B GIO DC V O TO

TRNG I HC S PHM TPHCM

KHA LUN TT NGHIP

C NHN HA HC
CHUYN NGNH: HA HU C

XY DNG TIU CHUN KIM NGHIM

NGUYN LIU V CAO CHIT
T DU TM (Morus alba L.), H DU TM (Moraceae)
V VIN CH (Polygala japonica Houtt),
H VIN CH (Polygalaceae)

Gio vin hng dn khoa hc: ThS. DNG TH MNG NGC

Sinh vin thc hin: PHM TH KIM KHUYN

TP.HCM Nm 2013
 MC LC
DANH MC CC K HIU, CH VIT ................................................................. ii

DANH MC CC BNG ......................................................................................... iii

DANH MC CC HNH V V S ..................................................................iv

Phn 1:M U.............................................................................................................

Phn 2: TNG QUAN ....................................................................................................

1. GII THIU V CC DC LIU ...................................................................1

1.1. Cy Du tm (Morus alba L.) [1,4,5,10,11,12,15,16,17,19] ..........................1

1.2. Cy vin ch (Polygala japonica Houtt) [5,9,10,11,14,18,20] .......................8

2. C S L THUYT V PHNG PHP CHIT: [4,8] ................................13

2.1. Chit lng - lng ...........................................................................................13

2.2. Chit siu m ................................................................................................14

2.3. Chit Soxhlet ................................................................................................14

3. PHNG PHP SC K LP MNG [2,3,8,9] ..............................................14

4. NH TNH BNG PHN NG HA HC [1,4,8] .......................................15

5. PH T NGOI KH KIN (PH UV - VIS) [1,2,9] ....................................15

5.1. Khi nim......................................................................................................15

5.2. c im .......................................................................................................15

Phn 3: VT LIU V PHNG PHP NGHIN CU ...........................................

1. VT LIU NGHIN CU .................................................................................18

1.1. i tng nghin cu ...................................................................................18

1.2. Dung mi, ha cht, trang thit b ................................................................18

2. PHNG PHP NGHIN CU .......................................................................19

2.1. Xy dng tiu chun kim nghim nguyn liu ...........................................19

 2.2. Xy dng tiu chun kim nghim cao ........................................................27

Phn 4: KT QU V BN LUN ..............................................................................

1. XY DNG TIU CHUN KIM NGHIM NGUYN LIU ......................36

1.2. Kho st tinh khit ca nguyn liu.........................................................36

1.3. nh tnh cc hp cht ha thc vt trong nguyn liu ................................37

1.4. nh lng cc hp cht ha thc vt trong nguyn liu .............................42

2. XY DNG TIU CHUN KIM NGHIM CAO CHIT............................43

2.1. Kho st tinh khit ca cao chit .............................................................43

2.2. nh tnh cc hp cht ha thc vt trong cao .............................................44

2.3. nh lng cc hp cht ha thc vt trong cao. .........................................50

Phn 5: KT LUN V KIN NGH ...........................................................................

1. KT LUN .........................................................................................................52

2. KIN NGH ........................................................................................................54

2.1. Xy dng tiu chun kim nghim nguyn liu ...........................................54

2.2. Xy dng tiu chun kim nghim cao ........................................................56

TI LIU THAM KHO.............................................................................................v

 LI CM N

Trong sut qu trnh thc hin kha lun tt nghip ny, c s hng dn tn
tnh ca qu thy c v cc anh ch, cc bn cng thc hin kha lun, ti Trung
tm Sm v Dc Liu TP.HCM to iu kin gip em hon thnh tt kha lun
ny. Em xin gi li cm n chn thnh n:
o Gia nh ng h tinh thn, vt cht v to iu kin cho em trong nhng nm
hc va qua v trong sut qu trnh lm kha lun.
o Cc thy c Khoa Ha hc trng H S Phm TP.HCM tn ty dy d em
trong sut kha hc ti trng.
o PGS.TS. Trn Cng Lun, Gim c Trung tm Sm v Dc liu to iu
kin cho em hon thnh kha lun.
o C ThS. Dng Th Mng Ngc v ch Nguyn Th Ngc an l ngi trc tip
hng dn truyn t kin thc, kinh nghim v to iu kin tt nht em thc hin
tt kha lun ny.
o C Nguyn Th Hoa, ch Lm Th Bch Tho, anh Bi Th Vinh v qu thy c,
anh ch trong trung tm Sm v Dc Liu ht lng gip v to iu kin tt nht
trong thi gian em thc hin kha lun ca mnh.
o Cc bn sinh vin trng H Khoa Hc T Nhin, H Nng Lm TP.HCM v
cc bn sinh vin cng lp gip , chia s kinh nghim hc tp v vui bun trong
sut thi gian cng em lm kha lun ti Trung tm.

Sinh vin thc hin

PHM TH KIM KHUYN
 DANH MC CC K HIU, CH VIT

NL: Nguyn liu

dd: Dung dch
TT: Thuc th
SKLM: Sc k lp mng
MeOH: Methanol
n-BuOH: n- butanol
EtOAc: Ethyl acetat
HCl: Acid chlohydric
H 2 SO 4 : Acid sulfuric
CHCl 3 : Chloroform
CH 3 COOH: Acid acetic
(CH 3 CO) 2 O: Anhydric acetic
NaOH: Natri hydroxyd
FeCl 3 : St (III) chlorid
AlCl 3 : Nhm chlorid
H 2 O: Nc ct
R f : Ratio of flow
UV: Ultraviolet
LD 50 : Lethal Dose 50
DVN: Dc in Vit Nam

ii
 DANH MC CC BNG
Bng 1. Cc ch phm t Du tm................................................................................... 7
Bng 2. Cc ch phm t Vin ch ................................................................................. 13
Bng 3. Dung mi- ha cht- trang thit b.................................................................... 18
Bng 4. nh tnh hp cht flavonoid trong nguyn liu Du tm ................................ 21
Bng 5. nh tnh hp cht ha thc vt trong nguyn liu Vin ch ........................... 21
Bng 6. nh tnh hp cht flavonoid trong cao Du tm ............................................. 28
Bng 7. nh tnh hp cht ha thc vt trong cao Vin ch ......................................... 29
Bng 8. m ca nguyn liu ..................................................................................... 36
Bng 9. tro ton phn ca nguyn liu ..................................................................... 36
Bng 10. tro khng tan trong acid ca nguyn liu ................................................. 37
Bng 11. Hm lng flavonoid tnh theo rutin trong nguyn liu Du tm .................. 42
Bng 12. Hm lng saponin ton phn trong nguyn liu Vin ch ............................ 42
Bng 13. m ca cao chit........................................................................................ 43
Bng 14. tro ton phn ca cao chit ....................................................................... 43
Bng 15. Hm lng flavonoid tnh theo rutin trong cao Du tm ............................... 50
Bng 16. Hm lng saponin ton phn trong cao Vin ch ......................................... 50

iii
 DANH MC CC HNH V V S
Hnh 1. Cy Du tm (Morus alba L.) ............................................................................. 1
Hnh 2. R Vin ch (Radix Polygala japonica Houtt) .................................................... 8
Hnh 3. Phn ng ha hc ca nguyn liu Du tm ..................................................... 38
Hnh 4. SKLM kim tra s hin din ca cht rutin trong nguyn liu Du tm .......... 39
Hnh 5. Phn ng ca nguyn liu Vin ch .................................................................. 40
Hnh 7.Phn ng ha hc ca cao Du tm ................................................................... 44
Hnh 8. SKLM kim tra s hin din ca nguyn liu trong cao Du tm .................... 45
Hnh 9. SKLM kim tra s hin din ca cht chun rutin trong cao Du tm ............. 46
Hnh 10. Phn ng ha hc ca caoVin ch ................................................................. 47
Hnh 11. SKLM kim tra s hin din ca nguyn liu trong cao Vin ch .................. 49

S 1. S xc nh hm lng flavonoid tng trong Du tm ......................... 24

S 2. nh lng saponin ton phn trong nguyn liu Vin ch ........................ 26
S 3. S xc nh hm lng flavonoid tng trong Du tm ......................... 32
S 4. nh lng saponin ton phn trong cao Vin ch theo phng php cn
Namba ....................................................................................................................... 34

iv
Phn 1:
M U
 Vit Nam l quc gia nm trn bn o ng Dng. Hin nay, cc vn v
mi trng khng kh Vit Nam ang c quan tm rt nhiu t cc thnh ph ln
n cc vng nng thn thnh th. Khng kh trong kh quyn m chng ta ang ht
th l khng kh m, bao gm hn hp cc cht dng kh c th tch gn nh khng
i v c cha mt lng hi nc nht nh ty thuc vo nhit v p sut kh
quyn. V vy, chng ta thng i mt vi cc bnh v h hp nh ho, nght xoang
v suynHin nay, iu tr cc loi bnh ny, cc dc phm ty y ang c s
dng rng ri, tuy nhin, bn cnh tc dng tr liu l nhng tc dng ph ng k
nh hng n th trng chung ca ngi s dng.

Trc tnh hnh T chc Y t Th gii (WHO) khuyn co cc quc gia

nn pht trin y hc c truyn trong vic chm sc sc kho cng ng, do , vic
s dng ngun dc liu trong thin nhin lm thuc phng v cha bnh ang l xu
hng hin nay ca y hc Vit Nam v y hc th gii.

Trong kho tng y hc dn gian c nhiu cy thuc v bi thuc, tuy nhin phn
ln cc cy thuc v bi thuc u ch mi c s dng theo kinh nghim dn
gian m cha c nghin cu, nh gi mt cch khoa hc. t rt lu dn gian ta
s dng Du tm trong bi thuc cha phong thp, cha cm mo, ho, hng au,
nhc u, mt , chy nc mt, pht ban, huyt p cao, m hi trm...Cn Vin ch
th c xem nh mt loi cy tr vim phi, ho c nhiu m, bch i, l, lo u,
mt ng...

Trc thc trng ny, Trung tm Sm v Dc liu TP.H Ch Minh ang

nghin cu sn phm iu tr bnh h hp t cc dc liu. Trong qu trnh nghin
cu mt sn phm iu tr bnh th vic nh gi cht lng ca nguyn liu v cc
bn thnh phm, m bo hiu qu s dng v tnh an ton ca nguyn liu v cc
bn thnh phm trong sn xut, qun l, phn phi v s dng th cng tc kim
nghim cn phi c quan tm nhiu hn.
 Chnh v vy, tip tc hon thnh h s nghin cu cc sn phm iu tr
bnh h hp t cc dc liu, ti Xy dng tiu chun kim nghim nguyn
liu v cao chit t Du tm (Morus alba L.), h Du tm (Moraceae) v Vin ch
(Polygala japonica Houtt), h Vin ch (Polygalaceae) c thc hin vi cc mc
tiu sau:

Xy dng tiu chun kim nghim nguyn liu Du tm (Morus alba L.) v
Vin ch (Polygala japonica Houtt).

