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B Y T

TRNG I HC DC H NI

DNG D THO

NGHIN CU NH TNH MT S

THNH PHN DC LIU V TC

DNG GIM HO, LONG M CA

CAO LNG V NGN

KHA LUN TT NGHIP DC S

H NI 2014

B Y T

TRNG I HC DC H NI

DNG D THO

NGHIN CU NH TNH MT S

THNH PHN DC LIU V TC

DNG GIM HO, LONG M CA

CAO LNG V NGN

KHA LUN TT NGHIP DC S

Ngi hng dn:

1. TS. Bi Hng Cng

2. DS. Nguyn Th Thu Trang

Ni thc hin:

1. B mn Dc hc c truyn

Trng i hc Dc H Ni

2. B mn Dc l

Trng i hc Y H Ni

H NI 2014

LI CM N

Li u tin ti xin by t lng knh trng v bit n su sc nht ti

TS. Bi Hng Cng, ngi thy tn tnh hng dn v ng vin ti trong sut

qu trnh thc thc hin kha lun tt nghip.

Ti xin chn thnh cm n DS. Nguyn Th Thu Trang to iu kin,

quan tm gip ti thc hin kha lun.

Ti xin cm n PGS.TS. Phm Th Vn Anh Trng B mn Dc l,

i hc Y H Ni cng cc anh ch B mn nhit tnh gip ti trong qu

trnh lm thc nghim.

Ti cng xin gi li cm n ti Ban gim hiu, thy c v cc anh ch B

mn Dc hc c truyn ch bo v gip ti trong thi gian nghin cu.

Cui cng, ti xin gi li cm n ti gia nh, bn b lun ng vin,

gip ti trong qu trnh hc tp v nghin cu hon thnh kha lun tt

nghip ny.

H Ni, thng 5 nm 2014

Sinh vin

Dng D Tho

MC LC

T VN 1

Chng 1 . TNG QUAN.2

1.1. Sinh l bnh vim hng ........................2

1.2. Phng thuc V Ngn..............3

1.3. Thng tin c bn v cc v thuc..4

1.3.1. X can..4

1.3.2. Kim ngn hoa..5

1.3.3. Hng chanh.7

1.3.4. Bc h..8

1.3.5. Cam tho.9

1.3.6. Nc nc..11

1.3.7. Ct cnh.....12

1.3.8. Huyn sm.13

1.3.9. Mch mn.14

1.3.10. Thin mn...15

1.3.11. Bng i hi.16

1.3.12. Sinh a16

Chng 2. I TNG V PHNG PHP NGHIN CU.....18

2.1. Nguyn liu.....18

2.2. Phng tin.....18

2.2.1. Thit b..18

2.2.2. Ha cht.19

2.2.3. ng vt th nghim..19

2.3. Ni dung nghin cu ..19

2.4. Phng php nghin cu.....19

2.4.1. nh tnh thnh phn ha hc ca mt s dc liu v cao lng V Ngn...19

2.4.2. Nghin cu tc dng gim ho, long m ca cao lng V Ngn.19

Chng 3. THC NGHIM, KT QU V BN LUN. 21

3.1. nh tnh cc nhm cht bng phn ng ha hc.. 21

3.2. nh tnh so snh cao v v thuc bng sc k lp mng.. 27

3.2.1. Kim ngn hoa....27

3.2.2. Cam tho...29

3.2.3. X can...30

3.2.4. Nc nc..32

3.2.5. Huyn sm.....33

3.2.6. Ct cnh, mch mn, thin mn35

3.3. Nghin cu tc dng sinh hc ....37

3.3.1. Tc dng gim ho..37

3.3.2. Tc dng long m ..39

3.4. Bn lun..42

3.4.1. Thnh phn ha hc..42

3.4.2. Tc dng sinh hc.44

KT LUN V KIN NGH.....45

TI LIU THAM KHO

DANH MC CC K HIU, CC CH VIT TT

CC : Ct cnh

CT : Cam tho

EtOAc Ethylacetat

EtOH Ethanol

HS : Huyn sm

KNH : Kim ngn hoa

MeOH Methanol

MM : Mch mn

NN : Nc nc

S : Sinh a

SKLM : Sc k lp mng

TB Trung bnh

TM : Thin mn

TT : Thuc th

XC : X can

YHCT : Y hc c truyn

DANH MC CC BNG

Bng Trang

Bng 3.1. Kt qu nh tnh ha hc cc nhm cht trong cao lng v dc liu....26

Bng 3.2. Kt qu phn tch sc k ca cao v KNH bc sng 366 nm.28

Bng 3.3. Kt qu phn tch sc k ca cao v CT bc sng 254 nm30

Bng 3.4. Kt qu phn tch sc k ca cao v XC sau khi phun thuc th31

Bng 3.5. Kt qu phn tch sc k ca cao v NN bc sng 254 nm33

Bng 3.6. Kt qu phn tch sc k ca cao v HS sau khi hin mu..34

Bng 3.7. Kt qu phn tch sc k ca cao v CC, TM, MM sau khi hin mu.36

Bng 3.8. nh hng ca cao lng V Ngn ln thi gian tim tng xut hin cn ho38

Bng 3.9. nh hng ca cao lng V Ngn ln tng s cn ho..39

Bng 3.10. nh hng ca cao lng V Ngn ln mt quang..40

DANH MC CC HNH V, TH

Hnh Trang

Hnh 2.1. H thng thit b sc k lp mng hiu nng cao Linomat 5...18

Hnh 3.1. Sc k cao-KNH bc sng 366 nm28

Hnh 3.2. Sc k cao-CT bc sng 254 nm...30

Hnh 3.3. Sc k cao-XC sau khi hin mu.31

Hnh 3.4. Sc k cao-NN bc sng 254 nm...33

Hnh 3.5. Sc k cao-HS sau khi hin mu..34

Hnh 3.6. Sc k CC- cao- TM- MM sau khi hin mu...........36

Hnh 3.7. Biu biu th thi gian tim tng xut hin cn ho chut.38

Hnh 3.8. Biu biu th tng s cn ho chut...39

Hnh 3.9. Biu biu th mt quang bc sng 265 nm41

1

T VN

Vim hng l mt bnh l vim nhim thng gp trong i sng. Vit

Nam, kh hu nhit i nng m, thi tit thay i tht thng, mi trng

nhim, l iu kin thun li cho bnh pht sinh v pht trin. Bnh gp mi

gii, mi la tui, c bit tr em, c th ti pht nhiu ln trong nm. Nguyn

nhn gy bnh ch yu do virus v vi khun. Vim hng c c trng bi cc

triu chng nh ph n nim mc hng, hng sng au, rt, km theo s tng tit

m, ho... Cc thuc c s dng trong iu tr thng l thuc khng sinh, chng

vim, gim au cng vi thuc gim ho, long m. Thc t, vic s dng thuc

tn dc trong iu tr vim hng, nht l i vi vim hng mn tnh, c th gy ra

nhiu tc dng bt li nh khng thuc, hi gan, thn, ri lon tiu ha, vim lot d

dy Do vy, vic nghin cu, s dng thuc ng dc iu tr vim hng

ngy cng c quan tm nhm hn ch tc dng ph, mang li ngha thit thc

i vi ngi bnh v gp phn pht trin nn ng dc Vit Nam.

Xut pht t iu , Cng ty Dc phm Hoa Linh nghin cu bo ch

cao lng V Ngn iu tr vim hng t phng thuc gm cc dc liu nh:

kim ngn hoa, x can, bc h, hng chanh, nc nc, cam tho, ct cnh, thin mn

Hin nay, cao lng V Ngn ang c tip tc nghin cu hon thin cho ra i

ch phm. Do , nhm gp phn xy dng tiu chun c s v nh gi tc dng

sinh hc ca ch phm, ti Nghin cu nh tnh mt s thnh phn dc liu

v tc dng gim ho, long m ca cao lng V Ngn c thc hin vi cc

mc tiu c th:

- nh tnh bng phn ng ha hc v sc k lp mng so snh cao lng v mt s

thnh phn dc liu, gp phn xy dng tiu chun bi thuc.

- Nghin cu mt s tc dng sinh hc ca cao lng V Ngn lin quan ti iu tr

vim hng: gim ho, long m.

2

CHNG 1. TNG QUAN

1.1. Sinh l bnh vim hng

Theo quan im Y hc hin i:

- Vim hng l tnh trng vim nhim nim mc hng, c nhiu nguyn nhn

nhng ch yu do virus hoc vi khun gy vim cc vng ln cn nh vim

mi, vim xoang mt lm dch tit chy xung hng hay do vi khun gy su

rng lan n hng. Ngoi ra, vic tip xc vi khi bi, ha cht trong thi gian

di, ht thuc l, ung ru, thay i thi tit gy tn thng nim mc hng,

lm hng mt chc nng bo v.

- Phn loi:

+ Vim hng cp: l vim cp tnh nim mc hng, nguyn nhn do virus (60-

80%) v vi khun (thng do bi nhim sau khi nhim virus) vi cc triu

chng: au hng km theo cm gic nng rt, nut au, st cao, mt mi,

+ Vim hng mn: l tnh trng vim hng ko di, do nh hng ca nght, tc

mi, phi th bng ming, nht l v ma lnh; vim xoang, vim amidan mn;

ht thuc l, ung ru, tip xc nhiu vi ha cht, khi bi; c a: d ng, i

tho ng, suy ganvi cc triu chng: cm gic hng kh, nng, rt; nga

hng, vng hng tng ln khi nut, c t nhy qunh, thng b ho vo ban

m, khi lnh [10], [22].

Theo quan im YHCT:

- Hu m nhim chc nng ht th thuc ph, hng l ca ng ra ca v, ca

ng ca cc tng. Vim hng do kh nhit gy m kt, l chng bnh sng,

nng, , au gi l hu t [20].

- Nguyn nhn: Ngoi t xm nhp, n ung khng iu , tnh ch b tn

thng, ph tng mt iu ha [17], do nhit c ph v xng ln v nhim

kh c dch l, tn thng ph m [6].

- Phn loi:

+ Th phong nhit xm nhp: hng kh, sng nng, au, nut kh, pht st,

kht nc, ho hen, ru li vng... iu tr: dng thuc s phong thanh nhit.

3

+ Th phong hn xm nhp: hng nht, khng sng, au mnh my, ru li

trng mng iu tr: dng thuc s phong tn hn [21].

Ngoi ra, cn kt hp iu tr cc triu chng bng cc thuc ha m, ch ho:

- Theo YHCT, m l cht dch nht, dnh, sn sinh trong qu trnh hot ng ca

lc ph, ng tng. m ngng ng ph th gi l m. m lm khng kh

vo ph kh khn, gy kh th, to mi trng pht trin cho cc loi virus, vi

khun; kch thch nim mc gy ho, kch thch c trn kh ph qun gy co tht.

Do , kh m l mt khu rt quan trng trong iu tr bnh ph. C 2 loi:

thuc ha m hn, thuc ha m nhit.

- Thuc ch ho c tc dng n ph, thanh ph, nhun ph, ging kh ph nghch,

ng thi c tc dng ha m, iu tr ho do nhiu nguyn nhn, c 2 loi: n

ph ch khi, thanh ph ch khi [16].

