong dch nc s c s c cc hp cht phn cc nh cc glycozit, tanin, cc ng, cc hidratcacbon phn t va nh pectin, cc protein thc vt, cc mui v c

1.2.4.2 Chit ly ton b thnh phn trong nguyn liu Dung mi thch hp nht l cn (metanol hay etanol) 80% trong nc. Cn, c bit l metanol c xem nh dung mi vn nng n c th ha tan cc cht khng phn cc cng nh cc cht phn cc khc Dch chit khi bay hi dung mi c cao ton phn cha hu ht cc hp cht trong nguyn liu

 Sau cn tch phn on cc cht trong cao th chun b mt dy cc cht khng tan trong nc c phn cc t yu n mnh nh v d dy ete- du ha, ete, cloroform, etyl axetat, butanol. Ha tan cao vo mt lng nc , cho vo bnh chit , ln lt chit vi cc dung mi trn. Dch chit mi phn on sau khi thu hi dung mi em i phn tch.

1.2.4.3 Cch chit

Chit nhit thng : c 2 cch l ngm kit v ngm phn on. Ngm kit l phng php tt hn v n chit c nhiu hp cht hn, t tn dung mi, nht l khi p dng phng php ngm kit ngc dng. Chit nng: Nu dung mi d bay hi phi dng phng php chit lin tc (trong soxhlet) hoc chit hi lu. Nu chit hi lu th t nht phi chit 2 ln.

 Phng php chit ngm kit ngc dng

A1 A2

B1 B2

C1 C2

D1 D2

Phng

php chit ngm phn on

1.2.4.4 Cch thu hi dung mi

Bng cch chng ct, trong tt nht l dng phng php chng ct p sut thp
khong khi

bom chan khong

1.3 PHNG PHP TCH BIT CC HP CHT THIN NHIN

1.3.1. CC PHNG PHP SC K 1.3.2 PHNG PHP CHIT

1.3.1. CC PHNG PHP SC K

1.3.1. 1. Khi nim v sc k Sc k l mt phng php vt l dng tch ring cc thnh phn ra khi hn hp bng cch phn b chng ra 2 pha: mt pha c b mt rng gi l pha c nh v mt pha kia l mt cht lng hay cht kh gi l pha di ng, di chuyn qua pha c nh. Phn loi: Chia thnh 2 loi: l sc k lng v sc k kh

a) Sc k lng : l sc k c pha ng l cht lng. Trong sc k lng c cc k thut: Sc k giy: pha tnh l giy Sc k lp mng: pha tnh l lp mng cht hp ph c tri bng trn tm thy tinh hoc kim loi Sc k ct: Pha tnh l cht rn c nhi thnh ct. Trong sc k ct ty thuc vo bn cht ca cht rn nhi ct m chia thnh cc loi -Ct c in: ct n gin vi cht hp ph l v c hay hu c -Ct trao i ion: Ct l cht trao i ion m hoc dng -Ct gel hay lc gel: Pha tnh l mt gel tng hp c l xp xc nh lc cc cht c kch thc khc nhau Sc k lng cao p: (sc k lng hiu nng cao-HPLC)

b) Sc k kh: l sc k c pha ng l cc cht kh. Da vo pha c nh ngi ta cn chia ra: Sc k kh- rn Sc k kh lng Trong thc t ngi ta thng dng SKK pht hin s lng cc cht trong hn hp, hm lng ca chng da vo din tch pic ca ph sc k

1.3.1. 2 Sc k giy
A. Cc bc tin hnh Chun b mu th Chm mu th Giy Dung mi Cch khai trin Pht hin vt

a) Chun b mu th Cht th c pha trong lng ti thiu dung mi. Dung mi thng dng l cc cht d bay hi nh axeton, metanol, etanol Tr trng hp nghin cu ton din v cy, thng thng ngi ta phi loi tp v c c trc khi chm. Cch loi tp thng dng l tch bng cc dung mi khc nhau, bng cch ta, ly tm, ng lnh

b) Chm mu th: Dng ng mao qun c ng knh t 0,5- 1mm. Thng thng nng cht chm t 0,1-1%, lng cht chm t 11000g ty thuc vo nhy pht hin chng. iu quan trng trong k thut chm vt l chm vt cng nh cng tt v cc vt trn cng mt ln sc k phi ng u nhau v kch thc v m c

c) Giy Nu dng sc k giy pht hin cht th c th dng loi giy mng, nhng nu dng tch cht th c th dng giy dy. Khi s dng cn trin khai ng chiu ca giy. Thng thng ngoi b giy c c mi tn ch chiu trin khai giy.

d) Dung mi: Vic chn dung mi thch hp l yu t chnh quyt nh kt qu th nghim + Cch chn dung mi: Th t phn cc ca dung mi ( theo E. Berg, 1963) Ete du ha < CCl4 < xyclohexan< cacbon disunfua < dietylete < benzen < cc este < Cloroform < dicloetan < cc ancol < nc < pyridin < cc axit hu c < cc axit v c v bazo Cng c th dng dung mi nguyn cht hoc hn hp dung mi vi t l thch hp. la chn h dung mi ta c th dng chy th vi chiu cao 10 cm l .

+ Cc loi h dung mi H dung mi c pha c nh l nc H dung mi c pha c nh l dung mi hu c phn cc (a nc) H dung mi c pha c nh l dung mi hu c khng phn cc (k nc)

e) Cch khai trin: Trin khai trn xung: u im ca phng php ny l tc chy tng i n nh v nhanh nh tc dng ca trng lc Trin khai di ln ( trin khai ngc): Phng php ny c u im l trin khai n gin, nhanh chng nhng c nhc im l khi dung mi chy c 25 mm th dung mi s chy chm li, do ngi ta thng s dng cch ny th tm h dung mi thch hp v thng cho chy n 25cm th ngng li. Trin khai ngang Trin khai vng Trin khai nhiu ln v trin khai qu c giy Sc k hai chiu

g) Pht hin vt Phng php ha hc: Dng cc cht hin mu c trng cho tng loi hp cht. Thuc th c pha c nng thch hp cho tc dng ln giy . C 2 cch: - Nhng giy: p dng cho cc sc nh,v thuc th khng ha tan cc vt - Phun : Phng php ny thng dng hn Phng php vt l: - Thng p dng vi cc hp cht hp th tia cc tm trong khong t 240-260 nm hnh thnh cc vt ti trn nn pht quang. Trong mt s trng hp c th phun ln giy dung dch Fluorescein (C20H18O2) vi mc ch lm tng pht quang ca nn nhn vt r hn. - Mt s cht c kh nng pht hunh quang khi chiu tia cc tm bc sng khong 360nm

B Cc yu t nh hng n gi tr Rf
a) Gi tr Rf Trong sc k biu th s di chuyn ca cc cht ngi ta dng khi nim Rf

Gi tr Rf khng phi l mt hng s c trng cho s di chuyn ca mt cht mi iu kin, do khi ghi gi tr Rf phi ghi y nhng iu kin th nghim km theo (loi giy, nng cht th, lng chm, h dung mi, lng dung mi, chiu trin khai, nhit tin hnh th nghim)

b) Phng php lm tng lp li trong SKG Gi nhit khng i trong khong nhit cho php 50C Trn u dung mi v gi nhit th nghim trong 1-2 ngy. Kim tra li dung mi bng cch chy th vi cht chun i chiu Giy chm cht th c a vo trc 24 gi to s cn bng gia giy v kh quyn trong bnh y bnh kn trong sut qu trnh trin khai ng di chuyn ca dung mi phi t 30-35 cm nu trin khai xui v 25 cm cho trin khai ngc Dng mt loi giy, tin hnh trong cng mt iu kin

C Hin tng vt d thng

a. Vt lan rng b.Vt c ui c.Hin tng hai ui d. Ngoi ra cn c cc hin tng: vt b ra hai bn , lan rng ra 2 bn, vt mo, vt khng di chuyn, mt vt hoc mt cht to nhiu vt

a. Vt lan rng ( Khng tp trung) khc phc cn phi Chm vt cng nh, trn Thuc hin mu khng qu c, pha va hin mu Chn h dung mi thch hp, nht l tc chy ca h dung mi. Ngoi ra nu hnh dng v kch thc l xp ca giy khng u vt cng lan rng

b.Vt c ui Vt c ui c th do Lng cht chm qu nhiu Dung mi chy qu nhanh S bin i khng thun nghch v t cc thnh phn trong cht tan trong qu trnh di chuyn S hp ph qu mnh ca b mt cht hp ph

b.Vt c ui Vt c ui c th do Lng cht chm qu nhiu Dung mi chy qu nhanh S bin di khng thun nghch v t cc thnh phn trong cht tan trong qu trnh di chuyn S hp ph qu mnh ca b mt cht hp ph

c.Hin tng hai ui Hin tng ny thng xy ra i vi cc cht mu ca dch chit. Do cc cht mu khng c kh nng hp ph mnh bng cc cht khng c mu c mt trong dch chit nn cc cht khng mu chim v tr ngay gia cn cc cht mu di chuyn ra ra khi chm. V vy khi trin khai th c hai ui do cc vt trn v di chy nhanh hn 2 bn nn vt c 2 ui

1.3.1.3. SC K LP MNG (SKLM) A. Nguyn l: B. Cht hp ph C. Hot nng D. Cht hp ph thng dng E. Dung mi

A. Nguyn l: L phng php phn tch trong dung dch cht phn tch di chuyn trn mt lp mng cht hp ph mn v c hay hu c theo mt chiu nht nh. Trong qu trnh di chuyn, mi cht chuyn dch vi tc khc nhau ty thuc vo bn cht ca chng v dng li nhng v tr khc nhau

B . Cht hp ph
Thng dng l cc oxit khng tan, cc oxit hidrat ha v cc mui Lc hp ph ca cc cht hp ph theo trt t tng dn (theo Strain) 1- Sacaroz 7- Magie cacbonat 2- Magie xitrat 8- Magie oxit 3- Tale ( tan, hot thch) 9- Axit silixic hot ha 4- Silicagel 10- Nhm oxit hot ha 5-Natri cacbonat 11- Than ng vt hot 6-Canxi cacbonat Trong phn tch nguyn liu, cht c dng thng dng nht l silicagel v oxit nhm

C. Hot nng
Hot nng ca cht hp ph ph thuc nhiu vo hm lng nc trong n v nc hp ph s chim v tr hot ng trn b mt lm gim kh nng hp ph ca cht hp ph, V vy c th iu chnh kh nng hp ph bng cch thm nc. Tuy nhin trong thc t khi trin khai thng thng ta phi sy kh kh nng hp ph ca cht hp ph tng ln v s hp ph n nh.

 D Cht hp ph thng dng

a. Silica gel: Dng trong bn mng l loi bt mn, v nh hnh c ng knh c ht t 10- 40 m, nu dng trong sc k ct thng thng th ng knh c ht 63-200 m (63-200 mesh) V mt ha hc silica gel v nh hnh l loi c cc nhm siloxan O-Si-O v cc nhm silinol O-Si-OH V tr hp ph trn b mt l cc nhm silinol, n c kh nng to lin kt cng ha tr vi cc hp cht phn tch. tng kh nng hp ph ca silicagel ngi ta sy nhit 110-1200C loi nc, nhng khng sy cao hn 150oC v khi cc nhm OH trong silica gel b mt nc v gim kh nng hp ph v khng ht nc Ngoi ra tng thm kh nng hp ph, ty theo bn cht ca cht cn tch ngi ta c th thm axit hay bazo s hp ph c th c tng ln hay gim xung.

b. Alumina Thnh phn ha hc, cha ch yu l -Al2O3. c 3 loi: trung tnh, baz v axit Alumin baz c cha khong 0,1-0,5 % NaOH bm trn mt alumin dng natri aluminat, c pH 10.N l cht trao i ion trong nc Alumin trung tnh, c pH t 6,5-7. y l loi dng tt cho sc k ct v khng gy phn ng vi cht th Alumin axit c iu ch t alumin vi HCl long, pH 4, c tc dng nh cht trao i ion Cc cht th c tnh axit nh cc phenol, axit cacboxylic s b gi li trn alumin baz hn loi trung tnh v axit, ngc li cc cht c tnh baz (amin, ankaloit...) s b gi li trn alumin axit hn loi trung tnh v baz Khc vi silicagel, cc alumin nu sy trn 2000C s hp ph la chn i vi cc cht phn cc v c th lm tng hp ph khi un nng n di 7000C Nu trn 7000C th kh nng hp ph gim.

E. Dung mi a) Tc di chuyn ca cht ph thuc vo dung mi Trt t tng dn lc phn hp ph dung mi nh sau ( Theo Trappe) Ete du ha < xyclohexan < CCl4 < tricloetylen < toluen < benzen < metylclorua < clorofoc < ete etylic < etyl axetat < pyridin < axeton < npropanol < etanol < metanol < nc Trong thc t ngi ta thng dng hn hp dung mi c phn cc vi t l ph hp to h cc dung mi mi

b) Phng php chn dung mi Nguyn tc chung: Nu cht th c i lc yu i vi cht hp ph th chn cht hp ph mnh vi h dung mi c lc phn hp ph yu. Ngc li nu cht th i lc mnh vi cht hp ph th chn cht hp ph yu v chn dung mi c lc phn hp ph mnh.
manh phan cuc manh (phan hap phu yeu) DUNG MOI

CHAT HAP PHU

yeu

phan cuc yeu (phan hap phu manh) ai luc manh CHAT THU

ai luc yeu

c) Cu to ha hc v i lc hp ph ca cht th Cc hidrocacbon no hu nh khng hp ph S c mt ca lin kt i tng thm lc hp ph, cng nhiu ni i, lc hp ph cng tng C thm cc nhm chc cng lm tng lc hp ph. Bng thc nghim, ngi ta thy lc hp ph gim dn theo dy sau: + i vi hidrocacbon thm: - COOH > -CO-NH2 > -OH > -NH-CONH2 > -NH2 > -OCO- CH3 > -CO-CH3> - COCH3 > -N(CH3)3 > -NO2 > -OCH3 > -H > Cl + i vi hidrocacbon mch thng : - COOH > -OH > -NH2 > -COOR(ankyl) > -CH3 + Cc hp cht cacbonyl hp ph yu hn hp cht c OH v amin tng ng

d) K thut sc k Trng knh, hot ha knh : hot ha 1100C 1 gi Bo qun knh trong bnh ht m Chm mu phn tch Trin khai bng dung mi Hin vt

1.3.1.4 Sc k ct
A. Khi nim C th ni sc k ct l mt dng ca sc k giy hoc sc k lp mng nhng y pha tnh c nhi vo ct, nh vy c th trin khai mt cch lin tc vi nhiu h dung mi khc nhau t phn cc yu n phn cc mnh Ty theo tnh cht ca cht dng lm ct m s tch c th xy ra ch yu theo c ch hp ph (ct hp ph) v d nh silicagel, oxit nhm hoc c ch phn b (ct phn b ) v d nh xenlulo.

B. Dng c - ha cht a) Ct Kch thc ct v lng cht hp ph: Thng thng lng cht hp ph gp 25- 50 ln lng cht cn tch v cao ca phn ct cht hp ph v ca phn mu th ln hn 8:1. Tuy nhin vi nhng cht kh tch th cn ct to v lng cht hp ph phi ln hn

Mu sc k Khi lng (g) cht hp ph (gam) 0,001 0,1 1,0 0,3 3 30

ng knh ct (mm) 3,5 7,5 16

Chiu cao c t (mm) 30 60 130

b) Ha cht lm ct
Ct phn b: xenlulo, kieselguhr (Cellite), gel ca axit silixic Ct hp ph : oxit nhm, silicagel, poliamit, CaCO3, MgO, than hot Mt s ha cht dng cho sc k ct c tiu chun ha + Oxit nhm trung tnh (Merck), c ht 0,063-0,200 mm (70- 230 mesh) + Oxit nhm bazo, c ht 0,063-0,200 mm ( 70230 mesh) + Silicagel 60 Merck, c ht 0,063-0,200 mm + Poliamit: c ht < 0,07 mm

 Khi cho cht hp ph vo dung dch cha cht hu c, th cc cht hu c s bm vo cht hp ph do nhiu loi lin kt khc nhau. Trt t cc loi lc ny thay i nh sau: S to mui > lin kt phi tr > lin kt hidro > tng tc lng cc > lc Van-der-Valls

 Dung mi: Cc dung mi thng dng cho sc k ct l hexan, benzen, CHCl3, axeton, etanol, metanol, butanol, nc. Th t phn cc tng dn ca dung mi: Ete du < hexan < xiclohexan < CCl4,< benzen < toluen < diclometan < CHCl3 < ete etylic < etyl axetat < axeton < pyridin < propanol < etanol < nc < axit axetic

 Hng cc phn on vo dng c hng v thu hi dung mi: Thng thng cc cht khng phn cc ra trc sau mi n cc cht phn cc yu v cui cng l cc cht phn cc mnh Vi dung mi khng phn cc: Gii ly ra sm Ankan Anken, ankin; Xicloankan, xicloanken; Aren; Xeton; Andehit Vi dung mi phn cc : Gii ly ra chm Este; Ancol, thiol; Amin; Phenol,axit cacboxylic

C K thut
a) Chun b ct: Yu cu l cht rn lm ct phi phn tn ng u mi im trong ct thnh mt khi ng nht. Ct hp ph: C 2 cch nhi ct + Nhi kh: + Vo ct bng dung mi (Nhi t): Ch l khng c kh dung mi trong ct. Tip tc cho dung mi hng c rt vo ct cho chy tip tc mt thi gian (t 5-10 gi) cho ct n nh hon ton Ct phn b: Cng nh sc k giy, tch bng ct phn b l thc hin s tch gia 2 pha l pha c nh v pha di ng do phi c giai on x l pha c nh.

b .a cht th vo ct:
Yu cu l phi phn tn cht th thnh mt lp mng ng u trn mt mt phng . C nhiu cch a cht th vo ct: Phng php dng a giy: Dng giy lc ct thnh a trn c ng knh nh hn ng knh trong ca ct khong 0,5 mm, dng kim khu m thnh nhng l thng nh cch u nhau khong 1,5 mm. Phng php cho thng dung dch th ln ct: Phng php trn u cht hp ph vi mu cht th Trong 3 cch trn cho thng mu th vo ct l nhanh hn c

c. Ra ct
c th ra ct trong cc iu kin sau: - p sut thng - p sut cao Ch : i vi ct nhi kh, c th cho chy ngc ct trc khi cho ra xui

D K thut sc k ct nhanh
a. c im K thut sc k ct nhanh l k thut m cc thng s ti u c xy dng t thc nghim vi mc ch lm cho vic ng dng c n gin, tit kim, nhanh nghin cu k thut ny, tc gi c nh mt s yu t v ch thay i mt s yu t cn thit. Cc yu t c nh l: - Cht hp ph dng cho mi trng hp l silicagel 60, c ht 40-60m - Chiu cao ct hp ph 15 cm - Tc chy: Mi pht chy c 5 cm chiu cao dung mi trong ct cho mi c ct vi mi cht th

b. Cc bc tin hnh b1) Chn dung mi: b2) Da vo Rf v Rf v lng cht th chn ct, th tch dung mi v n nh s phn on tp hp theo bng trn b3) Chun b ct: b4) Vo dung dch th: Ch yu bng phng php cho thng dung dch th vo ct. Sau khi cho mu th vo ct xong cn phi iu chnh p sut sao cho mi pht chy c 5cm chiu cao ca ct

c) u im ca k thut sc k nhanh Tit kim cht hp ph v dung mi, thi gian nhanh nhanh hn nhiu Nu cc cht c Rf t 0,1-0,2 u c th tch c bng phng php ny, nu chn c h dung mi thch hp n gin, d p dng nh cc thng s c xy dng sn.

