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B Y T
TRNG I HC DC H NI
CAO TH THU HNG
NGHIN CU PHN BIT C IM
THC VT, THNH PHN HA HC
CA HAI LOI NA THUC CHI
AMORPHOPHALLUS, H RY
(ARACEAE)
KHA LUN TT NGHIP DC S
H NI 2014
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B Y T
TRNG I HC DC H NI
CAO TH THU HNG
NGHIN CU PHN BIT C IM
THC VT, THNH PHN HA HC
CA HAI LOI NA THUC CHI
AMORPHOPHALLUS, H RY
(ARACEAE)
KHA LUN TT NGHIP DC S
Ngi hng dn:
1. PGS.TS. Nguyn Vit Thn
2. DS. Nguyn Th Thu Huyn
Ni thc hin:
B mn Dc liu
H NI 2014
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LI CM N
Trong qu trnh thc hin ti, em nhn c rt nhiu s quan tm, gip
, ng vin qu bu t cc thy, c gio, gia nh v bn b.
Vi lng knh trng v bit n su sc, em xin gi li cm n chn thnh ti
PGS. TS Nguyn Vit Thn ngi thy trc tip hng dn, ch bo tn tnh v to
mi iu kin gip em hon thnh kha lun.
Em xin gi li cm n chn thnh v su sc ti: DS. Nguyn Th Thu
Huyn, DS. Nguyn Thanh Tng, cc thy c gio ging vin v cc anh ch k
thut vin trong b mn Dc Liu tn tnh gip , ng vin v to mi iu
kin cho em trong sut qu trnh lm kha lun.
Em xin gi li cm n ti cc thy gio, c gio ging vin trng i hc
Dc H Ni tn tnh ch bo, ging dy, du dt v truyn nhit huyt cho em
trong sut 5 nm hc va qua.
Cui cng, em xin gi li cm n chn thnh ti gia nh v bn b lun
ng vin gip em trong sut qu trnh hc tp.
Sinh vin
Cao Th Thu Hng
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MC LC
DANH MC CC CH VIT TT
DANH MC CC BNG V HNH V
T VN ............................................................................................................ 1
CHNG 1. TNG QUAN ..................................................................................... 3
1.1. TNG QUAN V H RY (ARACEAE) ................................................... 3
1.1.1. V tr phn loi h Ry (Araceae) .............................................................. 3
1.1.2. c im thc vt v phn b ca h Ry (Araceae) ................................ 3
1.2. TNG QUAN V CHI AMORPHOPHALLUS ........................................... 4
1.2.1. c im thc vt v phn b chi Amorphophallus .................................. 4
1.2.2. c im thc vt ca mt s loi thuc chi Amorphophallus .................. 5
1.2.3. Cc nghin cu Vit Nam v chi Amorphophallus ............................... 10
CHNG 2: NGUYN LIU, PHNG TIN V PHNG PHP
NGHIN CU. ........................................................................................................ 13
2.1. NGUYN LIU, PHNG TIN NGHIN CU .................................. 13
2.1.1. Nguyn liu nghin cu ........................................................................... 13
2.1.2. Phng tin nghin cu ............................................................................ 13
2.2. PHNG PHP NGHIN CU ............................................................... 14
2.2.1. Nghin cu v mt cm quan ca hai loi Na ........................................ 14
2.2.2. Nghin cu v vi phu c v vi hc bt c ca hai loi Na ................... 14
2.2.2. Nghin cu v ha hc bt c ca hai loi Na ....................................... 15
CHNG 3. THC NGHIM, KT QU V BN LUN ............................ 16
3.1. NGHIN CU V C IM THC VT CA HAI LOI NA .... 16
3.1.1. M t v c im hnh thi ca hai loi Na .......................................... 16
3.1.2. Nghin cu vi hc bt ca hai loi Na ................................................... 17
3.1.3. Nghin cu vi phu c ca hai loi Na .................................................. 22
3.2. NH TNH S B CC NHM CHT TRONG C HAI LOI NA
BNG CC PHN NG HA HC ............................................................... 24
3.2.1. Phn ng nh tnh glycosid tim............................................................... 24
3.2.2. nh tnh saponin trong dc liu ............................................................ 26
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3.2.3. nh tnh anthranoid ................................................................................. 26
3.2.4. nh tnh tanin .......................................................................................... 27
3.2.5. nh tnh ng kh ................................................................................ 28
3.2.6. nh tnh polysaccharid............................................................................ 28
3.2.7. nh tnh acid amin .................................................................................. 29
3.2.8. nh tnh acid hu c ............................................................................... 29
3.2.9. nh tnh flavonoid .................................................................................. 29
3.2.10. nh tnh coumarin ................................................................................. 31
3.2.11. nh tnh alcaloid ................................................................................... 32
3.2.12. nh tnh cht bo, steroid, caroten ........................................................ 33
3.3. NH TNH DCH CHIT HAI LOI NA BNG SC K LP
MNG ................................................................................................................... 35
3.4. BN LUN .................................................................................................... 41
KT LUN V XUT .................................................................................... 43
1. KT LUN ....................................................................................................... 43
2. XUT.......................................................................................................... 43
TI LIU THAM KHO ...................................................................................... 44
TI LIU THAM KHO
PH LC
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DANH MC CC CH VIT TT
H: Hnh.
KH&CN: Khoa hc v cng ngh.
KM: Khoai mn.
Mu N1: Mu Na 1.
Mu N2: Mu Na 2.
Mu N3: Mu Na 3.
Mu N4: Mu Na 4.
Mu N5: Mu Na 5.
Mu N6: Mu Na 6.
Mu N7: Mu Na 7.
Mu N8: Mu Na 8.
Nxb: Nh xut bn.
P. : Phn ng.
TT: Thuc th.
UV: Ultra violet.
Tr: Trang.
: Bc sng.
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DANH MC CC BNG V HNH V
STT Tn cc cc bng v hnh v Trang
1 Bng 3.1. S khc nhau v c im hnh thi hai loi Na. 18
2 Bng 3.2. S khc nhau v c im vi hc bt hai loi Na. 18
3 Bng 3.3. Kt qu nh tnh cc nhm cht trong c ca hai
loi Na. 34
4 H 3.1.1. Cy Na mu N2. 19
5 H 3.1.2. C Na mu N2. 19
6 H 3.1.3. Cm hoa Na mu N2. 19
7 H 3.1.4. Cm qu Na mu N2. 19
8 H 3.2.1. Cy Na mu N6. 20
9 H 3.2.2. C Na mu N6. 20
10 H 3.2.3. Cm hoa Na mu N6. 20
11 H 3.2.4. Cm qu Na mu N6. 20
12 Hnh 3.3. Mt s c im bt mu N2. 21
13 Hnh 3.4. Mt s c im bt mu N6. 21
14 Hnh 3.5. S tng qut mu N2. 23
15 Hnh 3.6. S tng qut mu N6. 23
16 Hnh 3.7. Sc k dch chit thy phn hai mu Na vi h dung
mi toluen-ethyl acetat-acid formic (6:2:0,5). 36
17 Hnh 3.8. Sc k dch chit thy phn hai mu Na vi h dung
mi ether du ha-ethyl acetat-aceton-acid formic (3:1:0,5:0,05). 38
18 Hnh 3. 9. Sc k dch chit thy phn hai mu Na vi h dung
mi cloroform-aceton (9:1). 39
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T VN
Vit Nam l mt quc gia nm trong khu vc nhit i gi ma. V vy, nc
ta c thm thc vt phong ph v c nhiu tim nng v cy thuc. Theo thng k
s b ca vin dc liu nc ta c khong 14.000 loi thc vt, nm to trong
c gn 4000 cy thuc.
H Ry (Araceae) l mt h thc vt mt l mm, c c im hnh thi a
dng v phong ph. Theo PGS. TS Phm Hong H, Vit Nam c khong 30 chi,
135 loi Ry [12].
Chi Na (Amorphophallus) l mt chi thuc h Ry, c trng nhiu cc
tnh vng ni, trung du, min Nam v min Bc. Cy cho nng sut cao v chu c
iu kin khc nghit. Trc kia, mt vi a phng cy thng c trng ly c
v l dng lm lng thc, thc n trong chn nui. Hin nay, vi s pht trin ca
m hnh bnh tt tng cao, mt s loi trong chi Na c nghin cu v s dng
lm thuc. Nhiu nghin cu v thnh phn v vai tr c Na cho thy trong bt c
c cha glucomannan l hp cht c nhiu cng dng qu, c tc dng tt n bnh
ung th, tim mch, bo ph, cao huyt p... nhiu nc trn th gii c Na
c nghin cu, s dng v em li ngun li ln. nc ta cy Na t c
nghin cu v khai thc s dng hp l [13]. Mc d c rt nhiu loi Na nhng hu
ht cc nghin cu ch tp trung v hai loi Amorphophallus kojac C. Koch v
Amorphophallus peaoniifolius (Denst.) Nicols. Cc loi khc t c quan tm n,
c bit l v c im thc vt, thnh phn ha hc cng nh cch phn bit chng.