Xy dng tiu chun kim nghim cao chit t Du tm (Morus alba L.) v
Vin ch (Polygala japonica Houtt).

.
 Phn 2
TNG QUAN
1. GII THIU V CC DC LIU
1.1. Cy Du tm (Morus alba L.) [1,4,5,10,11,12,15,16,17,19]

Hnh 1. Cy Du tm (Morus alba L.)

- Tn ng ngha: Morus acidosa Griff
- Tn khc: Du, du ta, tang, du cang (HMng), my mn (Th), my b
(Ty), co mn (Thi), nn phong (Dao).
- Tn nc ngoi: Mulberry- tree
- H: Du tm (Moraceae)
1.1.1. M t thc vt
Cy nh, cao 6m hay hn trng thi hoang di, thng gim xung 1,5-2m
cy trng. Cnh mm, lc non c lng, sau nhn v c mu xm trng. L mc so le,
hnh bu dc, hnh tim hoc hnh trng rng, c mi nhn u, phin mng, mm,
di 3-7cm, rng 2,5- 4m, mp c rng ca nh u, i khi chia 3-5 thy, 3 gn
gc, hai mt c mu lc sng, cung di mnh, hi c lng, l km hnh di nhn.
Hoa n tnh, khng c cnh hoa; cm hoa c l ui sc di 1,5-2cm, hoa
c c 4 l di t, hi c lng, nh 4, xp i din vi l i v di gp i, ch nh
mnh, bao phn gn hnh cu; cm hoa ci l bng ngn hnh trng hoc gn hnh cu
di 1cm, hoa c 4 l i, bu c mt non.
 Qu b bao bc trong cc l i mng nc t hp thnh mt qu phc, khi
chn mu hay hng, sau en.
Ma hoa qu: thng 5-7.
1.1.2. B phn dng
L (Tang dip- Folium Mori), dng loi l bnh t (l cho tm n), loi b l
vng a, tp cht ri phi hay sy nh.
Cnh (Tang chi- Ramulus Mori), thu hi quanh nm, phi kh.
Qu (Tang thm- Fructus Mori), thu hi khi qu chn.
V r (Tang bch b- Cortex Mori), ly r ngm di t, phi hay sy kh.
Tm gi du (Tang k sinh- Ramulus Loranthi) l cy mc k sinh trn cy, thu
hi quanh nm.
T trng b nga trn cy du (Tang phiu tiu- Ootheca Mantidis), dng t
cha n, phi chn v sy kh.
1.1.3. Thnh phn ha hc
L du cha:
Cc thnh phn bay hi nh tinh du, gm cc phn on trung tnh, acid,
phenol, carbonyl, base. Phn on trung tnh cha isobutanol, alcol isoamylic,
isoamyl acetat v acetophenol. Phn on acid cha acid acetic, acid propionic, acid
butyric, acid isobutyric, acid isovaleric, acid caproic, acid isocaproic, acid lactic.
Phn on phenol cha o-, m-, p-cresol, guaiacol, eugenol, salicylat methyl. Phn
on carbonyl cha benzaldehyd v phenylacetaldehyd.
Cc thnh phn khng bay hi gm nhiu hp cht thuc nhiu nhm ha hc
khc nhau nh: protein, carbohydrat, flavonoid, coumarin, vitamin v mt s thnh
phn khc.
Cnh cha cc flavonoid, ngoi ra cn c tetrahydroxybenzophenon, maclurin.
V r cha: mulberin, cyclomulberin, mulberochromen, cc kuwanon...ngoi
ra cn c - tocopherol, umbeliferon, morin...
Qu cha lipid, acid hu c, alcol v tinh du, cyanidin.
B nga sng trn cy du cha albumin, cht bo, ng, cht x, nc, Ca,
Fe. Trng cha glycoprotein, lipoprotein. Ti trng cha acid amin ton phn.
1.1.4. Tc dng dc l
Cao nc v cao kim ca l c tc dng c ch cc vi khun gram dng v
cc men.
nghin cu v chng minh l du c tc dng gy chn tnh nh kiu
meprobamat.
L du c tc dng h huyt p yu, tc dng ny b o ngc bi atropin.
ng thi n c tc dng gin mch. Ch phm an thn Passerynum gm l du, lc
tin, vng nem, l sen, tho thuyt minh, ht t hng, ht keo giu, c sm i hnh,
th hin c tnh thp, gim c trng thi hng phn chut nht b kch thch
bi cafein, gy h huyt p, tng tn s v bin h hp ca th, c khuynh hng
lm gim nhp tim v sc co bp ca si c tim. p dng iu tr trn lm sng,
Passerynum c tc dng gy h thn nhit ca bnh nhn, khng lm thay i huyt
p, khng gy trng thi bun ng hoc ng g, ch lm bnh nhn ng d dng v
ngon gic. Thuc c tc dng an thn cho bnh nhn vi t l 16/25 ngi, so vi
placebo t l 6/15 ngi, chng t yu t tm l c vai tr quan trng trong mt ng.
V trong ca r du c tc dng gy h huyt p, tc dng ny hon ton b
i khng bi atropin. c ch tim ch c lp, gin mch ngoi bin trn tai th c lp,
co mch ni tng trn h mch chi sau ca ch, kch thch c t cung v rut th c
lp v gy co c thng bng ch, tc dng sau ny c tng cng bi physotigmin.
Ngoi ra cn c tc dng gy chn tnh trn chut nht trng.
1.1.5. Tnh v, cng nng
V r du c v ngt, hi ng, tnh mt, c tc dng thanh ph nhit, li
thng, gim ho, tr m, h suyn.
L du c v ng, ngt, tnh lnh, c tc dng tn phong, thanh nhit, lng
huyt, sng mt.
Cnh du c v ng nht, tnh bnh, c tc dng tr phong thp, li cc khp,
thng kinh lc, tiu vim.
Qu du c v ngt, chua, tnh mt, c tc dng b gan thn, b huyt tr
phong.
Tm gi du c v ng, tnh bnh c tc dng mnh gn ct, li huyt mch,
an thai, xung sa.
 Tang phiu tiu c v ngt, mn, tnh bnh, c tc dng ch thn, c tinh.
1.1.6. Cng dng
V r du cha ph nhit, ho c m, hen, khi huyt, tr con ho g, ph
thng, bng chng to, tiu tin khng thng, bng huyt, st, cao huyt p. Ngy
dng 4-12g, c khi n 20-40g dng thuc sc hay thuc bt.
L du cha cm mo, phong nhit, st nng, ho, vim hng, au rng, nhc
u, mt , chy nc mt, u lo, pht ban, cao huyt p, mt ng. Ngy dng 4-
12g, dng thuc sc.
Cnh du cha phong thp au nhc cc u xng, cc kh, sng l, chn
tay co qup. Ngy dng 6-12g, c khi n 40-60g, di dng thuc sc.
Qu du cha i tho ng, trng nhc (lao hch), mt m, tai, thiu mu,
au xng khp, km ng, ru tc bc sm, to bn. Ung lu khe ngi, ng
ngon, thnh tai sng mt, tr lu. Qu du vt ly nc c thnh cao mm, ngy ung
12-20g. Siro qu chn bi cha au hng, lot ming, l li.
Tang k sinh cha au lng, au mnh, chn tay t bi, ng thai, sau t sa.
Liu dng 12-20g dng thuc sc.
Tang phiu tiu cha i i nhiu ln, di tinh, lit dng, bch i, tr con i
dm. Liu dng 6-12g.
Su du cha bnh tr con b au mt, nhiu d, nhiu nc mt. C con su
hp chn vi mt ong hoc nng n.
1.1.7. Cc bi thuc c du tm
Cha khe mt b mng tht che lp trng (Hi Thng Ln ng):
L du, c mc u bng nhau, cho vo ni t, nc vo un, ri cho vo
mt t vi bt lu nm, bt ming ni li nu thm vi do, bc xung, xng 2-3
ln.
Cha chng cng sn khng c m (Hi Thng Ln ng):
V r du phi trong rm, tn nh, nu thnh cao ri ha vi nc bi.
Cha tr con au hng, ho khan, bch hu:
L du 20g, tm vi (bch cng tm) 10g, bc h 5g. Sc ung.
Cha sng phi, st, tr ho g, m suyn:
 V r du (phn non di t, b lp v ngoi, ly lp v trng trong, tm
mt sao qua), mch mn, ngu tt, mi v 10g, xuyn tm lin 5g. Sc ung.
Cha ho lu nm:
V r du 10g, v r chanh 10g. Sc ung trong ngy.
Cha ho, vim hng:
V trng r du 10g, bch b (b li sao vng) 10g, mch mn 10g, v qut
5g, x can 5g, cam tho dy 5g. Lm dng thuc phin, mi phin 3g, ngy ngm 4-
5 ln, mi ln 1 phin. Hoc dng cao lng, mi ln ung 1 tha cafe.
Cha phong nhit, st, ho nhiu, tc ngc, khc m vng:
L du 12g, kim ngn 12g, bc h 10g, ca hoa 10g, l ngi cu 10g, x can
8g. Sc ung ngy 1 thang trong 5 ngy lin.
Cha vim ph qun mn tnh:
V r du 16g, mch mn 16g, rau m 16g, bch b 10g, trn b 6g, bn h ch
6g. Sc ung ngy mt thang, ung trong thi gian di.
Cha ho ra mu:
V r du 12g, thin mn 12g, ca hoa 12g, c nh ni 12g, mch mn 12g,
qu dnh dnh 12g, sinh a 12g, trc bch dip 12g. Sc ngy ung 1 thang.
Cha nn ra mu:
L du cui ma, sao vng. Sc ung, mi ngy 12-16g.
Cha ho g:
V r du 12g, mch mn 12g, bch b 10g, rau sam 10g, hng chanh 10g.
Sc ung ngy 1 thang, ung lin tc 15-30 ngy. Hoc ch thnh siro, ngy ung 3
ln, mi ln 1 tha canh, tr em gim liu.
Cha phong thp, tay chn co qup, au nhc:
Cnh du sao 20g, huyt d 12g. Sc ung.
Cha vim a khp mn tnh dng thp:
Cnh du 16g, th phc linh 12g, m sao 16g, ngu tt 16g, sinh a 16g,
d 16g, en sao 16g, r l lt 12g. Sc ung ngy 1 thang.