1.3.2 PHNG PHP CHIT

1.3.2.1. Chit rn lng Chit gin on Chit lin tc: dng c soxhlet 1.3.2.2. Chit cht lng Yu cu dung mi: dng dung mi khng trn ln vi dung mi c v c kh nng tan tt hp cht hu c hn dung mi c ( thng l nc) Cc dung mi thng dng l ete etylic, ete du ha, xng, benzen, CHCl3...

1.4 PHNG PHP PHN TCH

1.4.1 PHN TCH NH TNH 1.Phng php sc k Sc k giy Sc k lp mng Sc k kh 2. Cc phng php ph( IR, UV, NMR, MS...) 3. Cc phng php vt l khc Nhit nng chy Nhit si Ch s khc x, gc quay cc

1.4.2 PHN TCH NH LNG 1. Phn tch khi lng 2. Phn tch th tch

CHNG 2 PHENOL AXIT PHENOL V DN XUT

 Phenol l nhm cc hp cht hu c trong phn t c nhm OH lin kt trc tip vi nhn thm. Cc hp cht phenol ni chung d tan trong nc v trong thin nhin chng thng tn ti dng glycozit Trong hng ngn hp cht phenol trong thin nhin bit r cu to th hp cht flavonoit l nhm hp cht quan trng nht. Ngoi hp cht phenol n chc mt vng, cc phenylpropanoit v quinon cng chim mt t l ng k. V cc poliphenol trong cy c lignin, melanin, tanin.

2.1 PHENOL V AXIT PHENOL

Cc phenol v axit phenol thng c nghin cu chung v chng thng song song tn ti vi nhau trong cy. Mt s phenol c trong cy l: hydroquinon (1,4-dihidroxibenzen), rezocxinol (1,3dihidroxibenzen), ocxinol (1,3-dihidroxi-5metylbenzen), phlorogluxinol (1,3,5trihidroxibenzen), catechol (1,2dihidroxibenzen), pyrogalol. (1,2,3trihidroxibenzen),..

 Cc axit phenol thng gp l:

axit p-hydroxibenzoic, axit protocatechic (axit 3,4-dihidroxibenzoic), axit vanilic (axit 4-hidroxi-3-metoxi benzoic), axit syringic (axit 4-hidroxi-2,3-dimetoxi benzoic),

2.1.1 S PHN B
Cc axit phenol tn ti dng kt hp vi lignin to thnh este hoc vi cc oza di dng glycozit. Cc axit phenol thng gp l: axit p-hydroxibenzoic, axit pyrocatechic, axit vanilic, axit syringic, cc axit t gp l axit salixylic, axit o-protocatechic (axit 2,3dihydroxibenzoic),. Ngc vi axit phenolic, cc phenol t do rt him thy trong cy. Hydroquinon l cht thng gp hn c, tip n l catechol, ocxinol, phlorogluxinol, pyrogalol

2.1.2 PHN TCH PHENOL V AXIT PHENOL Vic tch phenol tt nht bng SKLM. Thng thng nguyn liu ti hoc kh c nghin nh, thy phn vi kim hoc axit trong cn long 600 Thy phn axit: dng axit HCl 2M trong 30 pht, sau khi ngui, lc, chit bng ete. Gn lp ete, ra bng nc vi ln, lm khan, bc hi n kh. Ha cn kh vi metanol lm dung dch chm sc k Nu thy phn bng kim: NaOH 2M, thy phn trong 4 gi nhit thng. Dch thy phn em axit ha ri chit bng ete nh trn.

 Cht hp ph l silicagel G vi cc h dung mi c phn cc trung bnh. Cc h dung mi thng c dng l : A: Axit axetic : clorofoc (1 : 9) (Cht hp ph l silicagel G) B: Etyl axetat: Benzen (9:11) ( Cht hp ph l silicagel G) C: Benzen: metanol: axit axetic (45: 8: 6) (Cht hp ph xenlulo MN 300) D: Axit axetic : nc ( 6 : 94) (Cht hp ph xenlulo MN 300) Cht hin mu : Vanilin + HCl

2.2 PHENYL PROPANOIT

Phenyl propanoit l nhm hp cht phenolic t nhin gm mt mch nhnh 3 nguyn t cacbon gn vo nhn thm V mt sinh tng hp chng l dn xut ca axit amin thm phenylalanin

2.2.1 CC HP CHT THNG GP Hp cht ph bin nht l cc axit hydroxyxinamic

R
HO H3C-O

CH=CH-COOH

HO

CH=CH-COOH
R

R= H: axit p-cumaric R= OH: axit cafeic

R= H :axit ferulic R= OCH3: axit sinapic

2.2.2. PHN B
Cc axit hidroxixynamic tn ti trong cy ch yu dng este, d b thy phn bng kim cho axit t do. Axit cafeic thng tn ti dng este vi axit quinic, gi l axit clorogenic

2.2.3 CHIT XUT V PHN TCH

Axit hydroxixynamic thng c phn tch ng thi vi cc phenol v cc axit phenol sau khi thy phn dch chit thc vt bng kim v chit ly phenol bng ete hoc etyl axetat. Cn sau khi bc hi, ha tan trong metanol, chm trn SKG hoc SKLM xenluloz. Hin vt bng soi UV hoc UV + NH3. Dung mi: BAW: n-BuOH- HOAc - H2O (4: 1:5) lp trn BN: n-BuOH -NH4OH 2M (1: 1) lp trn BEW: n-BuOH EtOH- H2O (4: 1: 2,2)

2.2.4 CC POLYPHENOL C hai loi hp cht thng gp l lignin v lignan( xem ti liu)

2.3.1 I CNG Coumarin l nhm hp cht thin nhin c xem l dn xut lcton ca axit ortohydroxixynamic (I). Hu ht cc coumarin bit hin nay (khong 600 cht) tn ti ch yu di dng t do, mt s t tn ti dng glycozit nh cc glycozit ca psonolen (II). Coumarin ph bin nht trong cy l cht umbeliferin (III)
o

2.3 COUMARIN

HO

(I)

(II)

(III)

 V cu trc c th c cc loi khung isopren nh suberosin (IV), colombianetin (V)

CH3O

o
o
o
(V)

(IV)

 V phn b: Coumarin c trong nhiu h thc vt: Leguminosen, Umbeliferae, Ochidaceae, Rutaceae. Chng c mt trong hu ht cc b phn ca cy: r, l, hoa, qu. c tnh vt l Coumarin l cht pht quang mnh v rt nhy vi nh sng. Ngi ta li dng tnh cht ny tch cc hp cht coumarin

V tc dng dc l coumarin v dicoumarin dng lm thuc chng ng mu. Tnh cht ny c lin quan n nhm OH v tr C4 trong cu trc. Tnh cht chng ng mu bin mt hoc gim khi nhm OH v tr ny b bin mt hoc b thay th bi nhm khc. Ngoi ra mt s coumarin c tc dng lm gin ng mch vnh v mch ngoi vi, c tc dng chng co tht. Mt s coumarin c tc dng lm tng cholesterol trong mu. Mt s cht c tc dng c ch sinh trng thc vt, tc dng khng khun, dit nm v chng vim.

2.3.2 CHIT XUT

C th dng phng php chit phn on coumarin bng dy cc dung mi c phn cc tng dn. a s tan trong ete du ha hoc ete etylic. Nhiu trng hp c th thu c coumarin tinh th ngay trong dung dch ete hoc ete du ha khi chit soxhlet. Do c mt ca nhm lcton, mt s coumarin cn c nhm OH phenol nn chng tan c trong kim nng. V vy c th chit chng bng kim long nng , dch kim em axit ha thu c kt ta coumarin. Cc glycozit ca coumarin c th c chit bng dung dch metanol long hoc etanol long (60-80%). Nguyn liu c loi tp cht bo bng ete du ho sau chit bng cn. Dch chit cn sau khi lai tp c th cho tinh th coumarin khi lm bay hi dung mi

 C th loi tp bng dung dch ch axetat. Cc glycozit coumarin c th thy phn bng dung dch axit hoc bng enzym. Vic xc nh phn ng c th dng sc k giy. phn lp cc coumarin c th dng sc k ct polyamit, silicagel hoc sc k lp mng. C th tinh ch coumarin bng phng php thng hoa (nhng ch p dng vi cc coumarin bn vi nhit ) Nh c tnh pht quang mnh ca coumarin di n t ngoi nn vic theo di, pht hin chng trong qu trnh chit xut c d dng

2.3.3 PHN TCH COMARIN

2.3.3.1. nh tnh Hu ht c tnh pht quang mnh di n t ngoi. Cc coumarin c Oxi v tr s 7 c th pht quang nh sng thng nht l khi c axit sunfuric. Coumarin cn cho phn ng mu vi thuc th Emerson ( 0,5% Na2CO3, 0,9% 4-aminoantipyrin, 5,4 % feroxianua trong nc). Do c mt nhm lacton, coumarin tan c trong dung dch kim v lm dung dch c mu vng, mu ny bin mt khi thm axit. C th kim tra thm coumarin bng phn ng c trng ca vng lacton. xc nh vng furan trong furanocoumarin bng phn ng Erhlich (dung dch 0,5% p-dimetylaminobenzandehit trong etanol, sau cho tc dng vi kh HCl) to mu vng cam.

2.3.3.2 Sc k giy v sc k lp mng + Sc k giy: Cc h dung mi thng dng l : Axit axetic: Nc (98:2) Dimetyl focmamit : Etanol (4:6) Pht hin vt: bng n UV, bng dd KI + I2 hoc bng cc thuc th mu.

+ Sc k lp mng: Cht hp ph thng c dng l silicagel Cc h dung mi thng c dng l: Ete du ha : etylaxetat (100: 3) Toluen : etyl focmiat : axit focmic (5 : 4 :1) CHCl3: CH3OH ( 9 : 1) CHCl3 : etyl axetat (1 :1) Hexan : etyl axetat (3:1) Benzen : axeton (9:1) Benzen : etyl axetat (9 : 1)

2.3.3.3 Phn tch quang ph S dng cc ph:UV, IR, MS, NMR Do c cu trc lin hp nn ph UV s dng kh hiu qu,nh c cc bng c trng. Cc loi ph IR, NMR cng c s dng xc nh cu trc rt hiu qu

2.4 QUINON
Quinon l nhng dixeton cha no, khi kh chng to thnh poliphenol. Cc poliphenol ny d b oxi ha to li quinon Quinon l nhng hp cht c mu, chng gp phn to mu sc ca cy v ng vt. Nhiu hp cht ca quinon nh vitamin K... tham gia vo qu trnh h hp, vo s vn chuyn electron trong cc t chc thc vt V mt phn b, chng c ri rc thc vt n bo, nm, nhng t gp trong cy mt l mm

2.4.1 CU TRC- PHN LOI

Da vo sn phm kh ha ca quinon c th chia chng thnh cc nhm: - Benzoquinon - Naphtaquinon - Antraquinon - Phenantraquinon Nu 2 nhm xeton cnh nhau ta c octo quinon, cn nu n cch nhau 1 nhm vinyl ta c para quinon O
O

O O

octo-benzoquinon

para benzoquinon

2.4.1.1 Benzoquinon
Benzoquinon c trong nhiu loi nm v trong thc vt bc cao Thng gp chng trong r ca mt s loi h cc v mt s loi khc . V d
HOOC
O O CH3

HO CH3O
O

OH

CH2CH2CH2CH2COOH muscafurin

COOH Spinusolin

2.4.1.2. Naphtoquinon Cc naphtoquinon c t mu vng n . Cht quan trng nht l vitamin K

O CH3

CH2CH=C(CH3)-CH2-[CH2-CH2-CH(CH3)-CH3]3

Vitamin K

2.4.1.3. Anthraquinon
Phn ln cc anthraquinon c nhm OH C1 v C8 v tn ti dng glycozit. Cc glycozit ny d b thy phn trong qu trnh chit xut. Cng thc ca mt s anthraquinon
OH O OH OH O OH

HO O Emodin R=CH3

R1

R2

Aloe-emodin R1=H; R2=CH2OH Phiscion R1= OCH3, R2 = CH3

Axit emodic R=COOH

 i khi cc glycozit l cc cht kh, kt hp vi mt oza hoc hai quinon kt hp vi nhau qua nhm xeton
OH O OH OO OH

H H

CH2OH OH OH

R1 H

COOH COOH OO OH

bacbaloin

senozit

2.4.1.4. Phenantraquinon Nhm ny t thy trong t nhin, ch gp mt s hp cht . V d:

OH HOOC O HO O OH CH=CH-CH=CH-COOH CH3 O O O CH3

axit teleporic

tashinon

2.4.2 CHIT XUT

Do benzoquinon v naphtaquinon l nhng cht khng phn cc nn rt d tan trong du bo v cc dung mi khng phn cc (ete , benzen). Cc anthraquinon do c nhiu nhm OH nn ni chung l nhng nhm phn cc mnh, chng tan c trong nc nng, trong cn v cc dung dch kim v dung dch cacbonat kim. Cc anthraquinon c th chit bng cn. Dch chit cn em bc hi cn v kt tinh li trong hn hp axeton-nc s thu c glycozit anthranon ton phn.

 thy phn cc glycozit c th dng axit axetic hoc HCl 5% trong cn 700C trong 1 gi. Cc aglycon sau khi thy phn c chit bng benzen. Bc hi benzen v tinh ch li thu c anthranon tinh khit. tch hn hp cc quinon c th bng sc k ct vi cht hp ph l polyamit, silicagel, MgO hoc canxi photphat.

2.4.3 PHN NG NH TNH

Dng cht kh kh chng thnh polyphenol (mt mu) v oxi ha chng tr li hp cht ban u (c mu). Cht kh thng dng l bohidrua v oxi ha bng cch ngoi khng kh. Mt s p-benzoquinon v p-naphtaquinon cho mu xanh hoc tm vi thuc th xiano axetat etyl v amonoac hoc vi dung dch 0,2% pnitrophenyl axetonitrin vi natri hydroxit 0,1N. Dung dch 1,4-naphtaquinon trong benzen c mu vng chuyn sang mu khi cho thm kim. Cc 1,2-naphtaquinon c mu , khi cho thm kim chuyn sang mu tm xanh.

 Cc hydroxianthraquinon cho mu sc khc nhau khi un nng vi dung dich magie axetat 0,5% trong metanol. Phn ng Bortrager: Dng pht hin anthraquinon v naphtaquinon. +Trong ng nghim, cho vi mg cht th c ha tan trong 1 ml dung dch 2-5% KOH. un nng 10 pht, ngui. Axit ha dung dch v chit bng benzen. Tch lp benzen qua ng nghim khc. Cho vo ng cha benzen 1 ml dung dch KOH, lc. Nu lp benzen mt mu v lp kim c mu l phn ng dng tnh. +Nu c dn xut anthron th lp kim vn gi mu vng vi hunh quang lc, nhng s chuyn sang mu nu thm vo 2 git dung dch H2O2 3-6%.

2.4.4. SC K GIY V SC K BN MNG

Sc k giy: c s dng nhiu khi phn tch quinon. Giy c tm vi dung dch 5 du vazelin trong ete-du ha. Dng hn hp imetylfocmamit-H2O (97:3) lm pha di ng. Cc quinon cho vt pht quang mu lc hoc tm di n t ngoi i vi cc hydroxiantraquinon dung h dung mi ete du ha bo ha trong metanol 97%. hin vt dng dung dch 0,5% mage axetat trong methanol v h nng giy 900C trong 5 pht. Cc anthraquinon c OH meta cho mu da cam,, Nu OH v tr para cho mu mn chin v nu OH v tr octo cho mu tm.