Nhm gp phn pht hin, phn bit v nng cao gi tr s dng ca tng loi
Na thuc chi Amorphophallus v gp phn bo tn, pht trin ngun gen qu
pht trin ti Thu thp ngun gen v tri thc bn a cc ngun gen nghin
cu chng ti tin hnh thc hin ti: Nghin cu c im thc vt, thnh
phn ha hc ca hai loi Na thuc chi Amorphophallus, h Ry (Araceae)
vi mc tiu:
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To c s d liu, cc l lch v thc vt, hin vi, ha hc nhm tng bc
phn bit cc loi Na, cng nh mong mun gp phn vo cng tc kim nghim
dc liu.
thc hin c mc tiu trn, ti c tin hnh vi nhng ni dung sau:
V thc vt: M t c im hnh thi cy, vi phu, bt c ca hai loi Na v
so snh chng.
V ha hc: nh tnh cc nhm cht bng phng php ha hc, sc k lp
mng dch chit methanol, thy phn v so snh chng.
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CHNG 1. TNG QUAN
1.1. TNG QUAN V H RY (ARACEAE)
1.1.1. V tr phn loi h Ry (Araceae)
Theo t in cy thuc Vit Nam ca V Vn Chi (2012), trung tm d liu
thc vt Vit Nam v cc ti liu phn loi thc vt v tr phn loi ca h Ry
trong gii thc vt nh sau:
Gii thc vt (Plantae)
Ngnh Ngc Lan (Magnoliophyta)
Lp Hnh (Liloopsida)
Phn lp Ry (Aridae)
B Trch t (Alismatalus)
H Ry (Araceae) [7], [9], [24], [28].
1.1.2. c im thc vt v phn b ca h Ry (Araceae)
Cy c mc ni m, sng dai nh thn r. Thn nc hoc leo mang nhiu l
kh sinh thng xung. L thng n, c b, gn lng chim, chn vt hay song song.
Cm hoa bng mo nc, khng phn nhnh, mang nhiu hoa. Mo thng c mu sc
s, mt s c mi thm. Trc hoa c th mang hoa khp b mt hay kt thc bng
mt phn khng mang hoa, thng c hoa ci di, hoa c trn. Hoa nh
khng cung hay cung khng r rt, n tnh, lng tnh hay tp tnh. Hoa lng
tnh thng c 3 vng hoa, mi vng c 3 b phn; hoa n tnh c bao hoa tiu
gim hay trn. B nh c 2 vng, mi vng c 3 nh c khi cn 1 nh hoa n tnh.
B nhy thng c 2-3 l non, c khi ch 1 l non hoa n tnh, cha 1-nhiu
non o, cong hay thng [7], [24].
Qu mng mt hoc nhiu ht. Ht c ni nh nc [7], [24].
Theo sch thc vt, cy phn b vng nhit i, cn nhit i, mt s l n i.
Vit Nam c khong 30 chi, 135 loi mc hoang v c trng lm thc n cho ln (Ry,
Na), lng thc (Khoai s), rau n (cc loi Mn, S), cy cnh (Vn Nin Thanh). C
12 loi thng dng lm thuc trong c 4 loi thng dng trong cng nghip dc l
Bn h, Thch xng b, Thy xng b, Thin nin kin [7].
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Mt s chi in hnh ca h ry [7]:
Chi Acorus Chi Algaonema
Chi Amorphophallus Chi Aresonema
Chi Colocasia Chi Epipremnum
Chi Homalomena Chi Pinellia
Chi Pistia Chi Pothos
Chi Rhaphidophora Chi Typhonium
1.2. TNG QUAN V CHI AMORPHOPHALLUS
1.2.1. c im thc vt v phn b chi Amorphophallus
1.2.1.1. c im thc vt chi Amorphophallus
Cy a m, chu bng, thng mc di tn cy rng m trn ni t v ni
vi, t xp nhiu mu, pH t trung bnh n kim. Cy ra hoa hng nm ti sinh
ch yu bng ht. Cy mc t ht sau 2-3 nm mi c hoa. Phn trn mt t tn li
hng nm vo ma ng [1].
Theo t in thc vt thng dng, cy thn tho c dng r c. L c dng
lc, c phin chia nhiu hay t [10].
Cm hoa gm mt bng mo khng c hoa bt th, nh ca bng c mt
phn ph hnh nn, tt c c bao trong mt ci mo dng sng, lm m nu v
trng [9], [10].
1.2.1.2. Phn loi thc vt v phn b chi Amorphophallus
Chi Amorphophallus c khong 170 loi phn b ch yu vng nhit i
chu , chu Phi, Mardagascar, Australia. Vit Nam c khong 25 loi c thn
(c) to c nhiu tinh bt, n c v c 2 loi Amorphophallus konijac C. Koch v
Amorphophallus paeoniifolius (Denst.) Nicols l thng dng [10].
Na (chi Amorphophallus) phn b ch yu cc tnh vng ni (c cao
thng di 1000m) v trung du ca min Bc v min Nam. Vit Nam c ngun
Na phong ph. Tng lng c tnh c khong 1000 tn [10].
Mt s loi in hnh thuc chi Amorphophallus [12]:
Amorphophallus konijac C. Koch.
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Amorphophallus paeoniifolius (Denst.) Nicols.
Amorphophallus interrutuptus Engler & Gehrm.
Amorphophallus verticillatus Hett.
Amorphophallus mekongeasis Engler & Gehrm.
Amorphophallus panomensis Gagn.
Amorphophallus tonkinensis Engler & Gehrm.
Amorphophallus arnoutovil Hett.
Amorphophallus pilosus Hett.
Amorphophallus lanuginosus Hett.
Amorphophallus coeateus S. Y. Liu & S. J. Wei, Guihaia.
1.2.2. c im thc vt ca mt s loi thuc chi Amorphophallus
1.2.2.1. Amorphophallus konijac C. Koch
Tn khc: C Na, Khoai na, Khoai ngt.
c im thc vt v phn b
Cy tho ln, sng lu nm, cao 50-70cm. Thn c to, mt trn lm, mt
di li v mang nhiu u trn, v ngoi mu nu, rut trng vng. L mc thng t
thn c, sau khi cy ra hoa, thng ch c 1 (t khi 2 l) c cung di v mp, mu
nu, c nhng im m trng, phin l chia thnh 3 nhnh, cc nhnh li phn chia
tip v ch thy su hnh lng chim, hai mt nhn. Cm hoa mang trn cung mp
di, mc thng ng, cao 30-40cm, c mo to, mt ngoi mu lc, mt trong mu
ta, mp un ln, bng mo di gp i mo mang phn hoa ci di ngn, phn
hoa c gia di hn v phn ph trn di gp 3-4 ln phn kia. Hoa khng c
bao hoa, hoa c c nh ri, hoa ci c bu hnh trng. Hoa c mi kh ngi. Qu
mng [1], [14]. Cy mc hoang rng tha, ven sui, ch m, t nhiu mn. C
ni trng ly c n v nui ln [16].
Cc b phn u c dch nga. Cung l v c n c. C lm thuc cha
bnh ng tiu ha v trng c cm khu. Ma hoa t thng 3-5 [1], [14].
B phn dng
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Thn c, thu hoch vo thng 9-11 co sch v, v r con, chn hay sy kh.
Khi dng ngm cho mm, thi mng, ngm nc phn v gng (c 1kg c Na cn
300g gng ti v 50g phn chua) trong 24 gi ri sao thm cho ht nga [1].
Thnh phn ha hc
Tinh bt (thnh phn ch yu l Konjacmannan), cht gy nga [14], [16].
Cng dng
C Na c v cay v nga, c tc dng dit khun, tiu vim, h st, gii c.
Tinh bt dng nu n v nu ru [1], [16].
Tc dng dc l
Tc dng khng khun: Dch chit ethanol ca c Na trn mi trng nui
cy c tc dng c ch Bacillus diphtheria, Bacillus typhi, Streptococus
hemolyticus, nng c ch 3 chng vi khun trn ln lt l l 62,5g/l; 5,25g/l;
250g/l [1].
