Resveratrol A

7/22/2019 Resveratrol A

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Khoa Ha, i hc Khoa hc T nhin Thnh ph H Ch Minh

RESVERATROL vPhng php tng hp

GVHD: PGS.TS. Trn LQuanSV: Nguyn Tn Pht - 081415009-02-2012


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Nguyn Tn Pht (0814150)RESVERATROL v phng php tng hp 2

GVHD: PGS.TS. Trn L Quan i hc Khoa hc Tnhin Tp.HCM, 09-02-2012

MC L

C

PHN I - MU: .................................................................................................. 3PHN II - KHI QUT VRESVERATROL ....................................................... 3

1. PHT HIN:............................................................................................................... 32. TNH CHT: .............................................................................................................. 32.1. L tnh: ...................................................................................................... 42.2. Ha tnh: .................................................................................................... 42.3. Dc tnh: .................................................................................................. 43. TN TI TRONG TNHIN: ................................................................................. 54. THNG MI HA: ................................................................................................ 65. CC HOT CHT LIN QUAN: ............................................................................. 7

PHN III - TNG HP RESVERATROL: .............................................................. 91.CC PHNG PHP TNG HP: ......................................................................... 92.QUY TRNH CA JING LIU: ................................................................................. 102.1. iu ch3,5-diacetoxybenzoic acid: ............................................................... 102.2. iu ch3,5-diacetoxybenzoyl chloride:.......................................................... 112.3. iu chresveratrol triacetate:..................................................................... 122.4. Thy phn triacetate thu resveratrol:.............................................................. 13

PHN IV - THC NGHIM: ................................................................................... 141.IU CH3,5-DIACETOXYBENZOIC ACID: .................................................... 142.1. Tng hp li: ............................................................................................. 142.2. Kho st: .................................................................................................. 172.IU CH3,5-DIACETOXYBENZOYL CHLORIDE: ........................................ 20

PHN V - KT LUN V KHOCH: .............................................................. 241. KT LUN: .............................................................................................................. 242. KHOCH: ............................................................................................................. 24

PHN VI - PHLC: ............................................................................................... 25PHN VII - TI LIU THAM KHO: .................................................................... 28


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PHN I - M U:Cuc sng vnh hng l chni tri trong sut qu trnh tn ti v pht

trin vn minh loi ngi. Nhiu cuc tai bin, chin tranh ci cho n hnh ng

chy ua qun s v khoa hc k thut ca cng quc hin i u khng thkhng l hly ca vn . Con ngi ang u tranh vi nhau v hoc mt loithn dc ci thon ng, mt ci thing ling, thnh tch hay vt bo hmnhnh Chn Thnh, Cn Phng HPhch, Ngn La Vnh Cuhay Thnh a Ngc

Nhn (Shangri-La), ci Bng Laichtn ti trong tr tng tng(!) nhng tt cdng nh ngy mt kh khn. Ri, cc nh gikim n thut s, o s v ti ccnh khoa hc thi i ngh n vic dng thn vo con ng nghin cu iuchra phm thuc thay v u tranh v ngha trn chin trng. Trong scc thnhquthu c c resveratrol, mt hp cht c tin vo khnng ko di ssng vchng li bnh tt.

PHN II - KHI QUT V RESVERATROL

1. PHT HIN:Do vic pht hin resveratrol c bo co trong cc nghin cu ni bnn

cha r ngi u tin c lp c n. Mt nhm nh khoa hc phng Ty tuynbpht hin ln u vo nm1963 trong dch chit c c tcy ct kh (cn gil in tht, h trng tn khoa hc: Fallopia japonica, Polygonumcuspidatum, Reynoutria japonica) tuy c nhng c dng lm thuc trong dn

gian chu (li l mt loi cy xm hi chu u)... khng lu sau bbc bi nghincu vcy khuynh dip bn a c (Eucalyptus wandoo) nm 1959.[8]Ri svicli lp li bi c cc nghin cu trc .[8]Theo ghi nhn hin ti, bi bo caM. Takaoka nm 1939 l nghin cu sm nht. Takaoka c lp resveratrol tloitho dcVeratrum album.[6]

Tn gi resveratrol c th c ghp t rreessorcinol-vveerraattrrum-alcohooll. Bihp phn ca resveratrol l resorcinol, cht ny c tng hp tVeratrum albumv c mang ba nhmOH nh alcohol.[8]