 Sc k lp mng: Cht hp ph thng dung l polyamit v silicagel. Nu dung silicagel trng trn knh th c th dung cc h dung mi sau: -Benzoquinon: n-hexan etylaxetat (17:3) : benzene- axit axetic (9:1) : clorofom-axit axtic (9:1) -Naphtaquinon: Ete du ha-etyl axetat (7:3) : benzene-ete du ha (7:3) -Antraquinon: n-hexan etylaxetat (85:15) : benzene- axit axetic (10:2) Hin mu: Phun dung dch KOH 10% trong methanol cc vt mu vng nguyn thy ca ntaquinon s chuyn thnh mu , tm hoc lc. Cng c thhin vt bng cch h knh ln l ammoniac v soi pht quang UV 2.4.5 PHN TCH QUANG PH

CHNG 2: PHENOL- AXIT PHENOL V DN XUT

Chng 2 : PHENOL-AXIT PHENOL V DN XUT

2.1.Phenol v axit phenol 2.2.Phenylpropanoit 2.3.Coumarin 2.4.Quinon

2.1 PHENOL V AXIT PHENOL

Cc phenol v axit phenol thng c nghin cu chung v chng thng song song tn ti vi nhau trong cy.
OH

HO

OH

OH

OH

OH
hydroquinon

R
R= H Resoxinol R= CH3:ocxinol R=OH Florogluxinol R= H : catechol R=OH Pyrogalol

HO
COOH CH3O
axit isovanilic

HO

COOH

R= H axit.p-hydroxibenzoic R= OH: axit protocatechic R= OCH3 axit vanilic

2.1.1 S PHN B

Cc axit phenol tn ti dng kt hp vi lignin to thnh este hoc vi cc oza di dng glycozit. Cc axit phenol thng gp l: axit phydroxibenzoic, axit pyrocatechic, axit vanilic, axit syringic, cc axit t gp l axit salixylic, axit o-protocatechic (axit 2,3-dihidroxibenzoic),. Ngc vi axit phenolic, cc phenol t do rt him thy trong cy. Hydroquinon l cht thng gp hn c, tip n l catechol, ocxinol, phlorogluxinol, pyrogalol

2.1.2PHN TCH PHENOL V AXIT PHENOL Vic tch phenol tt nht bng SKLM Thng thng nguyn liu ti hoc kh c nghin nh,thy phn vi kim hoc axit trong cn long 600 + Thy phn axit: dng axit HCl 2M trong 30 pht, sau khi ngui, lc, chit bng ete. Gn lp ete, ra bng nc vi ln, lm khan, bc hi n kh. Ha cn kh vi metanol lm dung dch chm sc k + Nu thy phn bng kim: NaOH 2M, thy phn trong 4 gi nhit thng. Dch thy phn em axit ha ri chit bng ete nh trn. Sau chm trn SKLM silicagel, xenlulo vi cc h dung mi khc nhau v hin mu cc hp cht khc nhau cho cc gi tr Rf v mu khc nhau

Cht hp ph l silicagel G vi cc h dung mi c phn cc trung bnh. Cc h dung mi thng c dng l : A: Axit axetic : clorofoc (1 : 9) (Cht hp ph l silicagel G) B: Etyl axetat: Benzen (9:11) ( Cht hp ph l silicagel G) C: Benzen: metanol: axit axetic (45: 8: 6) (Cht hp ph xenlulo MN 300) D: Axit axetic : nc ( 6 : 94) (Cht hp ph xenlulo MN 300) Cht hin mu : Vanilin + HCl

2.2. PHENYL PROPANOIT Phenyl propanoit l nhm hp cht phenolic t nhin gm mt mch nhnh 3 nguyn t cacbon gn vo nhn thm V mt sinh tng hp chng l dn xut ca axit amin thm phenylalanin .

2.2.1 CC HP CHT THNG GP

Hp cht ph bin nht l cc axit hydroxyxinamic. Trong c bn axit ph bin nht l axit ferulic, axit sinapic, axit cafeic v axit p-coumaric. C bn axit trn v c kh nng pht quang mu xanh hoc lc di n t ngoi, v vy d dng pht hin trn SKG hoc SKLM H3C-O R
HO
OH
R= H: axit p-cumaric R= OH: axit cafeic

CH=CH-COOH

HO

CH=CH-COOH
R
OH

R= H : axit ferulic R= OCH3: axit sinapic

CHNG 3

FLAVONOIT

3.1 I CNG 3.2 CHIT XUT 3.3 TCH RING TINH CH 3.4. PHN TCH FLAVONOIT

3.1 I CNG
3.1.1 NH NGHA : Flavonoit l nhng cht mu thc vt c cu trc c bn nh sau:
8 7 2'

3' 4'

6 5

3 4

6'

5'

Ti cc vng c lin kt vi mt hay mt vi nhm hydroxyl t do hay thay th mt phn. V vy v bn cht chng l nhng poliphenol c tnh axit.

3.1.2 PHN LOI

Da vo s oxy ha ca vng pyran trung tm, ngi ta phn flavonoit thnh cc loi khc nhau 3.1.2.1 Flavon :
8 7 2'

O A C

3'

B
3 6' 5'

4'

6 5

Khung cu trc chung ca flavon

.V d
HO
7 8 2'

3'

HO OH
7

2'

3' OH 4'

OH

3 4

6'

5'

6 5

3 4

6'

5'

OH

OH

Apigenin

Luteolin

3.1.2.2. Flavanon khc flavon l khng c lin kt i C2-C3 Chng u c OH vng A hay vng B. Trong t nhin n thng c mt ng thi vi flavon tng ng.

 Cc cht thng gp:

2'

Glu-O
7

3'

Rh-Glu-O
7

2'

1 2

3' OH 4'

OCH3

3 4

6'

5'

6 5

3 4

6'

5'

OH

OH

prunnin

hesperidin

3.1.2.3. Flavonol khc vi Flavonon l c nhm OH C3 N l loi hp cht ph bin trong t nhin, c bit l trong cy ht kn

 Trong cy ht kn c cha nhiu cht sau: kaempferol, quecxetin v mirixetin

HO
7

2'

OH
3'

HO OH
7

2'

3'

OH
4'

OH

3 4

OH

6'

5'

6 5

3 4

OH

6'

5'

OH

quecxetin

mirixetin

3.1.2.4. Dihydroflavonol : Tng t nh flavonol nhng khng c ni i C2-C3 Cht ph bin nht l 7-hydroxidihydroflavonol
8 7 2'

3'

HO
4' 3 6' 5' 7

2'

3' 4'

6 5

OH

6 5

3 4

OH

6'

5'

(khung dihydroflavonol)

7-hydroxidihydroflavonol

3.1.2.5.Chalcon L loi flavonoit vng m, hai nhn thm lin kt vi nhau qua mch ba cacbon , khng no. Ph bin nht l butein
3' 4' 5' 6' 2' 2 3 3' 4 6 2 2' 3

OH
4

O
5

HO4'
5' 6'

6 5

OH

Khung Chalcon

OH O

Butein

3.1.2.6. Isoflavon l nhm isoflavonoit ph bin nht c nhiu tc dng

cha bnh V d daidzein c trong ct cn, hoc fomonometin c trong cam tho
8 7

R1O
7 3

6 5

OR2 O

(khung isoflavon)

daidzein R1 = R2 = H formonometin R1 = H, R2 = CH3

Ngoi ra cn c nhiu loi khc nh auron, rotenoit antoxyanidin, leucoantoxifanidin, neoflavonoit, biflavonoit.

3.1.3

PHN B

N l loi hp cht phn b rng nht trong thin nhin ( ln hn 2000 cht) Ni chung n khng c mt trong to v nm. Cc loi thc vt bc cao u c mt flavonoit, v n c mt lin kt cc b phn ca cy : thn, l, r, g v khu tr thnh t bo. Flavonoit tham gia vo s to thnh mu sc ca cy (c bit l hoa) l mt trong nhng chc nng c bn ca flavonoit i vi cy.

3.1.4 VAI TR CA FLAVONOIT TRONG CY

3.1.4.1 Cc phn ng sinh ha Cc nhm OH phenol ca flavonit c vai tr trong s ha tan cc cht v di chuyn d dng qua cc mng sinh l Mt s flavonoit c tc dng nh l cht chng oxi ha, bo v axit ascorbic trong cy Mt s c tc dng c ch cc enzym v cc cht c ca cy

3.1.4.2 Vai tr c ch v kch thch sinh trng ca cy Cc nhm OH c vai tr quyt nh vai tr c ch v kch thch sinh trng ca cy V d trong cy i cc flavonoit c cc nhm OH v tr 4 lm tng cng hot tnh ca enzim lm kch thch sinh trng, cn khi OH v tr 3v 4 li c ch... Flavonoit cn tham gia vo qu trnh h hp quang hp

3.1.4.3 Vai tr to mu sc To ra sc hp dn d ong bm n th phn gp phn cho cy tn ti & pht trin. Trong vic to mu cc flavonon, flavonol,chalcon cho mu vng trong khi cc autoxyanin cho cc mu hng , , tm hoc xanh thm 3.1.4.4 Vai tr bo v cy To v ng ng vt n c khng thch n cy

3.1.5 VAI TR CA FLAVONOIT TRONG Y HC

Nhiu flavonoit c tc dng c ch h hp v s ti sinh sn i vi nhiu chng vi khun Nhiu flavonoit c tc dng i vi mt s dng ung th (ung th vm hng) Nhiu flavonoit c tc dng nng cao tnh bn ca thnh mch mu c bit l cc flavonoit c nhm OH cc v tr 3, 4. Rutin l cht tiu biu ca tc dng ny Tc dng estrogen: Mt s flavonoit c tc dng gy sy thai ng vt Nhiu flavonoit c tc dng chng d ng, chng co git, gim au

3.2 CHIT XUT

Khng c phng php chung v chit xut.Tuy nhin c th nu nguyn tc chung nh sau : Do flavonoit c hu ht cc b phn ca cy (r, g, l, hoa, qu, ht, sp), trng hp chit lp sp ngoi l th phi dng ete ra ly sp, cc b phn khc ni chung sy kh, tn bt chit. Ni chung cc glycozit ca flavonoit c phn cc mnh hn aglycon tng ng. i vi aglycon m c t nhm OH , n phn cc yu th dng cc dung mi phn cc yu nh benzen, cloroform, etyl axetat chit.

 Cc flavonoit nhiu nhm OH nh flavonol cc glycozit.. l nhng cht phn cc mnh, nn c th chit chng bng cn, nc hoc l hn hp ca chng - Dung mi c th p dng cho hu ht flavonoit l cn 80%,60% ( c th l C2H5OH,CH3OH 60%,80%) - Chit cc glycozit : c th dng nc nng - Do c OH phenol nn c th dng dung dch kim long chit

3.3 TCH RING TINH CH

3.3.1.TCH S B: Nhm mc ch loi tp v tch phn on i vi hn hp cc flavonoit . C th p dng cc cch sau : 1. Dng hn hp cc dung mi C th dung hn hp cc dung mi c phn cc khc nhau, khng tan vi nhau phn on : V d : hn hp aglycon v glycozit : CHCl3-H2O, H2O etyl axetat , Butanol-ete du ha

2. Phng php c in: Dng ch axetat ta flavonoit (phenol ni chung) Ha tan cao hn hp vo cn long,cho dung dch ch axetat vo ta flavonoit. Lc, li cho ta ny tan trong cn long khuy u thnh dung dch ng nht. Sau cho 1 trong cc dung dch sau y : Na3PO4;Na2S kt ta ch. (Dng photphat hoc sunfat tt hn sunfua v cc ta ny hp ph flavonoit t hn sunfua ch )

3. Dng than hot tnh Cho vo dung dch cao hp ph flavonoit sau dng dung mi ra than ly li flavonoit Khi ra dung mi t khng phn cc => phn cc t => phn cc mnh V d : ete => methanol => nc nng

4. Tch phn on bng cc dung dch c kim khc nhau V d : u tin dng dung mi hu c CHCl3, etyl axetat; sau ln lt chit dung dch trn vi dung dch natriborat Na3BO3, NaHCO3 v Na2CO3 10% Axit ha cc dung dch kim, flavonoit s kt ta, lc thu kt ta hoc chit ly flavonoit bng dung mi hu c (CHCl3, etyl axetat )

3.3.2.SC K CT
L phng php tt nht tch ring flavonoit. - Cc cht hp th thng dng l poliamit, silicagel, xenluloza - Dung mi dng ra l : ete du, Benzen, CHCl3, ete, etyl axetat, axeton, ru etylic, ru metylic, H2O, p dng cho hu ht Flavonoit

3.3.3 TINH CH
Phng php tt nht l lc qua ct cht hp th ( poliamit, silicagel, bt xenlulo), hn ch dng than hot v than hot hp th rt mnh flavonoit Kt tinh flavonoit trong etanol , metanol hoc cn axeton, cn-etyt axetat.

3.3.4 THY PHN GLYCOZIT FLAVONOIT

C th dng axit hay enzym i vi enzym thng dng l -glycosidaza v hu ht cac glycozit flavonoit l -glycosit Cch thy phn bng axit: C th dng cc loi axit khc nhau (HCl, HCOOH, CH3COOH) vi nng thch hp, thm mt t cn ha tan glycozit, un si cch thy t vi pht n 1 gi hoc c th lu hn ty theo tng loi hp cht v iu kin tin hnh phn ng. ngui. Chit dung dch thu c bng ete ly phn aglycon. Phn dch nc cha cc ng, c th phn tch hn hp ny bng sc k giy

3.4. PHN TCH FLAVONOIT

3. 4.1 PHN TCH NH TNH 3. 4.1 .1 Cc phn ng mu flavonoit a/ Amoniac ( NH3): Khi c mt ca hi NH3, nhiu flavonoit thay i mu, c th quan st s thay i mu bng mt thng hoc di n t ngoi Cnh hoa mu trng + hi NH3 cho mu vng l flavon hoc flavonol Cnh hoa mu vng + NH3 cho mu vng cam hoc hng l Chalcon, auron Cnh hoa mu bt k ( vng , , tm) +NH3 cho mu thm l autoxyanidin

b/ Phn ng xianidin ca Wilstater -Benzopyron trong a s flavonoit c phn ng vi HCl +Mg bt Trong 3 ng nghim, cho vo mi ng 1-2 ml dung dch thuc th ng 1: lm i chng ng 2: thm 0,5 ml HCl c + 3-5 ht Mg. Quan st, nu sau 10 n t mu vng, , n xanh l c flavonoit. C th nhn nh thm - Vng cam- l flavon - thm l flavonol, flavanon - Mu ngay sau khi cho HCl l chalcon, auron ng 3: Thm 0,5 ml HCl c un cch thy 5 pht, nu c mu tm l leucoantoxyanidin

c/ Thuc th FeCl3 trong cn hoc nc => cho mu ca phenol v flavon d/ Thuc th SbCl5 1% trong tetraclorua cacbon => c ta mu hoc tm l ca Chalcon, mu vng hay da cam => flavon ( phn ng Marini Bettolo ) e/ Thuc th axit boric trong anhydrit axetic Cc hp cht 5- hydroxi flavonon cho mu cam hay

g/ H2SO4 Ha tan mt t cao ca dch chit flavon vo axit sunfuric c, nu cho mu: + vng ti : flavon, flavonol + mu cam : flavonon + hoc tm : chalcon, auron h/ Kim : Dch chit flavonoit, nu trong dung dch kim. Nu c: + Mu vng :flavon, flavonol, + Mu cam, : flavon, isoflavonon + Mu tm : Chalcon + Mu c ph: flavonol + Mu xanh: autoxyanin

3. 4.1.2 Sc k giy v sc k lp mng

Nu dng h dung mi thch hp ta c th tch c cc flavonoit khc nhau c Rf khc nhau

3.4.2 PHN TCH CU TRC BNG PHNG PHP QUANG PH Xem ti liu tham kho

CHNG 4

TINH DUHP CHT LACTON TECPENOIT

4.1 I CNG V TINH DU

Tinh du (cn c tn khc l cht thm, hng thm, tinh du) l hn hp ca nhiu cht bay hi, c mi c bit c ngun gc ch yu t thc vt, thu c bng cch chng ct hoc chit bng dung mi hu c N t tan trong nc, d tan trong ete, du bo, d bay hi, V phn b: Cy cha tinh du phn b kh rng trong nhiu h thc vt. Trong cy, tinh du c th khu tr ti l, hoa, qu, r,v, thn g Thnh phn tinh du ca mi b phn ca cy c th ging hoc khc nhau

4.2 THNH PHN HA HC

N l hn hp ca nhiu hp cht khc nhau: cc hydrocacbon bo hoc thm, cc dn xut ca chng nh ancol, andehit, xeton, este , ete Nhng chung qui li chng c hai nhm chnh l tecpenoit v cc dn xut ca phenol. Nhm tecpenoit ch yu l monotecpen v sesquitecpen

4.2.1 TECPENOIT
4.2.1.1. Monotecpen (C10) : Da vo c im cu trc, c th chia thnh 3 nhm chnh l: Loi khng vng ( nh geraniol), Loi 1 vng (nh limonen) v Loi 2 vng (nh pinen). Trong mi nhm c th l loi khng nhm chc hoc c th l nhm chc (ancol, andehit, xeton.. )

 Loi khng vng

OH OH

Geraniol

linalol

myrxen

 Loi mt vng

O OH

-tecpineol

limonen

tecpinolen

1,8- xineol

O OH

menthol

menthon

carvon

 Loi 2 vng
O O

pinen

thujon

camphor

fenchon

4.2.1.2 Sesquitecpen (C15)

Sesquitecpen lun c mt cng vi monotecpen trong tinh du. Sesquitepen trong tinh du c nhit si trn 200oC, do khi chng ct th chng c hm lng khng cao.

 Mt s sesquitecpen thng gp

Cadinen

Humulen

Selinen

Caryophyllen

c im cu to ca tecpenoit
Ng i ta coi tecpenoit l sn phm ca s kt hp cc phn t isopren vi nhau. l qui tc isopren ca tecpenoit Cch xc nh cu trc ca tecpen. Da vo cc phn ng c trng v qui tc isopren + X s c mt lin kt i: + Xc nh s c mt H linh ng + Xc nh s c mt -CHO + Xc nh v tr lin kt i: OXH, Ozon phn + Qui tc isopren: kt hp u vi ui

4.2.2 DN XUT PHENOL Nhm ny l thnh phn ch yu ca tinh du ca mt s cy trong h Umbeliferae, tiu hi, mi, tha l a s hp cht ca nhm ny l loi phenyl propanoit c cu trc C6-C3 c mt hay nhiu nhm OH t do hay b thay th H ca OH. V vy v bn cht chng l hp cht phenol

 Mt s hp cht thng gp
CH=CH-CH3
CH2-CH=CH2

CH2-CO-CHO

CH2-CH=CH2

OCH3 OCH3 OCH3 OCH3 OH

Anetol
CH=CH-CH3

Estragol
CH2-CH=CH2

Anison
CHO

Eugenol
CHO

OCH3 OH O O

OCH3 OCH3 OH

OCH3

Isoeugenol

Mirislixin

Anisandehit

Vanilin

 Thnh phn ca tinh du tuy khc nhau nhng thng c ngun gc sinh tng hp t mt s tin cht nht nh. V vy mi loi tinh du u c mt thnh phn ch yu v mt s thnh phn ph c cu trc tng t. V d + Tinh du bc h c 50-70% menthol, ngoi ra c menthon, limonene, -pinen, cadinen. + Tinh du tiu hi c 70-90% anetol, ngoi ra c cc thnh phn khc tng t

4.3 CHIT XUT

Tinh du khu tr trong nhiu b phn khc nhau ca cy: l, hoa , thn, v thn, r, quDo ty theo hm lng, thnh phn ha hc m ta c th tch ton b cy hay tng b phn ca cy Nguyn liu chit tinh du c th kh hoc ti, nhng nu c iu kin th dng ti tt hn v thnh phn khng b hao ht v phn hy trong qu trnh lm kh. Ty thuc vo cc dng tinh du c trong nguyn liu trng thi t do, hay ht hp glycozit m ngi ta dng cc phng php khc nhau tch chng.