Tc dng chng vim: Dch chit ethanol ca c Na th nghim trn chut
cng trng, tim thng vo d dy vi liu 15g/kg, dng 7 ngy lin tip, c tc
dng c ch ph chn chut do albumin gy nn [1].
Tc dng i vi tim mch: Dng dch chit ethanol t c Na (1:1) trn tiu
bn tai th c lp vi liu 2ml/ln cho vo dch truyn c tc dng gy gin mch.
Tc dng ny c lin quan ti th th 2 b kch thch. Trn tim ch c lp dng dch
chit vi nng 1:2 n 1:16 c tc dng c ch co bp tim, trn th gy m vi
liu 15g/kg tim tnh mch hoc tim phc mc u c tc dng h huyt p [1].
Tc dng h lipid mu: Th nghim trn chut cng trng c nng lipid cao
thc nghim, trn c Na vi thc n hng ngy ca chut vi t l 2,5; 5,0 v 10%
c tc dng gim cholesterol huyt thanh. Dng dch chit ethanol ca c Na cng
c tc dng lm gim lipid mu [1].
Tc dng khc: Dch chit ethanol t Na dng ng ung vi liu 15g/kg
c tc dng lm gim nht ca mu ton phn chut th nghim.
Ngoi ra c Na cn c tc dng i khng vi tnh trng thiu oxy sc vt
th nghim, ko di thi gian sng [1].
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Bi thuc dn gian thng dng
Cha lit na ngi (sau tai bin mch mu no, chn thng nng sau tht
lung): C Na sng 10g, u 1g, Ph t 1g, nc 600ml nu cn li 100ml, chia
ung nhiu ln trong ngy, ung sau khi n no (thng c c, cn thn khi dng).
Cha mn nht, sng ty, rn cn: C ti gi ra p ln mn nht v vt thng.
Cha st rt, n chm tiu: C Na phi kh 4-12g sc ung.
Cha ho, nhiu m: C Na, Trn b, Bn h nam mi th 40g sao thm, tn
mn, dng nc ct gng quy h, lm vin bng ht ng mi ln dng 30 vin. Tr
em di 10 tui mi ln dng 10-15 vin [16].
1.2.2.2. Amorphophallus paeoniifolius (Denst.) Nicols
Tn khc: Na chung, Khoai na.
c im thc vt v phn b
Cy tho sng hng nm c thn c nm trong t; c hnh bn cu rng, ng
knh ti 20cm, mt di li mang mt s r ph, v c nhng nt nh c khoai ty
xung quanh c 3-5 mu li. V c c mu nu, tht trng vng v cng. L mc sau
khi c hoa, thng ch c 1 l c cung cao ti 1,5m, mu xanh thm c m bt
trng; phin l chia lm 3 nom ta nh l u . Cm hoa gm mt mo khng cung
hoc cung ngn di khong 70cm, bng mo mu xanh c m trng, mt trong
thm, bao ly mt bng mo l mt phn mang phn hoa ci di phn hoa c
trn. Phn hoa ci c hnh tr, phn hoa ci hnh nn ngc. Hoa c mi kh ngi.
Qu mng, mu xanh, khi chn c mu vng cam ti [10], [24].
Cy thng phn b n , Mianma, Trung Quc, Vit Nam, Campuchia,
Malaixia v Philiphin. Vit Nam cy mc hoang v trng ph bin cc tnh
Lng Sn, Qung Ninh, Ha Bnh, H Ty, Ngh An, H Tnh vo ti Qung Tr,
Tha Thin Hu [9], [10].
B phn dng
Thn c thu hoch sau khi cy tn, co sch v ngoi thi mng [16].
Thnh phn ha hc
Tinh bt (thnh phn ch yu glucomannan), cht gy nga [16].
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8
Tc dng dc l
Tc dng chng oxy ha: Nghin cu cho thy, dch chit ethanol ca
Amorphophallus paeoniifolius (Denst.) Nicols c tc dng c ch peroxyd lipid
gim t 4,3% n 67,2% ph thuc vo liu. Kh nng peroxyd lipid ca dch chit
cn ca chng da trn gc t do 1,1-diphenyl-2-picrylhydrazyl-2 (DPPH) v kh
nng c ch t l phn trm da trn 2,2-azinobis-(-3-ethyl) benzo-thiozoline-6-
sulfonate (ABTS +) v H2O2. Dch chit ethanol ca Amorphophallus paeoniifolius
(Denst.) Nicols cho thy ti a l 68,6% hot ng peroxyd lipid l nh gc t do
DPPH v c ch ti a 74% v 67,2% trong trng hp tng ng l ABTS v
H2O2. Hiu qu chng oxy ha v c ch qu trnh oxy ha ca dch chit ethanol
ca Amorphophallus paeoniifolius (Denst.) Nicols c tm thy l ph thuc vo
liu nng th nghim ca 1-50 g/ml. Sc k lng hiu nng cao (HPTLC) cho
thy s xut hin ca cht polyphenol nh axit galic, resveratrol, quercetin v hai
hp cht khng xc nh. Nh vy kt qu cho thy dch chit ethanol ca
Amorphophallus paeoniifolius (Denst.) Nicols c hot tnh chng oxy ha mnh
trong phng th nghim v c th c s dng nh mt ngun cht chng oxy ha
an ton v hiu qu [22].
Tc dng ln h thn kinh trung ng: Khi nghin cu tc dng ca dch chit
ether du ha ca Amorphophallus paeoniifolius (Denst.) Nicols trn nhm 5 nhm
chut (chut bch), tim phc mc vi liu nh sau: nhm I liu 1500mg/kg, nhm
II vi liu 2000mg/kg, nhm III vi liu 2500mg/kg, nhm IV vi liu 3000mg/kg,
ng vt nhm V vi liu 3500mg/kg cho thy gy ra trm cm nhm ny, t l t
vong gy ra mc liu 2500 mg/kg v c th s dng an ton mc liu
1500mg/kg. Tc ng ca dch chit ny ln hot ng ca h thn kinh trung ng
l gy ra hot ng trm cm nh: bun ng v lm gim hot ng vn ng ca
ng vt th nghim. C ch ca hot ng trm cm ny cha c xc nh r
rng, nhng ngi ta cho rng l do nh hng trn v no. Hot ng trm cm
cao im c quan st thy 60 pht k t khi tim dch chit. Thi gian phc
hi hot ng trm cm ca dch chit ether du ha (1000mg/kg) v diazepam
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9
(1,5mg/kg) c tm thy tng ng l 24 gi v 20 gi. Phenobarbiton v
diazepam c bit n thuc gim au thn kinh trung ng v cng cho thy hot
ng trm cm thn kinh trung ng mt cch ph thuc vo liu. Mt tc dng
hip ng ng k ca cc dch chit ether du ha vi diazepam c tm thy
trong khi c rt t tc dng hip ng vi phenobarbiton. Dch chit ether du ha
ca Amorphophallus paeoniifolius (Denst.) Nicols gy ra hot ng trm cm ng
k ln thn kinh trung ng v hot ng ph thuc vo liu [26].
Tc dng chng vim: Dch chit methanol ca Amorphophallus paeoniifolius
(Denst.) Nicols c hot ng chng vim ni bt trong khi dch chit cloroform c tc
dng nh hn. Ba gi sau khi tim carrageenan, dch chit methanol vi liu 200 v
400mg/kg sn xut 37,5% v 45,83% c ch khi so snh vi nhm i chng [20].
Tc dng gim au: Pht hin dch chit methanol ca Amorphophallus
paeoniifolius (Denst.) Nicols liu 250 v 500mg/kg trng lng c th dng mng
bng c tc dng gim au ng k [21], [27].
Tc dng khng khun: Dch chit ethanol ca Amorphophallus paeoniifolius
(Denst.) Nicols c tc dng khng khun, khng nm, v cc tc nhn gy c t bo [23].
Tc dng dit giun: Dch chit methanol t c ca Amorphophallus
paeoniifolius (Denst.) Nicols c nghin cu chng li antihelmintic pheretima
posthuma v tubifex. Dch chit methanol vi nng 25, 50 v 100 mg/ml c
th nghim trong cc xt nghim sinh hc, c lin quan n xc nh thi gian t
lit v thi gian gy cht ca giun [21].
Cng dng
C Na chung c v cay, tnh nng, c c. C c tc dng li tiu ha b
dng hi phc sc khe, li trung tin, dng cha tr kit l. Nu dng ti c
tc dng kch thch lm long m v c th dng tr thp khp cp tnh. Na
chung c th dng vi cng dng nh c Na, c trng ch yu ly bt. Dc
Na chung cng n c, nhng phi ngm nc vo go cho ht nga [16].