2. TNH CHT:Resveratrol (CTPT: C14H12O3; cn gi l 3,4',5-trihydroxystilbene, 3,4',5-

stilbenetriol hay 5-(p-hydroxystyryl)resorcinol) c xp vo nhm stilbenoid,nhm phenol v nhm phytoalexin, trong thin nhin. C c tnh chng oxy ha,khng vim v em li nhiu li ch cho h huyt mch. Hp cht c mt trongnhiu loi thc vt thng xuyn b nm mc, vi khun quy ph nh nho, nhiuloi tri mng, u phng Gn y ru vang, nc p, scla v b u phngc cp l cc thc phm chbin cha di do lng hot cht ny.
http://en.wikipedia.org/wiki/Veratrum_albumhttp://en.wikipedia.org/wiki/Veratrum_albumhttp://en.wikipedia.org/wiki/Veratrum_albumhttp://en.wikipedia.org/wiki/Veratrum_album


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Hnh 1: trans-resveratrol Hnh 2: cis-resveratrol

2.1.L tnh:

Resveratrol tn ti dng bt trng c ln vng ng: nhit nng chy 259-261oC,[9] khi lng phn t 228.24 g/mol. Khng tan trong nc (0.03 g/L), tan

c trong DMSO (16 g/L) v ethanol (50 g/L).[6]

2.2.Ha tnh:

Resveratrol thhin y tnh cht ca mt phenol, v ca mt alkene.

Resveratrol c hai ng phn: trans-resveratrol v cis-resveratrol. Cu trngcisthu c tchuyn ha dng transtrong iu kin chiu xUV. Resveratrol bnvng trong mi trng tnhin, minh chng bi hm lng ca chng sau khi lnmen ru.[6]

2.3.Dc tnh:Tc ng ca resveratrol ln c thsinh vt ang gy nhiu tranh ci trong

gii khoa hc. Nhiu bo co cho thy resveratrol mang n mt li l ko di tuith, ngn nga ung th, u bu v khng vim song, li c bo co phn bcnhng cng bva nu.

Ko di tui th-Nm 2003 v khng lu sau , nhm nghin cu khoa hc ca nh sinh hc

David Sinclair v Konrad Howitz ng trn chuyn san Nature pht hin kdiu cah v kh nng hot cht m rng thi gian sng nm men (Saccharomycescerevisiae),[1]rui gim (Drosophila melanogaster) v giun trn C. elegans.[2]

-Mt nhm khc thc nghim li v xc nhn kt quca tin s Sinclair trnC. elegans nm 2007.[3]Nhm nghin cu th ba cng b phn bc theo kt quvng sng khng c g bin i cC. elegansvD. melanogaster.[4]


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-Kt qu li cng gy tranh ci khi gii khoa hc pht hin giun trn C.elegans mang trong ngi mt gene kim sot thi gian vn lu nay lm khoa hcgia mt mi tm kim chu ksinh hc loi ny. Giun trn C. elegans gy cch bi mi sinh vt u hot ng v nghngi theo chu kcnh, gi l nhp

sinh hc. Ring, chng l loi duy nht hot ng khng c nhp sinh hc. Chnglm vic lin tc ngy v m.[6][7]

Ngn nga ung thCc nh khoa hc c kt qu thc nghim trn ng vt. Cho thy c

khnng ca hot cht. Cha c nghin cu no c p dng ln ngi tnh nnay.[6]

Khc

Nhng li ch khc cng c bo co gm c mi lin quan trong cha tr

i tho ng, khng vim (bnh ph thng), li cho tim mch, tc ng bo vthn kinh (ca bnh nhn hi chng Alzheimer), khng virus, tng lngtestosterone (gip chut tng tinh dch) y l nhng bo co cha c chngthc.[6]

3. TN TI TRONG T NHIN:Bng di lit k hm lng trans-resveratrol trong cc ging cy trng ph

bin v cc thphm hay chnh phm ca ngnh cng nghip gii kht:[5]

Ngun Ging / Loi lng trans-resveratrol

Ct kh(Polygonumcuspidatum)

R. japonica 64R. bohemica 23R. sachalinensis 29HZ, MB 3770 , 2960

Nc nho Palomino fino 2.4Tri nho Muscadine 5.2-26.4

Ht nhoGamay 3.9Pinot Noir 588

VnhoPinot Noir 118Gamay 6.8fPalomino fino 15.7

B nhoPalomino fino 192Muscadine 22.1-84.2

Ru Muscadine 0.4-2.0Cc loi khc: Cabernet Sauvignon (n = 40),Tempranillo (n = 12), Cabernet Franc (n = 8), Liatiko

0.2-14.3l


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(n = 4), Xinomauro (n = 10), Muscat Bailey A (n =5), Zinfandel (n = 5), Agiorgitiko (n = 9), Marzemino(n = 3), Blaufrnkisch (n = 18), Portugieser (n = 9),Zweigelt (n = 10), Negroamaro (n = 3), Teroldego (n

= 3), Nero dAvola (n = 5)

Ru trngNhiu loi: 21 loi Php, 23 loi M, 18 loi Hungaryv Trung u, 20 loi , 7 loi Canada, 2 loi NamMv 8 loi ca Australia