 Nhn chung cc phng php c dng tch tinh du phi t yu cu c bn nh sau : - Gi cho sn phm tinh du c mi t nhin nh nguyn liu - Qui trnh k thut khai thc ph hp, thun tin, nhanh chng - Phng php tch tinh du tng i trit khai thc ht tinh du trong nguyn liu m c u t t tn nht Trong thc t ngi ta thng s dng cc phng php sau:

4.3.1 PHNG PHP C HC

Phng php ny thng dng tch tinh du nm v (qut, cam, bi). Khi tc dng mt lc ln cc t bo cha tinh du v, tinh du chy ra. Phng php ny c u im nhanh, tinh du gi c mi t nhin. Tuy nhin phng php ny khng s dng c cho nhiu nguyn liu cha tinh du hoc hm lng tinh du trong nguyn liu thp

4.3.2 PHNG PHP CHNG CT

y l thng dng nht v thit b khng phc tp lm m c kh nng ly kit c tinh du trong nguyn liu v da vo nhit si cc cht khc nhau c th ly c cc phn on vi thnh phn ch yu l mt s cht no . Tuy nhin c nhng nhc im l: Cc tinh du c thnh phn kh bay hi, hay mt s tinh du c thnh phn khng bn vng d thay i nhit cao th khng p dng c

4.3.3 PHNG PHP TRCH LY (ngm chit)

Phng php ny p dng cho cc nguyn liu tinh du m thnh phn ha hc ca n khng bn d thay i thnh phn v mi v nhit cao. Ngoi ra cc nguyn liu c hm lng tinh du qu t Phng php trch ly ngi ta thng dng cc dung mi hu c d bay hi nh ete du ha, ete, ancol, axeton hoc cc dung mi kh bay hi nh m ng vt, vazlin. Gn y ngi ta dng CO2 lng trch ly tinh du.

4.3.4 PHNG PHP HP PH

i vi mt s nguyn liu, nht l cc loi hoa, c th ko di thm thi gian to hng sau khi hi ri khi cy, ngi ta dng PP hp ph Trc y ngi ta dng m ng vt hp th. Nhng phng php ny mt nhiu thi gian, thao tc th cng, khng p dng c qui m cng nghip, gi thnh t. Hin nay ngi ta dng than hot tnh lm cht hp ph, bng cch cho lung kh m i qua lp hoa ti, nhm ko di thi gian sng ca hoa. Lung kh m khi cho qua lp hoa ti li cun theo cht thm i vo thit b hp ph Cht hp ph than hot s gi cc cht thm => sau trch ly => thu tinh du.

4.3.5 PHNG PHP LN MEN Phng php ny dng cho cc loi tinh du dng qu v cc dng m tinh du khng trng thi t do m cc dng kt hp khc ( dng glycozit l ch yu). Trc khi a s dng phng php tch tinh du thng thng, ngi ta phi dng men, enzym x l s b trc to tinh du dng t do

4.4 TNH CHT CA TINH DU

4.4.1 TNH CHT VT L

nhit thng , tinh du ch yu th lng, tr mt s trng hp c bit th rn Tinh du c tnh d bay hi, c th ct ko bng hi nc rt t tan trong nc, tan trong cn, cc dung mi hu c,cc du m. Ty theo thnh phn, tinh du tan trong cn vi nng khc nhau, c th tan mt phn trong dung dch kim (phenol), dung dch NaHSO3 (andehyt).

 Tinh du khng c mu hay c mu hi vng, c loi tinh du c mu xanh hay mu hoc mu nu thm Tinh du c v hi cay v hc. T trng : nm trong khong 0,850- 0,950. Mt s nng hn nc (tinh du qu, nh hng ). T trng ca tinh du ph thuc vo thnh phn ca tinh du + Nu t l cc hidrocacbon tecpenoit cao th t trng thp (<0.9). +Nu t trng cao hn 0,9 th thng cha nhiu hp cht cha oxi (este, ancol, andehit, xeton). +Nu t trng ln hn 1 thng cha nhiu thnh phn cha nhn thm (eugenol, andehit xinamic)

 N thng c nng sut quay cc : c th quay phi (+) nh tinh du long no hay tri quay tri (-) nh tinh du bc h Ch s khc x 1,450-1,560: ch s khc x ca tinh du ph thuc vo thnh phn ha hc ca tinh du. Nu tinh du c cha nhiu nhn thm hoc vng c nhiu nhm cha oxi th t trng v ch s khc x u cao. im si : tinh du c c im si cao 150-180oC nu thnh phn l cc tecpen 250-280oC nu l sesquitecpen; v trn 300 oC nu thnh phn l l politecpen . Mt s tinh du c th pht quang di n t ngoi (eugenol, andehyt ximamic)

4.4.2. TNH CHT HA HC Di tc dng ca nhit , nh sng, khng kh, nc. tinh du d b oxi ha v mt phn bin thnh nha. Khi b oxi ha, thnh phn ca tinh du b thay i: ancol chuyn thnh andehit, axit , andehit chuyn thnh axit Nhiu thnh phn c nhm chc khc nhau (ancol, andehit, phenol) cc nhm ny gip cho vic kim nghim, chit xut tinh du

4.5 PHN TCH TINH DU

4.5.1 PHN TCH S B Da vo t trng v ch s khc x ca tinh du ngi ta c th s b phn on c thnh phn ch yu ca tinh du. + Nu t trng di 0,9 v ch s khc x di 1,47 th tinh du c t l cao hidrocacbon tecpen hoc mch thng + Nu t trng cao hn 0,9 v ch s khc x di 1,47 th tinh du c nhiu hp cht mch thng c nhiu nhm chc cha oxi + Nu t trng nh hn 0,9 v ch s khc x ln hn 1,47 th tinh du c nhiu hp cht thm + Nu tinh du c nhiu hp cht thm hoc vng c nhiu nhm chc c oxi th t trng v ch s khc x u cao

 ha tan ca tinh du + Trong hn hp cn-nc: ty thuc vo t l ca cc cht c cha oxi + Nu tinh du tan nhiu trong dung dch KOH chng t chng c nhiu hp cht cha OH phenol +Hn hp tinh du vi cacbon disunfua vi lng bng nhau, nu c l tinh du c ln nc.

4.5.2 PHN NG MU Pht hin ancol trong tinh du: Cho 1 git tinh du, 0,3ml CS2, 100mg KOH nghin nh. Lc trong 5 pht. +Nu c xut hin mu vng hay ta vng l c ancol. + Nu khng c phn ng, thm 1-2 git amoni molipdat 1%, sau axit ha t t bng axit sunfuric, lm lnh hn hp v thm 3-4 git CHCl3. Lc mnh hn hp v gi yn, nu lp CHCl3 c mu tm l phn ng dng tnh

 Pht hin andehit xeton : Ha tan 1 git tinh du vo 2-3 git etanol tuyt i + 1 git thuc th 2,4-dinitro phenyl hydrazin ( ha nng 5 gam 2,4-dinitro phenyl hydrazin trong 60ml axit photphoric 85% c pha bng 39,5 ml etanol). + Nu c kt ta l c nhm cacbonyl thm + Nu c mu vng cam l c ni i v tr ,, +Nu c mu vng l c cacbonyl bo ha. Thm 1 git KOH 2N trong etanol mu s chuyn sang mu thm, mu mn chn hoc mu xanh Ngoi ra c th pht hin andehyt bng thuc th Schiff (axit fucsinsunfuro), Tollens.

 Pht hin phenol : dng FeCl3 Pht hin hp cht cha bo ha: cng Br2/CCl4, nu c hp cht khng bo ha th mu ca brom bin mt sau 1 pht Cc phenol v mt s hp cht thm c nhm metylen cng c phn ng dng tnh. phn bit nn lm thm phn ng Baeyer.

4.6 TCH CC THNH PHN TRONG TINH DU i vi cht rn nhit thng: lnh, lc ly tm (mentol ca bc h) Tch axit t do ra khi tinh du: nu khng c phenol hay nhm cacbonyl th lc vi kim long, ra tinh du bng nc, lm kh bng Na2SO4 Phng php tch c in l chng ct phn on p sut thng hay p sut thp. Sc k ct: Cc phn an thu c c th tip tc tch hoc tinh ch bng sc k .

4.7 HP CHT LACTON TECPENOIT

Cc hp cht lacton ca tecpenoit c nhiu ng dng quan trng trong y hc v nng nghip. c bit l sesquitecpen lacton, chng c tc dng c t bo, khng ung th, khng vi trng, dit cn trng, dit nm mc a s cc lacton tecpenoit u c v ng, mt s cht c mi v kh chu. c tnh c th l mt trong cc chc nng ca lacton l bo v thc vt Vng laton khng no trong cu trc c quan h n v ng v cc tc dng c v khng trng. Cc lin kt khng no trong vng lacton d dng tc dng vi nhm OH SH ,NH ca cc axit amin v protein trong vi khun, bin thnh cht c v lm ri lon cc qu trnh sng ca vi khun.

4.7.1 SESQUITECPEN LACTON

4.7.1.1.I CNG Sesquitecpen lacton c b khung c bn 15C (C15) :V l thuyt chng gm ba n v isopren, mt phn ca b khung l mt metyl butenoit Sesquitecpen lacton phn b ch yu dng cy tho v cy bi. c bit tp trung nhiu trong h cc (Asteraceae). Trong cy cy chng tp trung ch yu trong l, hoa v ni chung l cc b phn trn mt t.

 Phn loi : Sesquitecpen lacton c sp sp theo cu trc ca b khung c bn ca sesquitecpen. Chng c khong 20 dng cu trc ca sesquitecpen lacton khc nhau: Cc dng thng gp nht l :
eudesmanolit (I), guianolit (II), pseudoguianolit (III) germacranolit (IV), eremophilanolit (V), eudesmanolit (nh Santonin (VI))

 eudesmanolit (I), guianolit (II), pseudoguianolit (III)

14 14 1 2 3 4 9 10 8 7 6 5 3 4 11 12 13 15 2 1 5 6 7 10 9 8

15 (I)

O O
(II)

11 12

13

O O
(III)

germacranolit (IV), eremophilanolit (V),Santonin (VI) ,

14 1 2 3 4 9 10 8 7 6 5

11 12

13

15

O
(IV)

O
(V)

O
(VI)

 artemisinin (VII) l mt loi sesquitecpen lacton c mt trong l cy thanh cao hoa vng (Artemisia annua L. ) mc nhiu ni min Bc Vit Nam v nhiu nc trn th gii c tc dng chng vi trng st rt .
14 2 3 1

CH3
7

H
6 5

8 9 10

H3C
15

O-O O H
4

H H CH3
13

12

11

(VII) Artemisinin

4.7.1.2 CHIT XUT Ni chung sesquitecpen lacton l cht phn cc trung bnh, n rt t tan trong dung mi khng phn cc nh ete du v trong nc, Chng tan nhiu trong CHCl3, ete etylic, t tan trong metanol v etanol ngui, nhng tan nhiu khi nng.

a. Chit bng CHCl3 Bt nguyn liu c th loi cht bo bng ete du, sau chit CHCl3 t 6-12 gi. Ct thu hi CHCl3. Cn ha tan vo C2H5OH nng. Sau cho thm 1 lng tng ng Pb(CH3COO)2 5% v vi git HCl ( dung dch c mi trng axit yu). yn 1 ngy, lc b kt ta. Thu hi cn (cn li nc), chit nc ny bng CHCl3. Ly lp CHCl3 , ra bng nc, lm khan dch CHCl3 bng Na2SO4. Ct thu hi dung mi thu c cn sesquitecpen. Tinh ch bng sc k ct oxit nhm

b. Chit bng cn (metanol hoc etanol) Bt nguyn liu c chit bng cn nng. Thu hi cn cn li th tch. Thm lng tng ng nc. Dng ete du chit loi cht bo. Sau chit sesquitecpen lacton bng CHCl3. Dch chit CHCl3 ra nhiu ln bng nc, lm khan bng Na2SO4 . Thu hi CHCl3 . Tinh ch nh trn thu c sn phm

c. Chit bng cn - nc. Chit nguyn liu bng cn (ngm kit ) . B cn li cho vo nc un si 1-2 gi. Dch chit cn thu hi cn. Cn cn li ha vo dch chit nc Dch nc c chit bng CHCl3 hoc butanol, thu hi dung mi thu c cn. un cn ny vi benzen, lc nng, sau lnh, sesquitecpen s kt ta. Lc ly ta. Tinh ch bng ct oxit nhm

d. Chit bng dung dch kim nng Vng lcton trong mi trng kim un nng s m ra to thnh mui tan c trong nc. Dung dch ny nu trong dung dch axit s ng vng tr li v v kt ta trong nc. Phn ng:
OH O

+ NaOH
O OH + COONa HCl O + NaCl O + H2O COONa

4.7.1.3. PHN NG NH TNH Phn ng nh tnh ca sesquitecpen lacton da vo s c mt ca nhm lacton. Ngoi ra c th p dng cc phn ng ca tecpenoit. - Phn ng vi hidroxymat feric: Cho mu mn chn. - Cc hp cht c vng lacton , khng no cho phn ng vi cc thuc th: + Legan (natri nitroprusinat = natri nitroxianua): mu hng + Baljet (axit picric): Mu vng da cam + Kedde ( axit 3,5-dinitrobenzoic trong metanol): mu hng

- Cc hp cht lacton khng no (, v ,) u kh dung dch Tollen, trong cc hp cht lacton , cho phn ng mnh hn do c th kh dung dch AgNO3 trong mi trng trung tnh. Ngi ta c th dng phn ng vi thuc th Tollen phn bit lacton no v khng no - Ngoi ra sesquitecpen lacton cho phn ng dng tnh vi thuc th Libermann Burchat (1 ml anhidrit axetic + 1mlCHCl3 + 1 git H2SO4)dung dch SbCl3, Vanilin photphoric.

4.7.2. IRIDOIT
4.7.2.1. I CNG Iridoit l nhm hp cht thin nhin c cu trc monotecpen vi mt vng xiclopentan v mt vng -lacton ( hoc dn xut) thng l khng no
o o

4.7.2.2.CHIT XUT
C th chit glycozit iridoit bng cn hc bng nc nng. Dch chit cho bay hi dung mi, cn thu c ha tan vo t nc, lng, lc b cn. Dch lc loi tp bng cch lc vi ln bng benzen hoc etyl axetat (hoc c th loi tp bng ch axetat nh i vi sesquitecpen lacton)

4.7.2.3 PHN NG NH TNH V PHN TCH HP CHT IRIDOIT

a. Phn ng nh tnh
Phn ng c trng l phn ng Trim-Hill (10 ml CH3COOH+ 10ml dung dch CuSO4 0,2% v 0,5 ml HCl c) nu c iridoit th s xut hin mu xanh Ngoi ra iridoit c cc phn ng mu vi mt s thuc th tecpenoit (Liebermann Burchat; SbCl3 trong CHCl3) v vi dung dch anisandedit trong H2SO4

b. Phn tch
C th phn tch bng sc k giy, lp mng Sc k lp mng vi cht hp ph l silicagel G vi cc h dung mi l + CHCl3 : CH3OH : H2O (6 :4 :1) + Xiclohexan : etyl axetat (3: 7) + n-butanol : axit axetic : nc ( 4: 1: 5) Sc k giy vi h dung mi l + n-butanol : axit axetic : nc +isopropanol bo ha trong nc. Cht hin mu anisandehit trong axit sunfuric hoc SbCl3 trong CHCl3 hoc c th dng thuc th Trim Hill

CHNG 5

STEROIT

M U
Steroit l nhng hp cht dn xut ca xiclopentenophenantren. Chng gm nhiu hp cht thin nhin trong c sterol, axit mt, hocmon gii tnh, vitamin D Nhiu steroit c nhiu tnh cht quan trng c dng trong iu tr bnh (iu ha h thng ni tit, cha vim khp). Hin nay steroit cn dng iu ch thuc trnh thai. Nhiu ng dng ca steroit trong chn nui v nng nghip

5.1 CU TRC V CCH GI TN

5.1.1 CU TRC Steroit l nhng cht do hidro mt phn hay hon ton 1,2xiclopentenophenantren. thun li, cc vng ca h thng ngi ta nh s theo th t ch s latinh A, B, C, D. Trong cc cht ny R v R l nhng nhm metyl hay dn xut oxi ha ca chng, cng c khi cc nhm khng tn ti. Cn nhm R l nhng gc ankyl khc nhau. Cn nhm R l nhng gc ankyl khc nhau
R
R'

R''

C B

KHUNG STEROIT

 5.1.2.CCH GI TN Tn ca steroit c da vo mt s cu trc c bn thng l tn ca cc hidrocacbon tng ng (hidrua nn).sau thm tip u ng v tip v ng ch r bn cht ca nhm th. V tr ca nhm th c ch nh bi ch s ca nguyn t cacbon m n lin kt. Vic nh khung sn cacbon ca steroit nh sau:

21 18 13

20 17

22 23 24 16 27 15 25 26

12 19 2 1 10 11

C D
8 14

A B7 3 5
4 6

5.1.2.1 Cu hnh v S nh hng trong khng gian ca cc nhm th c k hiu bng ch v . ( nm trn mt phng vng: nt m. (nm di mt phng vng): nt t on k hiu l (xi) cha xc nh c cu hnh: hnh ngon ngoo Di y l mt s hidrua nn quan trng

 V d
CH3 CH3 CH3 CH3 H CH3

CH3

H H

Estran

Androstan

Pregnan

.
CH3

21

20

22

23

CH3 24

CH3 H

21 22 H3C 20 CH3

23 24 25 27 26

H
CH3

H H

Cholan

Cholestan

5.1.2.2 ng phn lp th: Lun ch nh ng phn lp th C5 bi tip u ng 5 v 5, tr phi n l lin kt i. Nu mt nguyn t cacbon chung cho 2 vng khc vi C5 v c cu hnh bt thng s c ch nh bi tip u ng thch hp.