Cha au dy thn kinh ta:

 Cnh du 12g, th phc linh 12g, thin nin kin 12g, ngu tt 12g, sinh a
12g, c gai leo 10g, en sao 10g, l lt 10g. Sc ung ngy mt thang, ung thi
gian di.
Cha mn nht lu ngy khng lin ming:
L du sao vng tn nh, rc vo mn ra sch.
Cha rng tc:
V r du gi dp, ngm nc ri un si na gi, lc ngui, gi u.
Cha ng thai, au bng:
Tang k sinh 60g, cao ban long (hoc cao xng) nng thm 20g, ngi dip
20g, nc 600ml. Sc cn 100ml, chia 2 ln ung trong ngy.
Cha ng thai, b tiu tin:
Tang phiu tiu, nng vng tn nh. Ngy ung 2 ln, mi ln 5g.
1.1.8. Cc ch phm t Du tm
Kem c tr nm Melanil
- Cng dng
Kem gip lm sng v loi b cc
m nm trn da, gy ra bi nh nng,
lo ha, mang thai, di truyn
- Thnh phn:
Du tm, l hi, cam tho, nm
men

Sa ra mt Sokruy: - Cng dng: c chit xut

Hng sm, du tm, tr xanh, nm
thng hong vng, qu lu v qu
lu cng lp vng go non gip loi
b cc thnh phn oxy ha c hi cho
da.
- Cch dng: S dng vo bui
sng, bui ti hng ngy.
Thc phm chc nng Morussan
Thnh phn: Cao kh du tm tiu
chun.
Liu lng v cch dng:
Dng 1-2 vin/ln, ngy 2 ln.
ung vo trc ba n khong 30 pht.

Bng 1. Cc ch phm t Du tm
1.2. Cy Vin ch (Polygala japonica Houtt) [5,9,10,11,14,18,20]

Hnh 2. R Vin ch (Radix Polygala japonica Houtt)

Tn khc:
Kh vin ch (Trn Nam Bn Tho), Yu nhiu, Cc quyn (Nh Nh), Nga
qun ch thng, Ch nhc, Ch thng, Vin ch nhc, Chch vin ch, Kh yu, D
lng, A ch tho, Tnh tm trng (Trung Quc Dc Hc i T in).
Tn khoa hc: Polygala japonica Houtt
H Vin ch (Polygalaceae).
1.2.1. M t thc vt
Cy Vin ch Polygala japonica Houtt, cn gi l nam Vin ch, hay Tiu
tho. Cy tho lu nm, cao 10-20cm, phn cnh t gc. Cnh mc ta rng, hi c
lng mn. L mc sole, rt a dng: l gc hnh elip, l pha trn hnh mc di 30mm,
rng 3-5mm, mp thng cun xung di, gn chy men theo mp l, gn ph r,
cung di 0,5mm.
Cm hoa mc k l hoc u cnh thnh chm mnh, c 1-3 hoa mu trng,
u nhum tm; l bc rt nh, sm rng, i 3 rng ngoi rt nh, 2 rng trong rng
hn, c lng mi, trng 5 cnh, 3 cnh ri, 3 cnh bn hn lin thnh cnh c, mo
lng mu lam hoc tm, nh nhn; bu thun nhn.
 Qu nang, c cnh bn; ht hnh trng, c lng, o ht chia 3 thy.
Ma hoa qu: thng 11-12.
1.2.2. Thu hoch
Vo ma xun, thu o ln, b thn tn, r con v t, phi cho v hi nhn,
rt b li g, phi kh l c.
1.2.3. B phn dng
R kh (Radix Polygalae). Th ng to, tht dy, b ht li g l th tt.
1.2.4. Thnh phn ha hc
Theo Trung dc i t in I, 1997, r Vin ch cha saponin triterpen, nha,
du bo v polygalitol.
- Tenuigenin A, B (Chou T Q v cng s, Am Pharm Assoc Sci Ed, 1947).
- Tenuifolin (Pelletier S W v cng s, Tetrahydron, 1971).
- Onjisaponin A, B, C, D, E, F, G (Sakuma S v cng s, Pharm Bull, 1981).
- Tenuifoliside A, B, C, D v a-D- (3-O-Sinapoyl) Fructofuranosyl--D- (6-O-
Sinapoyl) Glucopyranoside (Ikeya Y v cng s, Chem Pharm Bull, 1991).
- Tenuifoliose A, B, C, D, E, F (Miyase Y v cng s Chem Pharm Bull 1991).
1.2.5. Tc dng dc l
Tc dng gim ho: Trn m hnh thc nghim gy ho cho chut nht trng
bng cch phun xng amoniac, liu 0,75g/kg Vin ch cho ung di dng cao, c tc
dng gim ho r rt.
Tc dng li m: Th nghim trn th, Vin ch c tc dng lm tng dch tit
kh ph qun.
Tc dng gim au: Trn m hnh gy au biu hin bng cc phn ng vn
xon mnh khi tim trong mng bng dung dch acid acetic, Vin ch liu ung
0,8g/kg c tc dng gim au r rt chut nht trng.
Tc dng trn thi gian ng: Vin ch c tc dng hip ng, lm ko di thi
gian ng do thuc ng barbituric chut nht trng.
Tc dng trn thn kinh trung ng: Vin ch c tc dng c ch c mc
h thn kinh trung ng, nhng khng thy c tc dng i khng vi liu lng gy
co git do cafein gy nn chut nht trng.
 Tc dng trn t cung: Th tc dng ca cao lng Vin ch trn t cung th,
mo v chut cng trng invitro v insitu, thy thuc c tc dng kch thch co bp c
t cung c con vt c thai v khng c thai.
Tc dng khng khun: Cao mm Vin ch c tc dng khng khun, c ch
s pht trin ca cc vi khun Staphylococcus aureus, Bacillus subtilis, Streptococcus
hemolyticus, Diplococcus pneumoniae.
Tc dng tn huyt: Dch chit 5% ca r v b phn trn mt t ca cy ra
hoa c tc dng tn huyt.
Lu : Saponin Vin ch kch thch d dy gy bun nn v th khng nn
dng i vi nhng bnh nhn vim lot d dy.
c tnh:
Liu c LD 50 ca v r Vin ch cho chut nht ung l 10,03 1,98g/kg.
Liu LD 50 ton r l 16,95 2,01g/kg m r b li g i dng n 75g/kg th gy t
vong.
1.2.6. Tnh v, cng nng
Vin ch c v hc, ng, the, tnh hi m, vo 2 kinh tm v thn, c tc dng
an thn, ch tr, khu m, ch khi, ch tinh, hot huyt, tn , tiu thng, gii c.
1.2.7. Cng dng
Vin ch c dng cha ho, nhiu m, vim ph qun, hay qun, gim tr
nh, lit dng, yu sc, mng tinh, b cho nam gii v ngi gi, thuc lm sng
mt, thnh tai do tc dng trn thn. Cn cha au tc ngc, lao, ng km, suy nhc
thn kinh, c mng. Ngy 6-12g dng thuc sc hoc 2-5g cao lng, bt thuc hoc
cn thuc.
Dng ngoi, Vin ch phi kh tn bt, tm nc, p cha n ng tn
thng, mn nht, l lot, sng v au v, rn c cn.

1.2.8. Cc bi thuc c Vin ch

Cha ho c m:
 Vin ch 8g, ct cnh 6g, cam tho 6g, sc chia 3 ln ung trong ngy. Trng
hp ngi gi ho c m lu nm, m kt gy tc ngc, kh th, dng Vin ch 8g,
mch mn 12g, sc ung dn tng ngm, ngy 1 thang.
Cha thn kinh suy nhc, hay qun, n n, kinh s, hong ht, km n, t ng:
Vin ch, ng sm, bch trut, lin nhc, long nhn, to nhn (sao en), mch
mn, mi v 10g, sc ung. Hoc Vin ch, tm sen, ht mung (sao), mch mn,
nhn ht to (sao en), huyn sm, dnh dnh, mi v 12g, sc ung.
Cha tr st cao sinh co git:
Vin ch, sinh a, cu ng, thin trc hong (bt phn ng t cy na),
mi v 8-10g, sc ung.
Tr tm thng lu ngy:
Vin ch (b li), Xng b (thi nh) u 40g, tn bt. Mi ln dng 12g,
nc 1 chn, sc cn 7 phn, b b, ung m.
Tr ung th, pht bi, nht c:
Vin ch (b li), gi nt. Ru 1 chn, sc chung, ly b p vt thng.
Tr hng sng au:
Vin ch nhc, tn nhuyn, thi vo, m s tit ra nhiu.
Tr no phong, u au khng chu c:
Vin ch (b li), tn nhuyn. Mi ln dng 2g, ly nc lnh ngm trong
ming ri thi thuc vo mi.
Tr kh ut hoc c trng:
Vin ch nhc 160g (sao vi tru). Mi ln dng 20g, thm gng 3 lt, sc
ung.
Tr tiu c, nc tiu :
Vin ch kg (ngm nc cam tho, b li), phc thn (b li), ch tr nhn
u 80g. Tn bt. Ly ru chng vi min lm h, trn thuc bt lm thnh vin, to
bng ht ng ng ln. Mi ln ung 50 vin vi nc to sc.
Tr v sng (suy nh):
Vin ch chng vi ru, ung, b p vo vt thng.
Tr thn kinh suy nhc, hay qun, hi hp, m nhiu, mt ng:
Vin ch (tn). Mi ln ung 8g vi nc cm, ngy 2 ln.
 Tr tuyn v vim, tuyn v u x:
Vin ch 12g, thm 15ml ru 600 ngm 1 lc, cho nc 1 chn, un si 15-20
pht, lc cho ung. Tr 62 ca tuyn v vim cp, c kt qu.
Tr m o vim do trng roi:
Vin ch tn bt, thm glycerin lm thnh thuc n (t vo m o), mi
vin c hm lng thuc sng l 0,75g. Trc khi t thuc, dng bi thuc nc ra
ph khoa: (ngi dip, x sng t, kh sm, ch xc, u 15g, bch ch 9g), sc ly
nc xng v ra m h. t thuc vo m o mi ti 1 ln. Tr 225 ca, sau 3
12 ln, ht triu chng v kim tra trng roi m tnh c 193 ca khi, t l 85,8%.
Tr suy nhc thn kinh do tm huyt km c triu chng mt ng hay qun, hi
hp, m nhiu:
ng sm, Vin ch, mch mn, phc linh u 10g, cam tho 3g, ng quy,
bch thc, sinh khng, i to u 10g, qu tm 3g, sc, thm bt qu tm vo, ha
ung.
1.2.9. Cc ch phm t Vin ch
Ch phm Cng dng v cch dng
Eugica Syrup - Thnh phn:
B mm, nc nc, Vin ch, v
qut
- Cng dng:
Cha vim hng, ho lu ngy,
vim kh- ph qun, ho nhiu m,
cha ho mt ting, kch thch tiu ha.

An thn b tm - Thnh phn:

Sinh a, ng sm, ng qui,
mch mn, to nhn, b t nhn, an
sm, phc thn, huyn sm, Vin ch,
ct cnh, ng v t.
T dc va 1 vin nang.
- Cng dng: dng m, an thn, iu
ha nhp tim.
- Liu dng: ung mt ln 2 - 3 vin,
ngy 3 ln
Bng 2. Cc ch phm t Vin ch
2. C S L THUYT V PHNG PHP CHIT: [4,8]
2.1. Chit lng - lng
Phng php chit l phng php chuyn mt cht trng thi ha tan hay
huyn ph t pha lng (hoc pha rn) ny sang pha lng khc.
Chit nhng cht tan trong dung dch hoc dng huyn ph gi l chit cht
lng, chit nhng cht t hn hp rn gi l chit cht rn.
Chit lng-lng , cn c gi l chit bng dung mi, l mt phng php
tch cc hp cht ring bit da trn tnh tan ca hai cht lng khc nhau, thng l
nc v mt dung mi hu c.
2.2. Chit siu m
Chit siu m l phng php s dng sng vi tn s 20.000 Hz
Cch tin hnh: Bt nguyn liu c xay vi kch thc khong 2mm ri cho
vo becher, lm m bt nguyn liu ri thm mt lng dung mi mi thch hp. t
becher vo trong b siu m sau bt my ri iu chnh thi gian chit. Sau khi
chit xong, lc ly dch chit, thm vo b nguyn liu mt lng dung mi cho ln
chit sau.
2.3. Chit Soxhlet
Bt nguyn liu xay th hoc cao c t trong mt ti vi (hoc giy lc) ri
cho vo ng cha mu. Rt dung mi vo dung mi chn vo bnh cu bng cch
rt trc tip vo ng cha mu. Kim tra h thng kn, m sinh hn, cm vo bp
in v iu chnh sao cho dung mi trong bnh cu si nh.
3. PHNG PHP SC K LP MNG [2,3,8,9]
Phng php sc k lp mng c dng nh tnh, th tinh khit v i
khi bn nh lng hoc nh lng hot cht thuc.
Sc k lp mng l mt k thut tch cc cht c tin hnh khi cho pha
ng di chuyn qua pha tnh trn t hn hp cc cht cn tch. Pha tnh l cht
hp ph c chn ph hp theo tng yu cu phn tch, c tri thnh lp mng
ng nht v c c nh trn cc phin knh hoc phin kim loi.
Pha ng l h dung mi n hoc a thnh phn c trn vi nhau theo t l
quy nh trong tng chuyn lun. Trong qu trnh di chuyn qua cht hp ph, cc
cu t trong hn hp mu th c di chuyn trn lp mng, theo hng pha ng,
vi nhng tc khc nhau. Kt qu, ta thu c mt sc k trn lp mng. i
lng c trng cho mc di chuyn ca cht cn phn tch l h s di chuyn R f
c tnh bng t l gia khong cch dch chuyn ca cht th v khong cch dch
chuyn ca dung mi.

Rf =

Vi a: khong cch t im xut pht n tm ca vt mu th (cm)

b: khong cch t im xut pht n mc dung mi o trn cng ng
i ca vt (cm)
4. NH TNH BNG PHN NG HA HC [1,4,8]
Mi loi dc liu u c cc nhm hp cht xc nh. C th da vo cc
nhm hp cht ny nh tnh cc dc liu bng cc phn ng mu hoc kt ta
c trng.
V d:
Hp cht flavonoid: da vo phn ng vi NaOH, vi FeCl 3
Hp cht saponin: th tnh to bt, tnh ph huyt, phn ng
Liebermann Burchard, phn ng Kahlenberg
Yu cu cc phn ng nh tnh dc liu l cc phn ng ny cng c hiu
cng tt hay t nht l khi kt hp vi cc ch tiu khc ca tiu chun c sn c
th xc nh c dc liu v phn bit vi nhng dc liu tng t. Khi c
nhiu phn ng nh tnh cc cht c trong dc liu, khng nht thit phi a tt c
cc phn ng vo tiu chun. C th chn mt hay hai phn ng tiu biu nht gip
xc nh dc liu. Cc phn ng s dng cho tiu chun nn d thc hin, s dng
cc dung mi ha cht thng dng, cho kt qu r rng v d nhn bit.
5. PH T NGOI KH KIN (PH UV - VIS) [1,2,9]
5.1. Khi nim
Ph t ngoi - kh kin l cc dng ca bc x in t, c bn cht l sng
in t, chng ch khc nhau v di sng. Vng t ngoi - kh kin bao gm cc
bc x in t c di sng trong khong 200 - 800 nm (vng kh kin 360 - 800
nm).
Khi phn t hp th bc x t ngoi hoc kh kin th nhng electron ha tr
ca n b kch thch v chuyn t trng thi c bn ln trng thi kch thch. S hp
th bc x ca phn t c ghi li, ph thu c gi l ph t ngoi - kh kin v
cng c gi ph hp th electron.
5.2. c im
S hp th trong vng t ngoi v vng kh kin ph thuc vo cu trc in
t ca phn t. Ch mt s dng cu trc trong cc hp cht hu c mi c s hp th
nu trn, thc t vic ng dng ph t ngoi cng gii hn trong mt s hp cht, ch
yu l cc cht c cu trc dy ni i lin hp.
 c im ca ph t ngoi l ph ca mt cht phc tp c th rt ging vi
ph ca mt cht n gin nu 2 cht ny cng cha mt nhm cu trc ging nhau.
Hot ng ca ph t ngoi - kh kin
Vng t ngoi - kh kin bao gm cc bc x in t c di sng trong
khong 200 - 800 nm (vng kh kin 360 - 800 nm). S hp th c nhiu ng dng
trong quang ph t ngoi l trong vng t 200 - 380 nm, gi l vng t ngoi gn.
Trn ph t ngoi cc v tr bng hay cn gi l di hp th c o bng
di sng , n v l nanomet (nm) hoc Angstrom (A o ). Cng ca tia n sc
trc v sau khi i qua mi trng hp th c lin h vi nhau bi nh lut
Lambert - Beer.
Khc vi vng ph hng ngoi, vng ph t ngoi - kh kin, nh lut
Lambert - Beer c tun theo tt hn nhiu, do thng c xc nh chnh xc
v c tnh lp li.
Ph t ngoi - kh kin (Ph UV - Vis) thng cho ta ng cong ph thuc
hp th A (lg (I o /I)) vo bc sng . Theo nh lut Lambert - Beer mt quang
ph thuc vo nng ca cht hp ph.
 Phn 3:
VT LIU
V
PHNG PHP NGHIN CU
Kha lun tt nghip Vt liu v phng php nghin cu

1. VT LIU NGHIN CU
1.1. i tng nghin cu
- Cy Du tm (Morus alba L)
B phn dng: L
Thu mua ti Nh thuc YHCT Kim Anh, s 88 Hi Thng Ln ng, phng
10, qun 5, Tp.HCM.
- Cy Vin ch (Polygala japonica Houtt)
B phn dng: r.
Thu mua ti Nh thuc YHCT Kim Anh, s 88 Hi Thng Ln ng, phng
10, qun 5, Tp.HCM
- Cao Du tm, do Trung tm Sm v Dc liu Tp.HCM cung cp.
- Cao Vin ch, do Trung tm Sm v Dc liu Tp.HCM cung cp.
- Rutin chun, do Trung tm Sm v Dc liu Tp.HCM cung cp.
1.2. Dung mi, ha cht, trang thit b

Dung mi Ha cht Trang thit b

-Cn , MeOH, CHCl 3 , H 2 SO 4 , HCl - My KTS samsung

- Vanillin - My c quay gim p Buchi
- Toluen, EtOAc, HCOOH - Bnh chit Sohxlet
- n-BuOH, CH 3 COOH, H 2 O - L nung
- Diethyl ether, anhydric acetic, ether - Cn phn tch Sartorius
du ha - My o quang ph kh kin Thermo
- NaOH, FeCl 3 , AlCl 3 . Spectronic HEIOS
Bng 3.Dung mi- ha cht- trang thit b

SVTH: PHM TH KIM KHUYN 18 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

2. PHNG PHP NGHIN CU

2.1. Xy dng tiu chun kim nghim nguyn liu
2.1.1. Kho st tinh khit
2.1.1.1. Xc nh m
Theo tiu chun DVN IV, ph lc 12.16
Xc nh m bng phng php sy
Nguyn tc:
Bt nguyn liu c sy 105oC, nc trong mu s bc hi lm gim khi
lng mu, cn khi lng mu sau khi sy s tnh c m ca mu nguyn liu.
Tin hnh:
Cn chnh xc 1g nguyn liu vo chn cn dng xc nh m c
cn b trc. Chn cn cn c kch thc thch hp lp nguyn liu cho vo khng
dy qu 5 mm. Cho chn cha nguyn liu vo t sy, sy nhit 100- 1050C
trong 4 gi. Cho chn vo bnh ht m n khi ngui, y np v cn. Lm li nhiu
ln n khi trng lng gia 2 ln cn khng vt qu 0,5mg.
m (x %) ca nguyn liu c tnh theo cng thc sau:
p: S gam ca mu th trc khi sy (g)
a: S gam ca mu th sau khi sy (g)
2.1.1.2. tro ton phn
Theo tiu chun DVN IV, ph lc 9.8
Nguyn tc:
Bt nguyn liu c nung 550oC, cacbon trong mu s chy thnh tro lm
gim khi lng mu, cn khi lng mu sau khi nung s tnh c tro ton phn
ca mu nguyn liu.
Tin hnh:
Ly mt chn s nung ti trong 30 pht. ngui trong bnh ht m ri
cn. Ly 1g nguyn liu ri u vo chn nung, sau t ht hi nc ca nguyn
liu trn bp in ri em nung trong l nung 550oC trong 4 gi. Sau mi ln nung,

SVTH: PHM TH KIM KHUYN 19 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

ly chn nung cng cn tro em lm ngui trong bnh ht m ri cn. Nung n khi
lng gia hai ln cn khng qu 5mg.
tro ton phn c tnh theo cng thc:
X: tro ton phn (%)
X=
a: Khi lng nguyn liu sau khi nung (g)
b: Khi lng nguyn liu trc khi nung (g)
2.1.1.3. Xc nh tro khng tan trong acid
Theo tiu chun DVN IV, ph lc 9.7
Nguyn tc:
Dng acid HCl ha tan tro ton phn, lng cn khng tan nung 550oC, cn
khi lng cn sau khi nung s tnh c tro khng tan trong acid ca mu.
Tin hnh:
Cho 25ml dung dch acid HCl 2M vo tro ton phn, un si 5 pht, lc tp
trung nhng cht khng tan vo mt 1 ming giy lc khng tro, ra bng nc nng
ri em nung 550oC n khi lng khng i. Tnh t l phn trm tro khng tan
trong acid so vi nguyn liu lm kh trong khng kh.

Y: tro khng tan trong acid (%)

Y= a: Khi lng cn sau khi nung (g)
b: Khi lng nguyn liu ban u (g)

2.1.2. nh tnh cc hp cht ha thc vt trong nguyn liu

2.1.2.1. Nguyn liu Du tm
a. Da vo phn ng ha hc
Ly 5g nguyn liu Du tm chit soxhlet bng ether du ha (60o-90oC)
loi tp cht, loi b lp ether du ha, ly b bay hi ht dung mi n kh, chit
tip bng MeOH 70%, thu v c cn dch lc, sau ha MeOH c dung dch
A.

SVTH: PHM TH KIM KHUYN 20 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

Phn ng Cch tin hnh Yu cu xut hin

Vi TT NaOH 20% 2ml dd A + 3 git NaOH 20% Ta vng cam

Vi TT FeCl 3 5%. 2ml dd A + 2 git FeCl 3 5% Mu xanh ru

Bng 4. nh tnh hp cht flavonoid trong nguyn liu Du tm
b. Da vo sc k lp mng
Kim tra s hin din ca cht rutin trong nguyn liu Du tm
Bn mng Silicagel GF 254
Dung mi khai trin: EtOAc - HCOOH - CH 3 COOH - H 2 O (100:11:11:26)
Dung dch th:
Ly 5g nguyn liu chit soxhlet bng ether du ha (60o-90oC) loi tp
cht, loi b lp ether du ha, ly b bay hi ht dung mi n kh, chit tip bng
MeOH 70%, thu v c cn dch lc, sau ha MeOH c dung dch th.
Dung dch chun:
Ly 1mg rutin chun, pha vi 1ml MeOH c dung dch c nng
1mg/ml dng chm sc k.
Cch tin hnh: Chm ring bit ln bn mng 10l mi dung dch trn.
Cch pht hin: Phun thuc th AlCl 3 5% trong MeOH.
Quan st di nh sng thng
Quan st di nh sng t ngoi bc sng 365nm
2.1.2.2. Nguyn liu Vin ch
a. Da vo phn ng ha hc

Phn ng Cch tin hnh Yu cu xut hin

To bt Cn 0,1g nguyn liu+ 5ml
Bt bn t nht 10 pht
nc nng, lc mnh 5 pht.
Liebermann- Burchard Cn 0,5g nguyn liu + 2ml Vng ngn cch xut
(CH 3 CO) 2 O + 2ml CHCl 3 + t hin mu hng
t 2-3 git H 2 SO 4
Bng 5.nh tnh hp cht ha thc vt trong nguyn liu Vin ch

SVTH: PHM TH KIM KHUYN 21 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

b. Da vo sc k lp mng
Bn mng Silicagel GF 254
Dung mi khai trin: CHCl 3 - MeOH (9 : 1).
Dung dch th:
Ly 5g nguyn liu Vin ch, chit Soxhlet vi MeOH, ly dch chit em c
cn. Ha cn vi 50ml nc, ly dch nc em lc vi diethyl ether (20ml 3 ln).
Tip tc lc vi n-BuOH (20ml 3 ln). Dch n-BuOH em c cn sau ha vi
MeOH chm sc k.
Cch tin hnh: Chm ln bn mng 10l dung dch th.
Cch pht hin:
Quan st di nh sng t ngoi bc sng 365nm
Phun thuc th vanilin 2% trong ethanol v acid sulfuric, sy 1050C,
quan st di nh sng thng v di nh sng t ngoi bc sng 365nm
2.1.3. nh lng cc hp cht ha thc vt trong nguyn liu
2.1.3.1. nh lng hm lng flavonoid tnh theo rutin trong nguyn liu Du tm
- Thuc th AlCl 3 5% trong MeOH 70%
- Cht chun: 60g rutin chun
- Mu th: Cn chnh xc 5g nguyn liu cho vo bnh Soxhlet. Thm 250ml ether
du ha, chit n khi dch chit khng cn mu. ngui v gn b dch ether
du ha. Cn trong bnh Soxhlet c chit tip tc vi 250ml MeOH 70%, cho
n khi dch chit khng cn mu. Dch chit MeOH 70% c c cn, chuyn
vo bnh nh mc 100ml, b sung MeOH 70%va n vch, lc u, thu
dung dch A. Ly chnh xc 50l dung dch A bng micropipet, cho vo ng
nghim c np vn, c n cn, thm chnh xc 5 ml MeOH 70% v 75l dung
dch AlCl 3 5% trong MeOH 70%, lc u (tin hnh 3 mu th tng t ly
gi tr trung bnh).
- Mu trng: MeOH 70%

SVTH: PHM TH KIM KHUYN 22 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

- Mu trng th: Ly chnh xc 50l dung dch A cho vo ng nghim c nt vn,

c n cn, thm chnh xc 5,075ml MeOH 70%, lc u.
- Mu chun: Ly chnh xc 60g rutin chun cho vo ng nghim c np vn, thm
chnh xc 5ml MeOH 70% v 75l dung dch AlCl 3 5% trong MeOH 70%, lc
u.
Tt c cc mu u nhit phng lnh, y kn ng nghim, sau 30
pht tin hnh o, thnh thong lc nh cho phn ng xy ra hon ton.
Tin hnh o hp thu cc mu nh hp thu cc i max = 422nm-
424nm trn my UV Visible Spectrophotometre, dng MeOH 70% lm mu
trng chnh my v 0.
Hm lng flavonoid tnh theo rutin c trong nguyn liu Du tm c
tnh theo cng thc sau:
At A't 2000 10 4
X = mc d
Ac m

Trong : A t : hp thu mu th.

A t : hp thu mu trng th
A c : hp thu mu chun
m c : Khi lng cht chun (g)
d : tinh khit ca cht chun
m : Khi lng nguyn liu cn nh lng tr m (g).

SVTH: PHM TH KIM KHUYN 23 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

S 1. S xc nh hm lng flavonoid tnh theo rutin trong nguyn liu

Du tm
Bt nguyn liu

Chit Soxhlet vi 250ml ether du ha.

Loi dch ether du ha

B nguyn liu

Thm 250ml MeOH 70%

Dch chit MeOH 70%

C cch thy

Cn MeOH 70%

nh mc 100ml bng MeOH 70%

Dung dch A

Ht 50 l dung dch A.
nh mc 5ml bng MeOH 70%

Xc nh mt quang
OD

Hm lng flavonoid
tnh theo rutin

SVTH: PHM TH KIM KHUYN 24 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

2.1.3.2. nh lng hm lng saponin ton phn trong nguyn liu Vin ch:
Hm lng hp cht saponin ton phn c nh lng theo phng php
Namba (S 2): Cn 5g nguyn liu Vin ch cho vo bnh Soxhlet chit vi 250ml
MeOH cho n kit saponin. Dch MeOH c c gim p n cn. Ha cn vi
50ml nc. Dung dch nc c lc vi diethyl ether (20ml x 5 ln), b lp diethyl
ether. Lp nc c lc vi n-BuOH bo ha nc (20ml x 6 ln), chit ly lp n-
BuOH. Ra nc lp n-BuOH khong 3 ln. Dch n-BuOH c bc hi trn bp
cch thy cho n cn, lm kh trong t sy chn khng cho n khi lng khng
i, ta thu c saponin ton phn. Cn v tnh khi lng saponin ton phn trong
nguyn liu. (Thc hin 3 mu ly gi tr trung bnh).
Hm lng hp cht saponin ton phn c tnh theo cng thc:

M% =

Trong :
M%: Hm lng saponin ton phn
A: m ca nguyn liu
a: Khi lng saponin ton phn (g)
b: khi lng nguyn liu ban u (g)

SVTH: PHM TH KIM KHUYN 25 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

S 2. nh lng saponin ton phn trong nguyn liu Vin ch theo phng php
cn Namba

Bt nguyn liu

Chit theo phng php Soxhlet

Dch chit MeOH

C gim p

Cn MeOH

Thm 50ml nc

Dch nc

Lc vi diethyl ether (20ml x 5 ln)

Dch nc Dch diethyl ether

(b)

Lc vi n-BuOH bo ha (20ml x 6 ln)

Dch n-BuOH

C cch thy

Cn n-BuOH
(Saponin ton phn)

SVTH: PHM TH KIM KHUYN 26 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

2.2. Xy dng tiu chun kim nghim cao

2.2.1. Kho st tinh khit
2.2.1.1. Xc nh m
Theo tiu chun DVN IV, ph lc 12.16
Xc nh m bng phng php sy
Nguyn tc:
Mu th c sy 105oC, nc trong mu s bc hi lm gim khi lng
mu, cn khi lng mu sau khi sy s tnh c m ca mu.
Tin hnh:
Cn chnh xc 0,5g cao cho vo chn cn dng xc nh m c cn
b trc. Cho chn cha cao vo t sy, sy nhit 100- 1050C trong 4 gi. Cho
chn vo bnh ht m n khi ngui. Cn, lm li nhiu ln n khi trng lng gia
2 ln cn khng vt qu 0,5 mg.
m (x %) ca cao chit c tnh theo cng thc sau:
p: S gam ca cao trc khi sy (g)
a: S gam ca cao sau khi sy (g)
2.2.1.2. tro ton phn
Theo tiu chun DVN IV, ph lc 9.8
Tin hnh:
Ly mt chn s nung ti trong 30 pht. ngui trong bnh ht m ri
cn. Ly 0,5g cao vo chn nung, sau t ht hi nc ca cao trn bp in ri
em nung trong l nung 550oC trong 4 gi. Sau mi ln nung, ly chn nung cng
cn tro em lm ngui trong bnh ht m ri cn. Nung n khi lng gia hai ln
cn khng qu 5mg.
tro ton phn c tnh theo cng thc:

X: tro ton phn (%)

X= a: Khi lng cao sau khi nung (g)
b: Khi lng cao trc khi nung (g)

SVTH: PHM TH KIM KHUYN 27 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

2.2.2. nh tnh cc hp cht ha thc vt trong cao

2.2.2.1. Cao Du tm
a. Da vo phn ng ha hc
- Ly 0,5g cao Du tm chit soxhlet bng ether du ha (60o-90oC)
loi tp cht, loi b lp ether du ha, ly b bay hi ht dung mi n kh, chit
tip bng MeOH 70%, thu v c cn dch lc, sau ha MeOH c dung
dch B.