1.2. Phng thuc V Ngn

Cao lng V Ngn c bo ch t phng thuc V Ngn, vi cng thc

c Cng ty Dc phm Hoa Linh nghin cu v xut, c th nh sau:

V thuc Khi lng

Kim ngn hoa (Flos Lonicerae) 10g

X can (Rhizoma Belamcandae) 12g

Bc h (Herba Menthae) 10g

Hng chanh (Folium Coleus) 8g

Cam tho (Radix Glycyrrhizae) 8g

Nc nc (Cortex Oroxyli) 8g

Ct cnh (Radix Platycodi grandiflori) 4g

Mch mn (Radix Ophiopogonis) 4g

Thin mn (Radix Asparagi) 4g

Huyn sm (Radix Scrophulariae) 8g

Sinh a (Radix Rhemanniae glutinosae) 8g

Bng i hi (Semen Scaphii) 4g

4

Cng nng: Khng khun, tiu vim, thanh nhit, ha m, ch ho

Ch tr: Cc chng hu hng b vim, sng au, rt, nhiu m, khn ting, ho,

vim amidan, vim thanh qun, vim kh qun

1.3. Thng tin c bn v cc v thuc

1.3.1. X can

a, Tn khoa hc v thuc: Rhizoma Belamcandae

b, B phn dng: Thn r phi hay sy kh ca cy x can Belamcanda chinensis

(L.), DC. H La dn (Iridaceae) [11], [15], [16], [25].

c, Thnh phn ha hc

- Flavonoid thuc nhm isoflavonoid: tectoridin (1,5%), tectorigenin, 7-O-

methylirisolidon, irigenin, iridin, dimethyltectorigenin, irisflorentin, muningin, cc

iristectorigenin A v B, belamcandin, dichotomitin [4], [14], [25], 9-methoxy

dehydrodiconiferyl alcol [3].

- Cc thnh phn khc: belamcandal, desacetylbelamcandal, belamcandol A [4].

d, Tc dng sinh hc

- Tc dng khng khun: Cao chit ethanol thn r x can c ch cc chng vi

khun: ph cu, lin cu tan mu, trc khun ho g, Bacillus subtilis, c tc dng

yu i vi cc chng vi khun: t cu vng, Shigella dysenteriae, Sh.shigae,

Enterococcus [18], [25].

- Ngoi ra, x can cn c tc dng chng vim chut cng trng, chng co tht

gy ra bi histamine trn c trn chut lang; tc dng li tiu nh, gim au v c

c tnh thp [1], [9], [18], [25].

- Cao chit vi nc nng nng 0,05 mg/ml c hot tnh c ch aldose

reductase, enzym gy tch ly sorbitol trong t bo bnh i tho ng [25].

- TEC-01 chit xut t x can, thnh phn chnh l tectorigenin, c tc dng c

ch PGE2 v COX2 i thc bo phc mc chut [35], ngn hnh thnh khi u

[33], c ch bnh sinh ca x ha phi v cn [50], chng li s hnh thnh x gan

[48], chng t bin v chng oxy ha [47].

e, Tc dng v cng dng theo YHCT

5

- Tnh v: v ng, cay, tnh hn.

- Qui kinh: Ph v can [7], [15], [16], [25].

- Cng nng, ch tr:

+ Thanh nhit gii c: cha cc bnh v hng, vim amidan c m, vim

amidan cp tnh v mn tnh, ho nhiu m, khn ting, tc c hng, sng au,

vim hng cp tnh, vim hng ht, vim hng mn sng v, tc tia sa, ung

c, mn nht [7], [15], [16], [25]; cha st, i tiu tin khng thng [15], [16].

+ Ging ph kh, ha m, bnh suyn: dng trong bnh ho vi th nhit, m

nhiu, c, hoc kh th do co tht kh qun [4], [11], [16], [25].

+ Thng kinh hot lc, tn huyt: dng trong trng hp b kinh dn n bng

sn cng tc, y trng [16].

- Liu dng: Ngy dng 4 12g [16].

- King k: ngi t v h hn [7], [16], ph n c thai [11], [25].

1.3.2. Kim ngn hoa

a, Tn khoa hc v thuc: Flos Lonicerae

b, B phn dng: n hoa c ln mt s hoa phi hay sy kh ca cy kim ngn

(Lonicera japonica Thunb.) v mt s loi khc cng chi nh L. dasystyla Rehd.; L.

confusa DC. v L. cambodiana Pierre. H Kim ngn (Caprifoliaceae) [7], [16], [25].

c, Thnh phn ha hc

- Flavonoid thuc nhm euflavonoid: luteolin, luteolin -7- rutinosid [4], [25].

- Cc flavonoid khc gm: lonicerin, loniceraflavon v mt s iridoid

glycoside nh loganin, secoxyloganin, secologanin, secologanin dimethyl acetal,

vogelosid, epivogelosid [25].

- Tinh du: -pinen; hex-1-en; hex-3-en-1-ol; cis v trans -2-methyl-2-vinyl-5-

(-hydroxyisopropyl)-tetrahydrofuran; geraniol; -terpineol; alcol benzylic;

alcol -phenyl ethylic; carvacrol; eugenol; linalool; 2,6,6-trimethyl-2-vinyl-5

hydroxy tetrahydrydropyran [25].

6

- Acid clorogenic 6%, acid isoclorogenic gm 3 ng phn l acid

isoclorogenic a (acid 3,5-dicafeoyl quinic), isoclorogenic b v c (l 2 ng phn

ca acid 3,4-dicafeoylquinic [4], [25].

- Saponin, trong aglycon l acid oleanolic hoc hederagenin [25].

- Mt s cht carotenoid: -caroten, -cryptoxanthin, auroxanthin [4].

d, Tc dng sinh hc

- Kim ngn hoa c ph khng khun rng, c ch nhiu loi vi khun nh trc

khun l, trc khun thng hn, ph thng hn, i trng, ho g, m xanh,

phy khun t, bch hu, lao, t cu khun, lin cu khun, song cu khun vim

phi [16], [19], [25]. Ngoi ra cn c ch mt s nm ngoi da [16].

- Kim ngn hoa c dng ch yu tr vim nhim ng h hp trn nh

vim amidan, vim hng, vim thanh qun; vim da, mn nht, sng v, vim

rut tha; tr l trc trng, vim mng kt do siu vi, cm [4].

- Ngoi ra, kim ngn hoa cn c tc dng chng vim [12], [13], [19], [23], c tc

dng tt trong iu tr COPD [42], tng cng chuyn ha cc cht bo, chng

sc phn v v khng c [15], [25], khng mt s vius, bo v t bo gan [44],

[46], c ch enzym -Glucosidase, gim ng mu sau n [52].


- Tnh v: V ngt, hi ng, tnh mt [25], tnh hn [16].

- Quy kinh: Tm, ph, v v t [7], [11], [16], [25].

- Cng nng, ch tr:

+ Thanh nhit gii c: dng ring hay phi hp vi nhiu v thuc khc cha

mn nht, inh c, nht v, nht trong rut, d ng, my ay, mn nga, ban

si, t, l, ho do ph nhit [7], [11], [15], [16], [25].

+ Gii c st khun: dng trong bnh sng au ca hu hng, vim amidan,

vim thanh qun, au mt , phng bnh vim no [16]; cha cm cm, vim

phi, p xe phi, vim gan virus, vim gan mn tnh, vim cu thn cp tnh,

nhim khun huyt, vim phi tr em, co git tr em [25].

7

+ Thanh gii biu nhit: dng trong cc trng hp ngoi cm phong nhit, n

nhit s khi (st nng thi k u) [16].

- Liu dng: 12-20g/ngy [16]. Ngy dng 4 6g hoa di dng thuc sc, hm,

cao, ru thuc hoc hon tn [25].

- King k: Nhng ngi th h hn, khng thc nhit, m hi ra nhiu, mn

nht c m v lot [7], [11], [16], [25].

1.3.3. Hng chanh

a, Tn khoa hc v thuc: Folium Coleus

b, B phn dng: l dng ti ca cy hng chanh Coleus aromaticus Benth. H

Bc h (Lamiaceae) [24].

c, Thnh phn ha hc

Tinh du (0,05 0,12%) [5], vi thnh phn chnh l carvacrol [5], [15], [24].

Ngoi ra c -terpinen, -terpinen, thymol [24].

d, Tc dng sinh hc

- Tc dng khng khun: Tinh du hng chanh c tc dng c ch mnh i vi

cc vi khun: trc khun mycoides, trc khun subtilis, trc khun lao (gim

c), trc khun l Flexner, trc khun bch hu, trc khun ho g, trc khun

l Shiga, E. coli, t cu vng, lin cu khun tan mu, trc khun thng hn;

nm Candida albicans, dit amip Entamoeba moshkowskii [15], [24].

- Cao nc hng chanh c tc dng c ch s pht trin ca ph cu khun, t cu

khun vng. Nh vy, tc dng khng khun ca hng chanh khng ch do tinh

du, m cn c th do nhng thnh phn khc nh flavon, acid nhn thm [24].


- Tnh v: V the cay, hi chua, mi thm, tnh m [24].

- Quy kinh: Can, ph [24].

- Cng nng, ch tr: Li ph, tr m, gii cm, pht hn, thoi nhit, tiu c:

Dng cha cm cm, nhc u ngt mi, ho, hen, vim hng, khn ting, ho ra

mu, ho g, st cao, st khng ra m hi, nn ra mu, chy mu cam [16], [24].

- Liu dng: Ngy dng 10 16g l ti, hoc 4 8g l kh [24].

8

1.3.4. Bc h

a, Tn khoa hc v thuc: Herba Menthae

b, B phn dng: Phn trn mt t ca cy bc h Vit Nam Mentha arvensis L..

H Hoa mi (Lamiaceae) [7], [8], [16], [24].

c, Thnh phn ha hc

- Tinh du: Thnh phn chnh l menthol (trn 70%). Ngoi ra c menthol ester,

menthon, cc hp cht hydrocarbon monoterpenic [5].

- Bc h tm Vit Nam gm 32 thnh phn, trong xc nh: -pinen, -

pinen, menthol, menthyl acetat, myrcen, limonen, pulegon, piperiton, piperiton

oxyd, p.cymol[24].

- Tinh du Mentha arvensis di thc vo Vit Nam (NV.74) cha sabinen, myrcen,

-pinen, limonen, cineol, methyheptenon, menthon, isomenthol, menthyl acetat,

neomenthol, menthol, isomenthon, pulegon [24].

d, Tc dng sinh hc

- Vi liu nh, bc h c tc dng kch thch trung khu thn kinh, gin mch, thc

y bi tit m hi v h nhit.Vi liu ln, kch thch ty sng, lm t lit phn

x vn ng [16].

- Tinh du bc h v menthol bc hi nhanh, gy cm gic mt v t ti ch, dng

trong trng hp au dy thn kinh [8], [16], [24].

- Tc dng khng khun: ph khng khun rng, c ch nhiu vi khun nh:

Staphilo.aureus, Sal.typhy, Sh. Flexneri, Sh. Sonnei. Sh. Shiga, B. subtilis,

Strepto. D. pneumonie, H. perrtussis [16], c ch cc chng vi khun t Vibrio

cholera Eltor, Vibrio cholera Inaba, Vibrio cholera Ogawa [24].

- Menthol c tc dng chng co tht, gim au, kch thch tiu ha, cha hi

ming [5], [8].


- Tnh v: V cay thm, tnh mt [7], [8], [16].

- Qui kinh: Ph v can [7], [11], [16].

- Cng nng, ch tr:

9

+ Pht tn phong nhit: Tr ngoi cm phong nhit, st khng ra m hi, nhc

u, ngt mi, gim ho vim hng, mt , nga ni m ay, bng au, y

chng, tiu ha km, nn ma [5], [7], [11], [16], [24].