-
10
Bi thuc dn gian thng dng
Dng lm thc n ph cho ngi tiu ng: Thn c Na chung thu hoch
sau khi tn cy, co sch v phi hay sy kh. Khi dng nu chn nh hay phi hp
vi cc v thuc khc [16].
1.2.2.3. Amorphophallus coeataneus S. Y. Liu & S. J. Wei, Guihaia
c im thc vt v phn b
C trn, mu nu n en, kch thc 3x6cm, y chia nhnh t nhnh hnh
thnh 2-6 c mi, c mi c kch khong 3-5cm.
L thng c 1-3 cung mc t c, cung l mp mng nc, nhn, mu xanh
thm, c kch thc 60-120x1-2cm; phin l c kch thc khong 100cm chia
thnh 3 thy, thy x lng chim 2-3 ln, phin l thun nhn, mp l nguyn, mt
trn xanh bng, mt di xanh lc. Cm hoa bng mo di, c kch thc 10-50x0,6-
1,3cm; l mo mu trng xanh, phn trn hnh tam gic hp, phn di thun hnh
trng, c kch thc 7,5-15x3-7cm; hnh bao quanh cm hoa cung hoa xanh v
nhn nh cung l. Hoa c mi kh ngi [24].
Qu mng mu xanh m hnh bu dc.
B phn dng: Thn c [24].
1.2.3. Cc nghin cu Vit Nam v chi Amorphophallus
Nm 2012, khi Nghin cu trng v pht trin cy Na konjac
(Amorphophallus) v mt s loi khc trong chi Na (h RyAraceae) Vit Nam
hng ti vic ly c lm nguyn liu sn xut thc phm chc nng v thuc iu
tr bnh tiu ng, m mu v bo ph TS. Trn Vn D xc nh c bn loi Na
c cho l c trin vng lm ngun nguyn liu tch chit glucomannan l cc loi Na
konjac (Amorphhophallus konjac K.. Koch), Na u nhn (Amorphophallus
corrugatus N. E. Br), Na krausei (Amorphophallus krausei Engler, Pflanzenr) v Na
Vn Nam (Amorphophallus yunnanensis Engler, Pflanzenr). Nghin cu xc nh c
hm lng glucomannan c cy Na Chung (Amorphophallus paeoniifolius (Denst.)
Nicols) l 5% v cy Na u nhn (Amorphophallus corrugatus N. E. Br) l 10% trng
-
11
lng ti, tng ng 25% v 50% trng lng kh. ng thi, pht hin c qun th
loi Na konjac Vit Nam ti 2 huyn ng Vn v Qun B tnh H Giang [32].
Nm 2013, trong Nghin cu quy trnh tch chit, cu trc ha hc v hot tnh
sinh hc ca glucomannan t cy Na-Amorphophallus sp. (h ry-Araceae) ca TS.
Trn Th Nhi (Vin ha hc Vin Hn lm Khoa hc v Cng ngh Vit Nam)
nghin cu v xc nh c tn khoa hc ca 3 loi Na Vit Nam
l: Amorphhophallus konjac C. Koch ; Amorphophallus corrugatus N. E. Br v
Amorphophallus paeoniifolius (Dennst.) Nicolson. Trong loi Amorphophallus
konjac C. Koch mi c pht hin Vit Nam nm 2012. Hm lng glucomannan
trong thn (c) ca mi loi Na ti Vit Nam c nghin cu v xc nh bng
phng php so mu thuc th 3,5-DNS. Kt qu ca nghin cu l c s xy dng quy
trnh tch chit glucomannan quy m phng th nghim t 3 loi Na trn. Trong nghin
cu ny cu trc ha hc tnh cht ha l ca sn phm cng c xc nh [29].
Nm 2014, trong Nghin cu hm lng, cht lng, tc dng dc l v xy
dng quy trnh sn xut glucomannan trong c Na-Amorphophallus sp. (h Ry-
Araceae) trng ti tnh Tha Thin Hu thuc S KH&CN tnh Tha Thin Hu
xc nh c cc c im sinh hc, cc ch tiu sinh trng, iu kin sinh thi v
quy m trng trt ca loi Na hin c ti Tha Thin Hu. Mu nghin cu ly ti
x Qung Th. Ti hin nay c gn 10ha Na c trng trong v ng, em li
thu nhp 5-7 triu ng/so/v, bnh qun mi ha Na thu nhp trn 100 triu
ng/v. Cy Na l cy c kh nng chu c iu kin khc nghit, thi gian sinh
trng ngn, v c th trng tri v nn c ngi dn a vo trng v mang li
gi tr kinh t cao. ti lm sng t thnh phn ha hc, tnh cht ca
glucomannan, sn phm bt glucomannan thu c c dng ht mn, trng, hiu sut
tch chit t t l 30,4% trn khi lng c Na kh. ti cng nghin cu
c tc dng h ng huyt ca glucomannan t c Na trn chut b gy i tho
ng bng streptozocin, tc dng h cholesterol mu bng th nghim trn chut
gy tng cholesterol ngoi sinh v th gy tng cholesterol ni sinh v kh nng
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12
chng bo ph theo phng php Kobayasshi-xc nh lng m quanh thn, m
mo tinh hon trn chut cng trng [29].
Cng nm 2014, Nghin cu sinh trng ca cy Na Chung
(Amorphophallus Paeoniifolius (Denst.) Nicols) Tha Thin Hu ca trng
i hc Hu xc nh: Nng sut l thuyt ca c Na khong 92 tn/ha, nng
sut thc t t 82,8 tn/ha. Kt qu ny cho thy nng sut c Na cao hn rt
nhiu so vi khi trng nhng i tng khc nh khoai Lang v khoai Mn (nng
sut khoai Lang c coi l c trin vng t khong 11-23,6 tn/ha, nng sut c
trung bnh ca ging khoai Mn cao sn KM-1 khong 50-60 tn/ha (Nguyn Vn
Mi, 2001)) [15]. ti cng nghin cu hm lng glucomannan trong c Na,
kt qu nghin cu cho thy rng hm lng glucomannan trong c chnh chim
35,4%, trong c nhnh l 30,7% cao hn hm lng glucomannan m Liu v cng
s cng b c Na c trng Pinglan (t khong 20,1%) v cao hn so vi
Na c trng anyuan, Qijiang (12,5%) trong nghin cu ca cng tc gi trn
Trung Quc (Liu Pei-Ying, Wang Yulan, 1990; Liu Pei-Ying v nnk., 1998) [25].
Trong khi , hm lng tinh bt ca khoai ty dao ng t 8-30%, ca khoai lang
l 15-31%, ca sn l 20-25% (Nguyn Th Phng, 2011). Nghin cu xc nh
c tim nng cung cp tinh bt thc phm ca c Na l rt ln, c th m ra
mt hng mi trong vic dng c Na lm nguyn liu cho cc nh my sn xut
tinh bt [13].
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13
CHNG 2: NGUYN LIU, PHNG TIN V PHNG PHP
NGHIN CU.
2.1. NGUYN LIU, PHNG TIN NGHIN CU
2.1.1. Nguyn liu nghin cu
Nguyn liu nghin cu gm 8 mu dc liu sau:
- Mu Na 1 (k hiu mu N1) thu hi an Phng- H Ni vo 10/2013.
- Mu Na 2 (k hiu mu N2) thu hi Nho Quan-Ninh Bnh vo 1/2014.
- Mu Na 3 (k hiu mu N3) thu hi Nho Quan-Ninh Bnh vo thng 1/2014
(mu ny c ngun gc H Tnh v c trng Ninh bnh mt nm tnh n
thi im thu hi).
- Mu Na 4 (k hiu mu N4) thu hi Nho Quan-Ninh Bnh vo thng 1/2014
(mu ny c ngun gc Hu v c trng Ninh bnh mt nm tnh n thi
im thu hi).
- Mu Na 5 (k hiu mu N5) thu hi ti vn Quc Gia Ba V vo thng 2/2014.
- Mu Na 6 (k hiu mu N6) thu hi ti vn Quc Gia Ba V vo thng 2/2014.
- Mu Na 7 (k hiu mu N7) thu hi ti vn Quc Gia Ba V vo thng 2/2014.
- Mu Na 8 (k hiu mu N8) thu hi ti Ph Quc vo thng 4/2014.
Cc mu sau khi thu hi, mt phn c trng vn Thc Vt trng i
hc Dc H Ni v Nho Quan-Ninh Bnh lu mu tip tc nghin cu v
gim nh tn khoa hc ca cc loi.
Cc mu nghin cu c ra sch, co b lp bn, thi lt v sy kh, nghin
th bng thuyn tn ly mu nghin cu.
Sau khi nghin cu 8 mu Na trn, chn 2 mu N2 v mu N6 so snh v
phn bit trong ti.
2.1.2. Phng tin nghin cu
2.1.2.1. Ha cht v dng c
- Ha cht v thuc th trong nghin cu t tiu chun phn tch theo Dc in
Vit Nam IV [8].
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14
- Ha cht: Javen, acid acetic, xanh methylen, son phn, nc ct, cloral hydrat
25%, natri sulfat khan.
- Dung mi hu c: Cloroform, n-butanol, ethyl acetat, acid formic, ether du ha,
toluen
- Thuc th: Cc thuc th dng trong phn ng nh tnh v sc k.
- Bn mng trng sn silicagel 60F254 ca Merck.
- Dng c: Dng c thy tinh v cc dng c khc dng trong phng th nghim
(cc c m, bt s, chy, ci, thuyn tn, a thy tinh, lam knh, bnh nn)
2.1.2.2. Thit b dng trong nghin cu
- Knh hin vi Labomed.
- Cn k thut Sartorius.
- Cn phn tch Precisa.
- T sy.
- My chm sc k CAMAG LINOMAT 5.
- My chp nh sc k CAMAG REPROSTAR 3.
- My vi tnh vi phn mm winCATS v VideoScan.
- My nh Canon, Sony.
2.2. PHNG PHP NGHIN CU
Phng php xy dng tiu chun kim nghim v kim nh theo tiu chun
Dc in Vit Nam IV [8].
2.2.1. Nghin cu v mt cm quan ca hai loi Na
Quan st, m t dc liu v c im thc vt, hnh dng, mu sc, mi v,
kch thc bng mt thng v chp nh [4].
2.2.2. Nghin cu v vi phu c v vi hc bt c ca hai loi Na
c im vi phu: Mu Na (c) c ct vi phu bng dng c ct cm tay,
ty v nhum theo phng php nhum kp, quan st di knh hin vi xc nh
c im vi phu v chp nh cc c im vi phu di knh hin vi sau v s
tng qut [2], [4], [17].
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15
Soi bt: Mu Na (c) c thu hoch, sa sch, sy kh, c nghin nh
thnh bt bng thuyn tn v ci s, ry ly bt mn, ln tiu bn v quan st, chp
nh di knh hin vi xc nh cc c im bt [2], [17].
2.2.2. Nghin cu v ha hc bt c ca hai loi Na
Hai mu Na sau khi thu hi c ra sch, co v, sy kh v nghin th
bng thuyn tn lm cc phn ng kim nghim bng phng php ha hc nh
sau:
- nh tnh cc nhm cht hu c c trong dc liu bng phn ng ha hc theo
cc ti liu dc liu hc [5], [6], thc tp dc liu phn kim nghim bng
phng php ha hc [3], phng php nghin cu ha hc cy thuc [11], ha
hc saponin [16].
- Sc k lp mng [11]:
Chun b dch chit methanol v dch chit thy phn
S dng bn mng Silicagel 60F254 trng sn ca Merck, hot ha 110oC
trong 1 gi, bo qun trong bnh ht m.
Chm sc k bng my chm sc k CAMAG LINOMAT 5.
Hin vt cc bc sng 254nm v 366nm, phun thuc th hin mu Vanilin
1%/ H2SO4.
Chp nh sc k v phn tch sc k .
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16
CHNG 3. THC NGHIM, KT QU V BN LUN
3.1. NGHIN CU V C IM THC VT CA HAI LOI NA
3.1.1. M t v c im hnh thi ca hai loi Na
Mu N2: Cy c cao 1-1,5m, sng hng nm c thn c. C hnh cu c mt
trn lm mt di li, xung quanh c nhiu r con, trn thn c c nhiu nt sn
nh c khoai ty. L mc t thn c. Cung l hi sn c nhiu m trng trn nn
xanh nht. Phin l c mu xanh nht, ch thy 3, thy x lng chim nhiu ln.
Cm hoa mc t c, cm hoa mo mu xanh c m trng, mt trong thm,
bao ly mt bng mo l mt phn mang phn hoa ci di phn hoa c trn.
Qu mng hnh bu dc, mu vng cam.
Dc liu l thn c, c ng knh 20cm, mu nu nht, c non c tht trng,
c gi c tht mu trng vng. C gy nga nh, n khng nga ch hi t u li.
Hnh nh c im hnh thi ca mu N2 c trnh by cc hnh 3.1.1,
3.1.2, 3.1.3, 3.1.4 (tr.19).
Mu N6: Cy c cao 50-70cm, sng hng nm c thn c mt phn ni ln trn
mt t. C di, chia thnh tng t, mu nu n en, ng knh ca mt ct
ngang c khong 3-6cm. L mc t thn c. Cung l nhn. Cung v phin l c
mu xanh m. Phin l ch thy 3, thy x lng chim nhiu ln. Cm hoa mc t
c, cm hoa mo mu trng xanh. Qu mng mu xanh.
Dc liu l thn c di, khi ct b mt cc lt ct nht, mu nu vng chia
thnh tng khc hnh cu. C non c tht mu trng xm, c gi c tht mu trng
vng. C gy nga nh.
Hnh nh c im hnh thi ca mu N6 c trnh by cc hnh 3.2.1,
3.2.2, 3.2.3, 3.2.4 (tr.20).
Nhn xt: Sau khi quan st cc c im thc vt ca 2 mu Na ta c th s
b phn bit c cc c im thc vt ca 2 mu Na da vo cc c im c
trnh by bng 3.1 (tr. 18).
Sau khi phn tch cc c im hnh thi ca cc b phn ca hai loi Na mu N2
v N6, cn c vo cc ti liu hin c ti trng i hc Dc H Ni, cng vi s
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17
t vn ca TS. Nguyn Quc Huy, chng ti xc nh tn khoa hc ca 2 mu
nghin cu nh sau:
Mu N2: Amorphophallus paeoniifolius (Dennst.) Nicolson, h Ry (Araceae).
Mu N6: Amorphophallus coaetaneus S. Y. Liu & S. J. Wei, h Ry (Araceae).
3.1.2. Nghin cu vi hc bt ca hai loi Na
Tin hnh:
Sy kh dc liu nhit 60, nghin bng thuyn tn v chy ci s thnh
bt. Ry bt qua ry 180 m. Dng kim mi mc ly bt dc liu cho ln phin
knh nh sn mt git nc ct, t lamen ln v quan st trn knh hin vi. Chn
cc c im c trng chp nh. Cc c im c tp hp thnh nhm theo
phng i.
Kt qu v nhn xt:
Mu N2:
- c im bt: Bt mu trng hi nu, khng mi, khng v.
- Soi trn knh hin vi thy cc c im sau:
Tinh bt xp ring l hay thnh m. C tinh bt n v tinh bt kp bi. Tinh
bt c nhiu hnh dng (hnh trn, hnh a gic) kch thc t 0,008mm n
0,024mm (8), (9), (10). Mnh m mm c thnh mng cha tinh bt v tinh th
canxi oxalat (1), (2). C 2 loi tinh th canxi oxalat hnh kim ng ring r hoc
thnh m. Loi 1 c kch thc 0,148mm (4). Loi 2 c kch thc 0,048mm (5).
C nhiu mnh mch (5), (6), (7). (hnh 3.3 tr.21)
Mu N6:
- c im bt: Bt mu xm en, khng mi, khng v.
- Soi trn knh hin vi thy c cc c im sau:
Tinh bt xp ring l hay thnh m, c dng kp bi. Tinh bt n c kch
thc t 0,004mm n 0,006mm (9), (10). Mnh m mm c thnh mng cha tinh
bt v tinh th canxi oxalat (1), (2). C hai loi tinh th canxi oxalat hnh kim ng
ring r hoc thnh m. Loi 1 c kch thc 0,328mm (6). Loi 2 c kch thc
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18
0,036mm (8). Tinh th canxi oxalat hnh cu gai c kch thc 0,024 mm (7). V c
nhiu mnh mch (3), (4), (5). (hnh 3.4 tr.21)
Nhn xt:
Khi quan st hai bt trn knh hin vi, ta c bn phn bit c 2 bt da vo
cc c i m c thng k trong bng 3.2 (tr.18).
Bng 3.1. S khc nhau v c im hnh thi ca hai loi Na
c im Mu N2 Mu N6
Cung l Hi sn, c nhiu m trng. Xanh nhn.
C C n hnh cu. Gm nhiu t
Hoa Bng mo mu thm, l mo xe ngang.
Bng mo mu trng xanh, l mo m ly trc hoa.
Qu Mng, mu cam hoc Mng, mu xanh
Bng 3.2. S khc nhau v c im vi hc bt ca hai loi Na
c im Mu N2 Mu N6
Cm quan Bt c mu trng nu Bt c mu xm en
Tinh bt
C nhiu hnh dng (hnh trn,
hnh a gic, tinh bt kp i,
kp ba v kp nhiu ln) v
c kch thc ln hn
C hnh a gic, t tinh bt n,
thng l tinh bt kp v c kch
thc nh hn
Tinh th
canxi oxalate
C 2 loi tinh th canxi
oxalate hnh kim. Loi 1 c
kch thc 0,148mm. Loi 2
c kch thc 0,048mm.
C 3 loi tinh th canxi oxalat
gm 2 loi hnh kim. Loi 1 c
kch thc 0,328mm. Loi 2 c
kch thc 0,036mm. Tinh th
canxi oxalat hnh cu gai c kch
thc 0,024 mm.
Mu
Mu
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19
Ch thch: H 3.1.1: Cy Na mu N2 H 3.1.2: C Na mu N2
H 3.1.3: Cm hoa Na mu N2 H 3.1.4: Cm qu Na mu N2
H 3.1.1 H 3.1.2
H 3.1.4 H 3.1.3
-
20
Ch thch: H 3.2.1: Cy Na mu N6 H 3.2.2: C Na mu N6
H 3.2.3: Cm hoa Na mu N6 H 3.2.4: Cm qu Na mu N6
H 3.2.1 H 3.2.2
H 3.2.3 H 3.2.4
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21
Hnh 3.3. Mt s c im bt mu N2
1.Mnh m mm 2. Mnh m mm mang ht tinh bt
3,4. Tinh th canxi oxalat hnh kim (loi 1) 5,6,7. Mnh mch 8,9. Tinh bt kp
10. m tinh bt n 11. Tinh th canxi oxlat hnh kim (loi 2)
Hnh 3.4. Mt s c im bt mu N6
1.Mnh m mm 2. Mnh m mm mang b tinh th 3,4,5. Mnh mch
6. Tinh th canxi oxalat hnh kim (loi 1) 7. Tinh th canxi oxalat hnh cu gai
8.Tinh th canxi oxalat hnh kim (loi 2) 9,10. Tinh bt kp
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22
3.1.3. Nghin cu vi phu c ca hai loi Na
Tin hnh:
- Mu c hai loi Na c ct bng dng c ct vi phu cm tay, chn cc lt ct mng.
- Ty cc lt ct bng cloramin B pha bo ha ti khi lt ct trng hon ton.
Ra nhiu ln bng nc.
- Ngm trong dung dch acid acetic 5% trong 15 pht. Ra li nhiu ln bng nc.
- Nhum xanh methylen trong vng 30 pht. Ra li nhiu ln bng nc.
- Nhum son phn trong vng 10 pht. Ra li nhiu ln bng nc.
- t vi phu vo mt git glycerin trn phin knh, y lamen, soi trn knh hin vi
v quan st cc c im. V s tng qut.
- Sau khi quan st vi phu hai mu c Na trn knh hin vi ta c th m t c
cc c im sau:
Mu N2: T ngoi vo trong gm c: Lp bn gm 4-5 hng t bo hnh ch
nht xp u n thnh dy xuyn tm. Tip theo lp bn l lp m mm v gm
nhiu t bo hnh a gic thnh mng xp u n. Trong lp m mm v c
nhiu b tinh th canxi oxalat hnh kim. Ri rc trong m mm l cc b libe g.
Nm su bn trong l lp m mm rut gm nhng t bo hnh trn hay a gic
thnh mng. (hnh 3.5 tr.23)
Mu N6: T ngoi vo trong gm c: Lp bn gm 5-7 hng t bo hnh ch
nht xp u n thnh dy xuyn tm. Tip theo lp bn l lp m mm v gm
nhiu t bo hnh a gic thnh mng xp u n. Trong lp m mm v c
nhiu b tinh th canxi oxalat hnh kim v nhiu tinh th canxi oxalat hnh cu gai.
Ri rc trong m mm l cc b libe g. Nm su bn trong l lp m mm rut
gm nhng t bo hnh trn hay a gic thnh mng. (hnh 3.6 tr.23)
Nhn xt: Sau khi quan st c im vi phu ca hai mu dc liu ta thy v c bn
chng c cc c im ging nhau. Tuy nhin s lng tinh th canxi oxalat ca mu N6
nhiu hn so vi mu N2 v mu N6 c nhiu tinh th canxi oxalat hnh kim nm tp
trung trong m mm v st vi lp bn.
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23
Hnh 3.5 S tng qut mu N2
1. Bn 2. M mm 3. B tinh th canxi oxalat hnh kim 4. B libe g
Hnh 3.6 S tng qut mu N6
1. Bn 2. M mm 3.Tinh th canxi oxalat hnh cu gai
4. B tinh th canxi oxalat hnh kim 5. B libe g
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24
3.2. NH TNH S B CC NHM CHT TRONG C HAI LOI NA
BNG CC PHN NG HA HC
3.2.1. Phn ng nh tnh glycosid tim
Cn 5g bt c Na cho vo bnh nn dung tch 250ml. Thm 100ml ethanol
25% ri ngm trong 24 gi. Gn dch chit vo cc c m dung tch 100ml. Thm
vi ml ch acetat 30%, khuy u. lng, lc dch vo mt cc c m dung tch
100ml. Nh vi git dch lc u tin vo mt ng nghim, thm mt git ch
acetat. Nu xut hin ta th ngng lc, thm tip ch acetat 30% vo dch chit,
khuy u, lc li v tip tc th n khi dch lc khng cn ta vi ch acetat.
Chuyn dch lc vo mt bnh gn dung tch 100ml. Lc dch chit vi hn
hp cloroform ethanol (4:1), lc 2 ln, mi ln 8ml. Gn lp dch chit cloroform
trong ethanol vo mt cc c m c sy kh. Gp cc dch chit cloroform v
loi nc bng natri sulfat khan.
Chia u dch chit vo 4 ng nghim nh c sy kh. em c cch thy
n kh. Cn thu c em tin hnh lm cc phn ng nh tnh.
Phn ng ca khung steroid: Phn ng Liebermann Burchardt
Tin hnh:
Cho vo ng nghim c cha cn trn 1ml anhydrid acetic, lc u cho tan
ht cn. Nghing ng 450. Thm t t theo thnh ng 0,5ml acid sulfuric c, trnh
xo trn cht lng trong ng.
Kt qu:
Mu N2: mt tip xc gia hai lp cht lng xut hin mt vng mu tm
(phn ng dng tnh).
Mu N6: mt tip xc gia hai lp cht lng xut hin mt vng mu tm
(phn ng dng tnh).
Phn ng ca vng lacton 5 cnh
- Phn ng Baljet
Tin hnh:
-
25
Cho vo ng nghim c cha cn trn 0,5ml ethanol 90%. Lc u cho tan
ht cn. Nh tng git thuc th Baljet mi pha (1 phn dung dch acid picric 1%
v 9 phn dung dch NaOH 10%). So snh mu sc vi ng chng l ng khng c
cn glycosid tim.
Kt qu:
Mu N2: ng th c mu cam khng m hn ng chng (phn ng m tnh).
Mu N6: ng th c mu cam khng m hn ng chng (phn ng m tnh).
- Phn ng Legal
Tin hnh:
Cho vo ng nghim c cha cn trn 0,5ml ethanol 90%. Lc u cho tan
ht cn. Nh 1 git thuc th natri nitroprussiat 0,5% v 2 git dung dch NaOH
10%. Lc u s xut hin mu cam. So snh mu sc vi ng chng l ng
khng c cn glycosid tim.
Kt qu:
Mu N2: ng th c mu cam khng m hn ng chng (phn ng m tnh).
Mu N6: ng th c mu cam khng m hn ng chng (phn ng m tnh).
- Phn ng ca phn ng 2,6 desoxy: Phn ng Keller Kiliani
Tin hnh:
Cho vo ng nghim cha cn trn 0,5ml ethanol 90%. Lc u cho tan ht
cn. Thm vi git dung dch st (III) clorid 5% pha trong acid acetic. Lc u.
Nghing ng 450. Cho t t theo thnh ng 0,5ml acid sulfuric c, trnh xo trn
cht lng trong ng.