Phn ng Cch tin hnh Yu cu xut hin

Vi dd NaOH 20% 2ml dd B + 3 git NaOH 20% Ta vng cam
Vi dd FeCl 3 5%. 2ml dd B + 2 git FeCl 3 5% Mu xanh ru
Bng 6. nh tnh hp cht flavonoid trong cao Du tm
b. Da vo sc k lp mng
Mc ch: xc nh c cc thnh phn ha thc vt trong nguyn liu
Du tm cng phi hin din trong cao Du tm v s hin din ca cht chun rutin
trong cao Du phi c kim tra nh gi qua SKLM.
Kim tra s hin din ca nguyn liu trong cao Du tm
Bn mng Silicagel GF 254
Dung mi khai trin: EtOAc - HCOOH - H 2 O (50:4:10), lp trn.
CHCl 3 - MeOH (9:1)
Dung dch th:
Ly 5g nguyn liu chit siu m vi MeOH (30ml x 3 ln), thu v c dch lc
cn 2ml chm sc k
Ly 0,5g cao Du tm, chit siu m vi MeOH (30ml x 3 ln), thu v c dch
lc cn 2ml chm sc k.
Cch tin hnh:
Chm ring bit ln bn mng 10l mi dung dch trn.
Cch pht hin:
Phun thuc th NaOH 10% trong cn

SVTH: PHM TH KIM KHUYN 28 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

Quan st di nh sng thng

Quan st di nh sng t ngoi bc sng 365nm
Phun thuc th H 2 SO 4 10% trong cn, sy 1050C, quan st di nh sng
thng
Kim tra s hin din ca hp cht chun rutin trong cao Du tm
Bn mng Silicagel GF 254
Dung mi khai trin: EtOAc - HCOOH - CH 3 COOH - H 2 O (100:11:11:26).
Dung dch th:
Ly 0,5g cao Du tm chit soxhlet bng ether du ha (60o-90oC) loi tp
cht, loi b lp ether du ha, ly b bay hi ht dung mi n kh, chit tip bng
MeOH 70%, thu v c cn dch lc, sau ha MeOH c dung dch th.
Dung dch chun:
Ly 1mg rutin chun, pha vi 1ml MeOH c dung dch c nng
1mg/ml dng chm sc k.
Cch tin hnh:
Chm ring bit ln bn mng 10l mi dung dch trn.
Cch pht hin: Phun thuc th AlCl 3 5% trong MeOH.
Quan st di nh sng thng
Quan st di nh sng t ngoi bc sng 365nm
2.2.2.2. Cao Vin ch
a. Da vo phn ng ha hc

Phn ng Cch tin hnh Yu cu xut hin

Liebermann- 0,5g cao + 2ml (CH 3 CO) 2 O Vng ngn cch xut hin
Burchard + 2ml CHCl 3 + t t 2-3 mu nu .
git H 2 SO 4
Bng 7. nh tnh hp cht ha thc vt trong cao Vin ch
b. Da vo sc k lp mng
Kim tra s hin din ca nguyn liu trong cao Vin ch

SVTH: PHM TH KIM KHUYN 29 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

Bn mng Silicagel GF 254

Dung mi khai trin: CHCl 3 - MeOH (9 : 1).
n-BuOH - CH 3 COOH - H 2 O (7:1:2).
Dung dch th:
Ly 5g nguyn liu Vin ch, chit Soxhlet vi MeOH, ly dch chit em c
cn. Ha cn vi 50ml nc, ly dch nc em lc vi diethyl ether (20ml
3 ln). Tip tc lc vi n-BuOH (20ml 3 ln). Dch n-BuOH em c cn sau
ha vi MeOH chm sc k.
Ly 1g cao Vin ch, ha vi 50ml nc, lc vi diethyl ether v n-BuOH nh
tng t nh nguyn liu. Thu c dung dch th.
Cch tin hnh: Chm ring bit ln bn mng 10l cc dung dch th.
Cch pht hin:
Quan st di nh sng t ngoi bc sng 254nm
Quan st di nh sng t ngoi bc sng 365nm
Phun thuc th H 2 SO 4 10% trong cn, sy 1050C, quan st
di nh sng thng
2.2.3. nh lng cc hp cht ha thc vt trong cao
2.2.3.1. nh lng hm lng flavonoid tnh theo rutin trong cao Du tm
o Thuc th AlCl 3 5% trong MeOH 70 %
o Cht chun: 60g rutin chun
o Mu th: Cn chnh xc 0,5g cao cho vo bnh Soxhlet. Thm 250ml ether du,
chit n khi dch chit khng cn mu. ngui v gn b dch ether du. Cn
cao trong bnh Soxhlet c chit tip tc vi 250ml MeOH 70%, cho n khi
dch chit khng cn mu. Dch chit MeOH 70% c chuyn vo bnh nh
mc 100ml, b sung MeOH va n vch, lc u, thu dung dch A. Ly chnh
xc 40l dung dch A bng micropipet, cho vo ng nghim c np vn, c n
cn, thm chnh xc 5 ml MeOH 70% v 75l dung dch AlCl 3 5% trong MeOH
70%, lc u (tin hnh 3 mu th tng t ly gi tr trung bnh).

SVTH: PHM TH KIM KHUYN 30 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

o Mu trng:MeOH 70%
o Mu trng th: Ly chnh xc 40l dung dch A bng micropipet, cho vo ng
nghim c np vn, c n cn,, thm chnh xc 5,075ml MeOH 70%, lc u.
o Mu chun: Ly chnh xc 60g rutin chun cho vo ng nghim c np vn,
thm chnh xc 5ml MeOH 70% v 75l dung dch AlCl 3 5% trong MeOH 70%,
lc u.
Tt c cc mu u nhit phng lnh, y kn ng nghim, sau 30 pht
tin hnh o, thnh thong lc nh cho phn ng xy ra hon ton.
Tin hnh o hp thu cc mu nh hp thu cc i max = 422nm-
424nm trn my UV Visible Spectrophotometre, dng MeOH 70% lm mu trng
chnh my v 0.
Lng flavonoid ton phn tnh theo rutin c trong cao Du tm c tnh
theo cng thc sau:
At A't 2500 10 4
X = mc d
Ac m

Trong : A t : hp thu mu th.

A t : hp thu mu trng th
A c : hp thu mu chun
m c : Khi lng cht chun (g)
d: tinh khit ca cht chun
m: Khi lng cao cn nh lng tr m (g).

SVTH: PHM TH KIM KHUYN 31 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

S 3. S xc nh hm lng flavonoid tnh theo rutin trong Du tm

Cao Du tm

Chit Soxhlet vi 250ml ether du.

Loi dch ether du

Cn Du tm

Thm 250ml MeOH 70%

Dch chit MeOH 70%

C cch thy

Cn MeOH 70%

nh mc 100ml bng MeOH 70%

Dung dch A

Ht 40l dung dch A.

nh mc 5ml bng MeOH 70%

Xc nh mt quang
OD

Hm lng flavonoid
tnh theo rutin

SVTH: PHM TH KIM KHUYN 32 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

2.2.3.2. nh lng hm lng saponin ton phn trong cao Vin ch:
Hm lng hp cht saponin ton phn c nh lng theo phng php
Namba (S 4):
Cn 1g cao Vin ch cho vo bnh Soxhlet chit vi MeOH cho n kit
saponin. Dch MeOH c c gim p n cn. Ha cn vi khong 50ml nc.
Dung dch nc c lc vi diethyl ether (20ml x 5 ln) cho n khi lp diethyl
ether khng mu hoc rt nht, b lp diethyl ether. Lp nc tip tc c lc vi n-
BuOH bo ha nc (20ml x 8 ln), chit ly lp n- BuOH. Ra nc lp n- BuOH
khong 3 ln. Dch n- BuOH c bc hi trn bp cch thy cho n cn, lm kh
trong t sy chn khng cho n khi lng khng i, ta thu c saponin ton
phn. Cn v tnh khi lng saponin ton phn trong cao (thc hin 3 mu ly gi
tr trung bnh).
Hm lng hp cht saponin ton phn c tnh theo cng thc:

M% =

Trong :
M%: Hm lng saponin ton phn
A: m ca cao Vin ch
a: Khi lng saponin ton phn (g)
b: khi lng cao ban u (g)

SVTH: PHM TH KIM KHUYN 33 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

S 4. nh lng saponin ton phn trong cao Vin ch theo phng php cn
Namba

Cao Vin ch

Chit theo phng php Soxhlet

Dch chit MeOH

C gim p

Cn MeOH

Thm 50ml nc

Dch nc

Lc vi diethyl ether (20ml x 5 ln)

Dch nc Dch diethyl ether

(b)

Lc vi n-BuOH bo ha (20ml x 8 ln)

Dch n-BuOH

C cch thy

Cn n-BuOH
(Saponin ton phn)

SVTH: PHM TH KIM KHUYN 34 ThS. DNG TH MNG NGC

Kha lun tt nghip Vt liu v phng php nghin cu

Phn 4
KT QU
V
BN LUN

SVTH: PHM TH KIM KHUYN 35 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1. XY DNG TIU CHUN KIM NGHIM NGUYN LIU

1.2. Kho st tinh khit ca nguyn liu
1.2.1. m
Du tm Vin ch
Ln 1 Ln 2 Ln 3 Ln 1 Ln 2 Ln 3
Khi lng b (g) 5,2600 4,9170 5,0948 4,6195 4,2956 5,0536
Khi lng nguyn liu (g) 1,0406 1,0951 1,0764 1,1993 1,0021 1,0434
Khi lng b v nguyn
6,2204 5,9309 6,0872 5,7339 5,2344 6,0311
liu sau khi sy (g)
m (%) 7,71 7,41 7,80 7,08 6,32 6,32
Trung bnh 7,64 6,57
Bng 8. m ca nguyn liu
Nhn xt:
T kt qu trn cho thy nguyn liu c cc gi tr v m nm trong gii hn
an ton cho php ng theo tiu chun Dc in Vit Nam IV l m ca nguyn
liu Du tm <15% v ca Vin ch <14%.
1.2.2. tro ton phn
Du tm Vin ch
Ln 1 Ln 2 Ln 3 Ln 1 Ln 2 Ln 3
Khi lng b (g) 31,2718 39,0600 38,7460 34,2415 36,7750 36,3080
Khi lng nguyn liu
1,0103 1,0056 1,0087 1,0099 1,0186 1,0123
(g)
Khi lng b v nguyn
31,4209 39,2072 38,8900 34,2724 36,8057 36,3400
liu sau khi nung (g)
tro (%) 14,76 14,64 14,28 3,06 3,04 3,09
Trung bnh 14,56 3,07
Bng 9. tro ton phn ca nguyn liu