+ Tr phong gim au: tr au u, au mt do phong nhit, hng , sng

au [7], [16], [24].

+ Kin v, ch t, gii c, tng tit mt, kch thch tiu ha: dng trong cc

trng hp n ung khng tiu, nn lm, chua, au bng, i t [8], [16].

+ Ha m, h tch, tiu sng, ch nga [24].

- Liu dng: L v ton cy, ngy ung 4 8g di dng thuc hm [24], 2

12g [16]. Tinh du v menthol: 0,02 0,2ml/ln, 0,06 0,6ml/ngy [24].

- King k: Ngi kh h huyt to, can dng thnh biu h, m hi nhiu,

ngi gy yu, suy nhc ton thn, to bn, huyt p cao, tr em di 1 tui

khng nn dng [7], [11], [16], [24].

1.3.5. Cam tho

a, Tn khoa hc v thuc: Radix Glycyrrhizae

b, B phn dng: r phi hay sy kh ca cy cam tho bc Glycyrrhiza glabra L

hoc Glycyrrhiza uralensis Fisch ex DC. H u (Fabaceae) [4], [8], [16], [24].

c, Thnh phn ha hc

- Glycyrrhizin l mt saponin nhm olean (10 14% trong dc liu kh).

- Cc dn cht triterpenoid khc nh acid liquiritic, acid 18--hydroxy-

glycyrrhetic, acid 24-hydroxyglycyrrhetic, glabrolid, desoxyglabrolid,

isoglabrolid, acid liquiridiolic

- Cc flavonoid (3 4%): c 27 cht c bit, quan trng nht l liquiritin (hay

liquiritirosid) v isoliquiritin (hay isoliquiritirosid).

- Ngoi ra cn c nhiu flavonoid thuc cc nhm khc: isoflavan (gla-bridin),

isoflavon (glabron), isoflaven (glabren).

- Cc hot cht estrogen steroid, dn cht coumarin (umbelliferon, herniarin,

liqcoumarin), tinh bt, glucose, saccharose [4], [8], [15], [24].

d, Tc dng sinh hc

10

- Dch chit cam tho c tc dng chng lot d dy, c ch tc dng gy tng tit

dch v ca histamin, chng co tht c trn rut (i khng vi histamin,

acetycholin), do thnh phn flavonoid [4], [8], [15], [16], [24].

- Tc dng long m do cc saponin, tc dng trn ho ca acid-18-glycyrrhitic

tng t codein [4], [8].

- Tc dng c ch enzym monoaminoxydase (MAO) ca liquiritigenin v

isoliquiritigenin [4], chng trm cm [31], bo v t bo thn kinh [49].

- Gy trn tnh, c ch thn kinh trung ng, h thn nhit, gim h hp, bo v

gan trong vim gan mn tnh, tng bi tit mt, chng vim gan, chng d ng

[8], [24].

- Cam tho cha cht glycyrrhizin c tc dng gii c nht nh vi ng c thc

n hoc trng c mt s thuc khc: morphin, cocain, strychnin, atropin; c t

bch hu, un vn (c ch: glycyrrhizin thy phn thnh acid glucuronic kt hp

vi cht c) [4], [16].

- Tc dng oestrogen, tc dng tng t cortison ca glycyrrhizin trn chuyn ha

cht in gii, gi natri v clorid, bi tit kali, chng vim, cha to bn [4], [24].

- Nghin cu gn y cn cho thy cam tho c tc dng nng cao kh nng min

dch ca c th [4], gim cholesterol mu [16].


- Tnh v: V ngt, tnh bnh.

- Quy kinh: can, t, thng hnh 12 kinh [11], [16], [24].

- Cng nng, ch tr:

+ ch kh, dng huyt: tr bnh kh huyt h nhc, mt mi thiu mu [11],

[16].

+ Nhun ph, ch ho: tr bnh au hu hng, vim hng cp, mn tnh, vim

amidan, ho mt ting, ho nhiu m [8], [11], [16].

+ T ha gii c: tr mn nht inh c sng au [11], [16].

+ Hon cp ch thng: tr au d dy, au bng, gn mch co rt [8], [16].

11

- Liu dng: Ngy dng 4 20g, di dng bt, thuc hm, nc sc, cao mm

[11], [24].

- King k: T v thp tr, si bng, y bng; dng lu d b ph n [16].

1.3.6. Nc nc

a, Tn khoa hc v thuc: Cortex Oroxyli

b, B phn dng: v thn cy nc nc Oroxylum indicum Vent., h Nc nc

(Bignoniaceae) [4], [11], [8].

c, Thnh phn ha hc

Flavonoid thuc nhm euflavonoid: chrysin, baicalein, oroxylin A [4], [15], [24],

baicalein -7 -O- glucoside, baicalein -7 -O- diglucoside, chrysin - diglucoside [29],

acid p-coumaric [24], cc hp cht phenolic [30].

d, Tc dng sinh hc

- V nc nc nghin cu thc nghim thy c tc dng chng d ng r rt v lm

tng sc khng ca c th i vi mt s tc nhn c hi; c ch giai on cp

tnh ca phn ng vim [24].

- Flavonoid ton phn chit t v nc nc dng iu tr cho bnh nhn mc bnh

vy nn, hen ph qun, t a, my ay cho kt qu tt (Ch phm Nunaxin) [24].

- Ngoi ra, cc nghin cu cn chng minh nc nc c tc dng chng t bin

(baicalein), c ch t bo ung th, kch thch min dch [24], [30], chng oxy ha,

c ch - glucosidase trong iu tr tiu ng [45], chng lot d dy [27].


- Tnh v: V ng, tnh mt [24].

- Quy kinh: bng quang, t [11].

- Cng nng, ch tr: Thanh can gii nhit, tiu c, st trng, li thp, nhun

ph, ch khi, ch thng: dng cha cc bnh vng da, d ng mn nga, vim

hng, ho khn ting, au d dy, l, vim ng tit niu, tr con ban si [8],

[11], [16], [24].

- Liu dng: Ngy dng 8 16g di dng thuc sc hoc cao lng [11], [24].

- King k: Ngi h hn au bng, y bng, tiu chy [11], [24].

12

1.3.7. Ct cnh

a, Tn khoa hc v thuc: Radix Platycodi grandiflori

b, B phn dng: r phi hay sy kh ca cy ct cnh Platycodon grandiflorum

(Jacd) A.DC. H Hoa chung (Campanulaceae) [4], [7], [8], [16], [24].

c, Thnh phn ha hc

Hot cht chnh l saponin triterpenoid nhm olean (platycodin A,C,D, D2, E;

cc polygalacin D, D2, 16-oxo-platycodin D) [8], [24], [40]; cc sapogenin l

platycodigenin, acid polygalacic [4], [24].

Ngoi ra cn c polysaccharid, phytosterol, tanin [24], inulin [4].

d, Tc dng sinh hc

- Trn lm sng, saponin ca ct cnh c tc dng tiu m r rt. Khi ung,

saponin gy kch thch nim mc hng v d dy, a n phn ng tng tit dch

ng h hp lm cho m long v d b tng ra ngoi [4], [15], [16], [24].

- Saponin ca ct cnh c tc dng ph huyt mnh. Ngoi ra, c tc dng gim

au, lm trn tnh, h nhit, gim ho, chng lot, chng vim [16].

- Tc dng khng khun: c ch Staphylococcus aureus, B. mycoides, D.

pneumonia [16].

- Tc dng chng oxy ha, chng ung th [39], [43], bo v t bo gan [37].


- Tnh v: V hi ngt, sau ng, hi cay, tnh bnh [2], [7], [8], [24], tnh m [16].

- Qui kinh: Ph [7], [24].

- Cng nng, ch tr:

+ Thng kh ph, li hu hng: dng khi kh ph b tc, hu hng sng au nh

vim hng, vim amidan, ngc sn au nh dao m.

+ Kh m ch ho: Cha ho c m hi tanh, kh khc, khn ting, hen suyn,

tc ngc, kh th.

+ Tr m, tiu ung thng: tr ph ung, ph c m, p xe phi [2], [8], [15], [24].

- Liu dng: Ngy dng 4 12g [16], 3-10 g [2], [7].

13

- King k: ngi m h ha vng, ho lu ngy, ho ra mu; dng lng ln qu

dn n au tim, bun nn [16]. Thn trng vi ph n c thai [24].

1.3.8. Huyn sm

a, Tn khoa hc v thuc: Radix Scrophulariae

b, B phn dng: r phi hay sy kh ca cy huyn sm Scrophularia

buergeriana Miq. H Hoa mm si (Scrophulariaceae) [11], [16], [24].

c, Thnh phn ha hc

- Cc iridoid: Scrophularin, harpagid, harpagosid, ningpogenin [15], [24]; O-Me

catalpol, angorosid C [24], buergerinin F1 v G2 [41].

- Ngoi ra c alcaloid, ng, steroid, acid amin, acid bo, carotene, 17 nguyn t

vi lng [24], cc ester phenylpropanoid ca rhamnose [36].

d, Tc dng sinh hc

+ Tc dng khng khun, an thn [24].

+ Tc dng iu tr tt i vi vim hng mn tnh [24].

+ Tc dng tng huyt p v cng tim nh trn th, liu cao c tc dng ngc li,

h ng huyt [16].

+ Tc dng ci thin nhn thc do c ch acetylcholinesterase, chng oxy ha liu

2mg/kg th trng trn chut [32], bo v t bo thn kinh [36].


- Tnh v: V ng ngt, hi mn, tnh hn.

- Qui kinh: Ph, v, thn [7], [16], [11], [24].

- Cng nng, ch tr:

+ Thanh nhit ging ha, sinh tn dng huyt: dng khi nhit c nhp vo

phn inh huyt, dn n st cao, m sng, tn thng tn dch.

+ Gii c, chng vim: dng vi bnh st pht ban chn, lm thuc gim st,

chng vim trong cc chng st nng, kht nc, pht ban, ming li l lot,

vim hng, vim amidan, vim thanh qun, vim ming, vim li, vim kt mc,

ho khan, to bn, mn nga, mn nht, au mt .

14

+ Tn kt, nhuyn kin, lm mm u, khi rn: dng trong bnh m kt hch

nh loa lch (trng nhc, lao hch).

+ B thn, t thn m: dng trng thy, ch ha.

+ Ch kht: tr tiu kht, dng trong bnh i ng [7], [11], [15], [16], [24].

- Liu dng: Ngy dng 4 12g, dng thuc sc hoc cn ngm [24].

- King k: Ngi c thp t v, t v h hn, i tin lng [11], [16], huyt p

thp [24].

1.3.9. Mch mn

a, Tn khoa hc v thuc: Radix Ophiopogonis

b, B phn dng: r c phi hay sy kh ca cy mch mn Ophiopogon

japonicus (L.f.) Ker. Gawl., h Hong tinh (Convallariaceae) [4], [11], [25].

c, Thnh phn ha hc

- Saponin steroid: Ophiopogonin A, B, C, D [4], nhm furostan v spirostan [51].

- Carbohydrat: glucofructan v mt s monosaccharide (glucose, fructose,).

- -sitosterol, stigmasterol, -sitosterol -D-glucosid, tinh du [4], [25].

d, Tc dng sinh hc

Tc dng chng vim r rt i vi c 2 giai on cp tnh v mn tnh ca phn

ng vim thc nghim. Tc dng c ch tng i kh trn ph cu, yu hn trn

cc chng: t cu vng, Shigella dysenteriae, Bacillus subtilis [25].