C2H5
CH3

CH3

H HO
. 3 ,

5-pregnan

CH3 OH CH3 H CH3 CH3 COOH

21 H3C CH3

20

22 23

24

25 27

26

H HO

OH

HO

axit 3 ,7 ,12 -trihidroxi5 -cholan-24-oic (axit cholic)

cholest-5-en-3 -ol (cholesterol)

 5.2 STEROL 5.2.1 CU TO Ngoi cholesterol, cc sterol thng gp l stigmaterol, -sitosterol, ergosterol

5.2.2 .PHN NG MU CA STEROL

Khng c phn ng mu ring cho sterol, ch c phn ng mu chung cho steroit. Nhng phn ng thng s dng l 5.2.2.1. PHN NG LIEBERMANN BURCHARD (anhidrit axetic 1ml + cloroform 1ml ( lnh O0) + 1 git axit sunfuric c) Cho 1 git dung dch thuc th vo dung dch thuc th. Nu c sterol xut hin mu xanh nht, lc, hng,cam, Mu ny bn vng trong mt thi gian. y l phng php th cholesterol trong mu. 5.2.2.2 PHN NG ROSENHEIM Dung dch th trong CHCl3 + dung dch axit tricloaxetic 90%. Nu c sterol vi nhn dien s cho mu tm. Mu ny chuyn sang xanh l sau 20 pht

5.2.2.3 PHN NG SALKOWSKI

( H2SO4/CHCl3)

Cho thuc th vo dung dch th, nu c mu th c cholesterol v cc sterol khc 5.2.2.4 PHN NG TORTELLI - JAFFE Ha tan 0,5 mg cht th bng 0,2 ml axit axetic v 0,2 ml CHCl3. Cho dc thnh ng 1,2 git dung dch nc brom trong CHCl3. Nu c sterol c lin kt i s c mu vng lc gia hai lp

5.2.3 PHN B
Sterol phn b rng, n thng c mt cng vi ankaloit hoc saponin steroit. Chng c mt trong tt c cc b phn ca cy, nhng ch yu l cc ht c du dng t do hoc este, mt s dng glycozit.

5.2.4 CHIT XUT

Do sterol l hp cht hu nh khng phn cc, rt t tan trong nc , tan nhiu trong dung mi km phn cc hoc khng phn cc, nn dng cc dung mi ny (ete du, ete, benzen, CHCl3) chit xut. Mun chit xut dng glycozit c th dng cn. Sn phm chit thng thng l hn hp cc este ca sterol vi lipit, carotennn phi x phng ha sau mi chit sterol bng cc dung mi hu c. Tinh ch bng kt tinh phn on

5.2.5 PHN TCH STEROL

5.2.5 1. SC K LP MNG Thng thng dng sc k lp mng vi cht hp ph l silicagel G hay oxit nhm. Cc h dung mi thng dng l + Benzen : etyl axetat ( 4:1) + Benzen : cloroform (9 :1) Hin mu bng dung dch SbCl3 trong CHCl3, H2SO4 20% trong cn hoc H2SO4 nc (1:1) v h nng Mt s sterol c th pht hin vt bng cch soi trn n t ngoi

5.2.5 2. SC K KH Thng c s dng rng ri phn tch sterol ni ring v steroit ni chung

5.2.5.3 XC NH CU TRC Thng dng cc ph UV, IR,NMR. MS xc nh cu trc

CHNG 6

SAPONIN

6.1 KHI NIM Saponin l thut ng ch mt nhm glucozit c c im chung l khi ho tan trong nc s c tc dng gim sc cng b mt ca dung dch v to nhiu bt. Di tc dng ca enzym thc vt hay vi khun, axit long saponin b thu phn to thnh genin (gi l sapogenin) v phn ng. Phn ng c th gm mt hay mt s phn t monosaccarit:ph bin nht l D-Glucoza,DGalactoza, L-Arabioza, L-Rammoza

 Phn sapogenin Da vo cu trc ca sapogenin ngi ta chia thnh 2 nhm ln + sapogenin tritecpen + sapogenin steroid. V phn b: Saponin phn b rt rng, tm thy trn 80 h thc vt. Saponin steroit tp trung ch yu cy mt l mm trong khi saponin tritecpen c nhiu cy hai l mm.

6.2 SAPONIN TRITECPEN Cu trc ca Saponin tritecpen rt a dng, da vo cu trc ca genin ngi ta chia thnh 4 nhm chnh: -Dn xut -amyrin - Dn xut - amyrin - Dn xut lupeol - Tritecpen: 4 vng, 5 vng. Di y l mt s khung c bn

Dn xut amyrin

25 2 1

26 27

16

28

HO

23

-amyrin

Dn xut lupeol v tritecpen

R HO
17 R' 2 1 16 2 1

R1

HO lupeol

HO R2 tritecpen 4 vong

H H COOR2

R1O

saponin tritecpen 5 vong

6.3 SAPONIN STEROIT

6.3.1 CU TRC 6.3.1.1. C IM CU TRC Tr hp cht Criptogenin (c cu trc 4 vng) tt c cc saponin steroit u c c im chung v cu trc l h thng vng spiroxetal (vng E v F) ni nhau qua C22. Cc hp cht ny ch khc nhau 3 im: - C5 vng A/B:Hidro c th c cu hnh v hoc dy ni i gia C5- C6 -Tu theo cu trc ca R1 v R2 (thng l H hoc CH3) m chng c th l ng phn iso (cu hnh C25 l R) hoc ng phn neo (C25 c cu hnh S). -V s nhm th: tt c cc saponin streroit u c nhm OH C3; mt s t cn c OH C2 hay C6 (hoc c hai). Vi sapogenin cn c thm C=O C12.

 V d
O CH2OH 19
2

21 22 18 E O C D 16 B 5 14

O F

R1 25 R2

5 HO criptogenin

HO

Khung sapogenin steroit

6.3.1.2 CU TRC CA MT S SAPOGENIN STEROIT

O R1 25 O R2

sarsapogenin : C25:neo, R1=CH3; R2=H smilagenin : C25:iso, R1=H R2= CH3;

5 HO H

.
O

R1 25 R2

Neotigogenin: C25:neo, R1=CH3; R2=H Tigogenin :: C25:iso, R1=H R2= CH3;

5 HO H
O R1 25 O R2

Yamogenin : C25:neo, R1=CH3; R2=H Dioagenin :: C25:iso, R1=H R2= CH3;

5 HO

.
O O O R1 25 R2

Hecogenin : C25:neo, R1=CH3; R2=H ,5 H Willaagenin : :C25:iso, R1=H R2= CH3; 5 H

5 HO
H

 Trong hng trm hp cht saponin streroit c bit cho n nay ch khong 10 cht c ngha kinh t ln v: - C ngun nguyn liu di do hoc trng d cho nng sut cao. - H thng spiroxetal trong cu trc d dng b ph v cho hp cht trung gian l ehydropregnenolon axetat (D.P.A) l khung c bn tng hp cc thuc steroit.Trong diosgenin c s dng nhiu nht.

 6.3.2 S THOI BIN DIOSGENIN THNH DEHDRO PREGNENOLON AXETAT (D.P.A)

O

CH3 CH3 anhydrit axetic O CH2OAc

AcO diosgenin axetat

AcO pseudodiosgenin diaxetat CrO3 CH3 CO CH3 CO H CH3 O-CO-(CH2)2-CH-CH2OAc

AcO dehidropegnenolon axetat (D.P.A)

AcO

6.4 PHN B- NG DNG CA SAPONIN 6.4.1 PHN B Saponin tritecpen phn b ch yu cy hai l mm, ng ch nht l tp trung h nhn sm. Saponin steroit tp trung ch yu cy mt l mm trong nhiu h khc nhau.

6.4.2 NG DNG - Tc dng bi b, tng cng sinh lc (nhn sm) - Tc dng long m, du ho (cam tho, cam tho t) - Gim au khp xng (ngu tt, c xc) - H cholesterol trong mu (ngu tt, c xc) Ring nhm saponin steroit l ngun nguyn liu quan trng bn tng hp cc thuc cocticoit v thuc hn ch sinh

6.5 CHIT XUT SAPONIN

Do saponin phn cc mnh tan tt trong nc hoc trong ancol thp, t tan trong dung mi hu c t phn cc hay khng phn cc (ete, axeton , benzen) Nu nguyn liu c nhiu cht bo, s b loi chng bng ete du hay benzen, cc sapogenin do khng cn phn ng gn vo nn ni chung l phn cc yu , nn t tan trong nc, tan nhiu trong dung mi hu c (ete du, n-hexan, ete,benzen , axeton ,cn) Tu thuc vo yu cu nghin cu v sn xut, cn chit chng dng saponin hay sapogenin m chn cch chit v dung mi thch hp.

6.5.1 CHIT SAPONIN

6.5.1.1 CHIT BNG NC Nguyn liu ti , xay bt ngm vi nc mt thi gian (6-12 gi). p, dng nc ra b. Dch p thu c lc vi benzen vi ln loi cht bo . Dch nc loi bo xong , chit saponin bng n-butanol bo ha nc. Dch chit butanol em ct thu hi dung mi. Cn thu c ha tan trong mt lng nh etanol. Cho vo dung dch ny mt lng gp 3-4 ln th tch dung dch saponin mt hn hp gm ete-axeton (1:4). Saponin s kt ta. lng, lc, thu ly kt ta. Dch lc cho thm mt lng ete-axeton khc ta ht saponin. Saponin th thu c em tinh ch v kt tinh trong cn + axeton Nu nguyn liu kh th lm bt th, chit nc nng v chit ly saponin nh trn.

6.5.1.2 CHIT BNG CN Bt nguyn liu kh chit nng hay ngm kit vi etanol hay metanol 80%. Dch chit ct thu hi cn cn nc;lc hn hp ny vi benzen loi cht bo. Sau chit saponin bng butanol bo ho nc. Ct thu hi butanol. Cn thu c ho tan vo mt t etanol v kt ta dung dch bng hn hp ete: axeton (1:4). Sau tinh ch cho saponin theo cch trn.

 Trong nghin cu, c th kt ta saponin t dung dch nc bng cholesterol hoc stearin. Ta c lc ring (thng p dng nh lng saponin) Cc saponin axit c th kt ta di dng mui amoni hoc natri. C th p dng tch saponin bng ct trao i ion: Dch chit saponin cho chy qua ct anion, sau y saponin ra khi ct bng dung dch anion (V d: dung dch HCl, axit axetic), cui cng ra, tch saponin bng cn.

6.5.2 CHIT SAPOGENIN chit sapogenin u tin phi c giai on thu phn saponin thnh ng v sapogenin. Nguyn liu xay nh, ngm trong nc nng 35-400C (nu cn c th cho thm enzym thu phn) trong vi ngy. Thi gian thy phn ty thuc vo tng loi saponin, thng l 48-72 gi.

 Nu thu phn trong axit (HCl, H2SO4) cho pH=1-2 (thng t 5-10%) un si 3- 6 gi ty thuc tng loi saponin., ngui, lc c b dc liu em sy kh. Sau b c chit bng dung mi hu c nng (hi lu, trong thit b soxhlet). Dung mi c th dng l nhexan , ete du, benzen hoc cn. Trong cng nghip c th dng xng cng nghip tinh ch li chit xut C th kt hp c hai cch trong vic thy phn saponin; thu phn bng enzym trc, sau thy phn bng axit.

6.6 PHN TCH SAPONIN

6.6.1 TC DNG PH HUYT Khi cho saponin vo mu s lm hu vch hng cu, lm cho hemoglobin khuych tn ra. Li dng tnh cht ny nh tnh saponin 6.6.2 PHN NG TO BT. i vi dch chit saponin trong nc hoc cn c kh nng to bt, nhng bn vng ca bt ty thuc vo bn cht ca saponin.Trong 2 ng nghim , ng th nht cho khong 5ml dung dch NaOH 0,1N (pH=13), ng th 2 cho 5ml dung dch HCl 0,1N (pH=1). Cho vo mi ng 5ml dung dch saponin. Lc mnh 2 ng. Nu c bt nhiu v bn vng ng ng NaOH ( ng 1) l saponin steroit, cn nu bt nhiu v bn vng trong mi trng axit l saponin tritecpen

6.6.3 PHN NG TO MU. - Phn ng Libermann-Burchard: Hn hp 1ml anhidrrit axetic + 1ml CHCl3 lm lnh 00C, sau thm 1 git H2SO4. Cht th c ha tan bng cloroformvaf cho vo hn hp trn. Nu xut hin mu xanh da tri, lc, hng, cam, v bn vng trong mt thi gian l c phn ng trung tnh - Phn ng Resenhein: Cho 1-2 git axit tricloaxetic 90% vo 1ml dung dch thtrong CHCl3 . Cc saponin tritecpen cho mu tm, chuyn sang mu xanh l sau 20 pht. -Phn ng Salkowski: 1-2 mg cht th ha trong 1ml CHCl3. thm 1ml H2SO4, c mu vng n hng l c saponin

-Phn ng Carr-Price: 1ml dung dch th + 1-2 git dung dch SbCl3 20% trong CHCl3. Nu c saponin steroit s cho mu hng hoc ( phn ng chung ca steroi) - Phn ng Rosenthaler: 1 mml dung dch th + 1-2 git dung dch vanilin 1% trong etanol, thm 1 git HCl c. Nu c saponin tritecpen hoc steroit th s c mu lc n tm. - Phn ng Molish: Cc saponin do c phn ng nn c phn ng vi thuc th Molish .1-2 mg saponin +1-2 git dung dch -naphtol trong etanol. Nh dc thnh ng nghim 1 git H2SO4 c. Nu xut hin mt vng mu tm l phn ng dng tnh. - Phn ng Tortelii-Jafe: ( pht hin lin kt i steroit): cho 0,5mg cht th cho ha tan trong 0,2ml CHCl3. Cho nh nhng khng ha ln 0,1 ml dung dch brom 0,2% trong CHCl3. Nu c steroit c lin kt i s c mt vng lc gi 2 lp.

6.6.4 PHNG PHP SC K GIy, LP MNG V QUANG PH ( xem ti liu)

CHNG 7

GLYCOZIT TIM V GLYCOZIT XIANOGEN

7.1 GLYCOZIT TIM

7.1.1 I CNG Glycozit tim l nhm glycozit c cu trc steroit, c tc dng c hiu i vi tim. Nhng liu cao chng tr nn gy c, dn n cht ngi. Trong cy glycozit tim tn ti dng glycozit ha tan trong dung dch t bo. Di tc dng ca men hoc axit long, cc glycozit b thy phn to thnh genin v cc ng. L glycozit nn chng d tan trong nc v cn long, rt t tan trong cc dung mi khng phn cc hoc phn cc yu nh ete, ete du ha, benzen...Trong dung dch nc cc glycozit tim lm gim sc cng b mt ca dung dch v to bt khi lc mnh. Glycozit tim tn ti trong khong 10 h thc vt, c bit l h Apocynaceae v h Asclepiadaceae

7.1.2 CU TRC HA HC 7.1.2.1. PHN GENIN Phn genin ca glycozit tim c cu trc 5 vng A, B, C, D, F. Vng F l mt vng lacton hoc gn vi C17. Da vo vng lacton ngi ta chia thnh hai nhm chnh: - Cacdenolit (I), genin c vng - lacton -, khng no ( cn gi l butenolit) - Butadienolit (II), genin c vng - lacton khng no vi 2 ni i (cn gi l pentenolit)

 Cu trc cc genin ca glycozit tim c th khc nhau 3 im: + Cu hnh C3, C5 hoc C17 ( hoc ) + C th thm nhm chc cha oxi, ch yu l nhm OH Ti C1, C2, C5, C11 ,C12 , C15 , C16 , v C19 + C th c cc lin kt i trong khung, thng l vng B

7.1.2.2. PHN NG Phn ng ca glycozit tim kh phc tp. C 3 loi ng ng thng thng ( v d Glucozo, Rhamnozo, Xylozo) ng 2-deoxy: l ng khng c oxi v tr s 2. Loi ny kh ph bin trong glycozit tim ( v d Digitosozo, Oleandrozo) ng c bit

 Trt t lin kt ca glycozit tim nh sau: Genin gn trc tip vi vi phn t ng 2-deoxy hoc ng c bit, tip n l mt vi phn t ng thng thng, nht l glucozo gn vo cui mch
Genin (ng 2-deoxy hay ng c bit)n -(glucozo)n

Tc dng ch yu ca glycozit tim ch yu do genin, nhng phn ng c nh hng n tan, s phn b c trng ca thuc trong c th, s tch ly v s thi tr thuc v c tnh.