SVTH: PHM TH KIM KHUYN 36 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

Nhn xt: Theo tiu chun Dc in Vit Nam IV tro ton phn
ca nguyn liu Vin ch t yu cu khi <6%. Vy tro ton phn ca Vin ch
nm trong gii hn cho php.
tro ton phn ca nguyn liu Du tm t 14,56%
1.2.3. tro khng tan trong acid
Du tm Vin ch
Ln 1 Ln 2 Ln 3 Ln 1 Ln 2 Ln 3
Khi lng b (g) 31,2718 39,0600 38,7460 34,2415 36,7750 36,3080
Khi lng nguyn
1,0103 1,0056 1,0087 1,0099 1,0186 1,0123
liu (g)
Khi lng b v
nguyn liu sau khi 31,3038 39,0890 38,7750 34,2517 36,7852 36,3139
nung (g)
tro (%) 3,17 2,88 2,80 1,01 1,01 1,09
Trung bnh 2,95 1,04
Bng 10. tro khng tan trong acid ca nguyn liu
Nhn xt
Theo tiu chun Dc in Vit Nam IV th tro khng tan trong acid ca
nguyn liu Du tm <4,5% v ca Vin ch <1,5%. C th kt lun nguyn liu t
tiu chun nm trong gii hn an ton.
1.3. nh tnh cc hp cht ha thc vt trong nguyn liu
1.3.1. Du tm
Cc hp cht hu c trong nguyn liu v cao c kim tra bng cc phn ng
ha hc da vo thuc th c trng cho mi cht v kt qu c trnh by nh sau:

SVTH: PHM TH KIM KHUYN 37 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1.3.1.1. Da vo phn ng ha hc

1 3
1 2

.
(A) (B)

Hnh 3. Phn ng ha hc ca nguyn liu Du tm

Ch thch: A. Phn ng vi TT NaOH 20%
B. Phn ng vi TT FeCl 3 5%
1. Dung dch i chng
2. Dung dch sau khi phn ng vi TT NaOH 20%
3. Dung dch sau khi phn ng vi TT FeCl 3 5%

SVTH: PHM TH KIM KHUYN 38 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1.3.1.2. Da vo sc k lp mng
Kim tra s hin din ca cht rutin trong nguyn liu Du tm
EtOAc - HCOOH - CH 3 COOH - H 2 O (100:11:11:26)

1 2 3 1 2 3

1. Nguyn liu 2. Trng 2 vt 3. Chun rutin

Phun thuc th AlCl 3 5% trong MeOH, sy 105oC n khi xut hin
vt
Quan st di n t ngoi bc Quan st di nh sng thng
sng 365nm
Hnh 4. SKLM kim tra s hin din ca cht rutin trong nguyn liu Du tm
Nhn xt: Mu sc v gi tr R f ca vt thu c trn sc dung dch
th chit t nguyn liu Du tm tng ng vi mu sc v gi tr R f ca vt
chun rutin. Nh vy, c s hin din ca chun rutin trong nguyn liu Du
tm.
.

SVTH: PHM TH KIM KHUYN 39 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1.3.2. Nguyn liu Vin ch

1.3.2.1. Da vo phn ng ha hc

1 2

(A) (B)
Hnh 5. Phn ng ca nguyn liu Vin ch
Ch thch: A. Phn ng to bt
B. Phn ng Liebermann- Burchard
1. Dung dch i chng
2. Dung dch sau phn ng

SVTH: PHM TH KIM KHUYN 40 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1.3.2.2. Da vo sc k lp mng
CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1)

Quan st di n t ngoi Phun thuc th vanilin 2% Phun thuc th vanilin 2%

bc sng 365nm trong cn v acid sulfuric, trong cn v acid sulfuric,
sy 105oC n khi xut sy 105oC n khi xut
hin vt. Quan st di hin vt. Quan st di
nh sng thng n t ngoi bc sng
365nm
Nhn xt: Trn sc k thy xut hin 4 vt pht quang mu xanh khi
quan st di n t ngoi bc sng 365nm. Bng thuc th hin mu xut hin 3
vt mu xanh khi quan st di nh sng thng v 1 vt mu hng m, 2 vt pht
quang mu xanh sng khi quan st di n t ngoi bc sng 365nm

SVTH: PHM TH KIM KHUYN 41 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

1.4. nh lng cc hp cht ha thc vt trong nguyn liu

1.4.1. nh lng hm lng flavonoid tnh theo rutin trong nguyn liu Du tm
Quy trnh nh lng hp cht flavonoid bng phng php ph t ngoi kh
kin da trn cht chun rutin. hp thu cc i ca rutin chun l 424nm. Tin
hnh o cc mu th bc sng trn ta thu c bng sau:
Ln 1 Ln 2 Ln 3
Khi lng nguyn liu (g) 5,0138 5,0079 5,0491
hp thu mu chun (A c ) 0,308
hp thu mu trng th (A t ) 0,163 0,167 0,169
hp thu mu th (A t ) 0,689 0,694 0,698
Hm lng flavonoid tnh theo rutin (%)
4,1961 4,2090 4,1905
Trung bnh 4,1985
Bng 11. Hm lng flavonoid tnh theo rutin trong nguyn liu Du tm
Nhn xt: Hm lng flavonoid tnh theo rutin trong nguyn liu Du tm t
4,1985%
1.4.2. nh lng hm lng saponin ton phn trong nguyn liu Vin ch
Ln 1 Ln 2 Ln 3
Khi lng nguyn liu (g) 5,1106 5,0901 5,0417

khi lng cn saponin ton phn (g) 0,9976 1,0339 1,0072

Hm lng cn saponin ton phn (%) 20,893 21,740 21,382
Trung bnh 21,338
Bng 12. Hm lng saponin ton phn trong nguyn liu Vin ch
Nhn xt: Hm lng saponin trong nguyn liu Vin ch t 21,338%

SVTH: PHM TH KIM KHUYN 42 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2. XY DNG TIU CHUN KIM NGHIM CAO CHIT

2.1. Kho st tinh khit ca cao chit
2.1.1. m
Cao Du tm Cao Vin ch
Ln 1 Ln 2 Ln 3 Ln 1 Ln 2 Ln 3
Khi lng b (g) 4,8425 4,9396 4,7337 4,7843 4,8188 4,8587
Khi lng cao (g) 0,5506 0,5365 0,5585 0,5130 0,5135 0,5002
Khi lng b v cao sau
5,3587 5,4413 5,2570 5,1906 5,2266 5,2549
khi sy (g)
m (%) 6,25 6,49 6,30 20,80 20,58 20,79
Trung bnh 6,35 20,72
Bng 13. m ca cao chit
Nhn xt: m ca cao Du tm t 6,35%.
m ca cao Vin ch t 20,72%.
2.1.2. . tro ton phn
Cao Du tm Cao Vin ch
Ln 1 Ln 2 Ln 3 Ln 1 Ln 2 Ln 3
Khi lng b (g) 39,0537 36,7667 31,2662 28,0682 23,8268 30,9338
Khi lng cao (g) 0,5650 0,5005 0,5469 0,5319 0,5218 0,5142
Khi lng b v cao
39,1095 36,8142 31,3180 28,0728 23,8309 30,9382
sau khi nung (g)
tro (%) 9,88 9,49 9,47 0,86 0,79 0,86
Trung bnh 9,61 0,84
Bng 14. tro ton phn ca cao chit
Nhn xt: tro ton phn ca cao Du tm t 9,61%.
tro ton phn m ca cao Vin ch t 0,84%.

SVTH: PHM TH KIM KHUYN 43 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2.2. nh tnh cc hp cht ha thc vt trong cao

2.2.1. Cao Du tm
2.2.1.1. Da vo phn ng ha hc

1 2 1 3

(A) (B)
Hnh 6.Phn ng ha hc ca cao Du tm
Ch thch: A. Phn ng vi TT NaOH 20%
B. Phn ng vi TT FeCl 3 5%
1. Dung dch i chng
2. Dung dch sau khi phn ng vi TT NaOH 20%
3. Dung dch sau khi phn ng vi TT FeCl 3 5%

SVTH: PHM TH KIM KHUYN 44 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2.2.1.2. Da vo sc k lp mng
Kim tra s hin din ca nguyn liu trong cao Du tm
EtOAc - HCOOH -
CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1) H 2 O (50:4:10), lp
trn.

1 2 1 2 1 2
1 2

1. Nguyn liu 2. Cao

Quan st di n Phun thuc th Phun thuc th Phun thuc th
t ngoi bc NaOH 10% trong NaOH 10% trong H 2 SO 4 10% trong
sng 365nm cn. Quan st di cn. Quan st di cn. Quan st di
n t ngoi bc nh sng thng. nh sng thng
sng 365nm sau khi sy 105oC
Hnh 7. SKLM kim tra s hin din ca nguyn liu trong cao Du tm
Nhn xt: Mu sc v gi tr R f ca cc vt thu c trn sc dung
dch th chit t nguyn liu Du tm tng ng vi mu sc v gi tr R f ca cc vt
thu c ca cao Du tm. Nh vy, chng t cc thnh phn ha thc vt trong
nguyn liu Du tm cng hin din trong cao Du tm.

SVTH: PHM TH KIM KHUYN 45 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

Kim tra s hin din ca cht chun rutin trong cao Du tm

EtOAc - HCOOH - CH 3 COOH - H 2 O (100:11:11:26)

1 2 3
1 2 3

1. Cao 2. Trng 2 vt 3. Chun rutin

Phun thuc th AlCl 3 5% trong MeOH, sy 105oC n khi xut hin
vt
Quan st di n t ngoi bc Quan st di nh sng thng
sng 365nm
Hnh 8. SKLM kim tra s hin din ca cht chun rutin trong cao Du tm
Nhn xt: Mu sc v gi tr R f ca vt thu c trn sc dung dch
th chit t cao Du tm tng ng vi mu sc v gi tr R f ca vt chun rutin.
Nh vy, c s hin din ca chun rutin trong cao Du tm.

SVTH: PHM TH KIM KHUYN 46 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2.2.2. . Cao Vin ch

2.2.2.1. Da vo phn ng ha hc
Phn ng Liebermann- Burchard

1 2

Hnh 9. Phn ng ha hc ca cao Vin ch

1. Dung dch i chng
2. Dung dch sau phn ng

SVTH: PHM TH KIM KHUYN 47 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2.2.2.2. Da vo sc k lp mng
Kim tra s hin din ca nguyn liu trong cao Vin ch
CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1) CHCl 3 - MeOH (9:1)

1 2 1 2
1 2

1.Nguyn liu 2. Cao

Quan st di n t Quan st di n t Phun thuc th
ngoi bc sng 254nm ngoi bc sng 365nm H 2 SO 4 10% trong cn, sy
105oC n khi xut hin
vt. Quan st di nh
sng thng

SVTH: PHM TH KIM KHUYN 48 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

n-BuOH-AcOH-H 2 O n-BuOH-AcOH-H 2 O n-BuOH-AcOH-H 2 O

(7:1:2) (7:1:2) (7:1:2)

1 2 1 2 1 2

Nguyn liu 2. Cao

Quan st di n t Quan st di n t Phun thuc th H 2 SO 4
ngoi bc sng 254nm ngoi bc sng 365nm 10% trong cn, sy
105oC n khi xut hin
vt. Quan st di nh
sng thng
Hnh 10. SKLM kim tra s hin din ca nguyn liu trong cao Vin ch
Nhn xt: Mu sc v gi tr R f ca cc vt thu c trn sc dung
dch th chit t nguyn liu Vin ch tng ng vi mu sc v gi tr R f ca cc vt
thu c ca cao Vin ch. Nh vy, chng t cc thnh phn ha thc vt trong
nguyn liu Vin ch cng hin din trong cao Vin ch.