0.1l

u phng

Jinpoong 3NC-7, om, Gazipaa, Florispan ,erezlik 5025, erezlik PI-355276

0.03-1.92d

Cc ging trng khc: Spanette, Pearl, NC-18016,Early Bunch, Whites Runner, Florunner C1, GA207-3-4, GA 207-2, NC-9, Florispan C1, NC-17291,Dixie Runner, PI-337396-FAV70, Florispan C3,

Small White Spanish

0.02-1.79d

Ru phngJinpoong 1.19Tainan 9, Tainan 11, Tainan 1215-1330

Du ty Allstar 0.09 , 0.83Vit qut(blueberry)

Highbush Michigan 0.03 , 0.02Lowbush "wild" Nova Scotia 0.01 , 0.02

Vit qut en(bilberry)

Ba Lan 0.02f, 0.02d

Ht dci(pistachio)

Ohadi, Uzun, Krmz, Halebi, Siirt 0.09-1.67d

Scla m mu 2.0

Hoa bia

Cc ging trng: Hersbrucker Spat, Spalter, Saphir,Hallertau Mittlefrher, Smaragd, Hallertau Tradition,Wye Target, Nugget, Hallertau Magnum, HallertauTaurus, Saaz, Sladeck, Premiant, Willamette,Cascade, Tomahawk, Simcoe, Warrior

0.10-2.28d

Tettnang (ci to gene) 13 , 2.7Rau dip(lettuce)

?? (ci to gene) 56.4f

Thu c l W38 (ci to gene) 9.3La m Florida, Combi (ci to gene) 35-190Kiwi Hayward (ci to gene) 182

dmg/kg kh fmg/kg ti lmg/L

4. THNG MI HA:Bt chp tranh ci ca gii khoa hc vcng nng xc thc ca hp cht.

Resveratrol c thng mi ha mnh di dng thc phm chc nng by bnrng khp thgii. Cc thtrng chnh ca n l M, Nht Bn, Hn Quc Hm


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lng resveratrol trong thc phm chc nng dao ng t50% n 99%.[6]Ngunsn xut rnht l tcy ct kh (Polygonum cuspidatum).[8]Cc cng ty qung bsn phm nh mt loi tin dc trc cc bnh nan y (tiu ng, ung th) vtt cho cnam ln ngii. i vi ngii, thc phm sgip trha ln da, duy tr

nt thanh xun. Cn vi nam gii th lm tng ni tit tnam, thch hp cho nhngngi tp luyn thhnh.

5. CC HOT CHT LIN QUAN:Nhng hp cht di y thuc cng nhm stilbenoid v c cu thnh t

cc n phn resveratrol vi nhau hoc resveratrol vi glucose:[6]

Hnh 3: Piceatannol

Hnh 4: Pallidol

Hnh 5: Hopeaphenol

Hnh 6: Piceid


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Hnh 7: trans-diptoindonesin BHnh 8: -viniferin


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PHN III - TNG HP RESVERATROL:

1. CC PHNG PHP TNG HP:[9][10]

Phng php tng hp u tin c Takaoka xut bng cch dng phn

ng Perkin ngng t sodium 3,5-dihydroxyphenylacetate vi 4-hydroxybenzaldehyde, v vi acetic anhydride. Sn phm thu c l 3,4,5-triacetoxystilbene--carboxylic acid. Sau , tip tc em kh carboxyl v thy

phn ta thu c resveratrol. Hiu sut cc giai on trong phng php ny khngc bo co cth.

Hnh 9: Phng php tng hp resveratrol u tin dng ngng tPerkin

Hnh 10: p dng phn ng WittigRi cc phng php tng hp da trn phn ng Wittig v ghp cp Horner-

Emmons (cn gi l phn ng Horner-Wadsworth-Emmons, phn ng Wittig-Horner) c cng bnhng khng nhn c tn thnh v cho ra sn phm lhn hp ng phnZ/Ev hiu sut t c khng c bit ni tri.

Phn ng Grignard cng c dng lm c snghin cu tng hp. Nguynliu ban u lp-anisaldehyde v 3,5-dimethoxybenzyl magnesium chloride.

Hnh 11: Tng hp da trn phn ng GrignardNi bt ti thi im hin ti l phng php tng hp bng phn ng ghp

cp Heck, nh khoa hc c trao gii Nobel Ha hc 2010 vi tm quan trng cbit trong pht hin, pht trin v nghin cu sdng Paladium lm tc nhn xcphn ng ghp cp to ni i cu hnh trans. Tng tphn ng Wittig, tc chtgc ca phn ng Heck c gi thnh cao (3,5-dihydroxybenzaldehyde). Aldehyde


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ny c bin i qua nhiu bc thnh alkene (styrene) v sau cho ghp cp vi4-acetoxyiodobenzene. Sn phm qua thy phn cng to ra resveratrol.