7.1.2.3. CU TRC CA MT S AGLYCON CA GLYCOZIT TIM

O O

O
OH OH OH

CH2

OH HO OH HO

OH

Ouabagenin

Digitoxigenin

O
OH

OH

OH HO HO

OH

Digoxigenin

Gitoxigenin

CHO

HO-CH2

OH HO OH HO OH

OH

Strophantidin

Strophantidol

O
O

HO

CHO

OH HO HO

OH

Calotropagenin

Scillarenin

7.1.2.4 Cng thc mt s ng ca glycozit tim

CHO HC-OH CH3OCH H COH H COH CH3

CHO HC-OH CH3OCH

H COH HO C-H

CHO HC-H HC-OCH3

HOC-H HOCH

CHO HC-H HCOH

H COH H COH

CHO HC-H HC-OCH3

HCOH H COH

CHO HC-OH HCOH

HOC-H H COH

CH3

CH3

CH3

CH3

CH3

D-Thevetozo

D-Digitalozo

L-Oleadrozo

L-Digitoxozo

D-Cymarozo

L-Rhamnozo

7.1.3. CHIT XUT

Ni chung cc sn phm c ng dng trong iu tr l cc glycozit c cha t 1 n 4 phn t ng. iu mu cht trong k thut l vic khng ch hoc dng men thy phn vi mc cn thit t c sn phm mong mun. Nu mun thu c glycozit nguyn sinh phi dit men thy phn trong nguyn liu bng cch un si nguyn liu vi cn 50-90% trong vi pht Nu cn ly glycozit th sinh th dng men thy phn trong nguyn liu hoc cho men khc thm vo

 Nu cn ly aglycon ( nghin cu) th thy phn bng axit long hoc thy phn men + thy phn axit Nguyn liu, c bit l ht phi tin hnh loi cht bo bng ete du trc khi chit Tnh phn cc ca cc glycozit ph thuc ch yu ca bn cht v s lng ca cc ng trong hp cht. Ni chung nu cng nhiu phn t ng th phn cc cng mnh. Dung mi thch hp chit glycozit tim l cn 5090%. loi tp (ch yu l cc hp cht phenol v tanin) c th dng ch axetat v cho chy qua ct hp ph. tch phn on cc glycozit c th dng cc h dung mi c phn cc khc nhau.

7.1.3.1 CHIT AGLYCON ( nghin cu) Nguyn liu c loi tp s b bng ete du ha. Ly b nguyn liu chit bng cn vi nng thch hp bng soxhlet hoc chit hi lu 2-3 ln. Dung dch thu c, ngui, cho dung dch ch axetat vo loi hp cht phenol (ta). Axit ha dch bng axit sunfuric n pH= 5-6. Lm bay hi bt dung mi v sau chit bng dung dch CHCl3-etanol (2:1) cho n khi ht glycozit. Cho bay hi dung mi, ha tan cn bng cn v thm mt t axit sunfuric v thy phn khong na gi khong 500C. ngui, chit bng CHCl3, ra dch CHCl3 bng NaHCO3, ra li bng nc , lm khan bng Na2SO4, ct thu hi dung mi thu c cn. Tch bng sc k ct thu c aglycon.

7.1.3.2. CHIT GLYCOZIT Nguyn liu c chit bng dung dch cn t 50-900, sau ct loi dung mi p sut thp v tch ring theo nh phng php chung nu trn.

7.1.4 PHN TCH 7.1.4.1. CC PHN NG MU Phn ng Baljet: 1 mg cht th thm 3-4 git thuc th (hn hp th tch tng ng axit picric 1% trong cn v dung dch 10% NaOH trong nc). nu c mu vng cam hoc hng xn l phn ng dng tnh Phn ng Kedde: thuc th l hn hp ng tch ca 2 dung dch: +Dung dch A: axit 3,5-dinitrobenzoic 2% trong cn +Dung dch B: KOH 5,7% trong nc. Cho 12 git thuc th vo 1mg cht th, nu c mu hng tm l dng tnh. Mu ny bin i sau 1-2 gi

 Phn ng Raymond: cho 1-2 git dung dch m-dinitrobenzen 1% trong cn v 2 git dung dch NaOH 20% trong nc vo 1mg cht th. Nu c mu tm hoc xanh l phn ng dng tnh ( cc nhm metylen hot ng cng cho phn ng ny) Phn ng Tollens: ng nghim cha 2-3 mg glycozit v 5 git pyridin, thm 4 git thuc th Tollens, nu dung dch c ta bc l c mt ca ng kh trong glycozit.

 Phn ng Keller- Kaliani: (Phn ng c trng cho ng 2-deoxy) . Trong ng nghim cha 0,1-0,2 mg cht th, thm 1ml dung dch thuc th mi pha gm 1ml feric sunfat Fe2(SO4)3 5% v 99 ml axit axetic bng. Sau cho thm 1-2 git axit sunfuric. Nu c mu xanh xut hin trong 1-2 pht l phn ng dng tnh Phn ng Xanthidrol: Ly 1-2 mg cht th , thm 3-5 git dung dch xanthidrol 0,125% trong axit axetic bng c thm 1% HCl c. Nu c mu chng t c ng 2-deoxy

7.1.4.2. SC K GIY V SC K LP MNG a) Sc k giy: Pha di ng dng cho sc k giy l h cc dung mi phn cc nh: + n-butanol bo ha hi nc + etanol-pyridin- nc (5:1:4)

b) Sc k lp mng: Cht hp ph l silicagel G hoc oxit nhm trung tnh, Cc h dung mi phn cc, cc h dung mi thng dng l: - etyl axetat Xiclohexan (4:1) - etyl axetat axeton (9:1) - metyl etyl xeton- CHCl3- focmamit (5:2:1) Hin mu trn giy hoc trn sc k lp mng bng cc dung dch sau: - m-dinitrobenzen 10% trong cn, tip n phun dung dch NaOH 10% trong metanol- nc, cc glycozit xut hin mu tm hoc xanh - Dung dch axit tricloaxetic 25% trong CHCl3, h nng 1100C, xem mu qua UV - Dung dch anisandehit trong axit axetic-H2SO4 h nng 1100C

7.1.4.3. QUANG PH: UV, IR, MS, NMR

7.2 GLYCOZIT XIANOGEN

7.2.1.CU TRC

Cu trc chung ca hp cht ny nh sau:

R1 C R2 CN O Glucozo

R1, R2: ankyl, aryl hay nhm khc

Khi b thy phn bi men chng gii phng HCN ( axit prussic, hidro xyanogen)
R1 C R2 CN R2 O Glucozo R1 C CN OH R1 -CO- R2 + HCN

7.2.2 PHN B
pht hin s c mt ca xyanogen glycozit trong khong 750 loi ca 250 chi h thc vt. Cc glycozit xyanogen ph bin nht l linamarin v lotaustralin v hai loi ny thng c mt ng thi vi nhau. Chng c trong mt s loi sn.

Ngoi ra cn c mt s loi khc.

7.2.3 PHN TCH

Pht hin HCN: Dng giy picrat + Giy picrat c iu ch bng cch dng axit picric 0,05M trong nc c trung ha bng NaHCO3 v lc, ct giy lc thnh tng gii nh, nhng vo dung dch picrat, lm kh, bo qun kn. + th ta dng 1 gam nguyn liu ti, thi nh, cho vo ng nghim, thm 1 git nc, 2 git toluen nghin nhanh bng a thy tinh. Dng nt bc y kn ming ng nghim, trong ng c t l lng 1 t giy picrat. Gi m ng nghim 400 trong 2 gi. Nu c HCN gii phng th giy picrat chuyn t mu vng sang mu . Mu cng m th HCN gii phng cng nhiu.

 Nu phn ng m tnh c th gi nguyn ng nghim 24-48 gi nhit phng sau lm li nh trn. Ngoi ra cng c th dng phng php sc k giy hoc lp mng phn tch xyanogen glycozit.

CHNG 8

ANKALOIT V CAROTENOIT

8.1 ANKALOIT
8.1.1 KHI NIM Ankaloit l nhng hp cht c cha d vng nit,c tnh baz, thng gp trong nhiu loi thc vt, i khi cn tm thy trong mt vi ng vt. Ngoi hp cht d vng, ngi ta cn thy mt s t ankaloit c nguyn t nit nm ngoi vng (nh colchixin, hordenin) Ankaloit c tnh cht hot ng sinh l cao i vi c th ngi v ng vt, nht l i vi h thn kinh.Vi mt lng nh c th l loi thuc c hiu, nhng lng tng i ln n l cht c gy cht ngi.

8.1.2 PHN LOI

V cu to, ankaloit c n 250 dng cu trc khc nhau vi gn 6000 cht t nhin (>5500).V vy, ngi ta phn loi da vo cu trc ca ankaloit thnh gn 20 nhm, nhng ngy nay, ngi ta cn ngh chia thnh cc nhm nh hn. Ankaloit thng c cu to khc nhau, nhng chung quy li n c cha mt s d vng nit

 V d

 Cc ankaloit c trong cng mt cy hoc cng mt h thc vt thng c cu trc gn ging nhau Cc ankaloit trong cy thng kt hp vi axit ( nh axit oxalic, tactric, lactic) to mui Thnh phn cc ankaloit trong cy ph thuc nhiu vo kh hu, ma trong nm

8.1.3 PHN B
C rt nhiu h, loi thc vt cha ankaloit trong i a s l cy 2 l mm. gii hn vi ngha thc tin, mt cy c coi l c ankaloit phi cha t nht 0,05% ankaloit so vi nguyn liu kh. iu ng lu : Cy c cha ankaloit u vng mt tinh du v ngc li. C kin cho rng chc nng ca 2 nhm ny i vi c cy l nh nhau.

- S phn b trong t chc cy: Trong cy ankaloit tp trung ch yu vo nhng c quan t chc hot ng nht : ni b v ngoi b, cc ti nha. Ni sn sinh ra ankaloit khng hn l ni tp trung nhiu ankaloit (v d thuc l: sn sinh r nhng tp trung l) iu ng lu l s thay i c chu k ca mt s loi c cha ankaloit, chu k c th l vi tunHm lng ca ankaloit trong cy bin i rt ln, v vy cn phi xc nh ng thi im thu hi nguyn liu.

8.1.4 SINH TNG HP ANKALOIT Cho n nay ngi ta tha nhn rng: i a s ankaloit l dn xut ca aminoaxit. Ngi cho rng: u tin aminoaxit c nhm cacbonyl b kh thnh mt amin, sau c s oxy ho thnh andehit bi men oxizada amin. Sau andehit ngng t vi nhm amin to vng c trng i vi ankaloit.

8.1.5 TNH CHT 8.1.5.1.TRNG THI Cc ankaloit c cha oxi thng trng thi rn Cc ankaloit khng cha oxi thng tn to trng thi lng.

 8.1.5.2.TNH TAN Hu ht ankaloit khng tan trong nc (tr mt s t nh nicotin, conin tan c trong nc), nhng tan trong dung mi hu c nh ete, CHCl3, cc ancol thp (metanol, etanol, propanol, butanol) Mt s ankaloit c nhm phn cc nn tan mt phn trong nc hay kim long (nh morphin, cephalin c OH phenol). Ngc li vi cc baz (ankaloit), cc mui ca ankaloit ni chung tan c trong nc v hu nh khng tan trong dung mi hu c (CHCl3, ete) Mt s hp cht ankaloit (c hp cht ngoi l) nh Ephindrin, colchixin cc baz ca chng tan c trong nc ng thi cng kh tan trong dung mi hu c v cc mui ca chng th ngc li.

8.1.5.3.TNH BAZO Ankaloit do c mt nit nn c tnh baz, nhng lc baz ca chng khng ging nhau, tu thuc bn cht ca cc nhm th v cc nhm chc trong phn t. - Hu ht c tnh baz mnh do lm xanh qu tm. - To mui vi axit. - Mt s to ta Al(OH)3, Fe(OH)3 trong dung dch ca mui kim loi ny.

 Do tnh baz khc nhau, nn mi trng axit yu, mt s baz mnh c th chuyn thnh mui, nhng baz yu th mt s ankaloit tn ti trong dung dch di dng baz (khng tc dng vi axit yu). Da vo c tnh ny, ngi ta p dng tch cc nhm ankaloit c pKa khc nhau ra khi hn hp ca chng.

8.1.5.4.TC DNG CA NH SNG Mt s ankaloit khng bn nh sang mt tri, tia t ngoi. 8.1.5.5.HOT TNH SINH L Tc dng ln h thn kinh mnh, c th dng lng nh cha bnh, nhng dng nhiu gy cht, gy nghin

8.1.6 CHIT XUT- TCH RING ANKALOIT

Trong cy, ankaloit tn ti dng mui vi cc axit hu c, nhng mt s kt hp vi tannin (nht l nhng cy c nhiu tannin). V vy i vi nguyn liu c nhiu tanin th cn dng dung mi c phn cc mnh hn hoc chit nng tch ankaloit ra khi tanin v ho tan vo dung mi.

 Mt s ankaloit tn ti dng este nh Atropin, Cocain, Heliotrin c th b thu phn trong qu trnh chit xut nn hn ch dng nhit cao. Ngc li mt s ankaloit tn ti dng glycozit (glycoankaloit) nh solasonin, solamacgin trong cc loi Solanum. chit cc ankaloit ny cn giai on thu phn.

 Ni chung cc ankaloit tng i bn vng so vi nhiu hp cht thin nhin khc, tuy nhin mt s ankaloit cha nhn indol t bn vng vi nh sng v cc tc nhn oxi ho- kh do nn ch cc yu t c th lm hng ankaloit . V vy tt nht chit trong kh quyn nit i a s ankaloit l nhng cht kt tinh khng mu, c nhit nng chy xc nh. Ch c mt s t hp cht ankaloit c mu vng (Becberin, Palmatin), c th dng tnh cht ny theo di chng trong qu trnh chit xut.

8.1.6.1.NGUYN TC CHUNG CHIT XUT

Ni chung ankaloit c th chit t nguyn liu kh tn bt. hn ch bt kh khn trong qu trnh chit tch , i vi nguyn liu nhiu cht bo, cht mu nn loi tp trc khi chit xut. C hai cch loi tp: Ngm bt nguyn liu vi ete-du ha trong vi gi n 1 ngy
Chit lin tc bng soxhlet hoc hi lu vi ete du trong 1-2 gi. Bt loi tp xong, cho kh t nhin.

C hai phng php chnh chit ankaloit: chit bng dung mi hu c chit bng dung dch nc axit hoc cn

a. Phng php chit bng dung mi hu c: Trc ht bt nguyn liu c tm kim chuyn ankaloit mui trong nguyn liu thnh dng bazo. Kim thng c dng l Ca(OH)2, NaOH, NH4OH. Dung mi dung chit c th l clorofoc, ete, benzene, etyl clorua, Clorofoc l dung mi thch hp nht cho hu ht ankaloit baz (tr ankaloit N bc 4, N-oxit c cch x l ring). .

Nh vy nguyn tc chung chit bng dung mi hu c nh sau: chuyn ankaloit dng mui thnh dng ankaloit bng cch tc dng vi kim sau chit ankaloit bng dung mi hu c CHCl3, ete, benzen

b.Phng php th 2: Chit bng dung dch axit v c hay hu c. Dng axit v c hoc hu c chit ankaloit dng mui ha tan. Cc axit thng c dng l: axetic, tactric, sunfuric, nitric, photphoric, ty theo loi ankaloit . Cch chn axit l da vo ha tan ca cc mui ankaloit trong nc.

 Trong sn xut hin nay : do cha c iu kin dung mi hu c chit ankaloit bazo th phng php thch hp nht l ta dung dch chit bng kim. Lc ly ta ha tan ankaloit kim vo dung dch axit, li ta bng kim. lp i lp li nh vy thu c ankaloit th, cui cng tinh ch v kt tinh trong cn.

Mt k thut cn lu l ta tt hn v d dng Nn ta nhit 60-700C. Kt ta xong khng lc ngay m nn yn 1-2 ngy. Ankaloit s ta trong thi gian ny Lc ly ta , dng dung dch NH3 hoc kim long ra ta v cui cng ra li bng nc. Hin nay nhiu nc ngi ta s dng nha trao i ion tch ankaloit ra khi dung dch.

8.1.6.2 TCH RING K thut tch ring ankaloit ty thuc vo thnh phn ca ankaloit trong hn hp quyt nh Nu hn hp ankaloit ton phn ch cha vi ba ankaloit, trong ch c mt ankaloit chnh th vic tch khng kh khn Phng php tch thng dng nht trong sn xut v nghin cu l l tch bng phn b gia hai pha dung mi c phn cc khc nhau v khng ha ln vo nhau. Mi phn on tch ring tip tc tinh ch bng kt tinh phn on.

V d: Li dng ha tan khc nhau ca bruxin v stricnin trong cn long 25% tch ring chng ra khi nhau. Ht m tin tn bt trn vi sa vi, sy kh lm bt. Chit nng vi CHCl3. Dch chit lc vi dung dch H2SO4 1%. Dch chit axit c kim ha bng dung dch NH3 thu c ta ankaloit ton phn. Chit ta ny vi dung dch cn 25% s ly c bruxin, cn li l stricnin.

 Dch chit cn c c cn bt, sau ta bruxin bng dung dch axit oxalic. Lc ly ta. Ra ta ny bng cn. Bc hi cn s thu c bruxin th, tinh ch li v kt tinh trong axeton thu c bruxin Cn khng tan trong cn 25% trn c em ha tan trong cn 950, tinh ch li thu c stricnin.

 Nhng vic tch ring s kh khn i vi trng hp c nhiu ankaloit c cu to tng t nhau v y cng l c im ca nguyn liu cha ankaloit. Trong trng hp ny phi phi hp nhiu loi phng php khc nhau. Chng hn u tin tch phn on bng dung mi c phn cc khc nhau. Sau mi phn on li tch rng bng dung mi c pH khc nhau. Cui cng, i vi cc phn on cn li l hn hp 3,4 cht th tch ring hn hp cc ankaloit bng phng php sc k ct hoc sc k tch lp.

8.1.7 PHN TCH ANKALOIT

8.1.7.1 THUC TH ANKALOIT C rt nhiu thuc th cho phn ng mu hoc kt ta vi ankaloit. Sau y l mt s thuc th thng dng Thuc th Dragendorff: Ha tan 8,0 gam bitmut nitrat kim trong 20 ml HNO3. Ha tan 27,2 gam KI trong 50ml H2O. Hn hp hai dung dch trn cho thm nc cho va u 100 ml. Phn ng cho ta mu vng cam. -Ngoi ankaloit, thuc th Dragendorff cn cho phn ng vi mt s hp cht hu c baz khc nh cholin v mt s cht khng phi l bazo hu c nh pyrol, Xinamandehyt, ninhydrin. Ngc li thuc th Dragendorff khng cho phn ng vi bt k mt amino axit no.