SVTH: PHM TH KIM KHUYN 49 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt qu v kin ngh

2.3. nh lng cc hp cht ha thc vt trong cao.

2.3.1. Cao Du tm
Quy trnh nh lng hp cht flavonoid bng phng php ph t ngoi kh
kin da trn cht chun rutin. hp thu cc i ca rutin chun o c l 424nm.
Tin hnh o cc mu th bc sng trn ta thu c bng sau:
Cao Du tm Ln 1 Ln 2 Ln 3
Khi lng cao (g) 0,5313 0,5348 0,5258
hp thu mu chun (A c ) 0,307
hp thu mu trng th (A t ) 0,294 0,296 0,299
hp thu mu th (A t ) 0,518 0,515 0,519
Hm lng flavonoid tnh theo rutin (%)
20,8967 20,2965 20,7382
Trung bnh 20,6438
Bng 15. Hm lng flavonoid tnh theo rutin trong cao Du tm
Nhn xt: Hm lng flavonoid tnh theo rutin trong cao Du tm l
20,6438%
2.3.2. Cao Vin ch
Hm lng saponin ton phn thu c nh sau:
Cao Vin ch Ln 1 Ln 2 Ln 3
Khi lng cao (g) 1,0576 1,0549 1,0439
Khi lng cn saponin ton phn (g) 0,3426 0,3394 0,3386
Hm lng cn saponin ton phn (%) 40,860 40,582 40,913
Trung bnh 40,785
Bng 16. Hm lng saponin ton phn trong cao Vin ch
Nhn xt: Hm lng saponin ton phn thu c trong cao l
40,785%

SVTH: PHM TH KIM KHUYN 50 ThS. DNG TH MNG NGC

 Phn 5:
KT LUN
V
KIN NGH
Kha lun tt nghip Kt lun v kin ngh

1. KT LUN
Qua qu trnh thc nghim v lm quen vi cng tc nghin cu khoa hc, em
thc hin c cc vn sau:
1. Xy dng tiu chun kim nghim c bn ca nguyn liu Du tm v Vin ch
theo chuyn lun Dc Liu trong Dc in Vit Nam nh:
m
tro ton phn
tro khng tan trong acid
nh gi c cht lng ca nguyn liu v mt nh tnh bng c hai
phng php:
Phn ng ha hc
Sc k lp mng
nh gi c cht lng ca nguyn liu v mt nh lng
Hm lng flavonoid tng trong nguyn liu Du tm bng
phng php quang ph t ngoi - kh kin.
Hm lng saponin ton phn trong nguyn liu Vin ch theo
phng php cn Namba.
2. Xy dng tiu chun kim nghim ca cao Du tm v cao Vin ch theo
chuyn lun trong Dc in Vit Nam nh :
m
tro ton phn
nh gi c cht lng ca cao Du tm v cao Vin ch v mt inh
tnh bng c hai phng php:
Phn ng ha hc
Sc k lp mng
nh gi c cht lng ca cao Du tm v cao Vin ch v mt nh
lng

SVTH: PHM TH KIM KHUYN 52 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt lun v kin ngh

Hm lng flavonoid trong cao Du tm bng phng php

quang ph t ngoi- kh kin
Hm lng saponin ton phn trong cao Vin ch theo phng
php cn Namba.

SVTH: PHM TH KIM KHUYN 53 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt lun v kin ngh

2. KIN NGH
Qua qu trnh xy dng tiu chun kim nghim nguyn liu v cao Du
tm, Vin ch em xin ngh cc bng tiu chun c s kim nghim nh
sau:
2.1. Xy dng tiu chun kim nghim nguyn liu
2.1.1. Nguyn liu Du tm
STT Cc tiu ch Yu cu Phng php th
1 m Khng c qu 15% DVN IV
2 tro ton phn Khng c qu 20% DVN IV
3 tro khng tan DVN IV
Khng c qu 4,5%
trong acid
4 nh tnh
Phn ng ha hc Mu th phi cho phn TCCS
ng vi TT NaOH 20%
v TT FeCl 3 5%.
Sc k lp mng Cc vt thu c trn
sc k ca mu th
phi c gi tr R f v
mu sc tng ng
vi cc vt thu c
ca cht chun rutin
5 nh lng Hm lng flavonoid TCCS
tnh theo rutin trong
khong 3,57% - 4,83%

SVTH: PHM TH KIM KHUYN 54 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt lun v kin ngh

2.1.2. Nguyn liu Vin ch

STT Cc tiu ch Yu cu Phng php th
1 m Khng c qu 14% DVN IV
2 tro ton phn Khng c qu 6% DVN IV
3 tro khng tan DVN IV
Khng c qu 1,5%
trong acid
4 nh tnh
Phn ng ha hc Mu th phi c phn DVN IV
ng to bt v phn
ng Liebermann-
Burchard
Sc k lp mng Mu th phi cho sc TCCS
k c vt pht quang
di n t ngoi bc
sng 366 nm v cc vt
khi phun thuc th
vanillin 2% trong
ethanol v acid sulfuric.
5 nh lng Hm lng hp cht TCCS
saponin ton phn trong
khong 19,2% - 23,4%

SVTH: PHM TH KIM KHUYN 55 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt lun v kin ngh

2.2. Xy dng tiu chun kim nghim cao

2.2.1. Cao Du tm
STT Cc tiu ch Yu cu Phng php th
1 m Khng c qu 10% DVN IV
2 tro ton phn Khng c qu 15% DVN IV
3 nh tnh
Phn ng ha hc Mu th phi cho phn ng TCCS
vi TT NaOH 20% v TT
FeCl 3 5%.
Sc k lp mng Cc vt thu c trn sc
k ca mu th phi c
gi tr R f v mu sc tng
ng vi cc vt thu c
ca
Nguyn liu du tm
Cht chun rutin
4 nh lng Hm lng flavonoid tnh TCCS
theo rutin trong khong
17,5% - 23,7%

SVTH: PHM TH KIM KHUYN 56 ThS. DNG TH MNG NGC

Kha lun tt nghip Kt lun v kin ngh

2.2.2. Cao Vin ch

STT Cc tiu ch Yu cu Phng php th
1 m Khng c qu 25 % DVN IV
2 tro ton phn Khng c qu 2% DVN IV
3 nh tnh
Phn ng ha hc Mu th phi c phn ng TCCS
Liebermann- Burchard
Cc vt thu c trn sc
Sc k lp mng k ca mu th phi c
gi tr R f v mu sc tng
ng vi cc vt thu c
ca nguyn liu Vin ch
4 nh lng Hm lng hp cht TCCS
saponin ton phn nm
trong khong 36,7% -
44,9%

SVTH: PHM TH KIM KHUYN 57 ThS. DNG TH MNG NGC

 TI LIU THAM KHO
Ting Vit
1. Dc in Vit Nam IV, Nh xut bn Y hc, 2009.
2. B mn Dc Liu, Gio trnh phng php nghin cu Dc liu, Trng i
hc Y Dc TP. H Ch Minh, 2004.
3. B Y t, Kim nghim dc phm, Nh xut bn Y hc, 2005.
4. Cy thuc v ng vt lm thuc Vit Nam - tp I, NXB KH & KT, H Ni,
182-184, 2003.
5. Tt Li, Nhng cy thuc v v thuc Vit Nam. NXB Y Hc, TP.HCM, 90-
92, 2004.
6. Hong Tr, Ti u ha quy trnh chit xut 10-deacetyl baccatin III (10-DAB) v
taxol t cnh l thng l di (Taxus wallichiana Zucc.), Kha lun tt nghip
c nhn ha hc, 2009.
7. Nguyn Khc Qunh C, Bi ging chit xut dc liu, Ti liu ging dy sinh
vin dc 5, Trng H Y Dc TP.HCM, B mn dc liu.
8. Nguyn Kim Phi Phng, Phng php c lp hp cht hu c, NXB i hc
quc gia TP. H Ch Minh, 2007.
9. Ng Vn Thu, Bi ging dc liu, tp I. Trng i hc Dc H Ni, 2011.
10. Phm Thanh K, Bi ging dc liu, tp II. Trng i hc Dc H Ni,
1998.
11. Vin Dc Liu, Cy thuc v ng vt lm thuc Vit Nam, Tp II, NXB
Khoa hc k thut, 2004.
Ting nc ngoi
12. Camila B. Pereira, Aline Marin, Tiago D. T. Maki, Raquel M. M.Necchi,
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polyphenols, flavonoids of Morus alba Leaves (MORACEAE), Sade (Santa
Maria), Ahead of Print, v.37, n.2, pp.5768, 2011.
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And The Determination Of Flavonoids Via MetalFlavonoid Complexing

v
 Reactions, Institute of Physical Chemistry, Faculty of Pharmacy, University of
Belgrade, Serbia, pp. 921-928, 2007.
14. Honglan Wang. Jie Gao, Danni Zhu and Boyang Yu, Two new Triterpenoid
Saponins isolated from Polygala japonica, China Pharmaceutical University,
2006.
15. Jun Wang, Fu An Wu, Hui Zhao, Li Liu and Qiu Sheng Wu, Isolation of
flavonoids from mulberry (Morus alba L.) leaves with macroporous resins,
African Journal of Biotechnology Vol. 7 (13), pp. 2147-2155, 4 July, 2008.
16. Kai Nie, Zhonghai Tang, Xu Wu, Xiaona Xu, Yizeng Liang, Huang Li and
Liqun Rao, Optimization of total flavonoids extraction from mulberry leaf using
an ethanol-based solvent system, Journal of Medicinal Plants Research Vol.
6(12), pp. 2373-2380, 30 March, 2012.
17. Mrghita , L. Al, D. Dezmirean, Laura Laslo, Adela Moise, Olimpia Popescu,
O. Maghear, Validated Method For Estimation Of Total Flavonoids, in
Romanian Propolis, Romania, Bulletin USAMV-CN, pp. 63 -64,2007.
18. Qing-Chun Xue, Chuang-Jun Li, Li Zuo, Jing-Zhi Yang and Dong-Ming Zhang,
Three new xanthones from the roots of Polygala japonica Houtt, Journal of
Asian Natural Products Research, Vol.11, No.5, 465469,May 2009.
19. Shreeda S. Adhyapak, Comparison of extraction techniques for quantitative
determination of Rutin from morus Alba Linn by reverse phase high
performance liquid chromatography, International Journal of Pharma and Bio
Sciences, vol 2, issue 1, pp. 751-752, 2011.
20. Walter A. Jacobs and Otto Isler, The Sapogenins Of Polygala Senega,
Laboratories of The Rockefeller Institute for Medical Research, New York,
pp. 155-157, 1937.

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