2. QUY TRNH CA JING LIU:Nghin cu ny tp trung kho st tng hp resveratrol theo phn ng ghp

cp Heck c skhcarbonyl c bo co bi gio s Jing Liu. Quy trnh tng hpgm bn bc:[9]

2.1.iu ch 3,5-diacetoxybenzoic acid:bc u, bng phn ng ester ha Steglich 3,5-dihydroxybenzoic acid

c ester ha bo vhai nhmOH vtr 3 v 5. y l phn ng ca cc tccht phenol ( nu), acetic anhydride trong dung mi ethyl acetate. Pyridine c

thm vo hn hp ng vai tr nh mt xc tc Steglich (tng t DMAP: 4-dimethylaminopyridine). C chxc tc phn ng c trnh by theo gin :

OH

OH

HOOC

H3C O CH3

OO

O

OH

HOOC

O

O

H3C

H3C

O

O

OH

HOOC

CH3

O

H3C

O

O

O

O

HOOC

CH3

O

O

CH3

Hnh 12: C chphn ng ester ha phenol


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H3C O CH3

OO

N N

CH3

O

O

O

CH3

N O

O

H3C

N H

OH

O CH3

O

H

O

O

CH3

Hnh 13: C chc chp nhn ca xc tc pyridine[11]

u tin, pyridine (hoc DMAP) to viacetic anhydride mt cp ion acetate

v acetylpyridinium bt n nh. Ion phenolate tc kch vo nhm acetyl sinh estertrong giai on hai, dn dn lm lin kt pyridinium v acetyl yu i cho ti khi tla. Trc , i ion acetate v/hoc pyridine de-proton ha phenol cho ra aceticacid v ion phenolate. Acetic acid mi sinh stip tc proton ha pyridine theo mtcn bng. Nhvo trbase (triethylamine), pyridinium c de-proton bc vovng xc tc mi.

2.2.iu ch 3,5-diacetoxybenzoyl chloride:

Sau khi bo vhai nhmOH phenol, gio s tip tc tin hnh phn ngto dn xut acid chloride t3,5-diacetoxybenzic acid. Cc ha cht cn cho phnng bao gm 3,5-diacetoxybenzoic acid, v SOCl2(thionyl chloride) vi sc mtca DMF trong vai tr tc nhn Vilsmeier. Sn phm sinh ra l mt acid chloride rtnhy nc v km bn trong iu kin khng kh m ca Vit Nam.
http://en.wikipedia.org/wiki/Acetic_anhydridehttp://en.wikipedia.org/wiki/Acetic_anhydride


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Cl

S

Cl

O

R

OH

O SO

Cl

O

ClR

O

S

O

Cl

O

O

RSO2Cl O

R

Cl

Hnh 14: Cchphn ng to acyl chloride

2.3.iu ch resveratrol triacetate:

Phn ng c tin hnh ksau bc trn do snhy nc ca tc cht vatng hp. Phn ng iu chresveratrol l sghp cp MizorokiHeck. Bao qut,

phn ng ghp cp Mirozoki-Heck s dng tc cht l mt halide bt bo ha valkene, Palladium xc tc kh hydride ca alkene ng thi to ni n carbon-carbon gia nhnh hydrocarbon ca halide vi phn cn li ca alkene, cho raalkene mi cu hnh trans. Mt bin thca phn ng Heck l phn ng Heck c skh trc tip nhm carbonyl, tc cht u ca phn ng l acid halide v alkene.

Nh to phc bn vi Palladium, carbon monoxide thot ra sau phn ng cngun gc t nhm carbonyl ca acid halide bn cnh trans-alkene mi sinh. Shnh thnh phc cng gia tng hiu sut phn ng i cht. Xc tc palladium

thng gp dng mui acetate, hn l chloride.Chu trnh xc tc c m hnh ha trong mt vng khp kn:


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Hnh 15: Chu trnh xc tc ca Palladium(II)Giai on A: Palladium xen vo gia phn tacid chloride, to ni vi aryl v vi

chlorocarbonyl. Alkene c bsung, tip tc to hp cht phi tr (phc ) vi tmPalladium.Giai on B: Xy ra qu trnh cng syn ca alkene, trong alkene chen vo niPdC hnh thnh ni CCAlkeneCAlkenePd mi. Do sc cng Pitzer, ni CCAlkeneCAlkenePd stxoy sang cu trng trans.Giai on C: tch/loi -hydride sinh phc mi ca alkene v PalladiumGiai on D: phc hi xc tc Pd, kt thc chu trnh cho ra sn phm transalkene,CO v HCl

2.4.Thy phn triacetate thu resveratrol:Bc cui cng ch n gin thy phn resveratrol triacetate trong mitrng acid mnh thu c resveratrol.