 Thuc th Mayer: Ha tan 1,36 gam HgCl2 trong 60 ml nc. Ha tan 5 gam KI trong 50 ml nc . H hp hai dung dch thm nc vo va u 100 ml Thuc th Wagner: Ha tan 1,27 gam Iot va 1 gam KI trong 20 ml nc. Thm nc va u 100ml. Phn ng cho kt ta nu Axit silicotungstic 5% , axit photphomolipdic

8.1.7.2 QUANG PH HP TH CA ANKALOIT

UV IR NMR

8.2 CAROTENOIT
8.2.1 I CNG Carotenoit l nhng cht mu vng,da cam, ...c nhiu trong nhiu loi ng thc vt. Ngi ta cn gi n l cht mu lipocromic ( mu trong cht lipit) v chng tan trong du bo Trong c th ng vt, carotenoit c ha tan trong m hoc ha hp vi protein pha nc Chng l nhng cht mu chnh trong mt s hoa mu vng, da cam, ca nhiu loi vi sinh vt.

8.2.2. PHN LOI V TNH CHT Cc carotenoit gm hai loi chnh -Nhm hydrocacbon: tan trong ete du -Nhm xanthophyl l cc dn xut oxi trong carotene.Chng bao gm andehyt, xeton, axitchng tan trong etanol

8.2.2.1 NHM CAROTEN HYDROCACBON

Ph bin l caroten c nhiu trong c rt

-caroten

-caroten

-caroten

b) 8.2.2.2 NHM XANTOPHIN

c th xp thnh nhiu loi khc nhau. V d: Nhm carotenoit c nhm OH (hoc este ho nhm OH)
OH

HO

xantophin

OH

HO

zeaxanthin

 Nhm carotenoit metoxyl ho (c nhm CH3O-). V d nh Rhodovibrin Nhm carotenoit oxi ho (oxicarotenoit) Nhm epoxi carotenoit: c nhm epoxi Nhm cacboxi carotenoit:c nhm COOH. V d bixin: cht mu c trong ht iu Bixa orellana
COOH COOCH3

bixin

 Hu ht cc carotenoit trong t nhin u l tetratecpenoit gm 8 n v isoprene (C40) Hu ht cc ni i u c cu hnh trans, lin kt gia vng cui v mch trung tm l lin kt n. Tuy nhin khi dehydro ho c th chuyn ni n thnh ni i trong trng hp ngi ta gi l retro xc nh cu trc ca carotenoit ngi ta c th dng cc phng php: -Hydro ho -Cng halogen: xc nh ni i -Oxi ho vi axit cromic bin CH3 mch nhnh thnh hp cht cacboxyl -Ct t lin kt i bng KMnO4 hay ozon phn. Phng php sc k (ct, giy, lp mng) l phng php ch yu phn lp v xc nh cu trc ca carotenoit

8.2.3 TNG HP CAROTENOIT

Thng thng ngi ta tng hp bng cch ghp: mt hp cht gm 2 nhm chc c kh nng phn ng vi 2 hp cht khc nm hai tn cng c 2 kiu 1) Kiu cng hp C19+C2+ C19 V d: iu ch -caroten
H 2 O + Br-MgC C-MgBr -caroten

2) Ho hp cc n v C16+C8+C16 V d:
C 2 OH C-MgBr + O O -caroten

3) Ho hp C14+C12+C14
O 2 HO C C-MgBr + O -caroten

CHNG 9

GLUXITAMINOAXIT-PROTEIN

9.1 GLUXIT
Gluxit (cacbohydrat) l nhm cc hp cht hu c ph bin trong t nhin, c bit trong thc vt (chim trn 80% khi lng kh) PHN LOI - Monosaccarit (monoz): l gluxit khng c kh nng thy phn thnh cc gluxit n gin hn. - Oligosaccarit (oligoz) l nhng gluxit khi thy phn cho ta t 2-10 monosaccarit. - Polisaccarit (polioz) gm trn 10 n v monosaccarit to nn. Loi ny gm 2 loi: + Homopolisaccatrit: khi thy phn cho trn 10 monosaccarit cng loi. V d nh tinh bt, xenlul + Hetoropolisaccarit: khi thy phn cho cc monosaccarit khc loi.V d nh hemixenlulozo, aga-aga....

9.1.1 MONOSACCARIT
( Gluxit n gin, ng n) 9.1.1.1 CU TRC: c 2 dng cu trc l mch h v mch vng a) Cu trc mch h: Nh vy xt v cu to n thuc loi polihydroxicacbonyl . Ty thuc vo bn cht ca nhm cacbonyl m chng c chia thnh 2 loi: + andoz ( nhm andehyt CH=O) + xetoz ( nhm xeto CO-)

b) Cu hnh ca monosaccarit. ng phn quang h c Dy D v L: Nu nhm OH ca monosaccarit c cu hnh ca nguyn t C xa nhm C=O nht nm bn phi trong cng thc Fischer pha phi thuc dy D, cn pha tri thuc dy L ng phn epimer: Hai monosaccarit c cu hnh ch khc nhau 1 nguyn t C th l ng phn epimer. V d Dglucozo v D- mannozo l hai ng phn epime

 ng phn quang hc thng. Trong cc ng phn quang hc ca D- Glucozo ch c ng phn L-gluccoz l ng phn i quang, cc ng phn cn li l ng phn quang hc thng ca D- glucozo. ng phn i quang: Hai ng phn i quang c cu hnh hon ton tri ngc nhau, c nng sut quay cc bng nhau nhng tri nhau v du, cn cc ng phn quang hc thng c nng sut quay cc khc nhau.

c-Cu trc dng vng ca monosaccarit. ng phn anomer

S c mt ng thi nhm C=O v OH trong phn t, do c kh nng tc dng ca nhm HO vi C=O to hp cht dng vng khi c nhm OH hemi-axetal. V d i vi D- Glucoz
6 CH2OH H 5 H O H H 1 OH 2 OH OH 3 H OH -D-glucopiranozo CH=O H HO H H CH2OH OH H OH OH H CH2OH H OH OH H
O

OH 1 H

OH

-D-glucopiranozo

 ng phn anomer: Hai monosaccarit c cu trc vng l ng phn anomer ca nhau khi cu hnh ca chng ch khc nhau nguyn t C cha nhm OH hemiaxetal. V d v - Dglucopiranozo l hai ng phn anomer, chng ch khc nhau v cu hnh ca cacbon s 1 (c nhm OH-hemiaxetal).

 i vi glucozo, cu trc ch yu l dng vng ( chim hn 99,9%), trong ng phn - D- gluccopiranozo l ch yu v c cu dng bn vng hn ( v cc nhm OH v tr bin nhiu hn ng phn )
CH2OH
O

CH2OH HO HO

O OH OH

HO

HO OH dang OH

dang

9.1.1.1 TNH CHT

a) TNH CHT VT L Cc monosaccarit u trng thi rn, khng mu, d tan trong nc, t tan trong ancol, hu nh khng tan trong cc dung mi hu c thng thng (benzen, ete...) cc ng phn anomer ca monosaccarit c nhit nng chy khc nhau v gc quay cc ring khc nhau. Cc monosaccarit u c v ngt, ngt cng khc nhau: fructozo ngt hn glucozo khong 2,4 ln v ngt hn galactozo khong 5,5 ln.

Nhit nng chy (0C) v quay cc [] ca mt s monosaccarit

Monosaccarit D- glucozo D- mannozo

T0nc T0nc p p
146 133 150 132

[]D p
+1120 +29,30

[]D p
+18,70 -170

[]D
+52,70 +140

D-galactozo

167

167

+150,70

+54,40

+810

b) TNH CHT HA HC b1) Phn ng ca nhm OH Phn ng vi Cu(OH)2 to hp cht phc Phn ng to este: khi tc dng vi clorua axit, anhydrit axit
CH2OH
O

CH2OAc
O

OH OH OH OH

+ (CH3CO)2O

HCl

OAc OAc OAc OAc

+5 AcOH

 Phn ng to thnh glicozit: + Tc dng vi CH3OH c HCl khan ch th OH hemiaxetal to thnh glucozit bn trong kim, do khng cn kh nng m vng nn khng c tnh kh, nhng d thy phn trong axit hoc enzim to hp cht u:
CH2OH
O

CH2OH
O

OH OH OH OH

+ CH3OH

HCl

OH OH OCH3 OH

metyl- -Dglucopiranozit

CH2OH
O

CH2OH
O

OH OH OH OCH3

+ H2 O

H+

OH OH OH OH -Dglucopiranozo

+CH3OH

 Phn ng to thnh ete: Trong iu kin xt HCl ch c OH hemiaxetal phn ng to glicozit, chuyn cc nhm OH khc thnh ete cho glicozit phn ng vi dimetyl sunfat trong kim
CH2OH
O

CH2OCH3
O

OH OH

+ 2(CH3)2SO4 + 4NaOH OCH3 OH

OCH3 OCH3

+ 2Na2SO4 + 2H2O OCH3

OCH3 Metyl-2,3,4,6-tetra-O-metyl- -D-glucopiranozit

 Phn ng to thnh axetal vng v xetal vng

CH2OH
O

CH2OH
O

OH OH OH OH

+ CH3-CO-CH3 OH

OH O O C

+ H2O CH3 CH3

1,2-isopropyliden- -D-glucopiranozit

b2) Phn ng ca nhm cacbonyl ) Phn ng oxi ha Phn ng oxi ha gi nguyn mch cacbon: + Phn ng vi thuc th Tollen ( trng bc): v phn ng xy ra trong OH- nn c xetozo cng phn ng v trong kim xy ra qu trnh ng phn ha ando xeto qua giai on trung gian l enol V d c D-Glucozo v D-fructozo u c phn ng trng gng

+Phn ng vi dung dch brm: to axit andonic, trong mi trng axit , cc andonic d dng chuyn thnh vng lacton

+Phn ng vi axit HNO3: To axit saccaric (axit polihydroxidicacboxylic), sau chuyn thnh dilacton

 Phn ng oxi ha ct mch cacbon: + Tc dng vi HIO4

Cc metyl glicozit cng b oxi ha bi HIO4 nhng ch xy ra lin kt CHOH-CHOH-

) - Phn ng kh: bng H2/Ni, t0, hoc NaBH4 to poliancol

CH2OH C=O HO H H H2,Ni, t0 CHO H HO H H D-Glucozo OH H H2,Ni, t OH OH CH2OH
0

OH OH CH2OH D-fructozo H2,Ni, t0 CH2OH HO HO H H H H OH OH H2,Ni, t

0

CH2OH H HO H H OH H OH OH CH2OH D-Gluxitol ( D-Sobitol)

CHO HO HO H H H H OH OH

CH2OH D- Mannitol

CH2OH D-Mannozo

) - Phn ng to phenylosazon : tc dng vi phenyl hydrazine trong mi trng axit axetic , u tin to phenyl hydrazon sau tip tc phn ng cho sn phm cui cng l osazon ) - Phn ng ni di v rt ngn mch cacbon ( xem ti liu) Cht tiu biu: Glucozo, mannozo, galactozo ( xem ti liu)

9.1.2 OLIGOSACCARIT (OLIGOZO) Oligosaccarit thng gp nht l isaccarit v trisaccarit. Nhng isaccarit quan trng l saccarozo, mantozo v lactozo. Cc oligosaccatrit thng gp di dng kt hp trong glycozit ( nht l flavonoit) l rutinozo (6--ramnosylglucozo), Neohesperidozo (2--ramnosylglucozo)... Trisaccarit nh: rafinozo

9.1.2.1 CU TRC PHN T Cu trc v tn gi h thng ca saccarozo v manto:

6CH2OH 5 OH HO OH
O

1CH2OH O 1 O OH 2 HO 5

6CH OH 2 5 OH
O

6CH OH
2

5 1 4 OH O

1 OH OH

CH2OH 6

HO

OH

2-O-( -D-Glucopiranozyl)- -D-Fructofuranozit ; 4-O-(-D-Glucopiranozyl)- -D-Glucopiranozo ( saccarozo) (mantozo)

 Mt s disaccarit khc
6CH3 5 OH HO OH OH HO OH
O

6CH OH 2 5 1 O CH2
O O

6CH OH 2 OH 1 HO HO HO 3 1 O O 5 OH
O OH

OH HO 6CH OH O 2 1 5 4 OH HO OH
O

4 OH

CH3 Neohesperiozo

Rutinozo

sophorozo

9.1.2.2 TNH CHT VT L Cc oligosaccarit u trng thi rn , c nhit nng chy v nng sut quay cc khc nhau. Saccaro c gc quay cc ring l + 66,50 (quay phi). Khi thy phn hon ton ( nh axit hoc enzim) thu c hn hp ng phn t DGlucozo v D-fructozo c nng sut quay cc l -200( quay tri). Hn hp sn phm thy phn ca saccarozo c gi l ng nghch o v lm i gc quay t phi sang tri.

9.1.2.3 TNH CHT HA HC a) Phn ng thy phn: c xc tc axit hoc enzim to thnh monosaccarit. V d: + Saccarozo b thy phn cho D- glucozo v DFructozo + Mantozo cho D- glucozo + Lactozo thy phn (nh men Galactoziada) cho D- galactozo v D- glucozo

6CH2OH 5 OH HO OH
O

1CH2OH O 1 O OH 2 HO 5 CH OH 6 2 H2O (H+) 6CH2OH 5 OH HO 1CH2OH

O

2-O-( -D-Glucopiranozyl)- -D-Fructof uranozit ; ( saccarozo)

1 OH OH

O 2 HO OH D-f ructof uranozo HO 5 CH OH 6 2

D-glucopiranozo

b) Phn ng ca nhm OH Tng t nh monosaccarit chng c kh nng to phc vi Cu(OH)2, to ete, este...

c) Phn ng ca nhm cacbonyl: Cc oligosaccarit cn nhm hemiaxetal nh mantozo, lactozo...u c kh nng m vng to ra nhm andehyt, v vy chng c th c cc phn ng sau: Phn ng oxi ha: nh phn ng trng gng, vi Cu(OH)2 to Cu2O, vi dung dch nc brom... Phn ng to thnh osazon: Tc dng vi phenylhydrazin

9.1.2.4 PHN TCH OLIGOSACCARIT

Sc k giy c th dng phn tch oligosaccarit nhng tc di chuyn chm hn Cc h dung mi c th dng l BAW: Butanol: axit axetic: nc ( 4: 5 : 1) lp trn BEW: n- butanol: etanol: nc ( 4 : 1: 2,2) BBPW: n-butanol: benzen:piridin: nc (5 : 1 : 3 : 3) Hin mu cc vt bng dung dch anilin phtalat

9.1.3 POLISACCARIT (poliozo)

Phn t do trn 10 n v monosaccarit kt hp vi nhau nh cc lin kt glycozit, to nn mch nhnh hoc khng phn nhnh Cc homopolisaccarit tiu biu l tinh bt, xenlulozo, glicogen Cc heteropolisaccarit tiu biu l hemixenlulozo, aga-aga...

3.1.3.1 TINH BT
Tinh bt l polisacarit d tr thc vt, chng c tch ly ch yu c khoai, sn...) ht ( go, ng...). Go cha nhiu tinh bt nht (khong 80%) m khong 70%, cc loi c qu khc t hn.

a) Cu trc Tinh bt l hn hp ca 2 loi polisaccarit: amilozo v amilopectin. C hai loi ny u do cc gc -DGlucopiranozo kt hp vi nhau to nn. Phn t amilozo c cu to khng phn nhnh gm cc gc -D-Glucopiranozo kt hp vi nhau nh lin kt -1,4-glycozit Phn t ca amilopectin c cu trc phn nhnh, gm mt s mch amilozo lin kt vi nhau nh lin kt -1,6-glycozit Phn t khi ca amilopectin c th t ti 1 triu v.C, cn amilozo ch khong 200.000 v.C Tinh bt cha khong 10-20% amilozo v 80-90% amilopectin, tn ti dng ht, amilopectin c phn b bn ngoi, pha trong ht l amilozo.

6CH 5

2OH

CH2OH
O
1

6CH 5

2OH

CH2OH
O
1

6CH 5

2OH

CH2OH
O
1

5 1 4

O
1 4

5 1 4

OH

OH

O OH OH

4 OH

OH

O OH OH

OH

OH

O OH OH

O-

Cu trc ca amilo
6

CH2OH
O
1

6 CH 5

2OH

OH

4 O

OH HO O
6

OH

CH2
O
1

6 CH 5

2OH

CH2OH
O
1

5 1 4

Cu trc ca amilopectin

OH

O OH

OH

OH

O OH OH

b) TNH CHT VT L Tinh bt tn ti trng thi rn, khng tan trong nc lnh, tan trong nc nng. Khi un nng cu trc ca amilopectin c nhiu thay i v gy ra hin tng h ha tinh bt. Tinh bt c gc quay cc ring []D t + 180 n + 2100

c) TNH CHT HA HC Tinh bt khng tham gia phn ng trng bc v vi thuc th Fehling v phn t cha mt khi lng rt ln gc -glucozo nhng ch rt t gc cui mch c -OH hemiaxetal. Tinh bt c tnh cht ha hc ch yu sau: c1) Phn ng thy phn -Xc tc axit: cho sp cui l glucozo -Trong c th ng thc vt do xc tc enzim

c2) Phn ng vi Iot nhit phng, tinh bt tc dng vi dung dch iot cho mu xanh tm. Mu ca dung dch ch yu l mu ca hp cht bc c to thnh bi phn t amiloz dng vng xon vi cc phn t iot nm pha trong ng xon.