- Tnh v: V ngt, hi ng, tnh mt [25].

- Quy kinh: Tm, ph, v [11], [16].

- Cng nng, ch tr:

Nhun ph, gim ho, cm mu, thanh nhit: Cha ho khan, vim hng, lao phi

nng m v chiu, st cao, tm phin kht nc, th huyt, khi huyt, chy mu

cam, hen ph qun, kh ng [11], [15], [25].

- Liu dng: Ngy dng 6 20, dng thuc sc [11], [15], [25].

- King k: T v h hn, tiu chy, n ung chm tiu [11].

15

1.3.10. Thin mn

a, Tn khoa hc v thuc: Radix Asparagi

b, B phn dng: c ca cy thin mn ng Asparagus cochinchinensis (Lour)

Merr. H Thin mn ng (Asparagaceae) [4], [7], [8], [16], [25].

c, Thnh phn ha hc

- Hot cht chnh l saponin steroid nhm furostan [4], [25], [53].

- Ngoi ra c carbohydrat, cc amino acid t do: aspargin, citrulin, serin, threonin,

prolin, glycin, alanin, valin, methionin, leucin, phenylalanine, thyroxin, acid

aspartic, acid glutamic, arginin, histidin, lysin [4].

d, Tc dng sinh hc

- Tc dng khng khun: c ch s pht trin ca cc chng vi khun Bacillus

anthracis, Streptococus hemolyticus A v B, B. diphtheria, Diplococcus

pneumonia, Staphylococcus aureus, Staphylococcus citreus, B. subtilis [25].

- Tc dng li tiu, li m, gim ho, h nhit [25].

- Tc dng chng vim, iu tr vim da [38], khng vim trong h thn kinh

trung ng [34].


- Tnh v: V ngt, ng, tnh i hn.

- Qui kinh: Ph, thn [7], [11], [16], [25].

- Cng nng, ch tr:

+ Thanh nhit ha m, dng m, thanh ph: tr ph m h, chc nng suy

nhc, ho lu ngy, ho c m kh khc, vim phi, ho g.

+ Dng v sinh tn: dng sau khi m dy, tn dch hao tn, ming kht.

+ Dng tm m: tr bnh tm huyt khng , hi hp, lon nhp tim, ngn hi,

v lc; ming li mn nht, m hi nhiu; an thn.

+ Nhun trng: cha i tin b to do ho kht [7], [8], [11], [15], [16], [25].

- Liu dng: Ngy dng 4 12 g [11], [16], [25].

- King k: Ngi t v h hn, a chy khng nn dng. [7], [25].

16

1.3.11. Bng i hi (Li i)

a, Tn khoa hc v thuc: Semen Scaphii

b, B phn dng: ht phi hay sy kh ca cy li i Scaphium lychnophorum

(Hance) Kost. H Trm (Sterculiaceae) [25].

c, Thnh phn ha hc

Ht c 2 phn: phn nhn cha cht bo, tinh bt, sterculin; phn v cha cht bo,

bassorin, cht nhy, tanin. Phn ng trong ht ch yu l galactose, pentose v

arabinose [25].

d, Tc dng v cng dng theo YHCT

- Tnh v: V ngt, nht, tnh hn [25].

- Cng nng, ch tr:

Thanh ph nhit, li yt hu, nhun trng, thng tin, gii c: Dng trong

trng hp c th nhit, to, st m , ho khan, au hng, nhc rng, i tin ra

mu, mn l. Ht li i l loi thuc b mt, sinh tn dch, tr cc chng au

rut, bnh v ng tiu ha. Dng nhiu v lin tc khng gy c hi [25].

1.3.12. Sinh a

a, Tn khoa hc v thuc: Radix Rehmanniae glutinosae

b, B phn dng: r c dng ti hoc phi, sy kh ca cy sinh a hong

Rehmannia glutinosa Gaertin. H Hoa mm si (Scrophulariaceae) [16], [24].

c, Thnh phn ha hc

- Iridoid glycosid: Catalpol, rehmaniosid A, B, C, D, ajugol, aucubin, melitosid,

rehmaglutin A, B, C, D [4], [24].

- Ionon glucosid: rehmaionosid A, B, C. Ngoi ra c monoterpen glucosid l

rehmapicrosid [24].

- Carbohydrat: D-glucose, D- fructose, sucrose, maninotriose, rafinose, stachyose,

D- manitol [24].

- Cc thnh phn khc: acid amin (t nht 15 acid amin t do), ester ca acid bo,

-sitosterol, daucosterol, acid palmitic, acid sucinic[24].

d, Tc dng sinh hc

17

- Tc dng an thn, li tiu, chng oxy ha [24].

- Tc dng khng khun, c ch nm ngoi da [16].

- Tc dng cm mu, cng tim, h ng huyt (do cht Catapol) [16].

- Tc dng cha lnh tn thng do i tho ng [28].


- Tnh v: V ngt, ng, tnh hn [16], [24].

- Qui kinh: Tm, can, thn, tiu trng [16], [24].

- Cng nng, ch tr:

B m, thanh nhit lng huyt, sinh tn dch, ch kht: Cha m h, pht nng

v chiu, kht nc nhiu, i tho ng, thiu mu, suy nhc c th, tng

chy mu, th huyt, chy mu cam, vim hng au, tn dch kh [11], [16],

[24].

- Liu dng: Ngy dng 12 40g [11], [16].

- King k: t h, bng y, i tin lng, dng h, nhiu m thp nhit [16].

18

CHNG 2. I TNG V PHNG PHP NGHIN CU

2.1. Nguyn liu: Dc liu v cao lng (cao 5/1) c bo ch t phng thuc

V Ngn, do Cng ty Dc phm Hoa Linh cung cp (Ph lc 2.1).

2.2. Phng tin

2.2.1. Thit b

- T sy: Memmert, Shellab.

- Cn phn tch Sartorius BP 221S (Germany).

- Cn k thut Precisa (Switzeland).

- My ly tm Hetich.

- My o quang ph UV-VIS U-1800 Hitachi.

- My siu m.

- n t ngoi.

- My nh Nikon.

- H thng thit b sc k lp mng hiu nng cao Linomat 5 (Camag - Switzeland)

(Hnh 2.1).

- Bp un cch thy, ng nghim, bnh nn, pipet, lng chut, dao, ko m,

(a) (b)

Hnh 2.1. H thng thit b sc k lp mng hiu nng cao Linomat 5.

(a) Thit b phun mu Linomat 5

(b) Thit b chp nh Camag

19

2.2.2. Ha cht

- Ha cht kim nghim ha hc t tiu chun phn tch, c mua ti Cng ty

TNHH Sela, A2, l 9, nh Cng, Hong Mai, H Ni.

- Ha cht th tc dng gim ho, long m: N-acetylcystein (bit dc Mucomyst

gi 200 mg), phenol dng bt tinh khit (Hng Sigma), nc mui sinh l v

cc ha cht khc t tiu chun phng th nghim trng i hc Y H Ni..

2.2.3. ng vt th nghim

Chut nht trng khe mnh c 2 ging, trng lng 18-22 g do Vin v sinh dch

t trung ng cung cp.

2.3. Ni dung nghin cu

- nh tnh thnh phn ha hc ca mt s dc liu v cao lng V Ngn.

+ nh tnh cc nhm cht trong cao lng v dc liu bng phn ng ha hc.

+ nh tnh bng sc k lp mng so snh gia cao lng v dc liu.

- Th tc dng gim ho, long m ca cao lng V Ngn.

2.4. Phng php nghin cu:

2.4.1. nh tnh thnh phn ha hc ca mt s dc liu v cao lng V Ngn.

Mu nghin cu: Cao lng V Ngn, cc dc liu: x can, kim ngn hoa, cam tho,

nc nc, ct cnh, thin mn, mch mn, huyn sm, sinh a.

- Chit xut: Xay th cc dc liu, chit cao lng v dc liu bng dung mi

thch hp nh tnh cc nhm cht hu c bng thuc th chung ca tng nhm

cht.

- nh tnh so snh cao lng v mt s dc liu bng sc k lp mng.

2.4.2. Nghin cu tc dng gim ho, long m ca cao lng V Ngn

2.4.2.1. Tc dng gim ho

Nguyn tc

Gy ho cho chut nht trng bng kh amoniac trong bnh thy tinh chuyn dng.

Xc nh thi gian tim tng (t) l thi gian tnh t khi th chut vo bnh n khi

chut xut hin cn ho u tin v m s cn ho trong mi mt pht cho n ht

20

pht th 10. So snh thi gian xut hin cn ho u tin v s lng cn ho trung

bnh trong 10 pht ca l th vi l chng v l i chiu.

2.4.2.2. Tc dng long m

Nguyn tc

Dch ph qun tit ra lm tng kh nng tr m ng h hp. Kh nng bi tit

dch ph qun c nh gi thng qua nng phenol thu c khi ra kh

qun bng NaHCO3 5 %.

nh lng phenol c tit ra theo dch tit ng h hp ca hng chut c

trong dch ra bng phng php o quang. o hp th quang (ABS) ca dch

ra bc sng 265 nm. hp th quang cng ln th kh nng long m cng

tt. So snh hp th quang trung bnh ca cc l th so vi l chng v l i

chiu [26].

X l s liu

Cc s liu nghin cu c x l thng k theo phng php t-test Student bng

phn mm Microsoft Excel 2010.

Kt qu th nghim c biu th bng tr s trung bnh cng/tr sai s chun

(= SEX ). S khc bit c ngha thng k khi P 0,05.

21

CHNG 3: THC NGHIM, KT QU V BN LUN

3.1. nh tnh cc nhm cht bng phn ng ha hc

3.1.1. nh tnh Flavonoid

* Chit xut: (Lm phn ng nh tnh Flavonoid v Saponin)

+ Ly 0,5g cao lng, thm 10 ml ethanol 90%. un cch thy v lc k cho tan, lc

nng qua giy lc, dch lc thu c dng tin hnh cc phn ng nh tnh.

+ Ly 5g bt dc liu, thm 20 ml ethanol 90%. un cch thy, lc nng qua giy

lc, dch lc thu c dng tin hnh cc phn ng nh tnh.

- Phn ng Cyanidin

Cho vo ng nghim nh 1 ml dung dch th. Thm mt t bt magnesi kim loi

(khong 10 mg). Nh tng git HCl c (3 5 git). yn mt vi pht.

Kt qu: Mu cao v cc dc liu kim ngn hoa, x can, nc nc, cam tho: dung

dch c mu thm (Phn ng dng tnh).

- Phn ng vi kim long

Cho vo ng nghim nh 1 ml dung dch th. Thm vi git dung dch NaOH 10%.

Kt qu: Mu cao v cc dc liu kim ngn hoa, x can, nc nc, cam tho: xut

hin ta vng (Phn ng dng tnh).

- Phn ng vi hi amoniac

Nh mt git dch chit ln giy lc. H kh ri ln ming l ammoniac c

c m nt. Nh mt git khc lm chng.

Kt qu: vt dch chit ca cao v cc dc liu kim ngn hoa, x can, nc nc, cam

tho: chuyn mu vng m hn (Phn ng dng tnh).

- Phn ng vi FeCl3

Cho vo ng nghim nh 1ml dung dch th. Thm vi git dung dch FeCl3 5%.

Kt qu: Mu cao v cc dc liu kim ngn hoa, x can, nc nc, cam tho: xut

hin ta mu xanh en (Phn ng dng tnh).

- Phn ng diazo ha

Cho 1 ml dung dch th vo ng nghim, kim ha bng dung dch NaOH 10%,

thm vi git thuc th diazo mi pha, lc u, un nng nh.

22

Kt qu: Mu cao v dc liu kim ngn hoa, x can, nc nc, cam tho: c kt ta

mu cam (Phn ng dng tnh).

Nhn xt: Cao lng v cc dc liu: kim ngn hoa, x can, nc nc, cam tho c

flavonoid.

3.1.2. nh tnh coumarin

- Phn ng m ng vng lacton

Cho vo 2 ng nghim, mi ng 1-2ml dch chit:

+ ng 1: Thm 0,5 ml dung dch NaOH 10%.

+ ng 2: yn.

un c 2 ng nghim n si, ngui, quan st. Thm vo c 2 ng nghim, mi

ng 2ml nc ct, lc u, quan st. Thm vo ng 1 vi git HCl, quan st.

Kt qu: Tt c cc mu: ng 1: ta c, ng 2: trong sut; sau khi thm nc ct

v HCl: khng c hin tng g (Phn ng m tnh).

- Phn ng diazo ha

Cho vo ng nghim 1ml dch chit, 2ml dung dch NaOH 10%. un cch thy 10

pht, ngui. Thm vi git thuc th diazo mi pha.

Kt qu: Khng c kt ta (Phn ng m tnh).

- Quan st hunh quang di nh sng t ngoi khi tc dng vi dung dch kim

Nh 1 git dch chit ln t giy lc, nh tip 1 git dung dch NaOH 10%, sy nh

cho n kh. Che 1 na din tch bng mt mu kim loi, soi n t ngoi trong vi

pht, b ming kim loi ra, tip tc chiu sng b phn che sng.

Kt qu: Khng thy s khc bit v mu sc gia 2 phn (Phn ng m tnh).

Nhn xt: Cao lng v cc dc liu khng c coumarin.

3.1.3. nh tnh Saponin

- Quan st hin tng to bt: Ly 0,1g cao lng hoc 1g bt dc liu vo ng

nghim to, thm 10 ml nc, un nng nh, lc mnh trong 2 3 pht, yn.

Kt qu: Mu cao lng v cc dc liu ct cnh, kim ngn hoa, mch mn, thin

mn, cam tho: ct bt cao 2-3 cm, bn vng hn 15 pht (Phn ng dng tnh).

23

Chit xut: Ly 2g bt dc liu, thm 10 ml nc, un si 10 pht, lc nng, dch

lc thu c tin hnh cc phn ng.

- Phn ng Salkowski

+ Ly 0,5 g cao lng, c to cn, thm t t 2 ml dung dch H2SO4 c.

+ Ly 2 g bt dc liu, thm 10 ml nc, un si 10 pht, lc nng, c dch lc

n cn, thm 2 ml dung dch H2SO4 c.

Kt qu: Mu cao lng v cc dc liu ct cnh, kim ngn hoa, mch mn, thin

mn, cam tho: xut hin mu m (Phn ng dng tnh).

- Phn ng Liebermann Burchard

Ly 1-2 ml cao lng hoc 2 ml dch lc, c to cn, thm vo 1 ml anhydrid acetic,

lc u cho tan ht, nghing ng nghim 450 , thm t t tng git H2SO4 c.

Kt qu: Mu cao lng v cc dc liu cam tho, ct cnh, mch mn, thin mn,

kim ngn hoa: c vng mu ta mt phn cch (Phn ng dng tnh).

Nhn xt: Cao lng v cc dc liu: cam tho, kim ngn hoa, ct cnh, mch mn,

thin mn c saponin.

3.1.4. nh tnh Alcaloid

Chit xut: Ly 0,5 g cao lng hoc 5g dc liu, cho vo bnh nn dung tch 100

ml. Thm 15ml dung dch acid sulfuric 1N. un n si. ngui. Kim ha dch

lc bng dung dch ammoniac 6N n pH=9-10. Chit Alcaloid base bng

chloroform (3 ln x 5ml). Gp cc dch chit chloroform, loi nc bng natrisulfat

khan.

Tin hnh : Ly mt phn dch chit chloroform, lc vi acid sulfuric 1N 2 ln, mi

ln 5ml. Gp cc dch chit. Chia u vo cc ng nghim nh, mi ng 1ml. Nh

vo tng ng nghim 2 3 git cc thuc th sau:

+ ng 1: Thuc th Mayer.

+ ng 2: Thuc th Bouchardat.

+ ng 3: Thuc th Dragendorff.

Kt qu: C 3 ng ca tt c cc mu: khng xut hin ta (Phn ng m tnh).

Nhn xt: Cao lng v cc dc liu khng c alcaloid.

24

3.1.5. nh tnh Glycosid tim

Chit xut: Ly 5ml cao lng hoc 4 g bt dc liu, cho vo bnh nn 100 ml,

thm 30 ml ethanol 25%, ngm 24h. Gn ly dch chit, thm 3ml dd ch acetat 30%

cho n d (ta ht), khuy u, lc qua giy lc gp np (th li dch lc n khi

khng cn ta vi ch acetat). Lc dch lc vi CHCl3 (2 ln x 5ml), chit ly lp

CHCl3, cho vo cc ng nghim nh, c ly cn.

- Phn ng Liebermann Burchard

Thm vo 1 ml anhydrid acetic, lc k, nghing 45o, cho t t theo thnh ng 0,5

ml H2SO4 c.

Kt qu: Khng xut hin vng mu tm v cht lng pha di mu hng, pha

trn mu xanh l (Phn ng m tnh).

- Phn ng Baljet

Thm vo 1ml ethanol 90%, lc u cho tan ht, nh tng git thuc th Baljet mi

pha (So snh vi ng chng khng c cn).

Kt qu: Gia 2 ng khng c s khc bit (Phn ng m tnh).

- Phn ng Keller Kiliani

Thm vo 0,5 ml ethanol 90%, lc u cho tan, thm vi git dd FeCl3 5% trong

acid acetic, lc u, nghing ng 450, thm t t theo thnh ng 0,5 ml H2SO4 c.

Kt qu: Khng xut hin vng mu tm v lp cht lng pha trn mu xanh l

(Phn ng m tnh).

Nhn xt: Cao lng v cc dc liu khng c glycosid tim.

* Chit xut: Ly 0,5g cao lng ha tan trong 20ml nc ct nng, hoc ly 2g

dc liu, thm 20 ml nc, un si cch thy 15 pht, lc ly dch, ngui.

Dch lc lm cc phn ng nh tnh tanin, acid hu c, polysaccharid, ng

kh, acid amin.

3.1.6. nh tnh tanin

Ly 3 ng nghim, cho vo mi ng 2 ml dch lc:

+ ng 1: Thm 2 git FeCl3 5% .

25

Kt qu: Mu cao lng v dc liu: nc nc, kim ngn hoa, ct cnh: xut hin ta

mu xanh en (Phn ng dng tnh).

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


bng trng (Phn ng dng tnh).

+ ng 3: Thm 5 git gelatin 1%.


bng trng (Phn ng dng tnh).

Nhn xt: Cao lng v dc liu: nc nc, kim ngn hoa, ct cnh c tanin.

3.1.7. nh tnh acid hu c

Ly 2ml dch lc cho vo ng nghim to. Thm mt t Na2CO3, lc nh.

Kt qu: Tt c cc mu: khng c bt kh (Phn ng m tnh).

Nhn xt: Cao lng v cc dc liu khng c acid hu c.

3.1.8. nh tnh polysaccharid

Cho 2ml dch lc vo ng nghim, thm 2 git thuc th Lugol.

Kt qu: Tt c cc mu: xut hin ta mu xanh en (Phn ng dng tnh).

Nhn xt: Cao lng v cc dc liu c polysaccharid.

3.1.9. nh tnh ng kh

Cho vo ng nghim 2 ml dch lc, thm 1 ml dd Fehling A (TT) v 1ml dd Fehling

B (TT), lc k, un cch thy.

Kt qu: mu cao lng v dc liu sinh a, thin mn, huyn sm, cam tho, ct

cnh: xut hin ta gch (Phn ng dng tnh).

Nhn xt: Cao lng v dc liu: sinh a, thin mn, huyn sm, cam tho, ct

cnh c ng kh.

3.1.10. nh tnh acid amin

Ly 2ml dch lc, thm 3 git thuc th Ninhydrin, un cch thy 2-3 pht.

Kt qu: Mu cao lng v dc liu: huyn sm, sinh a, thin mn: dung dch c

mu tm sm (Phn ng dng tnh).

Nhn xt: Cao lng v dc liu: huyn sm, sinh a, thin mn c acid amin.

26

Kt qu nh tnh cc nhm cht trong cao lng v dc liu bng phn ng ha hc

c tm tt Bng 3.1.

Bng 3.1. Kt qu nh tnh ha hc cc nhm cht trong cao lng v dc liu.

TT Nhm cht Phn ng Kt qu

Cao XC CT KNH NN MM TM CC HS S

1 Flavonoid - Cyanidin +++ +++ +++ +++ +++ - - - - -

- Vi FeCl3 5% +++ +++ ++ +++ +++ - - + - -

-Vi NaOH

10%

+++ +++ +++ +++ ++ - - - - -

- Vi hi NH3 +++ +++ +++ +++ ++ - - - - -

- Diazo ha +++ +++ +++ +++ ++ - - - - -

2 Saponin - To bt +++ - +++ +++ - +++ +++ ++ - -

- Salkowski +++ - +++ +++ - +++ +++ ++ - -

-Libermann-

Burchard

+++ - ++ ++ - ++ ++ ++ - -

3 Tanin -Vi FeCl3 5% +++ +++ ++ +++ +++ - - + - -

-Vi ch acetat +++ - - ++ ++ - - ++ - -

-Vi gelatin1% ++ - - + + - - ++ - -

4 Glycosid tim - - - - - - - - - -

5 Alcaloid TT chung ca

acaloid

- - - - - - - - - -

6 Coumarin - - - - - - - - - -

7 Acid amin -Vi TT

Ninhydrin

+++ - - - - - +++ - ++ ++

8 Polysaccharid -Vi TT Lugol +++ +++ ++ ++ + + + ++ + +

9 ng kh - TT FehlingA,

Fehling B

+++ - + - - ++ ++ ++ ++ ++

10 Acid hu c -Vi Na2CO3 - - - - - - - - - -

Ch thch:

(-): phn ng m tnh

(+): phn ng dng tnh

(++): phn ng dng tnh r

(+++): phn ng dng tnh rt r

27

Nhn xt:

- Cao lng c flavonoid, saponin, tanin, acid amin, polysaccharid, ng kh.

- X can c flavonoid, polysaccharid.

- Cam tho c flavonoid, saponin, polysaccharid, ng kh.

- Kim ngn hoa c flavonoid, saponin, polysaccharid, tanin.

- Nc nc c flavonoid, polysaccharid, tanin.

- Mch mn c saponin, polysaccharid, ng kh.

- Thin mn c saponin, acid amin, polysaccharid, ng kh.

- Ct cnh c saponin, tanin, polysaccharid, ng kh.

- Huyn sm c acid amin, polysaccharid, ng kh.

- Sinh a c acid amin, polysaccharid, ng kh.

3.2. nh tnh so snh cao v v thuc bng sc k lp mng:

3.2.1. Kim ngn hoa

Chun b

- Bn mng: Silica gel 60 F254, 4x10 cm, hot ha 1100 C trong 60 pht.

- Dung mi khai trin: Kho st trn cc h:

(I) Chloroform methanol (9:1)

(II) Toluen ethylacetat acid formic (5:4:1)

(III) Ethylacetat acid formic nc (8:1:1)

(IV) Chloroform methanol acid formic (7:2:1)

(V) Toluen - Chloroform- aceton- acid formic (8:5:7:0,2)

- Thuc th hin mu: dung dch KOH 5%/EtOH 96%.

Chit xut

- Dung dch th: Ly 5g cao lng, thm 30ml methanol, un trn cch thy 15 pht,

lc. Chit nh vy thm mt ln na. Gp dch chit methanol, c trn cch thy

n cn. Khuy k cn vi n butanol 2 ln, mi ln 10 ml. Gp dch chit

butanol, c trn cch thy n cn. Ha tan cn trong 1ml methanol c dung

dch chm sc k.

28

- Dung dch i chiu: Ly 2 g kim ngn hoa tn nh vo bnh nn, y np,

thm 20ml methanol, un cch thy trong 20 pht, lc, c dch lc n cn. Lm

tng t dung dch th bt u t khuy k cn.sc k.

Tin hnh

Chm ring bit ln bn mng 4 l dung dch th v 6 l dung dch i chiu. Sau

khi trin khai sc k, ly bn mng ra, kh nhit phng, quan st vt nh sng

thng v di n UV bc sng 254 nm, 366 nm, phun thuc th hin mu, sy

1200 C cho n khi hin r vt. Chp nh bng thit b chp nh Camag, x l kt

qu bng phn mm Wincats, Videoscan.

Kt qu: Trong cch h dung mi trn, h (V) cho kt qu tch vt v hin mu tt

nht bc sng 366 nm (Bng 3.2, Hnh 3.1).

Bng 3.2. Kt qu phn tch sc k ca cao v kim

ngn hoa bc sng 366 nm.

STT Rf Mu sc Cao KNH

1 0,08 Xanh ++ +++

2 0,25 Xanh ++ +++

3 0,28 Xanh +++ +++

4 0,32 Xanh +++ +++

5 0,38 Xanh nht +

6 0,40 Tm nht ++

7 0,45 Xanh nht +

8 0,45 Tm nht ++

9 0,52 Xanh +++

10 0,52 Tm nht ++

11 0,59 Xanh ++ +++

12 0,66 Xanh ++

Hnh 3.1. Sc k cao -

KNH bc sng 366 nm.

Ch thch: (+): C vt (++): vt m (+++): vt rt m

29

Nhn xt: Sc k bc sng 366nm cho thy: C 5 vt tng ng gia cao

v kim ngn hoa.

3.2.2. Cam tho

Chun b

- Bn mng: Silica gel 60 F254, 4x10 cm, hot ha 1100 C trong 60 pht.

- Dung mi khai trin:

(I) Ethyl acetat acid acetic bng acid formic nc (15:1:1:2)

(II) Toluen ethylacetat acid formic (5:6:1,5)

(III) Chloroform methanol acid formic (7:2:1)

- Hin mu: Dung dch acid sulfuric 10% / EtOH 96%.

Chit xut

- Dung dch th: Ly 5 g cao lng, thm 20ml nc, un nng nh v lc k cho tan.

ngui, chit bng 20ml n butanol, gn ly lp dch chit n butanol v c thu

hi dung mi n cn. Ha tan cn vo 1ml methanol c dung dch chm sc k.

- Dung dch i chiu: Ly 1g bt cam tho, thm 40ml ether, un hi lu cch thy

trong 1 gi, b dch chit ether. Thm 30ml methanol vo phn b cam tho, un

hi lu cch thy trong 1 gi, lc, bc hi dch lc n kh, ha tan cn trong 1ml

methanol c dung dch chm sc k.

Tin hnh

Chm ring bit ln bn mng 4 l mi dung dch th v 6 l dung dch i chiu.

Sau khi trin khai sc k, ly bn mng ra, kh nhit phng, quan st di

n t ngoi bc sng 254 nm, 366 nm, phun thuc th hin mu, sy 1200 C

n khi r vt. Chp nh bng thit b chp nh Camag, x l kt qu bng phn

mm Wincats, Videoscan.

Kt qu: Trong cc h dung mi, h (I) cho kt qu tch vt v hin mu tt nht

bc sng 254 nm (Bng 3.3, Hnh 3.4).

30

Bng 3.3. Kt qu phn tch sc k ca cao v cam

tho bc sng 254 nm.

STT Rf Mu sc Cao CT

1 0,14 en ++

2 0,20 en + ++

3 0,28 en ++ ++

4 0,35 en +

5 0,38 en ++

6 0,48 en +++ +++

7 0,54 en +

8 0,63 en +

9 0,80 en +

10 0,95 en ++

11 0,97 en +++

Hnh 3.2. Sc k

cao-CT bc sng

254 nm.


Nhn xt: Kt qu phn tch sc k bc sng 254 nm cho thy: c 3 vt tng

ng gia cao v cam tho.

3.2.3. X can

Chun b

- Bn mng Silica gel 60 F254 , 4x10 cm, hot ha 1100 C trong 60 pht.

- Dung mi trin khai: Kho st cc h:

(I) Ethylacetat acid formic nc (8:1:1)

(II) Chloroform methanol acid formic (7:2:1)

(II) Toluen ethylacetat aceton acid formic (5:2:2:1)

(IV) Toluen ethylacetat acid formic (5:4:1)

(V) Toluen ethylacetat acid formic (5:6:1,5)

- Hin mu: Dung dch KOH 5%/ EtOH 96%.

Chit xut

31

- Dung dch th: Ly 5g cao lng, thm 60ml nc, un nng nh, lc k cho tan,

ngui, lc. Lc vi ethyl acetat 2 ln, mi ln 20ml. Gp dch chit ethyl

acetat, c trn cch thy n cn. Ha tan cn trong 1ml ethanol 96% c dung

dch chm sc k.

- Dung dch i chiu: Ly 2 g x can tn nh, thm 40ml nc, un si nh

20 pht, ngui, lc. Lm tng t nh vi dung dch th, bt u t Lc vi

ethyl acetat n sc k.

Tin hnh

Chm ring bit ln bn mng 4 l dung dch th v 6 l dung dch i chiu.

Sau khi trin khai sc k, ly bn mng ra, kh nhit phng, quan st vt

nh sng thng v di n UV bc sng 254 nm, 366 nm; phun thuc th hin

mu, sy 1200 C, quan st di nh sng thng. Chp nh bng thit b chp nh

Camag, x l kt qu bng phn mm Wincats, Videoscan.

Kt qu: Trong cc h dung mi trn, h (V) cho kt qu tch vt v hin mu tt

nht sau khi hin mu bng thuc th KOH 5%/EtOH 96% (Bng 3.4, Hnh 3.3).

Bng 3.4. Kt qu phn tch sc k ca cao v x can

sau khi hin mu.


STT Rf Mu sc Cao XC

1 0,10 Nu nht ++

2 0,26 Vng ++

3 0,28 Nu +++

4 0,33 Nu +++

5 0,48 Tm nht ++

6 0,55 Tm nht ++ ++

7 0,64 Nu nht ++ ++

8 0,70 Vng nht +

Hnh 3.3. Sc k cao-

XC sau khi hin mu.

32

Nhn xt: Sc k sau khi phun thuc th cho thy: c 2 vt tng ng gia

cao v x can.

3.2.4. Nc nc

Chun b

- Bn mng: Silica gel 60 F254 , 4x10 cm, hot ha 1100 C trong 60 pht.


(I) Chloroform methanol (9:1)

(II) Toluen ethylacetat acid formic (5:5:1)

(III) Ethylacetat acid formic nc (8:1:1)

(IV) Chloroform methanol acid formic (7:2:1)

(V) Chloroform - Ethylacetat acid formic (2:2:1)

(VI) Ethylacetat acid formic acid acetic nc (10:1:1:1)

- Thuc th hin mu: dung dch KOH 5%/EtOH 96%.

Chit xut

- Dung dch th: Ly 5g cao lng, thm 60ml methanol, un trn cch thy 15 pht,

lc. Chit nh vy thm mt ln na. Gp dch chit methanol, c trn cch thy n

cn. Khuy k cn vi n butanol 2 ln, mi ln 20 ml. Gp dch chit butanol, c trn

cch thy n cn. Ha tan cn trong 1ml methanol c dung dch chm sc k.

- Dung dch i chiu: Ly 4g nc nc tn nh, thm 20 ml EtOH 70%, ngm lnh,

lc, c dch chit n cn, ha tan cn vi 1ml methanol c dung dch chm sc k.

Tin hnh

Chm ring bit ln bn mng 4l dung dch th v 6l dung dch i chiu. Sau khi

trin khai sc k, ly bn mng ra, kh nhit phng, quan st vt nh sng

thng v di n UV bc sng 254 nm, 366 nm phun thuc th hin mu, sy

1200 C cho n khi hin r vt. Chp nh bng thit b chp nh Camag, x l kt qu

bng phn mm Wincats, Videoscan.

Kt qu: Trong cch h dung mi trn, h (V) cho kt qu tch vt v hin mu tt

nht bc sng 254 nm (Bng 3.5, Hnh 3.4).

33

Bng 3.5. Kt qu phn tch sc k ca cao v nc nc

bc sng 254 nm.

STT Rf Mu sc Cao NN

1 0,13 en + ++

2 0,18 en +

3 0,25 en ++ ++

4 0,30 en ++

5 0,53 en ++

6 0,60 en +++

7 0,67 en ++

8 0,69 en +

9 0,75 en +++ +++


Hnh 3.4. Sc k cao-

NN bc sng 254 nm.

Nhn xt: Kt qu phn tch sc k bc sng 254 nm cho thy: c 3 vt tng

ng gia cao v nc nc.

3.2.5. Huyn sm

Chun b

- Bn mng Silica gel 60 F254 , 4x10 cm, hot ha 1100 C trong 60 pht.

- Dung mi trin khai: Kho st cc h:

(I) Toluen ethylacetat acid formic (5:6:1)

(II) Chloroform methanol nc (65:35:10), lc k, ly lp di.

(III) Chloroform methanol - acid formic (9:1: 0,2)

- Hin mu: Thuc th vanilin sulfuric (100 ml Vanillin 1% /EtOH 96%+ 2 ml

dung dch H2SO4 c).

Chit xut

- Dung dch th: Ly 5g cao dc liu vo bnh nn, thm 60ml methanol, y

np kn, un cch thy 15 pht, lc. Chit 2 ln nh trn. Gp dch chit

methanol, c trn cch thy n cn. Khuy k cn vi n butanol 2 ln, mi

34

ln 20ml, khuy trong 5 pht. Gp dch chit butanol, c trn cch thy n cn.

Ho tan cn trong 1ml ethanol 96% c dung dch chm sc k.

- Dung dch i chiu: Ly 1g huyn sm ct nh, thm 20ml butanol, un si

hi lu trong 1 gi, ngui, lc. C dch lc trn cch thy n cn. Ha cn

trong 1ml ethanol 96% c dung dch chm sc k.

Tin hnh

Chm ring bit ln bn mng 6l mi dung dch th v dung dch i chiu. Sau

khi trin khai sc k, ly bn mng ra, kh nhit phng, quan st di n

UV; phun thuc th hin mu, sy 1200C cho n khi hin r vt. Chp nh bng

thit b chp nh Camag, x l kt qu bng phn mm Wincats, Videoscan.

Kt qu: Trong cc h dung mi, h (III) tch vt v hin mu tt nht sau khi

hin mu bng thuc th vanilin sulfuric (Bng 3.6, Hnh 3.5).

Bng 3.6. Kt qu phn tch sc k ca cao v

huyn sm sau khi hin mu.

STT Rf Mu sc Cao HS

1 0,05 en +++ +++

2 0,09 en +++ ++

3 0,14 Xanh en +++

4 0,18 Xanh l ++ +

5 0,27 Tm ++

6 0,30 Tm +++

7 0,36 Nu ++ +++

8 0,42 Xanh l +++ +++

9 0,49 Hng nht +

10 0,56 Hng nht +

11 0,68 Hng ++ ++

12 0,72 Tm +++

13 0,95 Tm ++

Hnh 3.5. Sc k

cao-HS sau khi hin

mu.

35

Nhn xt: Kt qu phn tch sc k sau khi phun thuc th cho thy: c 6 vt

tng ng gia cao v huyn sm.

3.2.6. Ct cnh, Thin mn, Mch mn

Chun b

- Bn mng: Silica gel 60 F254 , 6x10 cm, hot ha 1100 C trong 60 pht.


(I) Chloroform - methanol (10:1)

(II) Chloroform - methanol - acid formic (19:1)

(III) Toluen ethylacetat ( 9:1)

(IV) Toluen ethylacetat acid formic (5:5:1)

- Hin mu: Thuc th vanilin sulfuric (100 ml Vanillin 1% /EtOH 96% + 2 ml

dung dch H2SO4 c).

Chit xut:

- Dung dch th: Ly 5 g cao lng, thm 10ml methanol, un hi lu cch thy 20

pht, lc ly dch chit, thy phn dch chit bng HCl 5% khong 1 gi, chit

bng 5ml chloroform x 2 ln. C dch chit chloroform n cn, ha tan cn trong

1ml ethanol 96% thu c dung dch chm sc k.

- Dung dch i chiu: Ly 4 g dc liu mi loi, ct nh, lm tng t nh vi

dung dch th, bt u t thm 10ml methanol sc k.

- Cch tin hnh: Chm ring bit ln bn mng 6l dung dch th v 5l dung

dch i chiu. Sau khi trin khai sc k, ly bn mng ra, kh nhit phng,

quan st di n t ngoi bc sng 254, 366 nm; phun thuc th hin mu, sy

1200 C cho n khi hin r vt. Chp nh bng thit b chp nh Camag, x l kt

qu bng phn mm Wincats, Videoscan.

Kt qu: Trong cc h dung mi trn, h (IV) cho kt qu tch vt v hin mu tt

nht sau khi hin mu bng thuc th vanilin sulfuric (Bng 3.7, Hnh 3.6).

36

Hnh 3.6. Sc k CC- cao- TM- MM sau khi hin mu

Bng 3.7. Kt qu phn tch sc k ca CC-cao-TM-MM sau khi hin mu

STT Rf Mu sc Ct cnh Cao Thin mn Mch mn

1 0,08 en +

2 0,15 n +

3 0,22 Tm ++

4 0,25 Tm ++ + +++ ++

5 0,30 Xanh dng ++

6 0,32 Tm nht +

7 0,40 Xanh l +++ ++ +++

8 0,51 Tm ++ ++ ++ ++

9 0,56 Tm nht +

10 0,60 Vng ++

11 0,73 Tm ++ ++ +++

12 0,76 Hng m +++

13 0,79 Tm + +++

14 0,85 Tm ++ ++ ++ ++

15 0,95 Tm ++ ++ ++ +++


37

Nhn xt: Kt qu phn tch sc k cho thy:

- Gia cao v ct cnh c 6 vt tng ng.

- Gia cao v thin mn c 6 vt tng ng.

- Gia cao v mch mn c 4 vt tng ng.

3.3. Nghin cu tc dng sinh hc:

Mu nghin cu: Cao lng V Ngn (cao 5/1), pha long vi nc ct theo t l

thch hp cc mc liu khc nhau.

3.3.1. Tc dng gim ho:

Tin hnh:

40 chut nht trng c chia ngu nhin thnh 4 l, mi l 10 con:

- L 1 (chng): ung nc ct, th tch 0,2ml/10g.

- L 2 (i chiu): ung codein phosphat liu 20 mg/kg.

- L 3: ung cao lng V Ngn liu 63,36 g dc liu/kg.

- L 4: ung cao lng V Ngn liu 190,08g dc liu/kg.

Chut c ung thuc th lin tc trong 3 ngy vo cc bui sng. Ngy th

3 sau khi ung thuc th 1 gi, tin hnh gy ho cho c 4 l chut bng kh amoniac

liu 0,5ml/bnh thy tinh chuyn dng. Xc nh thi gian tim tng (t) l thi gian

tnh t khi th chut vo bnh n khi chut xut hin cn ho u tin v tng s

cn ho trung bnh trong 10 pht, so snh mc tng (t) v mc gim s cn ho

ca cc l th so vi l chng v l i chiu.

Kt qu:

Trn m hnh gy ho cho chut nht trng bng kh amoniac, thi gian tim tng

xut hin cn ho v tng s cn ho c trnh by trong bng 3.8, 3.8; hnh 3.7; 3.8.

* Khc bit so vi nhm chng vi P 0,05

** Khc bit so vi nhm chng vi P 0,01

*** Khc bit so vi nhm chng vi P 0,001

38

Bng 3.8. nh hng ca cao lng V Ngn ln thi gian tim tng xut hin cn ho.

L chut N

Thi gian tim

tng (giy)

( SEX )

Mc tng

thi gian tim

tng so vi

chng (%)

P

L 1: Chng 9 118,00 22,05

L 2: Codein

phosphat 20mg/kg 9 226,00* 44,43 91,53 P 2-1 < 0,05

L 3: V Ngn liu

63,36 g dc liu/kg 10 240,40 55,19 103,39

P3-1 > 0,05

P 3-2 > 0,05

L 4: V Ngn liu

190,08 g dc liu/kg 9 268,89* 59,45 127,11

P4-1 < 0,05

P4-2 > 0,05

P4-3 > 0,05

Hnh 3.7. Biu biu th thi gian tim tng xut hin cn ho chut.

Nhn xt: Cao lng V Ngn mc liu 190,08 g dc liu/kg c tc dng ko di

thi gian tim tng xut hin cn ho trn chut nht trng so vi l chng vi mc

tng 127,11%, s khc bit c ngha thng k (P 0,05).

39

Bng 3.9. nh hng ca cao lng V Ngn ln tng s cn ho.

L chut N

Tng s cn ho

trong 10 pht

( SEX )

Mc gim s

cn ho so vi l

chng (%)

P

L 1: Chng 9 30,33 6,30

L 2: Codein

phosphat 20mg/kg 9 9,33* 3,75 69,24 P 2-1 < 0,05

L 3: V Ngn liu

63,36g dc liu/kg 10 14,30* 3,34 52,85 P3-1 < 0,05

P 3-2 >0,05

L 4: V Ngn liu

190,08 g dc liu/kg 9 7,00* 3,39 76,92

P4-1 = 0,05

P4-2 > 0,05

P4-3 > 0,05

30.33

9.33 *

14.30 *

7.00 *

0

5

10

15

20

25

30

35

40

L 1 L 2 L 3 L 4

S l

n h

o

L th nghim

Hnh 3.8. Biu biu th tng s cn ho chut.

Nhn xt: Cao lng V Ngn c 2 mc liu u c tc dng gim s cn ho

chut so vi l chng. Mc gim s cn ho cao nht l chut ung V Ngn liu

190,08 g dc liu/kg l 76,92%. S gim s cn ho 2 mc liu c ngha thng

k (P 0,05).

3.3.2. Tc dng long m:

40 chut nht trng c chia ngu nhin thnh 4 l, mi l 10 con:

40

- L 1 (chng): ung nc ct vi th tch 20ml/kg/ngy.

- L 2 (i chiu): N-acetylcystein 120mg/kg (Gi 200 mg pha va 30ml, ung

0,2 ml/10g).



Cho chut ung thuc th vi liu nh trn trong 3 ngy. Ngy th 3 (ngy

cui cng) sau ung thuc 1 gi, tim mng bng cho tt c cc chut dung dch

phenol 0,25% vi liu 25ml/kg. Sau 30 pht tim nhc li mt liu nh vy.

Ch 30 pht sau, git chut bng gy ngt hi chloroform, bc l kh qun

v lun vo mt kim t. Ra kh qun bng cch dng bm tim ly 0,5ml dung

dch NaHCO3 5% bm vo kh qun, ht nh nhng ri chuyn vo trong mt ng

nghim c nh du t trc.

Lm nh vy 3 ln i vi mi con, tp trung dch ra ca cng mt con

trong mt ng nghim (th tch dch ra thu c ca cc chut bng nhau). Ly tm

dch ra ly dch trong. o hp th quang (ABS) ca dch ra kh qun bc

sng 265 nm.

Kt qu: Kt qu mt quang c trnh by trong bng 3.10, hnh 3.8.

Bng 3.10. nh hng ca cao lng V Ngn ln mt quang.

L chut N Mt quang

( SEX )

Mc tng mt quang

(%) so vi chng P

L 1: Chng 10 0,406 0,036

L 2: N-acetyl cystein

liu 120 mg/kg 10 0,528* 0,023 30,05 % P 2-1 < 0,05

L 3: V Ngn liu

63,36 g dc liu/kg 10 0,447 0,035 10,10 %

P3-1 > 0,05

P 3-2 > 0,05

L 4: V Ngn liu

190,08g dc liu/kg 10 0,664*** 0,015 64,28 %

P4-1< 0,001

P4-2

41

Hnh 3.9. Biu biu th mt quang bc sng 265 nm.

Nhn xt:

Mc hp th quang bc sng 265 nm ca dch ra ph qun l

chut ung cao lng V Ngn liu 190,08g dc liu/kg tng 64,28 % so vi l

chng. S tng mt quang c ngha thng k (P < 0,001).

42

3.4. Bn lun

3.4.1. Thnh phn ha hc

Kt qu nh tnh bng phn ng ha hc cho thy trong cao lng c cc thnh

phn c bn tng t nh thnh phn ca cc v thuc, bao gm: flavonoid,

saponin, tanin, acid amin, polysaccharid, ng kh.

Nh vy, hai nhm cht chnh trong cao lng l flavonoid v saponin. Do ,

chng ti tin hnh chit chn lc theo tng nhm cht i vi cc dc liu ch

c thnh phn chnh l mt trong hai nhm cht ny, nhm flavonoid: x can, nc

nc; nhm saponin: ct cnh, mch mn, thin mn. Phng php ny tn dung mi

v nhiu cng on nhng c u im l loi c nhiu tp cht v cc nhm cht

khc, v vy s cho kt qu chnh xc hn. i vi cc dc liu c c hai thnh

phn trn l cam tho, kim ngn hoa, chng ti tin hnh chit khng chn lc:

dng dung mi methanol, n-butanol chit nhiu thnh phn c trong dc liu v

cao lng.