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PHN IV - THC NGHIM:Chng ti thc hin tng hp resveratrol theo quy trnh ca Jing Liu. Trong bo co

ny trnh by chhai giai on u ca quy trnh.

1.IU CH 3,5-DIACETOXYBENZOIC ACID:

Hnh 16: Phn ng iu ch

2.1.Tng hp li:

Thit b:Bnh cu 3 c20 mL, c nt siliconBp khuy t

C tBong bng cha kh nitrogen

Ha cht:-Resorcylic acid IUPAC: 3,5-dihydroxybenzoic acid

CTPT: C7H6O4PTK (g/mol): 154.12, rn

MP (oC): ---d (g/mL): ---

pKa: ---Ngun gc: Aldrich-Sigma

Acetic anhydride IUPAC: Acetic anhydride / Ethanoicanhydride

CTPT: C4H6O3PTK (g/mol): 102.09, lng

BP (oC): 139.8d (g/mL): 1.082

pKa:Ngun gc: Merck

Ethyl acetate IUPAC: Ethyl acetate / Ethyl ethanoateCTPT: C4H8O2

PTK (g/mol): 88.105, lng


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BP (oC): 77.1d (g/mL): 0.897

pKa: ---Ngun gc: Trung Quc

Pyridine IUPAC: Pyridine / AzabenzeneCTPT: C5H5N

PTK (g/mol): 79.1, lngBP (oC): 115.2

d (g/mL): 0.9819pKa: 5.25 (acid lin hp)

Ngun gc: MerckFormic acid IUPAC: Formic acid / Methanoic acid

CTPT: CH2O2PTK (g/mol): 46.03, lng

BP (oC): 100.8d (g/mL): 1.22

pKa: 3.77Ngun gc: Trung Quc

n-Hexane IUPAC: HexaneCTPT: C6H14

PTK (g/mol): 86.18, lngBP (oC): 68-69

d (g/mL): 0.6548pKa:

Ngun gc: MerckNaCl (khan) IUPAC: Sodium Chloride

CTPT: NaClPTK (g/mol): 58.44, rn

MP (oC): 801d (g/mL): 2.165

pKa: 6.7-7.3Ngun gc: Trung Quc

Na2SO4 (khan) IUPAC: Sodium sulfateCTPT: Na2SO4

PTK (g/mol): 142.04, rnMP (oC): 884d (g/mL): 2.664

pKa: ---Ngun gc: Trung Quc

Tin hnh phn ng:


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Cho 3,5-dihydroxybenzoic acid (~0.5 g, 3.2 mmol) vo bnh cu, sau ln ltthm EtOAc (7.1 mL), anhydride acetic (1.5 mL, 16 mmol) v pyridine (0.5 mL, 6.4mmol). ui ht khng kh bng dng kh nitrogen lin tc. Sau khuy nhit thng trong 4 gi. Sau thi gian , thm formic acid (0.3 mL, 3.8 mmol) vo, tip

tc khuy trong 1 gina. hn hp phn ng vo nc ri ra v lng pha huc bng nc ct 3 ln, nc mui (NaCl ~10%) 3 ln loi trmui piridinium,

pyridine tha, v formic acid. Dng Na2SO4hoc MgSO4lm khan, em i c quayv sau kt tinh li trong hdung mi n-hexane/EtOAc (1:1) thu c tinh th.[9]

Hnh 17: Shthng thc nghim

Nhn xt sn phm: Tinh thsn phm c mu trng hoc i khi ngmu vngkhng c dng hnh hc nht nh, c mi ln ca pyridine v formic acid. Cthtip tc lm sch bng cch dng phu lc v bnh Buchner, cho tinh thti kt tinh ln mt phu, xi ra bng dung mi EtOAc lnh. Sau qu trnh ny,mi m smt i.[10]Ngoi ra trong trng hp tinh thmu ng, kt qu thuc s l tinh th mu trng tinh khit. Hiu sut phn ng khng c nh,thng bin thin trong khong 75-90%.