3.1.3.2 XENLULOZO
a) CU TRC Phn t xenlulozo c cu trc khng phn nhnh, gm cc gc -D-glucopiranozo lin kt vi nhau bng cc lin kt -1,4glycozit:
CH2OH
O

OH O
1 4

CH2OH
O

OH O
1 4

CH2OH
O

OH

OH O

4 O

OH

OH O

OH OH

OH

CH2OH

OH

CH2OH

 -D-1,4-glucopiranozo trong phn t xenlulozo nm cu dng gh:

H CH OH
2

H H
O

H O H CH2OH

H HO O
O

H CH OH
2

H H
O

H O H CH2OH

H HO O
O

HO H

O OH H

O H H

O OH H

b) TNH CHT VT L Xenlulozo trng thi rn, mu trng , tn ti dng si vi di khc nhau. Xenlulozo khng tan trong nc v trong cc dung mi hu c thng thng. Tan c trong phc ng(II)-amoniac, ng (II)-etylendiamin, dung dch tetrametylamoni hydroxit

c) TNH CHT HA HC Tng t tinh bt, xenlulozo khng c kh nng tham gia phn ng trng bc v kh Fehling. Nhng phn ng xy ra i vi xenlulozo ch yu l lin kt -1,4- glycozit hoc cc nhm OH

c1) Phn ng thy phn: c xc tc axit hoc enzim xenlulaza cho sn phm cui cng l D- Glucozo
CH2OH
O

OH OH n

+ n H2O

H+, t0 hay enzim

n C6H12O6

c2) Phn ng ca cc nhm hydroxyl - Phn ng vi axit nitric: Xenlulozo tc dng vi HNO3 c nng nh xc tc H2SO4 to hn hp xenlulozo trinitrat v xenlulozo initrat. T l ca cc sn phm ny ph thuc vo t l HNO3: H2SO4 v thi gian phn ng
CH2OH
O

CH2ONO2 O

OH OH n

+3n HNO3

H2SO4 ,t

ONO2 ONO2 n xenlulozo trinitrat

-Phn ng vi anhydrite axetic: cho ta xenlulozo triaxetat (cn gi l triaxetyl xenlulozo) l polime nhn to iu ch t axetat, phim nh, mng cch in

- Phn ng vi dung dch NaOH v CS2

Xenlulozo tc dng vi dung dch NaOH c cho xenlulozo- kim. Ch ha xenlulozo -kimvi cacbon iunfua thu c xenlulozo angtogenat
CH2OH
O

CH2ONa O + n NaOH OH OH n n
O

O + n H2O

OH OH

xenlulozo kiem
S CH2ONa
O

CH2O-C-S-Na O + n CS2 OH OH n n
O

OH OH

xenlulozo xangtogenat

 Khc vi xenlulozo, xenlulozo xangtogenat tan c trong dung dch NaOH long to thnh dung dch rt nht c tn gi l visco. Thy phn xenlulozo xangtogenat bng dung dch H2SO4 thu c xenlulozo hydrat
S CH2O-C-S-Na
O

CH2OH
O

O + (n/2)H2SO4 OH n

O n

OH OH

+ (n/2) Na2SO4 + nCS2

OH

xenlulozo xangtogenat

+ Phn ng vi dung dch hp cht phc ng (II)-amoniac Xenlulozo phn ng vi [Cu(NH3)4](OH)2 (cn gi l dung dch Svayde) cho dung dch nht do c s to thnh hp cht phc gia xenlulozo vi ion ng. Khi thy phn , hp cht phc chuyn thnh xenlulozo hydrat dung sn xut t ha hc (t ng-amoniac)

9.2 AMINO AXIT PEPTIT- PROTIT

9.2.1 AMINO AXIT

9.2.1.1 KHI NIM Axit amino cacboxylic cn gi tt l axit amin hay aminoaxit, hay axit amin l nhng hp cht hu c trong phn t c ng thi nhm amino NH2 v nhm cacboxyl COOH, chng l n v cu to nn protein, enzym. Mt s axit amin m c th ng vt khng th tng hp c phi cung cp t thc phm ( gi l cc aminoaxit khng th thay th). Cc aminoaxit tham gia cu to protit u l -aminoaxit v c cu hnh L

a) Phn loi Ty thuc vo v tr tng i ca nhm amino v nhm cacboxyl m c cc loi -aminoaxit, aminoaxit, -aminoaxit.. Ty theo t l nhm COOH v NH2 trong phn t m ngi ta chia thnh: + Cc aminoaxit c tnh axit khi s nhm COOH nhiu hn NH2 + Cc aminoaxit c tnh bazo khi s nhm COOH t hn NH2 Ty thuc vo c im cu to ca gc hidrocacbon trong phn t m c cc loi aminoaxit mch thng v aminoaxit mch vng.

b) Cch gi tn. Tn thng thng. + Hu ht cc amino axit gi bng tn thng thng. + Ngoi ra ngi ta cn dng ba ch u tin ca tn aminoaxit, hoc mt mu t theo qui c quc t. V d nh glyxin, leuxin Tn quc t IUPAC. tng i t dng hn tn thng Tn gi theo dn xut ca axit cacboxylic tng ng.,nguyn t cacbon ca nhm COOH c nh s l s1, v tr ca nhm amino c nh s bng s t nhin 2,3,4 Ngoi ra ngi ta c th gi theo danh php hp l: gi theo dn xut ca axit. V tr ca cc nhm th c nh s bng ch Hylap ,,,...

9.2.1.2 IU CH
a) iu ch cc -amino axit
T axit - monohalocacboxylic. Cho axit -monohalocacboxylic tc dng vi amoniac. V d :
R CH Br COOH NH3 R CH NH2 COOH

Tng hp Strecker. Cng HCN v NH3 vo andehyt cho -aminonitrin. Thu phn c xt axit cho -amino axit.V d :
CH3 CHO + HCN + NH3 CH3 CH CN NH2 H3O CH3 CH COOH NH2

 Tng hp Gabriel. Cho phtalimit tc dng vi -haloeste to sn phm trung gian phtalimidoaxetatetyl. Thu phn hp cht ny cho glyxin v axit phtalic.
o N-K o Cl-CH2COOEt -KCl o N CH2COOEt o OH3+ COOH

COOH + NH2-CH2-COOH +C2H5OH

9.2.1.3 TNH CHT VT L

Cc aminoaxit l nhng cht rn kt tinh khng bay hi. Khi un nng mnh n nng chy ng thi b phn hy. C tnh cht l do trong phn t c nhm COOH v NH2 nn chng tn ti ch yu dng ion lng cc NH3+ -R-COO Tr Glyxin khng c cacbon bt i, cn cc aminoaxit khc u c tnh quang hot v cc aminoaxit tham gia cu to protit trong t nhin u c cu hnh L (nhm NH2 gn nhm COOH nht pha tri trong cng thc Fischer)

COOH H2N H CH3 L(+)-Alanin H

COOH NH2 CH3 D(-)-Alanin

9.2.1.4 TNH CHT HA HC

a) Tnh axit- baz. im ng in ca aminoaxit Axit amin l mt hp cht lng tnh. C th to mui vi axit v baz v c, do cc axit amin c th vit di 4 dng, tu theo pH ca mi trng m tn ti dng no .
NH2 H
CH 2 COOH

NH3

CH2 COOH

NH3

CH2 COO

OH

NH2

CH2 COO

Acid lien hp

Ion lng cc

Baz lien hp

 im ng in: l gi tr pH ca mi trng m trong phn t aminoaxit tng s in tch dng bng in tch m , ngha l phn t trung ha v in, tc l tng in tch ca phn t bng o. Nh vy ti im ng in phn t tn ti dng ion lng cc ( ch yu) hoc phn t trung ha (rt t). im ng n thng c k hiu l pI Ti im ng in, nu dung dch c t dng in 1 chiu th cc phn t aminoaxit khng di chuyn v pha in cc no.

 im ng in ca cc aminoaxit c s nhm amino bng s nhm cacboxyl nm khong 6,0-6,3 im ng in ca cc aminoaxit c s nhm amino ln hn cacboxyl nm khong 7,6-10,8 im ng in ca cc aminoaxit c s nhm amino nh hn cacboxyl nm khong 2,8-3,3 C th tnh c im ng in ca aminoaxit nu bit c pKa ca aminoaxit i vi hp cht c 1 nhm NH2 v 1 nhm COOH th pI= (pK1 + pK2)/2 V d: Glyxin H2N-CH2-COOH ; pK1= 2,35; pK2= 9,78; pI= (2,35+9,78)/2 = 6,06

b) Phn ng ca nhm COOH.

Aminoaxit c cc tnh cht ca nhm cacboxyl nh To ester.
H2 N CH2 COOH R EtOH H3O H3 N CH R COOEt + H2O

To clorua axit.
NH2 CH R COOH PCl5 NH2 CH R COCl

Decacboxyl ha ( tch CO2)

Ba(OH) 2

NH2

CH R

COOH

NH2

CH2

c) Phn ng ca nhm amino.

Nhm amino ca aminoait cho phn ng nh mt nhm amin thng thng. Phn ng ankyl ho. CH 3I H3C NH CH COOH NH 2 CH COOH R R Axyl ha: To amit.

Tc dng vi axit nitro. To hidroxyaxit v gii phng nit

NH 2 CH R COOH HONO HO CH R COOH + N 2

d) Phn ng chung ca nhm cacboxyl v nhm amino. Tch nc.

O COOH H2N CH R R CH + NH2 HOOC C R CH NH C O NH CH R

ixetopiperazin To phc vi mt s kim loi nng.V d

 Cc phn ng mu ca aminoaxit: +Ninhidrin

+ Phn ng xantoproteic

+ Phn ng milon, to sunfua ch

9.2.1.5 PHN TCH AMINO AXIT

phn tch amino axit c th dng sc k giy hoc sc k lp mng. -Sc k giy: chy 2 chiu, chiu chy 45-50cm Dung mi: Chiu 1: butanol- axit axetic-nc (BAW) (4:1:1) Chiu 2: Phenol- nc ( 3:1) -Sc k lp mng: Vi nhng hn hp nhiu axit amin nn chy sc k 2 chiu vi h dung mi nh i vi sc k giy Thuc th hin mu l dung dch ninhirin 0,1% trong axeton, h nng knh 1050C trong 10 pht

9.2.1.5 NH LNG AMINO AXIT

Trc y ngi ta thng nh lng bng cch sau: C th nh lng bng cch phi hp sc k giy hoc lp mng vi o mt quang .Dch chit vi mt lng xc nh c tch bng sc k giy hoc lp mng . Hin vt bng dung dch ninhidrin. Ct cc vt ring ra chit vi cn 500. cc dch chit to mu vi ninhidrin v o hp th trn quang ph k. Cc kt qu o c so snh vi ng cong chun c lp sn vi cc amino axit chun

 C th dng phng php chuyn thnh dn xut initrophenyl bng cch cho dch chit c cha aminoaxit vi dung dch 2,4initroflorobenzen trong nc v axeton. em dn xut ny tch bng sc kyd lp mng ( hoc giy) sau tin hnh nh lng nh trn. Nhng trong cch ny do sn phm initrophenyl c mu vng nn khng cn phi cht hin mu ninhidrin
NO2 O2N F COOH + H2N-CH-R O2N NO2 COOH HN-CH-R

( mau vang)

Phn tch bng phng php sc k in di Trong dung dch nc, cc aminoaxit c th tn ti 3 dng : anion, cation v ion lng cc.Ty theo pH ca mi trng m mt trong cc dng chim u th Do c im trn c th phn tch amino axit bng sc k in di Trong dung dch m mi trng axit, di tc dng ca in trng cc phn t amino axit dng anion s chuyn v cc m v ngc li nu phn tch mi trng kim th cc aminoaxit tr thnh anion v chuyn v cc dng. V mi amino axit c im ng in khc nhau v trong qu trnh phn tch nu c s thay i c kim sot pH ca dung dch in ly th c th tch chng ra khi nhau v mi cht c mt ng lng in tch khc nhau di chuyn vi mc khc nhau.

9.2.2. PEPTIT
Peptit l nhng polime aminoaxit cha t 2 n khong 50 gc aminoaxit trong phn t. 9.2.2.1 CU TRC V DANH PHP Cu trc: gm 2 hoc nhiu gc aminoaxit kt hp nh lin kt peptit

 Trong phn t peptit, u mch cha gc NH2( hoc NH3+) c gi l u N, cn u kia cha nhm COOH (hoc COO-) gi l ui C.

Khi vit cng thc cu to ca peptit u N c quy c vit pha bn tri trong cng thc phn t cn ui C c quy c vit v bn phi ca cng thc
H3N - CH - C - NH - CH - C- NH -CH - COO R1 O aminoaxit dau N R2 O R3 aminoaxit duoi C

b) Cch gi tn: Gi tn bng cch ghp tn gc axyl ca aminoaxit to nn t phn t peptit theo trnh t sp xp ca chng t u N sang pha ui C, ring aminoaxit ui C c gi nguyn tn:
H3N - CH - C - NH - CH - C- NH -CH2 - COO alanylphenylalanylglyxin CH3 O CH2 O C6H5 (Ala-Phe-Gly)

9.2.2.2 TNH CHT VT L

Cc peptit c khi lng phn t nh l cht kt tinh tan tt trong nc Cc peptit c phn t khi ln l cht rn khng nh hnh, to c dung dch keo vi nc.

9.2.2.3 TNH CHT HA HC

a) Tnh axit- bazo: c tnh lng tnh tng t nh amino axit b) Phn ng thy phn: Thy phn hon ton: trong axit nng hoc kim nng cho sn phm cui cng l aminoaxit hoc mui ca n. Thng thy phn trong khong 24-72 gi nhit 1100C, xc tc HCl 2N Thy phn khng hon ton: to cc peptit nh hn nh cc enzim c hiu

c-Phn ng vi 2,4-initrofluorobenzen: tng t nh aminoaxit , u N s phn ng to thnh dn xut 2,4-dinitrophenyl mu vng. d-Phn ng mu biure: cc peptit c t 2 nhm peptit tr ln phn ng vi dung dch CuSO4 long trong kim cho phn ng to thnh dung dch hp cht phc c mu tm hoc mu tm

9.2.2.4

TNG HP PEPTIT

a) Bo v nhm amino: Thng dng nhm C6H5-CH2-O-CO- ( cn gi l nhm cacbobenzoxi- vit tt Cbz) bng cch cho aminoaxit tc dng vi benzylclorofomiat C6H5CH2-O-CO-Cl trong mi trng kim. dng nhm benzyloxicacbonyl
H2N-CH2-COOH + C6H5-CH2-O-CO-Cl NaOH, H2O H3O

C6H5-CH2-O-CO-NH-CH2COO

C6H5-CH2-O-CO-NH-CH2COOH benzyloxicacbonyl glyxin (Cbz-Gly)

b)Bo v nhm cacboxyl: chuyn thnh dn xut benzyl hay metyl hoc etyleste
H2N-CH-COOH CH3 +PCl5 -POCl3-HCl H2N-CH-COOCl CH3 C6H5-CH2-OH -HCl

NH2-CH2COO-CH2C6H5 benzyleste cua alanin

c) Ngng t cc dn xut ca aminoaxit: qu trnh ny nh c cht xc tc l DDC (dixiclohexyl cacboiimit- C6H11-N=C=N-C6H11). Th d

CH3 C6H5-CH2-O-CO-NH-CH2COOH benzyloxicacbonyl glyxin + NH2-CHCOO-CH2C6H5 benzyleste cua alanin CH3 Cbz-Gly-Ala-CH2-C6H5 DDC -H2O

C6H5-CH2-O-CO-NH-CH2CO- NH-CH COO-CH2C6H5

d) Hydro phn dn xut ca peptit:

9.2.2.5 XC NH CU TRC PEPTIT

a) Xc nh thnh phn cc amino axit trong phn t peptit ngi ta thy phn hon ton peptit thnh aminoaxit (thng thy phn bng HCL 6N 1100C trong 24-72 gi), sau nhn bit cc aminoaxit bng phng php sc k. b) Xc nh trt t sp xp cc amino axit trong phn t peptit - Xc nh aminoaxit u N: c 2 phng php chnh: + Phng php Sanger + Phng php Edman: ( xem ti liu) - Xc nh aminoaxit ui C. Thy phn peptit nh enzim cacboxipeptiaza, aminoaxit xut hin u tin trong dung dch ( nhn bit bng phng php sc k chnh l aminoaxit ui C - Thy phn tng phn mch peptit (nh enzim th n ch ct cc lin kt peptit nhng v tr xc nh).

9.2.3. PROTEIN
9.2.3.1 PHN LOI V CU TRC a) Phn loi a1) Da vo thnh phn ha hc: chia thnh 2 nhm: - Protein n gin: khi thy phn hon ton ch cho hn hp cc aminoaxit -Protein phc tp: khi thy phn hon ton, ngoi sn phm l cc aminoaxit cn c cc hp cht khc khng cha aminoaxit gi l nhm prothetic a2) Da vo hnh dng phn t: chia thnh 2 nhm: -Protein hnh cu: phn t c dng hnh cu , tan trong nc ( anbumin, globulin...) - protein hnh si:

b) Cu trc Cu trc cp 1 (bc 1): l trt t sp xp ca cc gc aminoaxit trong phn t. Cu trc bc 2: l cu dng ca protein: Phn t c th tn ti mt trong hai dng ch yu l dng xon ( cn gi l keratin) v cu dng gp khc ( cn gi l dng -keratin) Cc cu dng ny c duy tr nh lin kt hydro N-H--O=C< gia cc nhm peptit vi nhau Cu trc bc 3: l hnh dng ca mch polipeptit cun li trong khng gian 3 chiu, cc nhm k nc nm pha trong cn cc nhm a nc nm trn b mt phn t. Cu trc bc 3 duy tr c nh tng tc Van der Vall, tng tc tc tnh in, lin kt i sufua S-S-, nhm este -Cu trc bc 4: l mt t hp 2 hoc nhiu i phn t polipeptit kt hp vi nhau nh lc ht Van der Vall v lin kt H gia cc nhm nguyn t phn b trn b mt cc i phn t protein.

9.2.3.2 TNH CHT

a) Tnh lng tnh: cng c tnh lng tnh, im ng in tng t nh aminoaxit v peptit b) Tnh tan: Tnh tan ca chng ph thuc nhiu vo cu to phn t, bn cht dung mi, pH ca dung dch, nhit ... c) S kt ta v s bin tnh: Khi thay i cc yu t nh nng , nhit ...th cc dung dch keo ca protein b kt ta. C hai loi kt ta: Kt ta thun nghch: Kt ta khng thun nghch: S bin tnh:Trong trng hp kt ta khng thun nghch, tnh cht ca protein khc i nhiu so vi cht ban u nn gi l s bin tnh ca protein.

d) Phn ng thy phn: Khi un nng dung dch protein nh xc tc axit, baz, hoc nh men, phn t protein b thy phn to thnh sn phm cui cng lad cc L--aminoaxit. e) Mt s phn ng nh tnh v nh lng protein: -Phn ng nh tnh: + Phn ng biure: phn ng vi Cu(OH)2 trong kim cho phc mu xanh tm + Phn ng xangtoproteic: phn ng vi HNO3 c cho kt ta mu vng -Phn ng nh lng: