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B Y T

TRNG I HC DC H NI

NGUYN TH BCH NGC

NGHIN CU THNH PHN HA HC

V TH MT S TC DNG DC L

CA CH PHM TA AN

KHA LUN TT NGHIP DC S

H NI 2014

B Y T

TRNG I HCDC H NI

NGUYN TH BCH NGC

NGHIN CU THNH PHN HA HC V TH MT S TC DNG DC L

CA CH PHM TA AN

KHA LUN TT NGHIP DC S

Ngi hng dn:

1. PGS.TS Phng Ha Bnh

2. DS. Nguyn Th Hi Yn

Ni thc hin:

1. B mn Dc hc c truyn

2. B mn Dc l

3. Vin kim nghim thuc Trung ng

H NI 2014

LI CM N

Sau mt thi gian nghin cu khoa hc, c s gip tn tnh ca cc thy

c gio, cc anh ch k thut vin, bn b, gia nh, ti hon thnh kha lun tt

nghip Nghin cu thnh phn ha hc v mt s tc dng sinh hc ch

phm Ta An.

Nhn dp ny, ti xin c by t lng cm n chn thnh v su sc ti

PGS.TSPhng Ha Bnh, ngi thy gio ng knh dnh nhiu thi gian, tm

huyt tn tnh hng dn, ch bo ti trong qu trnh thc hin ti.

Ti cng xin gi li cm n chn thnh ti DS. Nguyn Th Hi Yn nhit

tnh hng dn, trao i, gip ti trong qa trnh thc nghim.

ng thi, xin trn trng cm n cc thy c gio, cc anh ch k thut vin

thuc B mn Dc ha c truyn, B mn Dc lc Trng i hc Dc H

Ni, phng M phm Vin kim nghim thuc trung ng tn tnh gip , to

iu kin thun li ti c th hon thnh kha lun.

Cui cng, xin cm n gia nh, ngi thn, bn b khuyn khch, ng

vin ti c c kt qu hm nay.

H Ni, ngy 14 thng 05 nm 2014

Sinh vin

Nguyn Th Bch Ngc

MC LC

T VN ......................................................................................................... 1

CHNG I: TNG QUAN ................................................................................... 2

1.1. i cng v bnh tr ...................................................................................... 2

1.1.1. nh ngha ..................................................................................................... 2

1.1.2. Phn loi ........................................................................................................ 2

1.1.3. Nguyn nhn ................................................................................................ 3

1.1.4. C ch bnh sinh ......................................................................................... 3

1.1.5. Triu chng lm sng ................................................................................. 4

1.1.6. iu tr bnh tr ........................................................................................... 4

1.2. Tm tt v ch phm Ta An ..................................................................... 6

1.2.1. Ngun gc thnh phn ch phm Ta An .................................................. 6

1.2.2. Tm tt thng tin cc v thuc trong ch phm ........................................ 6

CHNG II: I TNG V PHNG PHP NGHIN CU ................ 20

2.1. Nguyn vt liu, phng tin...................................................................... 20

2.1.1. Nguyn liu .................................................................................................. 20

2.1.2. Phng tin .................................................................................................. 20

2.2. Ni dung nghin cu ................................................................................... 21

2.2.1. Xc nh tnh ng ca cc v thuc ......................................................... 21

2.2.2. Nghin cu thnh phn ha hc ch phm TA ........................................ 21

2.2.3. Th tc dng dc l .................................................................................. 21

2.3. Phng php nghin cu ............................................................................. 21

2.3.1. Xc nh tnh ng cc v dc liu .......................................................... 21

2.3.2. Nghin cu thnh phn ha hc ch phm Ta An.................................. 22

2.3.3. Th tc dng dc l .................................................................................. 23

CHNG III: THC NGHIM, KT QU, BN LUN ............................. 26

3.1. Thc nghim, kt qu................................................................................ 26

3.1.1. Xc nh tnh ng cc v thuc ............................................................... 26

3.1.2. Nghin cu thnh phn ha hc ch phm Ta An ............................... 37

3.1.3. Th tc dng dc l ................................................................................ 51

3.2. Bn lun ..................................................................................................... 54

3.2.1. Xc nh ng ca cc v thuc ................................................................ 54

3.2.2. Nghin cu thnh phn ha hc ch phm ............................................. 54

3.2.3. Th tc dng dc l................................................................................. 55

KT LUN V KIN NGH .............................................................................. 56

1. Kt lun ....................................................................................................... 56

2. Kin ngh...................................................................................................... 56

TI LIU THAM KHO

PH LC

DANH MC CC K HIU, CC CH VIT TT

AND: Acid deoxyribo nucleic

cAMP: AMP vng.

cGMP: GMP vng

BuOH: n Buthanol

Cyt P450: Cytochrom P450

DVN IV: Dc in Vit Nam IV

DTQ : c in Trung Quc

MeOH: Methanol

EtOAc: Ethyl Acetat

EtOH: Ethanol

MIC: Nng c ch ti thiu

NC: Nghin cu

SKLM: Sc l lp mng

TA: Ta An

TB: Trung bnh

TBD: trc bch dip

TM: hng ma

TNHH: Trch nhim hu hn

TT: Thuc th

TTM: Tim tnh mch

V: Vng en

HH: He hoa

DC: Dip c

C: i chiu

HK: Hong k

dd: Dung dch

HPLC: High Performance Liuid Chromatography

HIV: Human Immuno-deficiency Virus

HVS: Herpes Virus Simplex

ED50: Effective dose for 50 percent of the group

MTT: 3-(4,5-dimethylthiazol)-2-yl-2,5-diphenyltetrazolium

ACN: Aceton nitril

DANH MC CC BNG

Bng 3.1: Kt qu nh tnh Ta An v thng ma bng phn ng ha hc ........... 38

Bng 3.2. Kt qu sc k so snh TA, DC,quercetin.............................................. 39

Bng 3.3. Kt qu sc k so snhTA, TBD, quercetin ........................................... 41

Bng 3.4. Kt qu sc k so snh TA, TM .............................................................. 42

Bng 3.5. Kt qusc k so snh TA, HH, rutin .................................................... 44

Bng 3.6. Kt qu sc k so snh TA, HK, astragalosid IV ................................... 45

Bng 3.7. Kt qu sc k so snh TA,V ............................................................... 46

Bng 3.8. Hm lng cn phn on EtOAc ch phm TA theo khi lng 47

Bng 3.9. Thnh phn pha ng nh lng ng rutin, quercetin bng HPLC .... 49

Bng 3.10. Tnh thch hp h thng HPLC vi rutin ............................................. 49

Bng 3.11. Tnh thch hp h thng HPLC vi quercetin ..................................... 50

Bng 3.12. Hm lng rutin trong ch phm TA.................................................. 50

Bng 3.13 : Hm lng quercitin trong ch phm TA ........................................... 50

Bng 3.14. nh hng ca TA n thi gian chy mu ui chut ....................... 51

Bng 3.15. nh hng ca TA n ph chn chut .......................................... 53

DANH MC CC HNH V, TH

Hnh 1.1. Gii phu v tr tr ...................................................................................... 2

Hnh 2.1. Qui trnh th nghim tc dng chng vim cp trn m hnh gy ph bn

chn chut ............................................................................................................. 25

Hnh 3.1. nh dc liu dip c ............................................................................ 26

Hnh 3.2. nh c im bt dip c ....................................................................... 26

Hnh 3.3. nh dc liu trc bch dip .................................................................. 27

Hnh 3.4. nh c im bt trc bch dip ............................................................ 28

Hnh 3.5. nh dc liu thng ma .......................................................................... 28

Hnh 3.6. nh c im bt thng ma .................................................................... 29

Hnh 3.7. nh dc liu He hoa .......................................................................... 33

Hnh 3.8. nh c im bt He hoa ...................................................................... 34

Hnh 3.9. nh dc liu Hong k ......................................................................... 35

Hnh 3.10. nh c im bt Hong k ................................................................. 35

Hnh 3.11. nh dc liu Vng en ....................................................................... 36

Hnh 3.12.nh c im bt Vng en ................................................................. 36

Hnh 3.13. nh sc k so snh TA, DC, C quercetin ( = 254 nm) .................... 39

Hnh 3.14. nh sc k so snh TA, TBD, C quercetin ( = 366 nm) .................. 41

Hnh 3.15. nh sc k so snh TA v TM ( = 366 nm) ......................................... 42

Hnh 3.16. nh sc k so snh TA, HH, rutin ( = 366 nm ).................................. 44

Hnh 3.17. nh sc k so snh Hong k, TA v C astragalosid IV hin mu .... 45

Hnh 3. 18. nh sc k so snh TA v V ( = 366 nm) ...................................... 46

Hnh 3.19. nh hng ca To an n thi gian chy mu ui chut .................. 52

Hnh 3.20.nh hng ca Ta An n ph chn chut ..................................... 52

1

T VN

Vit Nam c T chc Y t th gii nh gi l mt trong nm nc hng

u trn th gii c h thng y hc c truyn pht trin lu i v ng gp tch cc

vo s nghip chm sc sc khe nhn dn. T nm 1955, ch tch H Ch Minh

nh hng pht trin nghnh y t Vit Nam theo hng hin i, khoa hc, kt hp

cht ch gia c truyn v hin i, tn dng trit ngun dc liu phong ph,

di do v cc kinh nghim cha bnh lu i. Nhiu bi thuc, phng php tr

liu ca Y hc c truyn c nghin cu, pht trin phc v cng tc phng

bnh, cha bnh, nng cao sc khe ngi bnh.

Ta An l ch phm h tr iu tr tr c B Y t Vit Nam cp php lu

hnh, sn xut bi cng ty TNHH Giai Cnh. Sn phm c ngun gc tho dc.

Cng thc Ta An da trn bi thuc gia truyn. Song, Ta An cha c bng chng

v tc dng trn thc nghim khoa hc. Mt khc, nhu cu ca cng ty Giai Cnh l

xy dng tiu chun ca sn phm nhm m bo tnh n nh. V vy, chng ti

thc hin ti Nghin cu thnh phn ha hc v th mt s tc dng dc

l ca ch phm Ta An nhm mc tiu kim nh cc thnh phn ha hc v th

mt s tc dng dc l ca ch phm Ta An.

t c mc tiu , kha lun tin hnh cc ni dung sau:

- Xc nh tnh ng cc dc liu trong thnh phn ch phm Ta An.

- Nghin cu thnh phn ha hc ca ch phm Ta An.

- Th mt s tc dng dc l lin quan ti tc dng iu tr ca ch phm: tc

dng cm mu v tc dng chng vim cp.

2

CHNG I. TNG QUAN

1.1. i cng v bnh tr

1.1.1. nh ngha

Bnh tr, hay cn gi l bnh li dom (theo dn gian), l bnh c to thnh

do gin qu mc cc m ri tnh mch tr (hay s phnh tnh mch) m xung

quanh hu mn; l bnh rt ph bin, ng hng u trong cc bnh l v vng hu

mn phi nhp vin [1],[5],[15].

1.1.2. Phn loi

Bnh tr l s phng ln ca mt hay nhiu tnh mch thuc h thng tnh

mch tr trn hoc tnh mch tr di. Da vo c im ny, c th phn loi bnh

tr thnh tr ni, tr ngoi hay tr hn hp [1], [15].

Tr ni: Cc xoang tnh mch tr trn

phng to, tr c hnh thnh trn

ng lc,thng nm trong ng hu

mn v khi phnh ln s sa ra ngoi. Tr

ni c phn chia lm 4 cp :

- Tr ni 1: Cc bi tr nm hon ton

trong ng hu mn, thng ch c biu

hin au rt v ra mu.

Hnh 1.1. Gii phu v tr tr

- Tr ni 2: Bi tr thp th hu mn khi i i tin, sau t co vo trong

ng hu mn.

- Tr ni 3: Bi tr sa hn ra ngoi sau khi i tin v khng t co vo trong ng

hu mn m phi dng tay y bi tr mi vo trong c.

- Tr ni 4: Bi tr sa ra ngoi hu mn v thng trc bn ngoi, dng tay

y nhng khng vo c hoc vo sau li sa ra ngay.

Tr ngoi: Cc xoang tnh mch tr di phng to, tr c hnh thnh

ding lc pha ngoi hu mn (ngay ra hu mn). Khc vi tr ni l hnh

thnh trong ng hu mn ri mi sa ra ngoi.

3

Tr hn hp: Tc l trn cng mt bnh nhn xut hin c tr ni v tr ngoi.

Thng thng, khi din tin lu ngy, phn tr ni v phn tr ngoi s lin kt vi

nhau, to thnh tr hn hp.

1.1.3. Nguyn nhn

Nguyn nhn bnh tr cha c xc nh r rng v chc chn. Nhng yu t

sau y c coi nh l nhng iu kin thun li cho bnh pht sinh:

- To bn kinh nin: Nhng bnh nhn ny mi khi i cu rn nhiu, khi rn p lc

trong lng ng hu mn tng ln gp 10 ln. To bn lu ngy lm xut hin cc bi

tr. Cc bi tr dn dn to ln v khi to qu s sa ra ngoi.

- Hi chng l: Nhng bnh nhn b bnh l mi ngy i tin nhiu ln, mi ln

i tin phi rn nhiu lm tng p lc trong bng.

- Tng p lc bng: Nhng bnh nhn vim ph qun mn tnh, nhng bnh

nhn gin ph qun, phi ho nhiu, nhng ngi lm lao ng nng nh khun

vc... lm tng p lc trong bng, d dng cho bnh tr xut hin.

- T th ng: Khi nghin cu p lc tnh mch tr, ngi ta ghi nhn p lc tnh

mch tr l 25cm H2O t th nm, tng vt ln 75cm H2O t th ng. V vy, t

l mc bnh tr cao hn ngi phi ng lu, ngi nhiu, t i li nh th k bn

giy, nhn vin bn hng, th may ...