Xc minh sn phm:

*o nhit nng chy: Tin hnh ba ln trong phng th nghim trn tng 2, dy Aca trng i hc Khoa hc T nhin. Tinh th c nhit nng chy 159160oC.*HRMS: Tin hnh phng th nghim B16, dy B ca trng i hc Khoa hcT nhin. Pht hin mi M+Na vi m/z = 261.0370 (tnh ton cho C11H10O6 l238.0477). [PHN VI - ]


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*1H-NMR (CDCl3, 500 MHz): 7.72 (d, J = 2 Hz), 7.20 (t, J = 2 Hz), 2.32 (s).Vi 7.72 v 7.20, l hai mi ca proton nhn hng phng c hng sghp J =2 Hz, do nhn hng phng c hai loi proton ghp metavi nhau. Vi tltch

phn ti ci mi 2.32 : 7.72 : 7.20 = 1 : 0.33 : 0.16 = 6 : 2 : 1, chng tnhn

thm c 3H v c 6H (ca mt loi proton) nhnh hydrocarbon ngoi nhn thm.Tng cng c 9H trong nhn hng phng v nhnh hydrocarbon, thiu 1H canhm carboxylic acid. Tn hiu ca proton acid khng pht hin c c ldo strao i nhanh vi vt nc. [PHN VI - ]

2.2.Kho st:Phn ng c kho st li da theo thi gian, theo nng acetic anhydride, vtheo cc cht phgia nhm nng cao hiu sut phn ng trong iu kin thc tca

phng th nghim Vit Nam.

Theo thi gian phn ngVc bn, hn hp phn ng gm cc cht nu theo ng tlthc trn phngtrnh phn ng. Trong smol acetic anhydride v smol xc tc pyridine lnlt gp i nng 3,5-dihydroxybenzoic acid; lng dung mi sdng theo ngtlgio s Jing Liu xut (tc c110 mL EtOAc cho phn ng ca 7.71 g 3,5-dihydroxybenzoic acid).

*Cch tin hnhCho a mol 3,5-dihydroxybenzoic acid vo bnh phn ng, cng vi 2a mol acetic

anhydride v 2a mol pyridine. Thm tip lng dung mi theo tl nu, ri dnga khuy hoc my khuy tlm ng nht ha hn hp phn ng. Thc hin uikhng kh bng dng nitrogen lin tc, nt kn bnh phn ng ynti nhit phng v ch thi gian. Sau thc hin li cc bcchit, lng, lm khan, v c quay (khng thc hin kt tinh li) c sn phm. Kt qu, sinh vin thc hin by phn ng theo thigian c tm lc bng th:

t (h) H%4 80.836 84.9113 83.5414 87.5415 88.0819 92.7725 85.98


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th kho sthiu sut theo thi gian (h)

80

82

84

86

88

90

92

94

0 5 10 15 20 25 30

h

H%

*Nhn xt: Theo thphn ng t hiu sut tt vi thi gian khong t14-20 ging h. Bn phn ng vi bn thi im (4h, 6h, 19h, 15h) c thc hin trongcng mt iu kin nhit , thi im thc hin, dng c ch khc nhau thigian. Ba th nghim cn li (13h, 14h, 15h) c thc hin trong mt hon cnhkhc (ngy khc, nhit phng khc) nn c thmang sai stheo b.

Theo nng acetic anhydride*Cch tin hnhThc hin tng t th nghim v thi gian, trong chthay i tlgia acetic anhydride vi 3,5-dihydroxybenzoic

acid (gi tt l phenol) t2.0 cho n 3.0. Thm tip lngdung mi theo t l nu, ri dng a khuy hoc mykhuy t lm ng nht ha hn hp phn ng. Thc hinui khng kh bng dng nitrogen lin tc, nt kn bnh

phn ng yn ti nhit phng v chthi gian. Sau thc hin li cc bc chit, lng, lm khan, v c quay(khng thc hin kt tinh li) c sn phm. Kt qu,sinh vin thc hin by phn ng theo thi gian c tmlc bng th:

anhydride/ henol H%

2.0 80.832.2 89.192.4 75.352.8 96.093.0 94.88


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th kho stt l acetic anhydride/phenol

70

75

80

85

90

95

100

2 2.2 2.4 2.6 2.8 3

H%

*Nhn xt: Nhm su phn ng c thc hin cng lc: cng nhit , cng bdng c th nghim vi tl2.4 t hiu sut thp hn c thdo mt mt snphm qu trnh chit lng. Khi sinh vin cha hai pha tch tt vi loi bpha nc v tip tc ra pha hu c vi nc. Theo th, tltt nht t hiusut cao c thl anhydride/phenol=2.6

Phn ng vi sc mt ca phgia:Cc ph gia c b sung vo phn ng ln lt l EtONa, Et3N v hn hpEtONa+Et3N cng t l vi acetic anhydride. Hiu sut phn ng thu c khngmy ni tri tng cho thm cc phgia ny xut pht tc chphn ng c

xc tc pyridine: c giai on chuyn dch cn bng pyridine & acetic acid vpyridinium & acetate. Thm EtONa v Et3N y chuyn dch cn bng theo chiusinh pyridine cho vng xc tc mi.


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2.IU CH 3,5-DIACETOXYBENZOYL CHLORIDE:

Hnh 18: Phn ng iu chThit b:


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Bnh cu 3 c20 mL, c nt siliconBp khuy tC tng hon lu

Bong bng cha kh nitrogenNi un ncPipette

Ha cht:3,5-diacetoxybenzoic acid IUPAC: 3,5-diacetoxybenzoic acid

CTPT: C11H10O6PTK (g/mol): 238.05, rn

MP (oC): 161-162d (g/mL): ---

pKa: ---Ngun gc: [tchun b]

DMF IUPAC: DimethylformamideCTPT: C3H7NO

PTK (g/mol): 73.09, lngBP (oC): 153

d (g/mL): ---pKa: ---

Ngun gc: Aldrich-SigmaThionyl Chloride IUPAC: Sulfurous dichloride

CTPT: SOCl2PTK (g/mol): 117.98, lng

BP (oC): 74.6d (g/mL): 1.638

pKa: ---Ngun gc: Merck

Toluene IUPAC: MethylbenzeneCTPT: C7H8

PTK (g/mol): 92.14, lngBP (oC): 110.6

d (g/mL): 0.8669pKa: ---

Ngun gc: Merck

*Cch tin hnh: Cho 3,5-diacetoxybenzoic acid (0.3664 g, 1.54 mmol), DMF (2git) vo bnh cu phn ng lp sn ng hon lu, ri thm vo thionyl chloride (0.7mL, 10 mmol), thc hin ui khng kh bng dng nitrogen lin tc. Dung dch


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c khuy nhit thng trong 15 pht trong kh nitrogen; sau tin hnhun hon lu 80oC trong 2 gi. Dung dch ttrong sut chuyn sang mu vng,ri m mu dn theo thi gian. Dung dch ng rn sau khi ngng un. Sau ,thc hin ui thionyl chloride bng rt chn khng. Cho dung dch toluene vo v

khuy ha tan cht rn cn li. Lc ny xut hin cht rn/vt du khng tantrong toluen mu nu, gn b. ui toluene, ta thu c cht rn mu vng nhtmi hc (0.3583g).[9]

Hnh 19: S hthng thc nghim

*Nhn xt sn phm: Sn phm c mu vng, tnh n mn mnh.

*Xc minh sn phm:+ HRMS: Tin hnh phng th nghim B16, dy B ca trng i hc Khoa hcTnhin. Khng pht hin thy mi cp mi M+/M+2 (do Chlorine c hai ng v35Cl v 37Cl t l3:1) hay M+Na/M+2+Na vi m/z ln lt l 253/255 v 276/278(tnh ton cho C11H9O5Cl l 256.0139). Chnhn thy mi 238.0561 ca cht acidgc. [PHN VI - ]+ 1H-NMR (CDCl3, 500 MHz): Khng c p dng do c bn cu trc ca acid vdn xut acyl chloride ca n c khung sn tng t, chkhc bit nhm OH


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carboxylic acid v nhm thCl dng nh khng lm thay i nhiu dchchuyn ha hc ca cc proton. Trong trng hp tng hp acid bn trn, ph1H-

NMR cng khng cho tn hiu proton acid.+ Trc nghim nhanh: phng php nh tnh nhm chc acyl chloride theo Vogel

[phin bn 5 tr. 1212, nm 1989[12]

] cho kt qusau:-Dung dch AgNO3: cho vo ng nghim dd AgNO3, nhnhcho chy ln thnhng nghim vi git dd acid chloride/toluene c hai lp dd trong sut trongng nghim. Lc ng nghim, lp tc lp dd nc bn di ha c. [DNGTNH viCl]-Thvi BaNO3: pht hin c hay khng SOCl2 tha. Cho vo ng nghim ddBaNO3, nhnhcho chy ln thnh ng nghim vi git dd acid chloride/toluene c hai lp dd trong sut trong ng nghim. Lc u v yn, quan st khng csvn c cc lp. [M TNH vi SOCl2, do SOCl2 trong mi trng nc sb

phn hy to HCl v H2SO3cho phn ng to kt ta trng vi BaNO3. Th nghim

c thcha ng do HCl to mi trng acid cn sto ta BaSO3. C thkim trali bng cch dng Ba(OH)2thay vai tr.]-Thvi alcohol tuyt i: cho vi tinh thacid chloride vo ng nghim tht kh,khong 1mL alcohol tuyt i c thm vo (Quan st xem c hay khng sxuthin ca bt kh HCl hay khng). ngay vo hn hp 2mL dd NaCl bo ha trongnc, thy hnh thnh mt lp hu c trn mt ng thi ngi thy mi thmngt/tri cy ca cc ester. [DNG TNH c trng vi acyl chloride, do SOCl2d khng thto ester c mi thm.]