Trn sc k ca cc dc liu, ngoi cc vt tng ng vi cao lng cn

xut hin nhiu vt khc, c th: ct cnh c thm 3 vt, thin mn c thm 4 vt,

mch mn c thm 3 vt, huyn sm c thm 4 vt, nc nc c thm 3 vt, x can

c thm 1 vt, cam tho c thm 4 vt, kim ngn hoa c thm 3 vt do trong qu

trnh bo ch, mt s thnh phn trong dc liu b gim hoc mt i. Ngoi ra

mt s vt xut hin gia cc dc liu kh ging nhau (ct cnh, thin mn, mch

mn) c th do cu trc gn ging nhau gia cc thnh phn trong cng nhm cht.

Mt s vt trong cao c mu m hn hoc nht hn so vi cc vt tng ng ca

dc liu c th do s bin i cc cht trong qu trnh bo ch cao, chit xut dc

liu, lng mu chm.

Qua qu trnh nghin cu v trin khai sc k lp mng xc nh du vn tay

ca cc v thuc, chng ti nhn thy rng, c kt qu tt th cn phi thc hin

cc bc kho st v khi lng mu nghin cu cn dng, h dung mi khai trin,

lng mu chm trn bn mng, v tnh lp li ca kt qu. T cc kt qu trn,

chng ti xut mt s ch tiu trong tiu chun k thut ca cao lng nh sau.

43

Bng 3.8. Mt s ch tiu xut trong tiu chun k thut ca cao lng V Ngn.

Ch tiu nh tnh Yu cu

Phn

ng

ha

hc

Flavonoid

Phn ng Cyanidin +

Vi FeCl3 +

Vi kim (NaOH 10%, NH3) +

Phn ng diazo ha +

Saponin

To bt +

Phn ng Salkowski +

Libermann- Burchard +

Sc

k

lp

mng

Cao- kim

ngn hoa

H dung mi: Toluen- CHCl3-

aceton- HCOOH (8:5:7:0,2)

Quan st bc sng 366nm

C 5 vt tng ng:

Rf = 0,08; 0,25; 0,28; 0,32; 0,59

(mu xanh)

Cao-

cam tho

H dung mi: EtOAc a.acetic

bng-HCOOH- H2O (15:1:1:2)

Quan st bc sng 254 nm

C 3 vt tng ng:

Rf= 0,20; 0,28; 0,48 (mu en)

Cao -

x can

H dung mi: Toluen-EtOAc-

HCOOH (5:6: 1,5)

Quan st: phun TT KOH/EtOH

C 2 vt tng ng:

Rf= 0,55 (tm nht)

Rf= 0,64 (nu nht)

Cao-

nc nc

H dung mi: CHCl3 - EtOAc

- HCOOH (2:2:1)

Quan st bc sng 254 nm

C 3 vt tng ng:

Rf= 0,13; 0,25; 0,75 (mu en)

Cao- huyn

sm

H dung mi: CHCl3 MeOH-

HCOOH (9:1: 0,2)

Quan st: TT vanilin sulfuric

C 6 vt tng ng:

Rf= 0,05; 0,09; 0,18; 0,36; 0,42;

0,68 (en, en, xanh l, nu, xanh

l, hng)

Cao- Saponin

(ct cnh,

thin mn,

mch mn)

H dung mi: Toluen-EtOA-

HCOOH (5:5:1)

Quan st: phun TT vanilin

sulfuric

Cao-CC,TM: c 6 vt tng ng:

Rf= 0,25; 0,40; 0,51; 0,73; 0,85;

0,95 (tm, xanh l, tm, tm, tm,

tm, tm)

Cao-MM: c 4 vt tng ng:

Rf= 0,25; 0,51; 0,85; 0,95 (u tm)

44

3.4.2. Tc dng sinh hc

Nguyn tc tnh liu

Liu bnh thng: 1 bi thuc 88g/ngi (50 kg)/ngy, tng ng vi liu 1,76

g/kg/ngy i vi ngi. Theo tng ng sinh hc, liu ca chut nht trng gp

12 ln ngi [26], do vy liu bnh thng ca chut l 21,12 g/kg/ngy, t lm

c s cho tnh liu th tc dng sinh hc. Qua kho st mt s mc liu khc

nhau, chn ra mc liu 63,36 v 190,08 g dc liu/kg th tc dng sinh hc.

Tc dng gim ho

Codein c tc dng gim ho do c ch trc tip trung tm ho, nhng lm kh

v tng qunh ca dch tit ph qun. Dng codein trong trng hp ho khan gy

kh chu, mt ng v trong cc chng au nh v va. y l thuc ho dng kh

ph bin nn trong nghin cu, codein c chn lm thuc i chng trn m hnh

gy ho bng kh amoniac chut nht trng.

Cc kt qu nghin cu cho thy cao lng V Ngn liu 190,08g dc liu/kg

c tc dng lm tng thi gian tim tng xut hin cn ho v lm gim s cn ho

ca chut nht trng trn m hnh gy ho bng amoniac.

Tc dng long m

N- acetylcystein: Dng lm thuc tiu cht nhy trong bnh nhy nht, cc

bnh l h hp c m nhy qunh nh trong vim ph qun cp hoc mn, lm

thuc gii c khi dng qu liu paracetamol. N- acetylcystein l mt trong nhng

thuc long m s dng ph bin nht trn lm sng nn c chn lm thuc i

chng trong m hnh thc nghim.

Cc kt qu nghin cu cho thy, cao lng V Ngn mc liu 190,08 g

dc liu/kg c tc dng long m thng qua lm tng nng phenol trong

dch ra kh ph qun chut nht trng.

Nh vy, cao lng V Ngn liu 190,08 g dc liu/kg c tc dng gim ho

v long m trn chut nht trng, tc dng ny ph hp vi tc dng ca cc v

thuc trong phng thuc.

45

KT LUN V KIN NGH

1. KT LUN

V thnh phn ha hc

nh tnh bng phn ng ha hc

- Cao lng V Ngn c cc thnh phn: flavonoid, saponin, tanin, acid amin,

polysaccharid, ng kh.

- X can c flavonoid, polysaccharid.

- Cam tho c flavonoid, saponin, polysaccharid, ng kh.

- Kim ngn c flavonoid, saponin, polysaccharid, tanin.

- Nc nc c flavonoid, polysaccharid, tanin.

- Mch mn c saponin, polysaccharid, ng kh.

- Thin mn c saponin, acid amin, polysaccharid, ng kh.

- Ct cnh c saponin, tanin, polysaccharid, ng kh.

- Huyn sm c acid amin, polysaccharid, ng kh.

- Sinh a c acid amin, polysaccharid, ng kh.

nh tnh so snh cao v dc liu bng sc k lp mng

- Sc k ca cao lng v kim ngn hoa bc sng 366 nm c 5 vt tng

ng: Rf = 0,08; 0,25; 0,28; 0,32; 0,59 , cc vt c mu xanh.

- Sc k ca cao lng v cam tho bc sng 254 nm c 3 vt tng ng:

Rf= 0,20; 0,28; 0,48, cc vt c mu en.

- Sc k ca cao lng v x can sau khi phun thuc th c 2 vt tng ng:

Rf= 0,55 (tm nht), Rf= 0,64 (nu nht).

- Sc k ca cao lng v nc nc bc sng 254 nm c 3 vt tng ng:

Rf= 0,13; 0,25; 0,75, cc vt c mu en.

- Sc k ca cao lng v huyn sm sau khi phun thuc th 6 vt tng ng:

Rf= 0,05 (en); 0,09 (en) ; 0,18 (xanh l); 0,36 (nu); 0,42 (xanh l); 0,68

(hng).

- Sc k ca cao lng v mch mn sau khi phun thuc th c 4 vt tng

ng: Rf= 0,25; 0,51; 0,85; 0,95 ( u mu tm).

46

- Sc k ca cao lng v ct cnh, thin mn sau khi phun thuc th c 5 vt

tng ng: Rf= 0,25 (tm); Rf= 0,40 (xanh l); Rf= 0,51 (tm); Rf= 0,73 (tm);

Rf= 0,85 (tm); Rf= 0,95 (tm).

Tc dng gim ho, long m

- Tc dng gim ho:

Cao lng V Ngn liu 190,08 g dc liu/kg/ngy c tc dng gim ho trn m

hnh gy ho bng kh amoniac chut nht trng.

- Tc dng long m:

Cao lng V Ngn liu 190,08 g dc liu/kg/ngy c tc dng long m trn

chut nht trng.

2. KIN NGH

T cc kt qu nghin cu bc u v cao lng V Ngn, chng ti ngh tip

tc nghin cu:

- Nghin cu bo ch dng thuc thnh phm tin li cho s dng.

- Nghin cu cc tc dng khc lin quan n iu tr vim hng: chng vim,

khng khun.

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53. Zhu G.L., Hao Q., Li R.T., Li H.Z. (2014), Steroidal saponins from the roots

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12(3), pp 213217.

PH LC

PH LC 1: S BO CH CAO LNG V NGN

Dc liu s ch

un si

Dch chit (2 ln)

Lc

C chn khng

Lng, c

Cao lng

ng gi cao

Kim nghim bn

thnh phm

Nc sinh hot

Acid benzoic/ cn 96o

PH LC 2

Ph lc 2.1 Thi gian tim tng v s lng cn ho ca chut nht trng

L 1 (Chng) L 2 (i chiu)

N=9 Thi gian tim

tng (giy) Tng s cn ho N=9

Thi gian tim tng (giy)

Tng s cn ho

1 23 52 1 530 6

2 127 15 2 120 13

3 107 67 3 184 6

4 105 28 4 247 4

5 70 35 5 69 38

6 130 13 6 305 4

7 120 27 7 225 2

8 270 9 8 160 8

9 110 27 9 194 3

TB 118 30,33 TB 226,00 9,33

SE 22,05 6,30 SE 44,43 3,75

P2-1 0,045 0,011

L 3 (liu thp) L 4 (liu cao)

N=10 Thi gian tim

tng (giy) Tng s cn ho N=9

Thi gian tim tng (giy)

Tng s cn ho

1 162 21 1 241 3 2 565 6 2 470 1 3 135 17 3 116 32 4 90 34 4 269 9 5 170 6 5 530 1 6 120 18 6 51 12 7 475 1 7 140 2 8 470 1 8 135 1 9 45 21 9 468 2 10 172 18 TB 240,40 14,30 TB 268,89 7 SE 59,19 3,34 SE 59,45 3,39 P3-1 0,081 0,033 P4-1 0,030 0,005 P3-2 0,851 0,335 P4-2 0,571 0,650

P4-3 0,739 0,144

Ph lc 2.2. Mt quang ca dch ra kh qun chut nht trng

N=10 L 1 (chng) L 2 (i chiu) L 3 (liu thp) L 4 (liu cao)

1 0,334 0,404 0,395 0,630

2 0,410 0,555 0,609 0,672

3 0,531 0,521 0,337 0,749

4 0,470 0,506 0,492 0,611

5 0,358 0,489 0,517 0,667

6 0,503 0,565 0,583 0,614

7 0,548 0,468 0,374 0.640

8 0,321 0,570 0,463 0,731

9 0,176 0,677 0,256 0,654

10 0,406 0,528 0,447 0,667

TB 0,406 0, 528 0,447 0,664

SE 0,036 0,023 0,035 0,015

P so vi l 1 0,010 0,416 3,14x10-6

P so vi l 2 0,672 9,46x10-5

P so vi l 3 1,91x10-5

1. Ba chnh.doc2. Ba ph.doc3. Li cm n..doc4. Muc luc VN.doc5. DANH MC k hiu..doc6. DANH MC CC BNG..doc7. DANH MC CC HNH V.doc8. KL V Ngn mi.doc9.TLTK V Ngn.doc10. ph lc.doc

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