- U bu hu mn trc trng v vng chung quanh: Ung th trc trng, u bu

vng tiu khung, thai nhiu thng khi to c th chn p v cn tr ng v tnh

mch hi lu lm cho cc m ri tr cng phng ln to thnh bnh tr. Trong

nhng trng hp ny, tr c to nn do nhng nguyn nhn c th, r rng nn

c gi l tr triu chng, khi iu tr ta phi iu tr nguyn nhn ch khng tr

nh bnh tr.

1.1.4. C ch bnh sinh

- Theo thuyt c hc:Cho rng m ri tnh mch nm mt phng su ca lp

di nim mc c gi ti ch bi cc si c c tnh cht n hi. Khi c hin

tng thoi ho keo th cc di ny trng nho, v nu p lc trong xoang bng tng

4

ln do to bn hoc do ri lon i tin th cc bi tr ni cng phng ln. Lc u

cc bi tr cn nm trong lng hu mn nhng khi cc di treo t hn th chng sa

ra ngoi hu mn.

- Thuyt huyt ng hc: Cho rng trong lp di nim mc ca phn thp trc

trng v ca ng hu mn c rt nhiu khoang mch. Vch cc khoang ny c ch

dy ch mng to nn t chc hang. y c s thng ni gia ng mch v tnh

mch. Hin tng chy mu trong bnh tr l do cc ri lon tun hon ti ch ca

chnh cc mch mu thng ni ny

1.1.5. Triu chng lm sng

- Chy mu: L triu chng c sm nht v cng l triu chng thng gp nht.

hnh thc chy mu rt khc nhau, lc u chy mu kn o v sau mi khi i tin

phi rn do to bn th mu chy thnh git hoc thnh tia. Khi nng bnh nhn

ngi xm, i li nhiu cng chy mu thm ch i tin ra mu cc do mu t bi tr

chy vo trong bng trc trng v ng .

- au: Hin tng au thng gp trong cc trng hp tc mch, nt hu mn,

c bit khi i tin. Cng c th gp khi c p xe i km khu tr di lp nim

mc hu mn. Khi tr sa ra ngoi v ph n, bi tr c th sng kh to v mc nght

khng th y ln c.

- Sa tr: Tr ni lc u khu tr hon ton bn trong hu mn, khi to ln th sa

xung nm ngoi hu mn.

1.1.6. iu tr bnh tr

Tr l mt trng thi sinh l bnh thng, ch khi no c s mt iu chnh mi

tr thnh bnh l. iu tr tr khi bnh nhn c nhng triu chng kh chu, au

n, nga, sng, chy nc hay chy mu. V vy, iu tr tr ch yu l iu tr cc

triu chng [7].

1.1.6.1.iu tr tr ni

Ch dinh dng: Ch n cn b sung nhiu cht x.

Thuc dng ton thn:

- Dng thuc lm bn thnh mch ngn cn chy mu: thuc cha vitamin PP,

5

cc flavonoid, rutin...[7], [15].

- Dng thuc gim au, tt nht l dng gim au bng paracetamol, khng dng

gim au opioid v c th gy to bn.

- Khi tr c vim, ph n hay tc mch th dng cc thuc chng vim: C

corticoid v chng vim khng steroid hay enzym chng vim nh

alphachymotrypsin

- Dng thuc nhun trng nu c to bn: Thuc nhun trng thng dng l

nhun trng c tc dng lm mm phn, ngoi ra nhun trng thm thu cng c th

dng.

Thuc dng ti ch

C thdng thuc bi ngoi gim cc triu chng, cc thuc bi ngoi

thng dng gmcc thuc [7], [15], [23]:

- Thuc t, gim au: gim au, nga bng rt, kh chu, kch ng xung

quanh hu mn: benzocain 5-20%, pramoxin 1%, dibucain 0.25-1%, tetracain 0.5-

1% lidocain 2-5%...Tc dng khng mong mun c th gp l phn ng qu mn

vi bng v nga.Ngoi ra c th dng menthol, campho... gim au, gim nga.

- Thuc co mch: gim chy mu, thuc cng lm gim nga v vim tm thi:

Dung dch ephedrin sulfat 0.1-0.125%, phenylephrin HCl 0.25%, epinephrin HCl

0.005 0.1%. Cc thuc ny chng ch nh dng cho bnh tim mch, tng huyt

p, cng gip, tiu ng, ph i tuyn tin lit.

- Cht bo v: To hng ro vt l bo v da, nim mc tm thi gim nga,

kh chu, bng rt. ng thi ngn kch ng cc m trc trng hu mn v s

mt nc lp sng biu b.

Cht hp ph: Ht cc cht dch, cht bi tit trn da hay nim mc.

Cht bi trn, lm du, lm mm: Kt hp vi nc to cht keo bo v da hoc

l cc cht du bi trn hoc lm mm da.

Mt s cht khc hay c kt hp do tnh cht lm du hoc lm sn nim mch

nh: mt s mui bismuth, km oxid, resorcinol, bm Peru, cao cy kim mai.

6

- Thuc chng vim ti ch: C th dng cccorticosteroid nh; hydrocortison

0.25-1% gim vim, gim nga, cc cht kt hp ny ch c tc dng ngn

hn.

- Dng khng sinh ti ch nu c nhim khun: Neomycin, framycetin

- Thuc lm lnh vt thng: Thuc kch thch s lm lnh vt thng hoc ti to

m quanh trc trng hu mn.

- Tim thuc lm x: Tim du phenol hoc polidocanol... vo chn bi tr gy

x ho lm bi tr teo nh li.

- Tht tr: Nhm gy thiu mu nui dng, bi tr s hoi t v rng i.

- Cc phng php khc: t in, chiu tia, phng php lnh...[15].

1.1.6.2. iu tr tr ngoi

Tr ngoi c huyt khi phi dng phu thut ct b bi tr.

1.2. Tm tt v ch phm Ta An

1.2.1. Ngun gc, thnh phn ch phm Ta An

Ta An l snphm h tr iu tr bnh tr c sn xut v phn phi bi

cng ty TNHHGiai Cnh.

- Thnh phn trong mi vin gmcao kh: Dip c 20%; Trc bch dip 20%;

Thng ma 20%; He hoa10%; Hong k10%; Vng en20%.

- Khi lng vin: 550 mg.

- Quy cch ng gi: Vin nang mu sm, ng gi v 10 vin, hp 6 v.

- S K: 289/2011/ YT CNTC.

- Cng nng: B kh, ch huyt, nhun trng.

- Ch tr: H tr iu tr bnh tr. Thch hp cho ngi b tr, sa bi tr, au rt hu

mn do tr, to bn.

1.2.2. Tm tt thng tin cc v thuc trong ch phm

1.2.2.1.Dip c(Ng tinh tho)

7

V thuc l b phn trn mt t phi hay sy kh ca cy Dip c (Houttuynia

cordata Thunb.), h L gip (Saururaceae)[10], [17], [30].

a) Phn b

Lc chu , t Nht Bn, Trung Quc, Thi Lan, n Vit NamLo v cc

nc ng Nam khc. nc ta, cy mc hoang khp ni m thp[6] [19].

b) Thnh phn ha hc

- Cc flavonoid: Afzelin, hyperin, rutin, isoquercitrin v quercitrin[6],[8],

[17],[19],[28], [30].

- Tinh du: Thnh phn ch yu l aldehyd v dn xut nhm ceton nh methyl-n-

nonyl ceton, l-decanal, l- dodecanal, 3oxododecanal. Nhm terpen: bao gm

camphen, myrcen, -pinen, limonen, linalolNgoi ra cn cha acid caprinic,

laurinaldehyd, benzamid, acidhexadecanoid, acid decanoic, acid palmetic, lipid v

vitamin K [6], [17], [19], [28], [30].

- Alcaloid : Cordalin[6], [17], [19], [28].

- Ngoi ra cn c nhiu cht khc : N ( 4-hydroxystyryl)-benzamid, aristolactam,

alcaloid nhn pyridin, 1,3,5-tridecanonylbenzen[6], [17], [19], [28].

c) Tc dng dc l

- Khng virus do tc dng ca quercitrin v tinh du(khng c decanonyl

acetaldehyd). Tinh du c ch c trc tip cc virus l: Virus gy Herpes chng

1, virus gy bnh cm v virus HIV chng 1 ngi.Th nghim o mu MTT cho

thy, dch chit dip c ton phn trong nc nng c tc dng c ch HVS ng k

nng 250 mg/ml (10.2% HVS-1, p< 0.05; 32.9% cho HVS-2,

p< 0.005). ED50 vi HVS-1 v HVS-2 ca dch chit dip c l 822.4mg/ml v

362.5 mcg/ml[17], [24],[30].

- Khng khun do tc dng ca decanonyl acetaldehyd: Dch chit dip c nng

1/40c tc dng c ch mnh nhiu chng vi khun nhStreptococcus aureus,

Pseudomonas aeruginosa, E.coli, Staphylococcus aureus... [17], [19],[29], [30],

[33], [34].

8

- Tc dng chng vim: Dch chit cloroform, dch chit nc, dch chit cn, dch

chit BuOH ca dip c u c tc dng chng vim trn chut cng c gy vim

bng Xylen. Dch chit nc cho tc dng tt hn dch chit cn, dch chit dc

liu ti tt hn dch chit dc liu kh.Th nghim trn chut lang cho

thy,quercitrin phn lp t dip c mc liu50, 100, v 200 mg/kg ng ung c

tc dng c ch ph trn chn chut c gy ph bi carrageenin, dextran,

histamin, serotonin, bradykinin [29], [35], [36], [42].

- Li tiu do hot cht quercitrin: Dung dch flavonoid ton phn nng 1mg/ml

c ch hot tnh men polyphenoloxydase huyt thanh ngi bnh thng mc

13.5%, nng 5mg/ml c ch 50% hot tnh men ny[9], [17], [19], [28], [29].

- c ch thn kinh trung ng: Nc sc, tim di da v phc mc chut nht

trng, c tc dng c ch vn ng t gic, ko di thi gian gy ng ca thuc ng

barbiturat, c ch co git do srtychnin. Tim tnh mch mo lm bin i v in

no tng ng trng thi an thn[17], [19].

- Tc dng chng i tho ng: Dch chit dip c c tc dng chng i tho

ng trong th nghim trn chut i tho ng gy ra bi Streptozotocin [45].

- Tc dng lm bn thnh mch do rutin, quercitrin c chng minh[17].

- Dn cht ca dioxyflavonon (3-4 dioxyflavonol) c tnh cht ca rutin: Tng sc

chu ng ca vi ti huyt qun lm cho huyt qun kh t v (Nht ch dc

1951) [13].

- c ch histamin v acetylcholin th hin trn s gim co tht c trn rut c

lp[19].

- Chng d ng: Dch chit dip c c tc dng c ch phn ng d ng trn chut.

Th nghim v tc dng ca nc rn h mang gy v dng bo v gii phng

histamin v mt s cht trung gian ha hc khc cho thy c mi lin quan gia tc

dng chng nc rn c v tc dng chng d ng ca dip c[19],[41].

- Kch thch d, gy phng do cordalin [13].

d) Tnh, v, quy kinh

V chua cay, hi c c, hi lnh, quy kinh ph [6], [10],[13], [18], [19].

9

e) Cng nng

Thanh nhit, gii c, li tiu, tiu thng, st trng[10], [13], [18], [19], [30].

f) Ch tr

To bn, tr, mn nht, l nga, tr con ln si, vim phi hoc phi c m, au

mt hoc au mt do nhim trc khun m xanh, vim rut, kit l, b tiu tin,

kinh nguyt khng u, st rt[6],[10], [13], [18], [19], [29].

g) Liu dng

Ngy dng 6 12g [6] [13].

1.2.2.2. Trc bch dip

V thuc l cnh v l phi hay sy kh ca cy Trc b (Platycladus orientalis

(L.) Franco), h Hong n (Cupressaceae)[6], [10], [13], [20].

a) Phn b

Phn b ch yu t Bc M ti ng Bc . Vit Nam, cy c trng khp ni

lm cnh v lm thuc[6], [20].

b) Thnh phn ha hc

- L trc bch dip cha tinh du (0.6 1%), flavonoid, lipid v acid hu c [6],

[20].

+ Tinh du c thnh phn ch yu gm fenchon, campho, borneol acetat,

terpincol [6], [20].

+ Flavonoid: quercetin, myricetin,hinokiflavon, amentoflavon [6], [20].

- Phn sp sau khi x phng ha cha 81% acid hu c, ch yu gm cc acid

juniperic, acid sabinic v 17% hexadecane-1,16- diol. Cc acid hu c dng

estolide [6], [20].

c) Tc dng dc l

- Cm mu: Th nghim in vivo trn ch v th thy nc sc trc bch dip c tc

dng ging nh vitamin K: Lm gim thi gian Quick, tc lm tng t l

prothrombin trong mu sau khi dng thuc chng ng mu. Trong th nghim

10

lm sng mt bi thuc c cha trc bch dip 66 bnh nhn tr chy mu, c

92.4% bnh nhn cm muhon ton, 4.6% gim chy mu [13], [20], [29].

- c ch in vitro mt s chng vi khun, siu vi khun[20].

- Tc dng trn thnh mch mu c lp (phng php Kravkov): Tin hnh th

nghim trn th chng 2kg. Dng dung dch 100% trc bch dip sao vng en, pha

long vi nc Ringer cho chy qua tai th. Nng 0.2%, 0.5%, 0.8%, 1% u

c tc dng co mch. Nng 5% - 10% c tc dng gin mch[13], [20].

- Tc dng ci thin s suy gim ca qa trnh thu nhn ca tr nh trn chut nht

trng liu 250 v 500 mg/kg/ngy [20].

- c ch mnh s gn ca cc yu t hot ha tiu cu vo tiu cu th [20].

- Tc dng trn t cung: Trn t cung c lp th thy nhp co bp ca t cung

mau hn, bin rt cao so vi bnh thng. Tc dng r rt nht mc liu 1%.

Vi nng 5%, trng lc co bp mnh nht[13].

- c tnh: th nghim nc sc trc bch dip sao vng en trn th, kh v

chut lang, thy: Vi th liu 100g/kg mt ln, th khng cht sau 4 ngy theo di.

Vi liu 30g/kg khng lm cht, sau na thng theo di (dung dch 200%). Vi

chut lang, liu 64g/kg (dung dch 400%) khng thy cht[13].

d) Tnh, v, quy kinh

V ng cht, hi hn, quy kinh ph, can, i trng[6], [10], [13], [20].

e) Cng nng

Lng huyt, cm mu, tr thp nhit[6], [10], [13], [20].

f) Ch tr

Ho ra mu, i tin ra mu, chy mu cam, xut huyt t cung, rong kinh. Cn dng

lm thuc cha l, st.Trong ng y, trc bch dip c dng lm thuc cm mu

di dng thn sao( sao chy) [6], [10], [13], [20], [29].

g) Liu dng

Ngy dng 6 12 g [10], [13], [20].

h) King k

Khng nn dng cho ngi i ngoi phn lng, nhiu m[6], [13].

11

1.2.2.3. Thng ma

V thuc l thn r phi hay sy kh ca cy i tam dip Thng ma (Cimicifuga

heracleifolia Kom.), Hng an Thng ma (Cimicifuga dahurica (Turcz) Maxim.),

hoc Thng ma (Cimicifuga foetida L.), h Hong lin (Ranunculaceae)[10].

a) Phn b

V thuc c nhp khu t Trung Quc. Cc v thng ma thng gp l:

- Bc thang ma: l thn r kh ca cy bc thng ma hay ng bc thng ma

(Cimifuga dahurica Maxim), h Mao lng (Ranunculaceae)[13], [20].

- Thin thng ma: l thn r kh ca cy i tam dip thng ma(Cimifuga

heracleifolia Komar), h Mao lng (Ranunculaceae) [13], [20].

- Ty thng ma: l thn r kh cu cy thng ma(Cimifuga foetida L.), h Mao

lng (Ranunculaceae) [13], [20].

- Qung ng thng ma: l thn r kh ca cy thng ma (Serratula sinensis S.

Moore.), h Cc (Asteraceae) [13], [20].

b) Thnh phn ha hc

- Thnh phn ch yu l cc triterpen, bao gm cimigenol, cimigenol 3- O-- D-

xylopyranoid, dahurinol, acid isoferulic... Hm lng triterpen trong thng mal

khong 4.3%[20],[30].

- Cc phenolic glycoside: isocimifugamid, cimidahurin v cimidahurinin [20],

[30].

- Cc hp cht furochromon nh: visamminol, visnagin, norvisnagin[20], [30].

c) Tc dng dc l

- Tc dng trn c trn: Visamminol v visnagin c tc dng chng co tht trn

hng trng c lp chut lang[20].

- Tc dng ln chuyn ha lipid: Lm gim nng cholesterol v triglycerid trn

chut cng trng c tng lypid mu gy ra bi vitamin D v cholesterol [20].

- Tc dng h ng huyt: Acid isoferulic c tc dng chng tng ng huyt

invivo[20].

- iu tr sa sinh dc [20].

12

- Tc dng trn gan: 3-O--D-xylopyranoid c tc dng d phng tn thng

gan[20].

- c: Chut TTM ximitin liu 100mg khng thy trng c. Ngi ung qu

liu thy u chong mt hoa, mch v hi th gim, kch thch d dy, nn ma,

nng c th gy chong vng, nhc u, suy nhc, pht cung nh[13].

d) Tnh, v, quy kinh

V ngt cay, hi ng, tnh bnh, hi c, quy vo 4 kinh: t, v, ph, i trng [13],

[20].

e) Cng nng

Thanh nhit, gii c, tn phong, thng dng [6] [20].

f) Ch tr

Cha au nhc rng, lot hng, st rt, nhc u, ban si, u ma, bch i, bng

huyt, sa t cung [6], [20],[29].

g) Liu dng

Ngy dng 4 10 g[13].

h) King k

Hen suyn, ho nc, nn e, ha bc nhc u [20].

1.2.2.4. He hoa

V thuc l n hoa phi hay sy nh n kh ca cy Ho (Styphnolobium

japonicum (L.) Schott, Syn. Sophora japonica L.), h u (Fabaceae) [3], [10],

[12].

a) Phn b

Ch yu vng nhit i v cn nhit i. Vit Nam, he c trng nhiu cc

tnh Thi Bnh, Ngh An, Hi Phng, H Nam, Lai Chu ...[6], [16],[19].

b) Thnh phn ha hc

- Thnh phn chnh l rutin, hm lng t 6 30%, c th ln ti 34%. Rutin l

mt loi glucozid, khi thy phn s cho quercetin, glucoza v ramnoza[6], [10],[13],

[19], [29].

- Cn c betulin, sophorin A, sophorin B v sophorin C[6], [13], [19].

13

c) Tc dng dc l

- Tng cng sc khng, gim tnh thm ca mao mch, hi phc tnh n hi

ca mao mch tn thng do tc dng ca rutin v quercetin.Trn th th

nghim, rutin TTM liu 1 mg/kg lm chm s khuch tn ca cc cht mu (xanh

trypan, xanh evans...) vo t chc di da khi chng c TTM [2], [13], [19],[29].

- Tc dng chng vim: Th nghim trn chut cng trng cho thy, rutin v

quercetin c tc dng c ch ph chn chut do albumin, histamin, serotonin, v

sng khp khuu do men hyaluronidase. Trn th c gy mn cm, rutin TTM

gip phng nga vim da d ng v hin tng Arthus. Trn ch gy vim tc

tnh mch, rutin c tc dng thc y qu trnh hi phc bnh[19], [30].

- Tc dng bo v c th chng chiu x: Chut nht trng tim di da rutin liu

2 mg/kg c tc dng gim t l t vong khi b chiu x[19].

- Tc dng h huyt p: Dch chit n he hoa TTM trn ch gy m c tc

dng h huyt p r rt. Trn chut cng trng cao huyt p di truyn, rutin TTM

liu 1 mg/kg c tc dng h huyt p [13], [19], [29].

- Tc dng h cholesterol mu: Quercetin tim di da liu 10mg/kg cho chut

cng trng c gy cholesterol mu tng caobng cch trn vo thc n hng

ngy cholesterol 30mg/kg v 6-methylthiouracil 90 mg/kg gip h cholesterol mu,

ng thi c tc dng iu tr v phng nga bnh x va ng mch thc nghim

[19], [29], [32].

- Tc dng cm mu:He hoa sao chy v nc sc he hoa, tim xoang bng liu

9g/kg c tc dng rt ngn thi gian chy mu.N hoa he sao chy nhit 190

195oC th hin tc dng mnh nht[19], [29], [30].

- Tc dng trn tim: Trn tiu bn tim ch c lp, quercetin th hin tc dng

cng tim. Trn tim ch c lp, dung dch quercetin nng 50 100 mol/l lm

hm lng cAMP tng 40%, hm lng cGMP khng thay i.Quercetin cn c tc

dng gin mch vnh, ci thin tun hon tim. Trn m hnh gy thiu mu c tim

thc nghim, quercetin TTM liu 10 ml/kg dung dch nng 0.5 mmol/l lm gim

14

thi gian xut hin ri lon nhp tim, h thp hm lng MDA trong t chc

tim[13],[19],[30].

- Bo v gan: Th nghim trn chut cng trng, dng carbon tetraclorid gy tn

thng gan th hot ng ca men Cyt P450 b c ch, dng rutin ng d dy liu

100 mg/kg c tc dng kch hot tr li men ny [19], [30].

d) Tnh, v, quy kinh

V ng nht, hi hn, vo hai kinh can v i trng[6], [10], [12],[18], [19].

e) Cng nng

Thu lim, thanh nhit bnh can, thanh ph chng vim, lng huyt, ch huyt[10],

[12], [19].

f) Ch tr

Dng trong cc trng hp xut huyt, phng tai bin do mch mu b x va,

xut huyt di da, xut huyt vng mc, tr chy mu, xch bch l; Tng huyt p

th nh v va[10], [12], [18], [19], [29], [30].

g) Liu dng

Ngy dng 6 12g, c khi n 20g[10], [12],[19], [30].

1.2.2.5. Hong k

V thuc l r phi hay sy kh ca cy hong k Mng C (Astragalus

membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao, hoc cy hong k

Mc Gip (Astragalus membranaceus (Fisch.) Bge.), h u (Fabaceae) [3], [10],

[14], [16], [21].

a) Phn b

Trng tng i ph bin Trung Quc[6], [13], [19].

b) Thnh phn ha hc

- Polysaccharid: saccharose, glucose, tinh bt, cht nhy, gm [6], [12],[19],[21],

[30].

- Saponin: c 9 astragalosid v isoastragalosid l astragalosid I, astragalosid II,

astragalosid III, astragalosid IV, astragalosid V, astragalosid VI, astragalosid VII,

15

isoastragalosid I, isoastragalosid II v 2 saponin kiu olean: astragalosid VIII,

soyasaponin I[6], [12], [19], [21], [30].

- Cholin, betain, nhiu loi acid amin, calycosin[6], [19], [21].

- Trong hong k cn c selenium [13].

c) Tc dng dc l

- Tc dng tng cng min dch: Hong k c tc dng hip ng vi interferon,

lm tng kh nng p ng ca interferon i vi nhim virus, tng kh nng thc

bo, tng khng th.

+ Th nghim in vitro: Polysaccharid nng 10 mg/ml chit t hong k lm

tng kh nng thc bo ca cc i thc bo v bch cu a nhn. Tch t bo

n nhn t bnh nhn ung th vi polysaccharid hong k s lm tng hot

tnh min dch, iu chnh li chc nng t bo T b suy gim.

+ Th nghim in vivo: TTM polysaccharid hong k khc phc c s c ch

min dch do cyclophosphamid chut cng trng. Nc sc hong k, cho

chut cng trng ung hng ngy hoc cch ngy trong 1 - 2 tun lm tng chc

nng thc bo ca h li ni m.

+ Nghin cu lm sng: Cao nc nng hong k, cho ngi ung liu

15.6g/ ngi/ ngy trong 20 ngy lm tng c ngha IgM, IgE, cAMP[6], [12],

[19], [21], [30].

- Tc dng trn tim: Cao cn hong k lm tng tnh co bp v bin co bp ca

tim ch c lp. Saponin hong k lm tng sc co c tim c lp ca chut cng

trng. Trn t bo c tim, hong k lm gim in th ngh v tng sc co. Trn

bnh nhn b au tht ngc, dng hong k c i chiu vi lidocain v mexiletin

lm gim VLP khong 10%[19].

- Tc dng gin mch h huyt p: TTM dch chit hong k cho ch, mo gy

m hoc th khng gy m thy gin mch v h huyt p ko di. Hong k cn c

tc dng bo v mao mch, chng gin, v do chiu x tia X [19], [30].

- Tc dng li niu: Nc sc 0.5g/kg TTM chut cng c tc dng li niu r rt.

Liu 0.2g/kg cng c tc dng li niu. Ch ung hong k liu 0.5 4g/ kg, gy

16

li niu. Th nghim trn th cng cho tc dng tng t. Nu cho ung ko di th

nhng ngy sau hiu qu khng r rt [13], [19].

- Tc dng trn gan: Hong k lm tng sinh tng hp ADN, tng ti sinh gan

chut nht trng ct mt phn gan. Hong k c tc dng bo v gan, ngn nga

s gim hm lng glycogen gan, lm tng hm lng protein v albumin ton

phn trong huyt thanh trn chut nht trng gy tn thng gan bng carbon

tetraclorid [19].

- Tc dng chng vim: Astramembrannin I chit t hong k, TTM liu 0.5

mg/kg hoc ung liu 50 mg/kg c ch s tng tnh thm mch chut cng trng.

Thuccn c tc dng c ch ph do carragenin chut cng trng[13], [19].

- Tc dng trn h sinh dc: Hong k c tc dng ko di thi k ng dc ca

chut bch, gy hng phn co bp t cung c lp chut cng c thai, nhng li c

ch s co bp ca rut th[13],[19].

- Tc dng c ch aldose reductase [19], [29].

- Tc dng khng khun: nc sc hong k 100% c tc dng khng cc chng vi

khun l Shigella, t cu vng, trc khun m xanh, thng hn,lin cu khun

dung huyt, ph cu[6], [13],[19], [29].

- c tnh: Hong k c c tnh thp. Cho chut nht trng ung liu 100g/kg, l

liu gp 500 ln liu thng dng cho ngi, khng thy c chut cht, khng c

biu hin tc dng ph c hi [19].

d) Tnh, v, quy kinh

V ngt, tnh n, vo hai kinh ph v t[6], [12], [13], [19].

e) Cng nng

B kh trung tiu, ch huyt, thng dng, c biu lim hn, li niu tiu ph, tr

tiu kht, sinh tn dch, Gii c tr m[12], [13], [14], [19].

f) Ch tr

- C th suy nhc, chn tay v lc, yu hi, chng mt, km n, cc bnh sa

ging tng ph, t cung, tr, l , t lu ngy; Huyt h, thiu mu, c bit l thiu

mu sau st rt; Ra nhiu m hi, m hi trm[12], [13], [14], [21].

17

- c dng sng cha i tho ng, i c, i but, ph thng, vim thn

mn, albumin niu, l lot, phong thp, au xng [6], [12], [19].

- Tm mt cha suy nhc lu ngy, ra nhiu m hi[6], [19].

g) Liu dng

Ngy dng 6 - 12g dng sng, 3 - 9g dng sao tm, thuc sc hoc cao [13], [19].

h) King k

Khng dng cho ngi m h, mu nng gy kh, bc ha, tai, chng mt, nhc

u, i tho ng, cao huyt p, cc bnh n nhit, xut huyt, vim no, cc loi

sng vim[6], [12].

1.2.2.6. Vng en

V thuc l ht phi kh ca cy vng en (Sesamum indicum L.) thuc h Vng

(Pedaliaceae) [14].

a) Phn b

Trng vng nhit i chu , ti cc tnh pha nam Trung Quc, n ,

Malaysia, Thi Lan, Vit Nam, Lo, Camphuchia... [6], [20].

b) Thnh phn ha hc

- Du vng: 40-55%, mu vng, gm acid oleic, acid linoleic, acid myristic, acid

palmatic, acid stearic, acid arachidic, acid haxexadeconoic, sesamin sesamol. Trong

du vng c cha cht sesamin 0.25- 1% v sesamol khong 0.1%[6], [13], [20],

[31].

- Protein: 20- 22%, gm -globulin v -globulin [6], [13], [20].

- Carbohydrat: glucose, sucrose, galactose, planteose, raffinose, pentosan, tinh bt

[6], [20], [31].

- Cc lignan glycoside: matairesinol [6], [20].

- Cc hp cht triterpen: 3-(cis-p-coumaroyloxy)-2-23-dihydroxyl-urs-12-en-

28-oic acid, 3-(trans-p-coumaroyloxy)-2-23-dihydroxyl-urs-12-en-28-oic acid[6],

[20], [31].

- Thnh phn v c: Cu, Zn, Fe, Mn, Ca, Mg, Se, Mo [6], [20].

c) Tc dng dc l

18

- Chng tng huyt p do tc dng ca sesamin trong m hnh gy tng huyt p

do bc p thn v chng s gin n to ca tim[20].

- Tc dng khng khun: Du vng c tc dng c ch nhiu vi khun nh

Streptococcus mutans,Lactobacilli acidophilus,Proteus vulgaris, Bacillus subtillis...

MIC khong 350- 500l/ml. Ring vi Salmonella typhi, du vng cho tc dng

khng khun tt nht, MIC l 10l/ml [38], [29].

- Tc dng c ch mnh s peroxid ha lipid do sesaminol [20].

- Tc dng h ng huyt trn chut cng trng[20], [37].

- Tc dng chng ung th do Glycosid ca vng c chng minh [20].

- Tc dng lm gim kch thch, chng vim khi bi ln nim mch[20].

- Du vng c tc dng nhun trng[13], [20],[29].

- B sung dinh dng cho c th [20].

- Gim cholesterol mu, phng tr x va ng mch [20], [25].

- Tc dng chng ung th: Th nghim in vivo cho thy dch chit cn ca vng

en nng 1000g/ml c thgy c cc t bo ung th dng He-2, AMN-3sau 72

gi[39].

d) Tnh, v, quy kinh

V ngt, bo, tnh bnh , vo 4 kinh ph, t, can, thn [6],[13].

e) Cng nng:

- B can thn, dng huyt; nhun trng thng tin; li sa [12], [14], [20].

- Cm mu [14], [20].

- ch kh lc, b no ty, bn gn ct, sng tai mt, thng sa, lm sng lu[3], [4].

f) Ch tr

- Tr ra mu [20].

- To bn, c bit l to bn sau sinh [14], [21].

- Thiu mu, chc nng can thn yu, huyt h, tc bc sm [13], [14].

- Xut huyt do gim tiu cu[14].

- Ph n sau t sa hoc khng c sa[14], [20].

19

- Tng cng dinh dng, u vng mt hoa, tr bng, rn cn, lot, nt n da,

vim thn, bng quang [6], [13], [20].

g) Liu dng

Ngy dng 12- 60 g [14].

20

CHNG II . I TNG VPHNG PHP NGHIN CU

2.1. Nguyn vt liu, phng tin

2.1.1. Nguyn liu

- Ch phm Ta An (Tham kho hnh nh ph lc 6).

- Cc v thuc: L nguyn liu sn xut ch phm Ta An.

n v cung cp: Cng ty TNHH Giai Cnh.

a ch: S 295 Nguyn Tri, phng 7, qun 5, Thnh ph H Ch Minh.

2.1.2. Phng tin

2.1.2.1. Thit b

- Knh hin vi LABOMED,Labo America. Inc (USA).

- H thng HPTLC CAMAG, Serial No. 181045 (Switzerland).

- H thng HPLC SHIMADZU LC Solution detector DAD,Serial No.

L20435002331 (Japan).

- n t ngoi Vilber Lourmat, Serial No.12 101537 (France).

- n t ngoi CAMAG, Serial No. 1812 (Switzerland).

- My li tm Universal, Gemmy Industrial Corp, Serial No. 717902 (Taiwan).

- Bp Wisd Therm, DAIHAN Scientific Co, Ltd., Serial No. 0406452128002

(Korea).

- Cn phn tch Sartorius BP 221S, Serial No. 23808521 (Germany).

- Cn k thut Precisa XB320C, Precissa Instruments Ltd.(Switzerland).

- Cn in t Shimadzu (Japan).

- Cn xc nh m Pressica, Precissa Instruments Ltd.(Switzerland).

- My ct quay thu hi dung mi Buchi Rotavapor R- 200, BUCHI Labortechnik

AG (Switzerland).

- Bp un cch thy BATH (China)

- B dng c Soxhlet.

- T sy Memmert, Shellab.

- My o ph chn chut 7140 Plethysmometer (UGO BASILE).

V cc thit b, dng c khc.

21

2.1.2.2. Ha cht

- Bn mng Silicagel60 GF 254 Merck (Germany).

- Dung mi phn tch, dung mi HPTLC, dung mi HPLC Merck (Germany).

- Cht i chiu: Rutin,quercetin, astragalosid IV, indomethacin do Vin kim

nghim cung cp.

2.1.2.3. ng vt th nghim

Chut cng trng khe mnh chng Wistar, c hai ging, trng lng 140 160g,

do Hc vin Qun y cung cp.

Chut sau khi mua v, c nui n nh trong iu kin phng th nghim B mn

Dc l, Trng H Dc H Ni 5 ngy trc khi thc hin nghin cu, c

nui dng bng thc n tiu chun do Vin V sinh dch t TW cung cp, ung

nc t do.

2.2. Ni dung nghin cu

2.2.1. Xc nh tnh ng ca cc dc liu

- M t hnh thi dc liu.

- M t c im bt.

- nh tnh bng phn ng ha hc.

2.2.2. Nghin cu thnh phn ha hc ch phm Ta An

- nh tnh bng phn ng ha hc v SKLM.

- nh lng cht chit c phn on EtOAc.

- nh lng rutin, quercetin trong ch phm bng HPTLC.

2.2.3. Th tc dng dc l

- Th tc dng cm mu.

- Th tc dng chng vim.

2.3. Phng php nghin cu

2.3.1. Xc nh tnh ng ca cc dc liu

- M t hnh thi dc liu: Quan st, m t c im bn ngoi, mi, v dc liu,

i chiu vi tiu chun DVN IV.

- Soi bt: Xc nh c im bt , i chiu vi tiu chun DVN IV.

22

- nh tnh bng phn ng ha hc.

2.3.2. Nghin cu thnh phn ha hc ch phm Ta An.

nh tnh:

- Kim tra s c mt ca mt s nhm cht chnh trong ch phm bng cc phn

ng ha hc.

- Kim tra s c mt ca dc liu trong ch phm bng SKLM.

nh lng cn phn on EtOAc bng phng php cn

Ly chnh xc mt lng bt thuc. Loi tp v chit kit vi EtOAc. Gp dch

chit v c cch thy ti cn. Sy kh cn nhit thp ti khi lng khng i,

cn cn v tnh kt qu hm lng cn theo cng thc (1).

% F=

. x 100 (1)

Trong : %H: hm lng cn chit c phn on EtOAc.

mcn: khi lng cht chit c.

mb.th: khi lng bt thuc.

Tin hnh song song 3 mu ly kt qa trung bnh .

nh lng rutin, quercetin trong ch phm Ta An bng HPLC

Tin hnh theo chng trnh HPLC nh lng cc flavonol trong vin Gingko

biloba [27].

Hm lng rutin, quercitin tnh trn vin tnh theo cng thc (2):

Trong :V l hm lng rutin, quercetin trong vin (kl/vin).

mc, mt ln lt l khi lng cn ca cht i chiu v cht th.

St, Scln lt l din tch peak ca mu th v mu i chiu

h l hm lng cht i chiu

f l t l h s pha long ca mu i chiu v mu th

mtbv : khi lng trung bnh vin thuc

23

Hm lng phn trm theo khi lng rutin, quercetin(X%) trong vin tnh theo cng thc (3):


2.3.3.1. Th tc dng cm mu

Chut nht trng c chia thnh 3 l[3], [22]:

- L chng: ung nc mui sinh l.

- L TA1: ung To An mc liu 1.25g/kg chut.

- L TA 2: ung To An mc liu 2.50 g/kg chut.

Cho chut ung nc mui sinh l hoc thuc nghin cu trong 5 ngy lin

tc.Ngy th 5, sau khi ung thuc 1 gi, gy m cho chut bng tim mng bng

thiopental liu 30mg/kg chut. Khi chut m, ct ui chut 2mm tnh t chp

ui, nhng ngay vo nc m 37oC, xc nh thi gian mu chy bng ng h

bm giy.

Thng s nghin cu:

- Thi gian chy mu: c tnh t khi mu bt u chy cho ti khi mu ngng

chy.

- Mc rt ngn thi gian chy mu ca l th so vi l chng theo cng thc

(4).

X (%) =

100 (4)

Trong : X (%): mc rt ngn thi gian chy mu ca l th so vi l chng.

Tc: thi gian chy mu trung bnh ca l chng.

Tt: thi gian chy mu trung bnh ca l th.

2.3.3.2. Th tc dng chng vim cp

24

nh gi tc dng chng vim cp ca vin nang To an, chng ti s

dng m hnh gy ph bng carrageenan theo Winter v cng s[3], [12], [22], [46],

[47].

Chut cng trng sau khi nui n nh 3 ngy trong iu kin phng th

nghim, c chia ngu nhin thnh 4 l, thit k nh sau:

- L chng: ung nc mui sinh l 1ml/100g chut.

- L indomethacin: ung indomethacin liu10mg/kg chut.

- L TA 1: ung cm ToAn 1.25g/kg chut.

- L TA 2: ung cm To An 2.5g/kg chut.

tt c cc l nghin cu, chut c cho ung nc mui sinh l, thuc i

chiu hoc ch phm nghin cu vi cng th tch 1 ml/100g chut lin tc trong 5

ngy. Ngy th nm, sau khi chut ung nc mui sinh l, thuc i chiu hoc

ch phm nghin cu, chut c tim 0,05ml carrageenan 1% trong nc mui

sinh l vo gan bn chn sau phi. S dng my o ph Plethysmometer (UGO

BASILE) o th tch bn chn sau phi ca tng chut ti thi im 1 gi, 3 gi,

5 gi, 7 gi sau khi gy vim.Quy trnh tin hnh th nghim c m t hnh 2.1.

K thut o ph bn chn chut

Dng bt nh du c nh mt bn khp gi chn sau phi ca chut. Nhng bn

chn sau phi vo dung dch o n ng v tr nh du, c kt qu hin th

trn thit b o. K thut o c thc hin bi cng mt k thut vin v l php o

m.

- Ch tiu quan st: Th tch bn chn sau phi ca chut.

- Ch tiu nh gi:

+ ph bn chn chut c tnh theo cng thc (5)

V (%)= 100 (5)

Trong :V: ph bn chn chut ti thi im t gi sau khi gy vim.

Vo, Vt: th tch bn chn chut trc khi gy vim v thi im t.

t o

o

V V

V

25

+ Phn trm c ch ph ca cc l th so vi l chng c tnh theo cng thc

(6):

I (%) = 100 (6)

Trong :I (%): phn trm c ch ph ca l th so vi l chng.

Vc: ph chn chut trung bnh ca l chng.

Vt: ph chn chut trung bnh ca l th.

Hnh 2.1. Qui trnh th nghim tc dng chng vim cp trn m hnh gy ph

bn chn chut

2.2.3.3. X l s liu

Cc s liu th nghim dc l ( 2.2.3.1, 2.2.3.2) cbiu din di dng

MSE ( M: l gi tr trung bnh, SE: sai s chun). So snh gi tr trung bnh gia

cc l bng one-way ANOVA, dng hu kim Dunnett test so snh gi tr trung

bnh ca cc l th so vi l chng. Phn tch thng k bng phn mm SPSS 16.0.

S khc bit gia cc l th c ngha thng k khi p< 0.05.

c t

c

V V

V

Nhn n

- Ung thuc

- Gy vim

V0 V1 V3 V5

---1,5 gi---

---------- o th tch bn chn gy vim ----------

26

CHNG III: THC NGHIM, KT QA, BN LUN

3.1. Thc nghim, kt qu

3.1.1. Xc nh tnh ng cc dc liu

3.1.1.1. Dip c

a) c im hnh thi

Thn hnh tr trn hay dt, cong, di 20

35cm, ng knh 2 3mm. Mt ngoi mu

vng nu nht, c vn dc nh v c mu r.

Cht gin, d gy. L mc so le, hnh tim,

u l nhn, phin l gp cun li, nhu nt,

cung nh gc ldi chng

2 3cm, gc cung rng thnh b mng.

Mi tanh c. V hi cht, se(Hnh 3.1).

Nhn xt: Ph hp vi m tdc

liu dip c trong DVN IV [10].

Hnh 3.1. nh dc liudip c

b) c im bt

Hnh 3.2. nh c im bt dip c

27

Bt mu lc vng, v hi mn, hi cay, mi tanh.Soi trn knh hin vi thy:

Mnh biu b trn v biu b di gm t bo hnh nhiu cnh thnh hi dy, mang

t bo tit (1,2). Biu b di c l kh, t bo tit trn, cha tinh du mu vng nht

hay vng nu, b mt c vn, xung quanh c 5-6 t bo xp to ra (2). L kh c 4-5

t bo km nh hn (2).Lng che ch a bo(6).Ht tinh bt hnh trng, hnh trn

hay hnh chung(3). Mnh thn gm t bo hnh ch nht thnh mng v t bo

tit(5). Mnh mch xon (4)(Hnh 3.2).

Nhn xt: Ph hp vi m t bt dip c trong DVN IV [10].

c) nh tnh

Quan st thn v bt l di n t ngoi thy pht quang mu nu hung.

nh tnh bng phn ng ha hc:

- Ly khong 1g bt dc liu vo ng nghim, dng a thu tinh n cht xung,

thm vi git dung dch fuchsin kh mu lm t bt pha trn, yn mt

lc.

Kt qu: Bt t c mu hng (phn ng dng tnh).

- Ly 1g bt dc liu, thm 10ml EtOH, un hi lu trn cch thu 10 pht, lc.

Ly 2ml dch lc, thm t bt Magnesi v 3 git HCl, un nng trn cch thu.

Kt qu: Xut hin mu (phn ng dng tnh).

Nhn xt: Cc hin tng ph hp vi m t trong DVN IV[10].

3.1.1.2. Trc bch dip

a)c im hnh thi

Dc liu phn thnh nhiu nhnh,

cc cnh nh dt, phng. Cc l hnh

vy nh, xp i cho ch thp, dnh

st vo cnh, mu xanh lc thm

hoc xanh lc hi vng. Cht gin,

d gy. C mi thm nh, v ng,

cht v hi cay (Hnh 3.3) Hnh 3.3. nh dc liu trc bch dip

28

Nhn xt: Ph hp vi m t dc liu trc bch dip trong DVN IV [10]

b) c im bt

Bt mu xanh lc hi vng. Soi di knh hin vi thy: Cc t bo biu b trn ca

l hnh ch nht, thnh hi dy ln (1,2).T bo biu b di hnh gn vung, c

nhiu l kh(4). Cc t bo m mm cha cc git du nh (5). Nhiu b si (3)

(Hnh 3.4).

Hnh 3.4. nh c im bt trc bch dip

Nhn xt: Ph hp vi m t bt trc bch dip trong DVN IV [10] .

3.1.1.3. Thng ma

a) c im hnh thi

V thuc l thn r, kch thc

khng u. Mt ngoi mu nu

en, th. Cht nh, cng, gin.

Mt b gy khng phng, c x,

mu trng c. Mi hc nh, v

hi ng (Hnh 3.5).

Nhn xt: Khng ph hp

vi m t dc liu thng ma

trong DVN IV [10].

Hnh 3.5. nh dc liu thng ma

29

b) c im bt

Bt mu vng nht. Soi di knh hin vi thy: Khi tinh bt b h ha(1). Mnh

mch (2,3). Ht tinh bt hnh a gic, hnh trn, c rn ht, ng ring l hay t li

thnh m (4,5). Si thnh dy (6) ( Hnh 3.6).

Hnh 3.6. nh c im bt thng ma

Nhn xt: Khng ph hp vi m t bt thng ma do c cc khi tinh bt b h

ha, tinh bt c rn ht [10].

c) nh tnh

Alcaloid [3].

Cn 5g dc liu, cho vo bnh nn dung tch 100ml. Thm 15ml dd H2SO4 1N.

un n si, ngui, lc. Kim ha dch lc bng dd NH3 6N n pH= 8 - 9. Cho

dch lc vo bnh gn. Lc, chit vi CHCl3 3 ln, mi ln 10ml. Gp cc dch chit

CHCl3, loi nc bng Na2SO4 khan. Lc, chit vi H2SO4 1N 3 ln, mi ln 10ml.

Chia dch chit vo 3 ng nghim, mi ng 1ml.

- ng 1: Thm 2 3 git thuc th Mayer.

Kt qu: Khng thy ta (phn ng m tnh).

- ng 2: Thm 2 -3 git thuc th Bouchardat.

Kt qu: Khng thy ta(phn ng m tnh).

- ng 3: Thm 2 -3 git thuc th Dragendorff

30

Kt qu: Khng thy ta(phn ng m tnh).

Flavonoid [3].

Cn 2g dc liu. Thm 20ml EtOH 90%. un si cch thy 10 pht, lc.Thu dch

lc lm phn ng:

- Phn ng Cyanidin: Cho vo ng nghim 2ml dch chit. thm mt t bt

Magnesi . Nh t t 5-6 git HCl. yn 2 pht.

Kt qu: Dung dch c mu vng (phn ng m tnh).

- Phn ng vi kim long: Cho vo ng nghim 2ml dch chit. Thm 3 git dd

NaOH 10%.

Kt qu: Xut hin ta mu vng. Thm 2ml nc ct, ta tan v mu vng ca

dung dch c tng nh(phn dng tnh).

- Phn ng vi NH3 : Nh 2 git dch chit ln t giy lc. Sy nh n kh. H

trn ming l NH3 c.

Kt qu: Mu vng vt dch chit khngtng thm (phn ng m tnh).

- Phn ng vi FeCl3: Cho vo ng nghim 2ml dch chit. Thm 2 git dd FeCl3

5%.

Kt qu: Khng thy xut hin ta (phn ng m tnh).

- Phn ng diazo ha: Cho vo ng nghim 2ml dch chit. Kim ha bng dd

NaOH 10%. Thm 2- 3 git TT diazo mi pha, lc u, un nng nh.

Kt qu: Xut hin ta gch (phn ng dng tnh).

Saponin [3].

- Hin tng to bt: Ly 2g bt dc liu, thm 10ml nc. Bt u ng nghim,

lc mnh 5 pht . yn v quan st hin tng to bt.

Kt qu: To bt khng bn vng(phn ng m tnh).

- Phn ng Salkowski: Ly 2g dc liu. Thm 5ml H2SO4 c.

Kt qu: Dung dch chuyn t mu vngvng sang nht (phn ngdng tnh).

- Phn ng Liebermann Burchardt: Ly 2g dc liu.Thm 10ml nc. un si,

lc. Cho dch lc vo ng nghim, c ti cn. Thm 1ml anhydrid acetic, lc u

31

cho tan ht. Nghing ng nghim 450, cho t t theo thnh ng 0.5ml H2SO4 c,

trnh xo trn cht lng trong ng nghim.

Kt qu: Khng thy xut hin vng mu mt tip xc gia hai lp cht lng(phn

ng m tnh).

Coumarin [3].

Chun b mu th: Ly 5g dc liu. Thm 30ml EtOH 90%. un si cch thy 15

pht, lc nng. Dch lc tin hnh cc phn ng:

- Phn ng ng m vng lacton: Cho vo 2 ng nghim, mi ng 2ml dch chit:

+ ng 1: Thm 0.5ml dd NaOH 10%

+ ng 2: nguyn

un c 2 ng nghim n si, ngui. Quan st thy:

+ ng 1: Ta c vng nht

+ ng 2: Trong

Thm vo mi ng nghim 2ml nc ct, lc u, thy:

+ ng 1: Trong sut

+ ng 2: Ta c

Thm vi git HCl vo ng 1, xut hin ta c tr li nh ng 2

Kt qu: Phn ng dng tnh.

- Phn ng diazo ha: Cho vo ng nghim 1ml dch lc. Thm 2ml dd NaOH

10%. un cch thy 10 pht, ngui. Thm vi git thuc th diazo mi pha.

Kt qu: Xut hin mu gch (phn ng dng tnh).

- Pht hunh quang di nh sng t ngoi: Nh 2 git dch lc ln giy lc. Nh

tip 3 git dd NaOH 10%, sy nh n kh. Che na vt bng ng xu. Soi di

n t ngoi vi pht, b ng xu ra, thy phn khng b che sng hn phn b che.

Tip tc chiu sng thy 2 phn c hunh quang ging nhau.

Kt qu: C hunh quang sng (phn ng dng tnh).

Tanin [3].

Chun b mu th: Cn 3g dc liu. Thm 20ml nc ct. un cch thy 3 pht,

lc ly dch. Dch lc cho vo 3 ng nghim, mi ng 2ml tin hnh phn ng:

32

- ng 1: Thm 2 git FeCl 5%.

Kt qu: Khng xut hin ta (phn ng m tnh).

- ng 2: Thm 2 git ch acetat 10%.

Kt qu: Xut hin kt ta bng trng (phn ng dng tnh).

- ng 3: Thm 5 git gelatin 1%.

Kt qu: Khng thy xut hin ta bng trng (phn ng m tnh).

Acid amin

Cn 5g dc liu. Thm 30ml nc ct, un si 10 pht, lc ly dch. Ly 2ml dch

lc, thm 3ml TT Ninhydrin 3%, un cch thy trong 2 3 pht.

Kt qu: Xut hin mu tm sm (phn ng dng tnh).

ng kh

Cn 5g dc liu. Thm 15ml nc ct. un cch thy 10 pht. ngui, lc. Cho

vo ng nghim 2ml dch lc, 1ml TT Fehling A v 1ml TT Fehling B . Lc k, un

si cch thy 10 pht.

Kt qu: Xut hin ta gch (phn ng dng tnh).

Polysaccharid

Cn 5g dc liu. Thm 30ml nc ct, un si 2-3 pht, lc ly dch, ngui.

Ly 2ml dch lc cho vo ng nghim, thm 2 git TT Lugol.

Kt qu: Dung dch chuyn mu xanh en(phn ng dng tnh).

Glycosid tim [3].

Cn 5g dc liu. Thm 30ml EtOH 25%. Ngm 24h, lc ly dch. Thm vo dch

lc 3ml dd ch acetat cho n d, khuy u, lc ly dch. Lc dch lc vi CHCl3 2

ln, mi ln 5ml. Chit ly lp CHCl3 , cho vo cc ng nghim nh, c ti cn.

- Phn ng Liebermann Burchard: Thm vo ng nghim 1ml anhydrid acetic,

nghing 450 , cho t t theo thnh ng 0.5ml H2SO4 c.

Kt qu: mt tip xc gia hai cht lng c khng thy vng mu (phn ng m

tnh).

- Phn ng Baljet: Thm vo 1ml EtOH 90%, lc u cho tan ht, nh tng git

thuc th Baljet mi pha.

33

Kt qu: Dung dch mu vng, khng m hn khi so vi ng chng (phn ng m

tnh).

- Phn ng Keller Kiliani: Thm vo 0.5ml EtOH 90%, lc u cho tan ht.

Thm vi git dd FeCl3 5%/ acid acetic, lc u, nghing ng nghim 450, thm t

t theo thnh ng 0.5 ml H2SO4 c.

Kt qu: mt tip xc 2 cht lng khng thy xut hin vng mu (phn ng m

tnh).

Acid hu c:

Cn 2g dc liu. Thm 20ml nc ct, uncch thy 5 pht. Lc ly dch,

ngui. Ly 2ml dch lc cho vo ng nghim. Thm mt lng nh Na2CO3, lc

nh.

Kt qu: Khng thy c bt kh (phn ng m tnh).

Nhn xt: Trong dc liu thng ma ny cha cc nhm cht: Coumarin,

ng kh, polysaccharid, acid amin (Bng 3.1. tr.38).

3.1.1.4. He hoa

a) c im hnh thi

N hoa hnh trng c cung nh,

ngn, mt u hi nhn, di 3 - 6 mm,

rng1 - 2 mm, mu vng xm. i hoa

hnh chung, mu vng xm, di bng

1/2 n 2/3 chiu di ca n hoa, pha

trn x thnh 5 rng nng. Hoa cha n

di t 4 - 10 mm, ng knh 2 - 4 mm.

Cnh hoa cha n mu vng. Mi thm,

v hi ng. (Hnh 3.7)

Nhn xt: Ph hp m t v thuc

he hoa trong DVN IV [10].

Hnh 3.7. nh dc liu he hoa

34

b) c im bt

Bt mu vng lc. Soi trn knh hin vi thy: Nhiu ht phn hnh cu, c 3 l

rnh (4,8). Lng che ch a bo(1,2). Mnh biu b cnh hoa gm nhng t bo

hnh nhiu cnh c nhiu vn nh, st nhau (9,5). Mnh cnh hoa mang mch

dn(6,9).Mnh biu b i hoa (7). Mnh mch xon (3) (Hnh 3.8).

Nhn xt: Ph hp vi m t bt he hoa trong DVN IV [10].

Hnh 3.8. nh c im bt he hoa

c) nh tnh

Nh 2 - 3 git dung dch A ln t giy lc, kh, soi di n t ngoi ( bc

sng 366 nm).

Kt qu: C hunh quang mu vng nu (phn ng dng tnh).

nh tnh bng phn ng ha hc

Ly 0,5g bt dc liu, thm 10ml EtOH. un si trong 3 pht, ngui, lc c

dch lc (dd A). Ly 2ml dd A pha long vi 10ml EtOH 90% ri chia vo 3 ng

nghim:

- ng 1: Thm 5 git HClv t bt magnesi.

35

Kt qu: Dung dch chuyn t mu vng sang mu hng ri tm (Phn ng dng

tnh).

- ng 2: Thm t t tng git dung dch NaOH 20%.

Kt qu: Xut hin kt ta cam. Thm thuc th n d, ta tan (phn ng

dng tnh).

- ng 3: Thm 2 git dung dch FeCl3 5%.

Kt qu: Dung dch c mu xanh ru (Phn ng dng tnh).

Nhn xt: Cc hin tng ph hp vi m t trong DVN IV [10].

3.1.1.5. Hong k

a) c im hnh thi

Mt ngoi mu nu, c

nhng vt nhn dc. Th

cht cng, dai, kh b. Mt

gy c nhiu si, nhiu tinh

bt, phn v mu vng v g

mu nu vng vi nhng vt

nt v tia hnh nan qut. Mi

thm nh, v ngt v hi tanh

( Hnh 3.9). Hnh 3.9. nh dc liu Hong k

Nhn xt: Ph hp m t dc liu hong k trong DVN IV [10]..

b) c im bt

Hnh 3.10. nh c im bt Hong k

36

Bt mu nu vng. Soi trn knh hin vi thy: Mnh m mm cha tinh bt (3).

Si nm ri rc hoc xp thnh tng b, trn cc si c tinh th calxi oxalat hnh

khi xp thnh hng(1). Tinh bt n, kp i, kp ba ng ring l hay to thnh

cc khi trong cc m(3,4). Tinh th calxi oxalat hnh khi (2). Cc mnh mch(5) (

Hnh 3.10).

Nhn xt: Ph hp vi m t bt hong k trong DVN IV [10]. Tuy nhin,

cc tinh th calxi oxalat khng thy c m t trong DTQ v D M[43],[44].

3.1.1.6. Vng en

a) c im hnh thi

Ht hnh trng dt, di khong 3

mm, rng 2 mm. V ngoi mu en,

trn hoc nhn hnh mng. Non c

chm nu nh. V ngoi ca ht

mng, 2 l mm mu trng c cha

du. Mi thm nh, v ngt(Hnh

3.11).

Nhn xt: Ph hp vi m t

dc liu Vng en trong DTQ

[43].

Hnh 3.11. nh dc liu Vng en

b) c im bt

37

Hnh 3.12.nh c im bt Vng en

Bt nu en. Soi trn knh hin vi thy: Cc lp t bo biu b v ngoi ca ht

c sp xp thnh nhm, cha cc sc t mu en, b mt nhiu cnh, cha cc

tinh th hnh cu, vch ngoi v vch na trn mng, hu ht b gy v vch na

di v vch pha trong dy (1,5). Tinh th canxi oxalate hnh lng tr nm ri rc

hoc thnh m trong cc t bo biu b v ngoi ht (2,3). Nhiu git du kch

thc khng u, ng t do hay trong cc m mm (4,6) (Hnh 3.12).

Nhn xt: Ph hp vi m t bt Vng en trong DTQ [43].

d) nh tnh

Nghin 1g dc liu thnh bt. Thm 10ml ether du ha, ngm lnh 30

pht. Gn phn cht lng pha trn cho vo ng nghim, cho thm 10ml HCl cha

0.1 g saccarose, lc trong 30 giy.

Kt qu: Xut hin mu hng lp acid v dn dn chuyn sang mu (Phn ng

dng tnh).

Nhn xt : Hin tng ph hp vi m t trong DTQ [43].

3.1.2. Nghin cu thnh phn ha hc ch phm Ta An

3.1.2.1. nh tnh

a) nh tnh bng phn ng ha hc

Tin hnh tng t nh nh tnh Thng ma vi 5g bt thuc.

Kt qu: Ch phm c phn ng ca cc nhm cht: Flavonoid, saponin, acid amin,

polysaccharid, ng kh (Bng 3.1).

38

Bng 3.1: Kt qu nh tnh Ta An v thng ma bng phn ng ha hc

STT Nhm cht Phn ng nh tnh TA TM

1.

Flavonoid

Cyanidin ++ - Vi NaOH 10% ++ ++ Vi NH3 m c ++ - Vi FeCl3 5% ++ - Diazo ha ++ ++

Phn ng ng m vng lacton - ++ 2. Coumarin Diazo ha + ++ Quan st hunh quang + + To bt ++ -

3. Saponin Salkowski ++ + Liebermann Burchardt ++ - Vi TT Mayer - -

4. Alcaloid Vi TT Bouchardat - - Vi TT Dragendorff - - Vi FeCl3 5% ++ -

5. Tanin Vi Ch acetat 10% ++ ++ Vi Gelatin 1% - -

6. ng kh Vi TT Fehling A+ Fehling B ++ ++ Baljet - -

7. Glycosid tim Keller Kiliani - - Liebermann Burchardt - -

8. Acid hu c Vi Na2CO3 - - 9. Polysaccharid Vi TT Lugol ++ ++ 10. Acid amin Vi TT Ninhydrin ++ ++

Ch thch: (-): m tnh (+): dng tnh (++): dng tnh r

b) nh tnh bng SKLM so snh vi dc liu

So snh vidip c.

Mu NC:

- Dch chit DC: Cn 1g bt dc liu. Thm 10ml EtOH 95%, un si cch thy

trong 5 pht, ngui, lc ly dch chm sc k.

- Mu i chiu: Ha tan quercetin i chiu trong EtOH 95% c dd c

nng 0.5 mg/ml.

39

- Dch chit TA : Ly 4 vin thuc. B v nang, ly bt thuc v tin hnh tng

t nh vi dc liu.

H dung mi khai trin: Th vi cc h sau:

- H 1: Toluen -EtOAc - acid formic (5: 4: 1)

- H 2: Toluen - EtOAc - acid formic (5: 2: 1)


- H 4: Toluen - EtOAc(9:1)

Bn mng: Silica gel 60 F254 hot ha 1100 C trong 1 gi, bo qun trong bnh

ht m.

Trin khai sc k: Chm 3 vt ring bit cc mu NC vi th tch nh nhau ln

bn mng, chy trn cng h dung mi n vch gii hn, ly ra, kh t nhin.

Pht hin vt: Soi di n t ngoi bc sng 254 nm v 366 nm.Hin mu:

KOH10% / EtOH.

Kt qu: Khai trin vi h 3 v quan st di bc sng 366 nm l r nht. Kt

qu c th hin bng 3.2 v hnh 3.14.

Bng 3.2. Kt qu sc k so snh TA, DC,

quercetin(= 366 nm)

STT Rf Mu sc TA DC CC

1. 0.16 Xanh + - -

2. 0.20 Vng + + -

3. 0.33 Xanh + + +

4. 0.42 Xanh + + -

6.. 0.62 Vng + + -

7. 0.70 - + -

8. 0.80 Xanh - + -

Nhn xt: Sc k ch phm TA c

4 vt c Rf tng ng vi dip c.

Trong s , c 1 vt trng vi quercetin.

a) = 366 nm b) = 254 nm

Hnh 3.13. nh sc k so snh TA,

DC, quercetin

40

So snh vi trc bch dip

Mu NC:

- Dch chit TBD: Ly 3g bt dc liu, cho vo bnh chit Soxhlext. Thm 50ml

ether, un hi lu cch thy 3 gi (n khi dch chit ht mu). Loi b dung mi

ether. Phn b dc liu sy nh n kh. Thm 50ml EtOH 70%, un hi lu cch

thy 1 gi. Lc nng. Bc hi dch lc ti kh. Thm 25ml nc v 3.5ml HCl

ha tan cn. un hi lu cch thy 30 pht. Lm mt nhanh. Chit vi EtOAc 2 ln,

mi ln 20ml. Gp dch chit EtOAc v ra li 3 ln, mi ln vi 10ml nc. Lm

khan bng Na2SO4 khan. Bc hi dch lc ti cn. Ha cn trong 5ml MeOH.

- Mu i chiu: Ha tan quercetin i chiu trong EtOH 70% c dd c nng

0.5 mg/ml.


t nh vi dc liu.

H dung mi khai trin: Th vi cc h sau:


- H 2: EtOAc- methanol (8:2)

- H 3: EtOAc - acid formic - nc (70: 15: 15)

- H 4: EtOAc -toluen - methanol (8:6: 1)

- H 5: Toluen - cloroform - aceton (8:5:7)

- H 6: EtOAc: n- butanol - acid acetic- nc (5:3:1:1)

Bn mng: Silica gel 60 F254 hot ha 1100 C trong 1 gi, bo qun trong bnh

ht m.

Trin khai sc k: Chm 3 vt ring bit cc mu NC vi th tch nh nhau ln

bn mng, chy trn cng h dung mi n vch gii hn, ly ra, kh t

nhin.

Pht hin vt: Soi di n t ngoi bc sng 254 nm v 366 nm. Hin mu: dd

KOH 10%/ EtOH.

Kt qu: Khai trin vi h 1 v quan st di bc sng 366 nm l r nht. Kt


41

Nhn xt: Sc k ch phm c 5 vt c Rf tng ng vi cc vt TBD. TBD c

vt c Rf tng ng vi cht i chiu quercetin.

Bng 3.3. Kt qu sc k so snh

TA, TBD, quercetin (= 366 nm)

ST

T

Rf Mu

sc

TA TBD CC

1. 0.22 Xanh + + -

2. 0.25 Xanh + + -

3 0.29 Tm - + -

4. 0.32 + - -

5. 0.36 + - -

6. 0.38 Xanh - + +

7. 0.44 Xanh + + -

8. 0.47 Xanh + + -

9. 0.55 Xanh + + -

10. 0.60 - - -

11. 0.73 - + -

a) = 366 nm b) = 254 nm

Hnh 3.14. nh sc k so snh

TA,TBD,quercetin

So snh vi thng ma

Chun b mu NC:

- Dch chit TM: Ly 1g bt dc liu, thm 50ml EtOH 95%, un si hi lu

trong 1h, lc. Bc hi dch lc ti cn. Ha tan cn trong 1ml EtOH 95% c dd

th.


t nh vi dc liu.

H dung mi khai trin: Th vi cc h dung mi sau:

- H 1: Toluen cloroform- acid acetic bng (6: 1: 0.5)

- H 2: cloroform- methanol- nc (2: 2: 1)

42

- H 3:cloroform - methanol (8:2)

- H 4: ethyl acetat- acid acetic- nc (65:35: 10)

- H 5:n- butanol-ethanol (10:2)

- H 6: cloroform- ethyl acetat(7:3)

Pht hin vt: Soi di n t ngoi bc sng 254 nm v 366 nm. Hin mu: dd

vanilin 10%/ EtOH.

Kt qu:Khai trin vi h 6 v quan st di bc sng 366 nm l r nht. Kt


Bng 3.4. Kt qu sc k so snh TA, TM

(=366 nm)

STT Rf Mu sc TA TM

1. 0.12 Xanh dng + +

2. 0.17 Xanh dng + +

3 0.20 Xanh dng + -

4. 0.28 Xanh dng - +

5. 0.32 Xanh dng + -

6. 0.37 Xanh dng + -

7. 0.46 Xanh dng + -

8. 0.48 Xanh dng - +

9. 0.61 Xanh dng + +

10. 0.66 Xanh dng + +

Nhn xt: Trn nh sc k, TA c 4 vt c Rf

tng ng vi cc vt ca TM.

Hnh 3.15. nh sc k so snh

TA v TM(=366 nm)

So snh vi he hoa

Mu NC:

- Dch chit HH: Cn 0,5g bt dc liu, thm 10ml EtOH 90%. un si trong 3

pht, ngui, lc

- Mu i chiu: Ho tan rutin trong EtOH 90% c dd nng 1 mg/ml.

43


t nh vi dc liu.

H dung mi khai trin: th vi cc h dung mi sau:

- H 1: Ethyl acetat- acid formic - nc (8: 1: 1)

- H 2: Ethyl acetat - acid formic - nc ( 3: 3: 1)

- H 3: n-butanol- acid acetic- nc (4: 1: 5 )

- H 4: n-butanol- acid acetic- nc (5: 4: 1 )

- H 5: Ethyl acetat - acid formic - cloroform(2: 1: 2)

- H 6: Ethyl acetat - acid formic - cloroform (3: 3: 1)

- H 7: Ethyl aceat methanol - nc (100: 17: 13)

- H 8:Toluen - ethyl acetat - acid formic (5: 5: 1)

Pht hin vt: Quan st di nh sng t ngoi bc sng 254 nm v 366 nm.

Hin mu: dd KOH 10% / EtOH.

Kt qu: Khai trin vi h 8 v quan st di bc sng 366 nm l nht. Kt qu

c th hin bng 3.5 v hnh 3.16.

Nhn xt: sc k TA c 4 vt c Rf tng ng vi cc vt ca HH. HH c vt

c Rf tng ng vi cht i chiu rutin.

44


TA, HH v rutin (=366 nm)..

STT Rf Mu

sc

TA HH C

1. 0.10 Xanh + - -

2. 0.16 Xanh + + -

3 0.23 Xanh + + -

4. 0.33 Xanh + - -

5. 0.37 Xanh + + -

6. 0.43 Xanh + - -

7. 0.46 Xanh - + +

8. 0.49 Xanh + - -

9. 0.50 - + -

10. 0.55 Xanh + + -

11. 0.60 Xanh + - -

12. 0.67 Vng + - -

13. 0.71 Xanh + - -

14. 0.78 - + -

a) =366 nm b) = 254 nm

Hnh 3.16. nh sc k so snh TA, HH,

rutin

Vi hong k

Mu NC:

- Dch chit HK: Cn 3g dc liu, thm 20ml EtOH, siu m 30 pht, lc. Thm

tip vo b 20ml EtOH, siu m 30 pht, lc. Gp dch lc. Bc hi dch lc trn

cch thy ti kh. Ha tan cn trong 25ml nc. Chit ln lt vi 15, 10, 10ml

BuOH bo ha vi nc. Gp cc dch chit BuOH, bc hi trn cch thy ti

kh. Cn ha tan trong 2ml MeOH lm dch chm sc k.

- Mu i chiu: . Ha tan cht i chiu astragalosid IV trong MeOH c dd

i chiu c nng 1 mg/ml.


t nh vi dc liu.

45


- H 1:Cloroform methanol - nc (65: 35:10)

- H 2:Toluen - ethyl acetat - acid acetic (8: 1.5: 1)

- H 3:Ethyl acetat - acid acetic - nc ( 8: 2: 1)

- H 4:Butanol - ethanol (10: 2)

- H 5:Butanol - acid acetic- nc (4: 1: 5)

- H 6:Dicloromethan - aceton (15: 1)

- H 7: Cloroform - methanol - nc(65: 30:7)

Pht hin vt: soi di n t ngoi bc sng 254 nm v 366 nm. Hin mu: dd

H2SO4 10%/ EtOH.

Kt qu:Khai trin vi h 7, hin mu bngdd H2SO4 10%/ EtOHl r nht.Kt



TA,HK,Castragalosid IV(hin mu)

STT Rf Mu

sc

TA HK C

1. 0.14 Tm + + -

2. 0.18 Tm + + -

3. 0.24 Nu + - -

4 0.39 Tm - - +

5. 0.49 Nu + - -

6. 0.54 Tm + + -

7. 0.62 Tm - + -

8. 0.71 Nu + + -

9. 0.77 Tm - + -

10. 0.84 Vng - + -

11. 0.88 Nu - + -

Hnh 3.17. nh sc kso snh HK, TA v

Castragalosid IV(hin mu v nhn di

snh sng thng)

46

Nhn xt: sc k TA c 4 vt c Rf tng ng vi cc vt ca HK. TA v

HK khng thy vt tng ng vi astragalosid IV.

Vi vng en

Mu NC[27]:

- Dch chit V: Nghin 0.5g dc liu thnh bt. Thm 10ml CHCl3 v ngm

lnh 2gi, lc, c dch lc ti cn. Ha tan cn trong 2ml MeOH, khuy u trong

30giy, ly tm 15 pht, tc 4000 vng/ pht. Thu ly phn dch pha trn lm

dch chm sc k.


t nh vi dc liu.


- H 1: Cyclohexan - ethyl ether - EtOAc(20:5.5: 2.5)

- H 2: Dicloromethan- acid acetic bng - aceton (2:4:5)

- H 3: Toluen - ethyl ether - EtOAc (20:5.5:2.5)

Pht hin vt : soi di n t ngoi bc sng 254 nm v 366 nm. Hin mu: dd

H2SO4 10%/ EtOH, sy 1050C trong 5 pht, quan st di nh sng thng.

Kt qu:Khai trin vi h 3 v quan st di

bc sng 366 nm l r nht. Kt qu c th

hin bng 3.7 v hnh 3.18.

Bng 3.7. Kt qu sc k so snh TA,V

STT Rf Mu

sc

TA V

1. 0.15 Xanh + +

2. 0.25 Xanh - +

3 0.38 Xanh + +

Hnh 3. 18. nh sc kTA v V ( = 366 nm)

47

Nhn xt:sc k TA c 2 vt c Rf tng ng vi cc vt ca vng en.

3.1.2.2. nh lng cht chit c phn on Ethyl acetat

Ly 10 vin TA. B v nang, thu ly bt thuc. Cn chnh xc khong 5.00 g

bt. Thm 20ml nc. Siu m 20 pht ha tan ch phm. Ly tm 15 pht, tc

4000 vng/pht. Lc, thu dch chit nc. Lm thm 2 ln nh vy. Dch chit nc

cho vo bnh gn. Thm 40ml ether du ha, lc u v tch lp. B lp ether

du ha. Lm li 4 ln (n khi lp ether mt mu). Dch chit loi tp lc vi

40ml EtOAc trong 5 pht. tch lp. Ly lp EtOAc. Lm li 5 ln (n khi lp

EtOAc khng phn ng vi NH3). Gp dch chit EtOAc. Cho vo bnh gn, tch

lp. Ly lp EtOAc. Ct thu hi dung mi 850C ti cn khong 10ml. Cho dch

sau khi thu hi vo cc c m bit khi lng. Trng bnh ct thu hi dung mi 2

ln, mi ln 5ml EtOAc. C cch thy ti cn. Tnh khi lng cn phn on

EtOAc ca TA.Kt qu c th hin bng 3.8.

Bng 3.8. Hm lng cn phn on EtOAc trong TA theo khi lng bt thuc

STT mb.th mcc mcc+cn mcn %F

1. 5.0060 109.8242 109.9351 0.1109 2.215

2. 5.0098 110.6349 110.7689 0.1340 2.675

3. 5.0078 84.3923 84.5195 0.1272 2.540

4. TB 2.476

3.1.2.3. nh lng rutin, quercetin trong ch phm TA bng HPLC.

Tin hnh theo chng trnh HPLC nh lng ng thi 5 flavonoid trong

dch chit Gingko biloba [27].

Xc nh khi lng trung bnh vin: Cn khi lng mt nang. Tho ri hai na

v nang, dng bng lau sch, cn khi lng v. Khi lng thuc trong nang l

hiu khi lng nang v khi lng v nang. Tin hnh tng t vi 19 nang khc.

Tnh khi lng trung bnh vin (Kt qu th hin ph lc 1).

Khi lng trung bnh vin: mtbv = 0.5701(g)

48

Chun b dung mi pha mu: MeOH: H2O (50:50)

Chun b mu NC:

- Mu th: Ly bt thuc trong khong 4 vin thuc vo bnh nn 100ml c nt

mi. Thm 75ml MeOH. Lc siu m 30 pht. Thm MeOH va n vch. Lc

u. Lc qua ly lc. Dch lc cho vo bt thy tinh 100ml. C cch thy n cn

1000C. Thm 100ml dung mi pha mu. Siu m n tan v chuyn dch chit vo

bnh nh mc 20ml. Trng bt thy tinh 3 ln, mi ln 3ml dung mi pha mu. Tp

hp nc ra vo bnh nh mc. Thm dung mi pha mu va n vch. Lc

u. Lc qua giy lc. Dch lc lc qua mng lc 0.45m.

- Mu chun:

+ Dung dch rutin: Cn chnh xc 10.8mg rutin chun (hm lng 87.12%)vo

bnh inh mc 25ml. Thm 15ml MeOH. Lc siu m n tan. Thm MeOH va

n vch.

+ Dung dch quercetin: Cn chnh xc 30.3mgquercetinchun (hm lng

81.60%)vo bnh nh mc 25ml. Thm 15ml MeOH. Lc siu m n tan. Thm

MeOH va n vch. Lc u. Ht chnh xc 1.0ml cho vo bnh nh mc

100ml. Thm MeOH va n vch.

+ Dung dchhn hp: Ht chnh xc 1.5ml dung dch rutin v 3.00 ml dung dch

quercetin cho vo bnh mc 20.0ml. Thm dung mi pha mu MeOH: H2O

(50:50)va n vch.

Pha tnh: Ct Phenomenex C18(250mm x 4.6mm, 5m).

Pha ng: Sau khi tin hnh kho st, chng ti chn pha ng chy theo ch

gradient dung mi (Bng 3.9).

Tc dng: 1.0 ml/ pht.

Th tch tim mu: 20 l/pht.

Detector: DAD bc sng 360nm.

Nhit : Nhit phng.

49

Bng 3.9. Thnh phn pha ng nh lng ng rutin, quercetin bng HPLC.

Thi gian

(pht)

Thnh phn pha ng

A (ACN) B(formic 0.3%)

0.01 15 85

8.00 15 85

21.00 80 20

21.01 15 85

29.00 15 85

Tnh thch hp h thng (Ph lc 2)

Tim 6 ln dd hn hprutin, quercetin vo my HPLC. Tnh thch hp h

thng c nh gi thng qua lch chun SD v lch chun tng i RSD

ca cc thng s: lp li din tch pic S, lp li thi gian lu tR, h s bt i

xng T, s a l thuyt N.

Bng 3.10. Tnh thch hp h thng HPLC vi rutin.

Mu tR(pht) S H T N

1 16.692 994113 147994 1.503 125150.2

2 16.700 993587 149245 1.491 124452.6

3 16.687 994501 148572 1.479 128594.9

4 16.694 993966 147313 1.491 125155.6

5 16.700 994739 148862 1.480 123329.3

6 16.719 993865 149024 1.506 130513.5

TB 16.699 994129 148502 1.492 126199.4

SD 0.0112 427.475 726.175 0.0110 2715.15

% RSD 0.067 0.043 0.489 0.740 2.180

50

Bng 3.11. Tnh thch hp h thng HPLC vi quercetin.

Mu tR(pht) S H T N

1 20.300 109832 12352 1.285 110578.9

2 20.284 109596 12397 1.280 111986.1

3 20.288 109143 12222 1.271 108956.1

4 20.303 109097 12332 1.280 113891.0

5 20.305 109187 12335 1.274 110525.4

6 20.311 108402 12381 1.275 114185.9

TB 20.298 109210 12336 1.277 111687.2

SD 0.0104 491.229 61.668 0.0051 2060.417

% RSD 0.0513 0.4498 0.4999 0.4008 1.8448

Kt qu hm lng rutin, quercetin trong ch phm Ta An c th hin bng

3.12 v 3.13(Hnh nh peak HPLC th v i chiu th hin ph lc 3)

Bng 3.12. Hm lng Rutin trong ch phm TA

Bng 3.13: Hm lng Quercitin trong ch phm TA

Quercitin M Spic V (g/vin) X (%kl/kl )

Chun 30.3 x 10-3 109210

Th 1 2.2409 85387 5.90 1.035 x 10-3

Th 2 2.2548 82239 5.65 0.991 x 10-3

Th 3 2.2076 80704 5.66 0.993x 10-3

TB 5.74 1.006 x10-3

Rutin M Spic V(mg/vin) X (%kl/kl )

Chun 10.8 x 10-3 994129

Th 1 2.2409 936143 0.135 0.0237

Th 2 2.2548 939308 0.135 0.0237

Th 3 2.2076 908480 0.133 0.0233

TB 0.134 0.0236

51

3.1.3. Thtc dng dc l

- Chun b mu NC: Tho v nang ly bt cm cha trong nang, pha long vi

nc c ch phm th ph hp cho ng vt th nghim.

- ng dng: ng ung.

- Cc mc liu th: c xc nh da vo liu dng trn ngi (theo liu dng

ghi trn t Hng dn s dng), qui i liu t ngi sang ng vt (c lm trn

s). D liu trn mt s t ng vt, bt u t liu tng ng, liu gp 2 v gp

3.Cui cng chng ti chn c 2 mc liu cho nghin cu tc dng dc l l

1,25g/kg v 2,5g/kg chut cng trng.

3.1.3.1. Th tc dng cm mu

S dng m hnh gy chy mu ui chut, xc nh thi gian chy mu ui

chut trn 2 mc liu 1,25g/kg chut v 2,50 g/kg chut cng. Kt qu thu c

trnh by trong bng 3.14 (S liu thc nghim c trnh by ph lc 4).

Bng 3.14. nh hng ca TA n thi gian chy mu ui chut

L chut

Thi gian

chy mu

(giy)

% gim thi

gian chy mu

so vi chng

p so vi

chng Liu dng

Chng

n=10

245.433.4

TA 1

n=10

159.826.9

1.25g/kg 34.9

52

ng l 34.9% v 37.7%. S khc nhau v nh hng trn thi gian chy mu gia 2

l To An 1 v To An 2 khng c ngha thng k (p > 0.05).

(*)p< 0.05 so vi l chng

Hnh 3.19. nh hng ca To an n thi gian chy mu ui chut

3.1.3.2. Th tc dng chng vim cp

S dng m hnh gy ph bn chn chut bng carrageenan, xc nh gim

ph trn2 mc liu 1,25g/kg chut; 2,5g /kg chut. Kt qu c trnhby trong

bng 3.15 v hnh 3.20 (S liu thc nghim c th c trnh by ph lc 5).

Hnh 3.20.nh hng ca Ta An n ph chn chut

Nhn xt:

245.4

159.8* 152.9*

0.0

50.0

100.0

150.0

200.0

250.0

300.0

L chng To An 1 To An 2

Th

i

gia

n c

h

y m

u

(g

iy

)

L nghin cu

0.0

5.0

10.0

15.0

20.0

25.0

1gi 3 gi 5 gi 7 gi

ph

c

h

n c

hu

t

Thi gian sau gy vim

Chng

Indomethacin

To An 1

To An 2

53

- Indomethacin mc liu 10mg/kg lm gim ph chn chut 3 thi im l 1

gi, 3 gi v 5 gi sau khi gy vim, c ngha thng k so vi l chng vi

p0.05

TA 2

2.5g/kg

(n=10)

6.74 1.95 11.57 2.22 10.94 2.24 8.92 2.47

I (%) 25.2 40.0 16.2 19.6

p so vi chng >0.05 0.05 >0.05

54

3.2. Bn lun

3.2.1. Xc nh tnh ng ca cc v thuc

Cng thc phng thuc Ta An do cng ty Giai Cnh thnh lp. Tn cc v

thuc do cng ty Giai Cnh cung cp. Tn c th l tn thng dng c ghi

trong cc ti liu chnh thng ca ngnh Dc (DVN IV, ti liu ging dy ti cc

trng y dc...). Tn cng c th l tn a phng. Do vy, vic xc nh tn

cc v thuc theo tn chnh thng v tn khoa hc l cn thit.

Qua kho nghim, thy rng cc v thuc c c im ph hp vi m t ca

DVN IV, DTQ l: dip c, trc bch dip, he hoa.