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PHN V - KT LUN V K HOCH:

1. KT LUN:Phn ng to 3,5-diacetoxybenzoic acid t3,5-dihydroxybenzoic acid c hiu

sut cao, trong khong t75%-90%. Phn ng c sdng cc tc cht l aceticanhydride, 3,5-dihydroxybenzoic acid (phenol), pyridine v dung mi ethylacetate. Hiu sut ti u khi t lmol anhydride:phenol = 2.6:1 trong khongthi gian t14-20h nu khng dng my khuy t. Sn phm thu c thng dng tinh thtrng hoc ngvng. Nhng tinh thny c ra li bng ethylacetate lnh trong phu lc Buchner stinh khit hn v mang mu trng.

Phn ng iu ch3,5-diacetoxybenzoyl chloride phi c tin hnh trong iu

kin khan nc. Cc dng ccn c lm kh sn. Sn phm thu c cn tinhnh ngay phn ng ghp cp Heck (giai on 3) nhm trnh sn phm bhhao do hi nc khng kh. bo qun sn phm, phi trtrong chai lkn vsc kh tr nh Nitrogen hoc tt hn lArgon, t vtr kh thong khng cnh sng.

2. K HOCH:

Trong iu kin c th, sinh vin s tip tc kho st phn ng iu ch 3,5-

diacetoxybenzoic acid theo nng pyridine v nng dung mi.

Tin hnh kho st chi tit phn ng iu ch3,5-diacetoxybenzoyl chloride nuc.

Bc n giai on thba l ghp cp Heck bng xc tc Palladium.

Sau thc hin qu trnh thy phn resveratrol triacetate c resveratrolthnh phm.


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PHN VI - PHLC:

Phlc 1: Ph1H-NMR (CDCl3, 500 MHz) ca 3,5-diacetoxybenzoic acid


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Phlc 2: PhHRMS ca 3,5-diacetoxybenzoic acid vi mi M+Na l 261.0370


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Phlc 3 Chy phHR-MS cho 3,5-diacetoxybenzoyl chloridechthu c mi M+ 238.0561 ca acid gc


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PHN VII - TI LIUTHAM KHO:

1. Howitz, K.T., et al., Small molecule activators of sirtuins extendSaccharomyces cerevisiae lifespan.Nature, 2003. 425(6954): p. 191-196.2. Wood, J.G., et al., Sirtuin activators mimic caloric restriction anddelay ageing in metazoans.Nature, 2004. 430(7000): p. 686-689.3. Gruber, J.A.N., S.Y. Tang, and B. Halliwell,Evidence for a Trade-Offbetween Survival and Fitness Caused by Resveratrol Treatment of

Caenorhabditis elegans.Annals of the New York Academy of Sciences, 2007.1100(1): p. 530-542.

4. Bass, T.M., et al., Effects of resveratrol on lifespan in Drosophilamelanogaster and Caenorhabditis elegans. Mechanisms of Ageing andDevelopment, 2007. 128(10): p. 546-552.5. Zhang, A., et al., Occurrence and estimation of trans-resveratrol inone-year-old canes from seven major Chinese grape producing regions. Molecules, 2011. 16(4): p. 2846-61.6. Resveratrol - Wikipedia English7. "Tm ra gen kim sot thi gian Giun trn C.elegans",

khoahoc.com.vn, ngy: 21-12-20108. "Resveratrol", www.biologie.uni-freiburg.de

9.

Jing Liu, et al., Synthesis of resveratrol and its analogs, phase-transfercatalyzed asymmetric glycolate aldol reactions, and total synthesis of 8,9-methylamido-geldanamycin. Department of Chemistry and Biochemistry,Brigham Young University, 2007: p.3-5, 101-104.10. Dustin Sprouse, The synthesis of Resveratrol. Department of Biologyand Chemistry, School of Science and Engineering, Oral Roberts University2009: p. 33-45.11. DMAP, Steglich's catalyst - Wikipedia English12. Vogels Textbook of Practical Organic Chemistry (Fifth Edition),John Wiley & Sons, Inc., New York, 1989: p. 1212.
http://en.wikipedia.org/wiki/resveratrolhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.biologie.uni-freiburg.de/data/bio2/schroeder/Resveratrol.html#Takaoka1939http://en.wikipedia.org/wiki/4-dimethylaminopyridinehttp://en.wikipedia.org/wiki/4-dimethylaminopyridinehttp://www.biologie.uni-freiburg.de/data/bio2/schroeder/Resveratrol.html#Takaoka1939http://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://www.khoahoc.com.vn/khampha/sinh-vat-hoc/vikhuan-contrung/30791_Dong-ho-sinh-hoc-Tim-ra-gen-kiem-soat-thoi-gian-o-Giun-tron-C-elegans.aspxhttp://en.wikipedia.org/wiki/resveratrol

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Resveratrol A