Kt qu:
Mu N2: mt tip xc gia 2 lp cht lng xut hin vng mu tm (phn
ng dng tnh).
Mu N6: mt tip xc gia 2 lp cht lng xut hin vng mu tm (phn
ng dng tnh).
Kt lun s b: C 2 loi Na khng cha glycosid tim.
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26
3.2.2. nh tnh saponin trong dc liu
Quan st hin tng to bt
Tin hnh:
Cho 1g bt c Na vo ng nghim ln. Thm 10ml nc. Lc mnh theo
chiu dc ng nghim trong 5 pht. yn v quan st hin tng to bt.
Kt qu:
Mu N2: Ct bt bn sau khi ngng lc 45 pht (phn ng dng tnh).
Mu N6: Ct bt bn sau khi ngng lc 45 pht (phn ng dng tnh).
Phn ng phn bit saponin steroid v saponin triterpenoid
Tin hnh:
Cho 1g bt c Na vo ng nghim ln (10ml). Thm 5ml cn un si cch
thy trong 15 pht, lc ly dch chit. Ly 2 ng nghim c bng nhau, cho vo ng
th nht 5ml HCl 0,1N (pH=1) v ng th hai 5ml NaOH 0,1N (pH=13). Cho thm
vo mi ng 2-3 git dung dch dch chit ri bt ng nghim, lc mnh c 2 ng
trong 15 giy. yn nu ct bt trong 2 ng cao bng nhau v bn nh nhau th s
b xc nh dc liu c saponin triterpenoid. Nu ng kim c ct bt cao hn ng
kia th s b xc nh l saponin steroid.
Kt qu:
Mu N2: ng kim c ct bt cao v bn hn ng acid.
Mu N6: ng kim c ct bt cao v bn hn ng acid.
Kt lun s b: Trong c ca hai loi Na u c cha saponin steroid.
3.2.3. nh tnh anthranoid
Phn ng Borntraeger
- nh tnh anthranoid dng t do
Tin hnh:
Cho 2g bt c Na vo ng nghim ln (10ml). Thm 5ml nc ct. un trc
tip vi ngun nhit cho n si. Lc dch chit cn nng qua mt lp bng mng
vo bnh gn dung tch 50ml. ngui dch lc. Thm 5ml cloroform. Lc nh.
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27
Gn b lp nc. Gi lp cloroform lm phn ng. Ly 1ml dch chit cloroform
cho vo ng nghim nh. Thm 1ml dung dch NaOH 10%, lc nh.
Kt qu:
Mu N2: Lp nc khng c mu sim (phn ng m tnh).
Mu N6: Lp nc khng c mu sim (phn ng m tnh).
- nh tnh anthranoid dng ton phn
Tin hnh:
Cho 2g bt c Na vo ng nghim ln. Thm 5ml dung dch acid sulfuric
1N. un trc tip trn ngun nhit n si. Tip tc lc v chit nh trn. Ly
1ml dch chit cloroform cho vo ng nghim nh. Thm 1ml dung dch NaOH
10%. Lc nh. Phn ng dng tnh nu lp nc c mu sim.
Kt qu:
Mu N2: Lp nc khng c mu sim (phn ng m tnh).
Mu N6: Lp nc khng c mu sim (phn ng m tnh).
Vi thng hoa
Tin hnh:
Tri bt dc liu thnh lp mng trong mt np chai bng nhm, t nh trn
n cn loi nc. Sau y ln np nhm mt ming lam knh, bn trn c ming
bng thm nc, tip tc un nng trong khong 5-10 pht. Ly lam knh ra
ngui, soi knh hin vi. Phn ng dng tnh nu xut hin tinh th trn lam knh.
Kt qu:
Mu N2: Khng xut hin tinh th trn lam knh (phn ng m tnh).
Mu N6: Khng xut hin tinh th trn lam knh (phn ng m tnh).
Kt lun s b: Trong c ca hai loi Na khng cha anthranoid.
3.2.4. nh tnh tanin
Tin hnh: Cho 2g bt c Na vo ng nghim ln, thm 10ml nc ct. un
si trc tip 5 pht. Lc qua giy lc gp np. Ly dch lc chia lm 3 ng nghim,
mi ng 1ml, em lm cc phn ng nh tnh.
Phn ng vi protein
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28
ng 1: Thm vo 1 git dung dch gelatin 1% mi pha.
Kt qu:
Mu N2: ng nghim khng xut hin kt ta bng trng (phn ng m tnh).
Mu N6: ng nghim khng xut hin kt ta bng trng (phn ng m tnh).
Phn ng vi ion kim loi nng
Thm vo vi git FeCl3 5%.
Kt qu:
Mu N2: ng nghim khng xut hin kt ta xanh en (phn ng m tnh).
Mu N6: ng nghim khng xut hin kt ta xanh en (phn ng m tnh).
ng 3: Thm vo vi git ch acetat 10%.
Kt qu:
Mu N2: ng nghim xut hin kt ta bng (phn ng dng tnh).
Mu N6: ng nghim xut hin kt ta bng (phn ng dng tnh).
Kt lun s b: Trong c ca hai loi Na khng cha tanin.
3.2.5. nh tnh ng kh
Tin hnh:
Cn khong 2g bt c Na cho vo ng nghim ln, thm 10ml nc ct, un
si vi pht. Lc qua giy lc vo 1 ng nghim khc. Thm 1ml dung dch thuc
th Felling A v 1ml dung dch Felling B. un cch thy si vi pht.
Kt qu:
Mu N2: ng nghim xut hin kt ta gch (phn ng dng tnh).
Mu N6: ng nghim xut hin kt ta gch (phn ng dng tnh).
Kt lun s b: Trong c ca hai loi Na c cha ng kh
3.2.6. nh tnh polysaccharid
Tin hnh: Ly 2g bt c Na cho vo ng nghim ln, thm 10ml nc ct. un
si cch thy vi pht. Lc ly dch. Cho vo 3 ng nghim:
ng 1: 4ml dch chit + 5 git thuc th Lugol.
ng 2: 4ml nc ct + 5 git thuc th Lugol.
ng 3: 4ml dch chit.
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29
Kt qu:
Mu N2: Mu ca ng 1 m hn mu ng 2, mu ng 2 m hn ng 3 (phn
ng dng tnh).
Mu N6: Mu ca ng 1 m hn mu ng 2, mu ng 2 m hn ng 3 (phn
ng dng tnh).
Kt lun s b: Trong c ca hai loi Na c cha polysaccharid.
3.2.7. nh tnh acid amin
Tin hnh:
Ly 2g bt c Na cho vo ng nghim ln, thm 10ml nc ct, un si vi
pht. Lc qua giy lc vo 1 ng nghim khc. Thm vi git thuc th Ninhydrin
3%. un cch thy si vi pht.
Kt qu:
Mu N2: ng nghim xut hin mu xanh (phn ng dng tnh).
Mu N6: ng nghim xut hin mu xanh tm (phn ng dng tnh).
Kt lun s b: Trong c ca hai loi Na c acid amin.
3.2.8. nh tnh acid hu c
Tin hnh:
Cho 2g bt c Na vo ng nghim ln, thm 10ml nc ct. un si trc tip vi
pht trn ngn la n cn. ngui v lc. Thm vo dch lc mt t bt Na2CO3.
Kt qu:
Mu 1: ng nghim xut hin bt kh CO2 bay ln (phn ng dng tnh).
Mu 2: ng nghim xut hin bt kh CO2 bay ln (phn ng dng tnh).
Kt lun s b: Trong c ca hai loi Na c cha acid hu c.
3.2.9. nh tnh flavonoid
Ly 2g bt c Na cho vo bnh nn dung tch 50ml. Thm 20ml ethanol 90%.
un cch thy si trong vi pht. Lc nng. Dch chit em tin hnh cc phn ng
nh tnh.
Phn ng Cyanidin (Phn ng Shinoda)
Tin hnh:
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30
Cho vo ng nghim nh 1ml dch chit. Thm mt t bt magnesi kim loi
(khong 10mg). Nh tng git HCl m c (3 - 5 git). yn mt vi pht.
Kt qu:
Mu N2: Dung dch khng chuyn t mu vng sang mu (phn ng m tnh).
Mu N6: Dung dch khng chuyn t mu vng sang mu (phn ng m tnh).
Phn ng vi kim
Tin hnh:
Bc 1: Cho 1ml dch chit vo ng nghim nh. Thm vi git dung dch
NaOH 10% sau thm 1ml nc ct.
Kt qu: Mu N2: Khng xut hin ta vng.