V thuc hong k c mt s c im ging m t trong DVN IV:Mu

trng hi vng, c cc b si, mnh mch, nhiu tinh bt. Tuy nhin, so snh vi

DTQ[43] v D M[44], thy c c im khc bit vi m t v thuc hong k

trong hai Dc in ny l c tinh th canxi oxalat. Mt khc, sc k dc liu

v ch phm u khng thy vt astragalosid IV. Cc c im ny ph hp vi m

t v thuc Hng k (tn khoa hc l Hedysarumpolybotrys Han.Mazz). May mn

rng, hng k c mt s tc dng tng t hong k [48]. Do , c th dng hng

k thay th cho hong k trong phng thuc.

V thuc thng ma: kt qu phn tch hnh thi v vi hc khng tng ng

vi m t v thuc thng ma theo DVN IV cc im: c cc khi tinh bt b h

ha, khng thy cc vt so li, cc mu b ngoi dc liu. Nh vy, thng ma

trong cng thc Ta An khng phi l thng ma c tn khoa hc l: Cimicifuga

heracleifolia Kom.,Cimicifuga dahurica (Turcz) Maxim., hocCimicifuga foetida

L., h Hong lin (Ranunculaceae)[9].

V vng en cha c trong DVN IV, chng ti xc nh c im hnh thi,

vi hc v thnh phn ha hc nhm gp phn tiu chun ha ch phm.

3.2.2. Thnh phn ha hc ch phm.

Mc ch nghin cu l kho st thnh phn ha hc ca ch phm hng ti

xy dng tiu chun ch phm Ta An. Thnh phn ha hc ca cc v thuc ch

mang tnh cht tham kho nh hng nghin cu v trong qa trnh bo ch, c

55

bit l giai on nu cao mi trng nc, nhit khong 1000C, chc chn s

xy ra nhiu phn ng ha hc nh oxy ha, thy phn, trng hp ha... nn thnh

phn cao thuc v ch phm c th khng tng ng vi thnh phn ha hc trong

cc v thuc. Ni cch khc, thnh phn ch khng th l php cng cc thnh phn

tng v thuc.

Qua nghin cu, nhn thy Ta An c cc thnh phn ha hc l: flavonoid,

saponin, acid amin polysaccharid, ng kh.

Rutin v quercetin l hai thnh phn c trong nhiu v thuc c tin hnh

xc nh hm lng nhm gip cho tiu chun v c xem l mt du im ch

ca Ta An. Kt qu cho thy trong Ta An rutin,quercetin c hm lng thp.

Nguyn nhn c th do tan ca hai cht ny trong nc thp hn trong cn. Hn

na, rutin c th b thy phn trong nc nhit cao.


Hin nay, phng php iu tr bnh tr ch yu l iu tr triu chng: vim

nhim v chy mu.

Kt qu nghin cu cho thy:

- To An c tc dng cm mu r rt cc liu 1.25 g/kg v 2.50 g/kg chut cng.

Kt qu ph hp vi d on v trong cng thc c nhiu v c tc dng cm mu

l: dip c,trc bch dip, he hoa. C ch cm mu l: co mch, tng tnh bn

thnh mch...

- Tc dng chng vim: Ta An liu 2.50 g /kg chut cng th hin tc dng

chng vim c ngha thng k ti thi im 3 gi sau khi gy vim, y cng

chnh l thi im vim mnh nht sau khi tim cht gy vim (p

56

KT LUN V KIN NGH

1. Kt lun

Cn c theo mc tiu ti, kt qu nghin cu c kt lun nh sau:

Tnh ng ca cc v thuc

- Cc v thuc ph hp vi DVN IV v DTQ 2010 gm: dip c, he hoa, trc

bch dip, vng en.

- V thuc c cc c im khng ph hp vi DVN IV: Thng ma (tn khoa hc

lCimicifuga heracleifolia Kom.,Cimicifuga dahurica (Turcz) Maxim. , hoc

Cimicifuga foetida L.), h Hong lin (Ranunculaceae).[9]

- V thuc hong k c cc c im tng ng vi vhng k theo m t D

M, DTQ 2010.

Thnh phn ha hc trong ch phm

- Ta An c cha cc nhm cht: Flavonoid, polysaccharid, saponin , acid amin,

ng kh.

- nh lng cn phn on EtOAc: Hm lng cn phn on EtOAc trong bt

thuc l 2.476%(kl/kl).

- Hm lng rutin trong ch phm l: 0.134mg/vin v hm lng quercitin l 5.74

g/vin. Hm lng phn trm ca rutin v quercitin trong ch phm ln lt l

0.0236% v 1.006 x 10-3%.

Tc dng sinh hc ca ch phm

- C tc dng trn cm mu trn chut theo ng ung c 2 mc liu dng 1.25

g/kg v 2.5g/kg .

- C tc dng chng vim cp trn chut theo ng ung mc liu 2.5g/kg.

2. Kin ngh

Do thi gian c hn, nhng nghin cu trn y ca chng ti ch l bc u.

Do vy, cn tip tc tin hnh nghin cu theo hng:

57

- Tip tc nghin cu thnh phn ha hc ca v thuc v ch phm: Cn chun

ha cc iu kin thc nghim hng ti xy dng tiu chun cht lng cho ch

phm.

- Tin hnh nghin cu thm cc tc dng dc l ca thuc c lin quan ti lm

sng bnh tr: Tc dng gim au, tc dng khng khun.

- Tuy cc v thuc trong phng thuc tng i an ton vi con ngi. Tuy

nhin, khi kt hp trong phng thuc c th c tc tc dng khng mong mun.

Do , cn tin hnh th c tnh cp, c tnh bn trng din.

TI LIU THAM KHO

Ting Vit

1. Nguyn Quc Anh, Ng Qy Chu (2011), Hng dn chn on v iu tr

bnh ni khoa, NXB Y hc , H Ni, tr. 558 - 559.

2. Nguyn Th Bay (2007), Bnh hc v iu tr ni khoa (Kt hp ng - Ty

y), NXB Y hc, H Ni, tr.22, H Ni

3. B mn Dc liu, Trng i hc Dc H Ni (2010), Thc tp dc liu

(Phn kim nghim bng phng php hin vi v phng php ha hc), Trung tm Thng tin Th vin HDHN, H Ni, tr. 29, 30, 61 76.

4. B mn Dc l, Thc tp dc l, Trung tm thng tin- Th vin i hc

Dc H Ni, H Ni, tr. 45, 41.

5. B mn y hc c truyn, Trng ai hc Y H Ni (2008), Y hc c truyn,

NXB Y hc, H Ni, tr. 579 - 583.

6. V Vn Chi (1997), T in Cy thuc Vit Nam, NXB Y hc, H Ni, tr.

1251-1252, 571 - 573, 564 - 565, 1338 - 1340, 406 - 407

7. Trn Th Thu Hng (2010),Dc l hc, NXB Phng ng, H Ni.

8. Trn Th Vit Hoa, L Th Kim Oanh (2008), Phn lp v xc nh cu trc

mt s hp cht t cy Dip c ( Houttuynia cordata Thumb) ca Vit Nam,

Tp ch pht trin Khoa hc v Cng ngh, 11(07), tr.73 -77.

9. nh H (2002), S tay xt nghim lm sng , NXB Y hc, tr. 46-47.

10. Hi ng Dc in (2010), Dc in Vit Nam, in ln th 4, NXB Y hc,

H Ni, tr.740 - 741, 921- 922, 897, 786 -787, 782, PL 11.3.

11. Nguyn Th Khnh, Phm T Dng (2001), Xt nghim s dng trong lm

sng, NXB Y hc, H Ni, tr. 112 - 162.

12. Khoa Y hc c truyn, i hc Y H Ni (2005) , Bo ch ng dc, NXB

Y hc, H Ni, tr. 107, 111, 210, 228.

13. Tt Li (2004), Nhng cy thuc v v thuc Vit Nam, NXB Y hc, H

Ni, tr. 40, 669 - 670, 887 - 889, H Ni.

14. Phm Xun Sinh (2004), Dc hcc truyn, NXB Y hc, H Ni, H Ni,

tr. 227- 228, 244, 296.

15. Phm Song, Nguyn Hu Qunh (1994), Bch khoa th bnh hc, tp 2,

Trung tm bin son t in bch khoa Vit Nam, H Ni,tr. 121-127.

16. Nguyn Vit Thn (2003), Kim nghim dc liu bng phng php hin

vi, tp 1, NXB khoa hc v k thut, H Ni, tr. 133- 135, 137- 139, 224-

227

17. Ng Vn Thu (2006), Bi ging Dc liu, tp 1, NXB Y hc, H Ni,

tr.290 292, 294-295.

18. Nguyn B Tnh (2004), Nam dc thn hiu, Tu Tnh ton tp,ln in th

4, NXB Y hc, H Ni, tr. 24,29,

19. Vin Dc liu (2006), Cy thuc v ng vt lm thuc Vit Nam tp I,

NXB khoa hc k thut, H Ni, tr. 672-675, 971- 976, 946- 949

20. Vin Dc liu (2006), Cy thuc v ng vt lm thuc Vit Nam tp II,

NXB bn khoa hc k thut, H Ni, tr. 997 1000, 844- 848, 1081- 1088

21. Vin Dc liu (2006), Nghin cu thuc t tho dc, NXB khoa hc k

thut, H Ni, tr. 288.

22. Vin Dc liu (2006), Phng php nghin cu tc dng dc l ca

thuc t dc tho, NXB Khoa v hc K thut, H Ni,tr. 140-149.

Ting Anh

23. Anne L. Mounsey; Jacqueline H., et.al (2011), Hemorrhoids, Am Fam

Physician, 84(2),pp. 204-210.

24. Chiang L.-C., Chang J., Chen C.-C., Tei N. (2003), Anti-Herpes Simplex

Virus Activity of Bidens pilosa and Houttuynia cordata, The American

Journal of Chinese Medicine, 31(3), pp 355.

25. Chinnala K.M., Elsani M.M. ,Mekala S. K. (2014), Evaluation of

antiobesity activity of Sesamun indicum L. in high fat diet induced obesity in

Rats, International Journal of Phytopharmacology, 5(3), pp.179-182.

26. Dong X.(2008), Study on Pharmacological Effects of Fresh and Dry

Houttuynia cordata Thunb, Li Shizhen Medicine and Materia Medica Re-

search,19(6), pp. 1315- 1316.

27. M.Durrer, I.Kanfer (2004), High performance liquid chromatography

determination of selected flavonols in Ginkgo biloba solid oral dosage

forms, J Pharmaceut Sci, 7(3), pp. 303 309.

28. Fu J., Dai L., Lin Z., Lu H. (2013), Houttuynia cordata Thunb: A Review

of Phytochemistryand Pharmacology and Quality Control , Chinese

Medicine, 4, 101-123.

29. Mary James A. Duke, with Mary Jo Bogenschutz-Godwin, Judi duCellier,

Peggy-Ann K. Duke (2002), Hanbook of Medicinal Herbs ,2nd ed, Library

of Congress Cataloging-in-Publication Data, pp. 253 -254 , 545 -546 , 549 -

550 , 394 -395 , 664- 665.

30. Huang K.C.(1991), Pharmacology of chinese Herbs, 2nd edition, USA, pp.

203, 350, 380, 385.

31. Amal El- Sayed Khailee, Mariam Husain Gonaid, Ramzia Ismail El- Bagry,

Amany Ameen Sleem, Marwan Shabana (2007), Chemical and Biological

Study of the Residual Aerial Parts of Sesamum indicum L., Journal of Food

and Drug Analysis, 15(3), pp.249- 257.

32. Park K.W., Lee J.-E.,Park K. (2009), Diets containing Sophora japonica L.

prevent weight gain in high-fat diet-induced obese mice, NutritionResearch,

Science direct, 29(11), pp. 819- 824.

33. Kwon H.-D, Cha I.-H., Lee W.-K, Song J.-H., Park I.-H. (1996),

Antibacterial Activity of Volatile Flavor Components from Houttuynia

cordata Thunb, Journal of Food Science and Nutrition, 1(2), pp. 208-213.

34. Lu H, Wu X, Liang Y, Zhang J. (2006), Variation in chemical composition

and antibacterial activities of essential oils from two species of Houttuynia

T., Chem Pharm Bull (Tokyo), 54(7), pp. 936-940.

35. Meng J., He D., Zhou Y., Dong X. (2008), Comparison on the

Pharmacological Effects of Different Extract Parts from Houttuynia cordata

Thunb, Li Shizhen Medicine And Materia Medica Research, 19(5), pp.

1050-1051.

36. Meng J., Zong X., Dong X.(2008), Study on Pharmacological Effects of

Fresh and Dry Houttuynia cordata Thunb, Li Shizhen Medicine and

Materia Medica Re- search, Vol. 19, No. 6, pp. 1315-1316.

37. Parvin Mirmiran PhD, Zahra BahadoranMSc, Mahdieh Golzarand MSc,

Asadolah Rajab MD, Feredoun Azizi MD (2003), Ardeh (Sesamum

indicum) could improves serum Triglycerides and Atherogenic lipid

parameters in type 2 diabetic patients: Arandomized Clinical Trial, Archives

of Iranian Medicine, 16(11), pp 652-656.

38. T.S. Mohamed Saleem (2011), Anti-microbial activity of sesame oil,

International Journal of Research in Phytochemistry & Pharmacology, 1(1),

pp. 21-23.

39. Noor A. Ghani, Mahfoodha A. Umran, Muayad S. Shawkat (2012),

CytotoxicActivity of Ethanol Extract of Sesamum indicum Seeds to Cancer

Cell Lines In Vitro,Iraqi Journal of Cancer and Medical Genetics, 5(1), pp.

28-3.

40. Romano M, Faggioni R, Sironi M, Sacco S, Echtenacher B, Di Santo E,

Salmona M, Ghezzi P (1997), Carrageenan-induced acute inflammation in

the mouse air pouch synovial model, Role of tumour necrosis factor.

Mediators Inflamm , 6(1), pp. 32-38.

41. Shim S.

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