Mu N6: Khng xut hin ta vng.
Bc 2: Nh mt git dch chit ln giy lc. H kh ri ln ming l
amoniac c c m nt. Nh mt git khc ln giy lc so snh.
Kt qu:
Mu N2: Mu vng ca vt dch chit khng tng ln (phn ng m tnh).
Mu N6: Mu vng ca vt dch chit khng tng ln (phn ng m tnh).
Phn ng vi FeCl3
Tin hnh:
Cho 1ml dch chit vo ng nghim nh. Thm vi git dung dch FeCl3 5%.
Kt qu:
Mu N2: Khng xut hin kt ta xanh m (phn ng m tnh).
Mu N6: Khng xut hin kt ta xanh m (phn ng m tnh).
Phn ng diazo ha
Tin hnh:
Cho 1ml dch chit vo ng nghim, kim ha bng dung dch kim (dung
dch NaOH, Na2CO3), thm vi git thuc th diazo mi pha,
lc u, un nng trn ni cch thy trong vi pht.
Kt qu:
Mu N2: Khng xut hin mu (phn ng m tnh).
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31
Mu N6: Khng xut hin mu (phn ng m tnh).
Kt lun s b: Trong c ca hai loi Na khng cha flavonoid.
3.2.10. nh tnh coumarin
Ly 2g bt c Na ri cho vo bnh nn dung tch 50ml. Thm 20ml ethanol
90%. un cch thy si trong vi pht. Lc nng. Dch chit em tin hnh cc
phn ng nh tnh.
Phn ng m ng vng lacton
Tin hnh:
Cho vo 2 ng nghim mi ng 1ml dch chit: ng 1 thm 0,5ml dung dch
NaOH 10%. ng 2 nguyn.
un c 2 ng nghim n si. ngui ri quan st:
ng 1 c ta vng.
ng 2 trong.
Thm vo c 2 ng nghim mi ng 2ml nc ct. Lc u ri quan st:
ng 1 trong.
ng 2 c.
Acid ha ng 1 bng vi git HCl c, quan st:
ng 1 c.
ng 2 trong. (phn ng m tnh).
Phn ng diazo ha
Tin hnh:
Cho 1ml dch chit vo ng nghim nh. Thm vo 2ml dung dch NaOH 10%.
un cch thy n si ri ngui. Nh vo git thuc th diazo.
Kt qu:
Mu N2: Khng xut hin mu (phn ng m tnh).
Mu N6: Khng xut hin mu (phn ng m tnh).
Quan st hunh quang ca cc vt coumarin di nh sng t ngoi khi tc
dng vi dung dch kim (phn ng chuyn t ng phn cis sang ng phn
trans di tc dng ca tia t ngoi)
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32
Tin hnh:
Nh vi git dch chit ln giy thm. Nh tip vi git dung dch NaOH 5%.
Sy nh. Che mt phn din tch dch chit trn giy lc bng mt ming kim loi
(cha kha, ng xu) ri chiu tia t ngoi trong mt vi pht. B ming kim loi
ra, quan st tip di n t ngoi.
Kt qu:
Mu N2: Phn khng b che c hunh quang khng sng hn phn b che
(phn ng m tnh).
Mu N6: Phn khng b che c hunh quang khng sng hn phn b che
(phn ng m tnh).
Kt lun s b: Trong c ca hai loi Na khng cha coumarin.
3.2.11. nh tnh alcaloid
Tin hnh:
Cn 5g bt c Na, cho vo bnh nn dung tch 50ml, thm m bng dung
dch ammoniac c. y kn khong 30 pht. Cho 15ml chloroform lc u
trong 1h. Lc ly dch chit cho vo bnh gn. Sau lc k 2 ln, mi ln vi
10ml dung dch H2SO4 1N. phn lp, gn ly dch chit acid, chia u vo cc
ng nghim nh, mi ng 1ml. Nh vo tng ng nghim 2-3 git ln lt cc
thuc th sau:
ng 1: Thuc th Mayer.
Kt qu:
Mu N2: Khng xut hin kt ta trng (phn ng m tnh).
Mu N6: Khng xut hin kt ta trng (phn ng m tnh).
ng 2: thuc th Bouchardat.
Kt qu:
Mu N2: Khng xut hin kt ta nu (phn ng m tnh).
Mu N6: Khng xut hin kt ta nu (phn ng m tnh).
ng 3: Thuc th Dragendorff.
Kt qu:
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33
Mu N2: Khng xut hin kt ta cam (phn ng m tnh).
Mu N6: Khng xut hin kt ta cam (phn ng m tnh).
Kt lun s b: Trong c ca hai loi Na khng cha alcaloid.
3.2.12. nh tnh cht bo, steroid, caroten
Tin hnh:
Ly 5g bt c Na vo bnh nn dung tch 50ml, ngp ether du ha, bc
kn, ngm 24 gi. Lc qua giy lc gp np ly dch lc. Chia dch lc lm 3 phn
nh tnh nh sau:
nh tnh cht bo
Nh 1 git dch chit ln mnh giy trng, sy nh cho bay ht hi dung mi.
Kt qu:
Mu N2 : Khng li vt m trn giy (phn ng m tnh).
Mu N6: Khng li vt m trn giy (phn ng m tnh).
nh tnh steroid
Cho 2ml dch chit vo ng nghim nh, c cch thy n cn. Thm vo ng
nghim khong 1ml anhydrid acetic, lc k cho tan ht cn. nghing ng nghim
450, thm t t H2SO4 c theo thnh ng nghim.
Kt qu:
Mu N2: Mt phn cch gia 2 lp cht lng c vng tm , lp cht lng pha
trn c mu xanh l (Phn ng dng tnh).
Mu N6: Mt phn cch gia 2 lp cht lng c vng tm , lp cht lng pha
trn c mu xanh l (Phn ng dng tnh).
nh tnh caroten
Tin hnh:
Cho 2ml dch chit vo ng nghim nh, c cch thy n cn, nh vi git
H2SO4 c vo cn.
Kt qu: Mu N2: Khng xut hin mu xanh l (phn ng m tnh).
Mu N6: Khng xut hin mu xanh l (phn ng m tnh).
Kt lun s b: Trong c ca hai loi Na khng cha cht bo, steroid v caroten.
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34
Bng 3.3. Kt qu nh tnh cc nhm cht trong c ca hai loi Na
STT Nhm cht Phn ng vi thuc
th
Kt qu Kt lun
Mu N2 Mu N6 Mu N2 Mu N6
1 Glycosid
P/ Liebermann + +
Khng
c
Khng
c
P/ Legal _ _
P/ Baljet _ _
P/ Keller-Kiliani + +
2 Saponin Hin tng to bt + + C C
3 Anthranoid
Vi thng hoa _ _ Khng
c
Khng
c P/ Borntraeger _ _
4 Flavonoid
P/ Cyanidin _ _
Khng
c
Khng
c
P/ vi dd NaOH _ _
Hi NH3 _ _
dd FeCl3 5% _ _
5 Coumarin
P/ m vng lacton _ _
Khng
c
Khng
c
P/ diazo ha _ +
P/ di chuyn dng
ng phn cis-trans _ _
6 Tanin
Ch acetat + + Khng
c
Khng
c FeCl3 _ _
dd Gelantin 1% _ _
7 Alcaloid
TT Mayer _ _ Khng
c
khng
c TT Bouchardat _ _
TT Dragendoff _ _
8 Acid hu c P/ vi bt Na2CO3 + + C C
9 ng kh TT Felling A,B + + C C
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35
10 Acid amin TT Ninhydrin 0,1% + + C C
11 Polysaccharid TT Lugol + + C C
12 Caroten TT H2SO4 m c _ _ Khng
c
Khng
c
13 Sterol P/ Liebermann + + C C
14 Cht bo vt m trn giy lc _ _ Khng
c
Khng
c
Ch thch: (-): phn ng m tnh, (+): phn ng dng tnh.
Nhn xt: C hai loi Na thu hi ti hai a im khc nhau u c thnh phn
ha hc tng i ging nhau. S b xc nh thnh phn ha hc chnh ca c hai loi
Na trn l saponin. Trn c s ta s a ra cc phng php kim nghim, nh
gi v phn bit hai mu dc liu trn, gp phn xy dng tiu chun kim nghim
cho tng dc liu.
3.3. NH TNH DCH CHIT HAI LOI NA BNG SC K LP MNG
Sau khi tin hnh sc k lp mng vi dch chit methanol c ca hai loi Na
trn nhiu h dung mi c phn cc khc nhau, nhn thy kh nng tch ca
chng km nn ta tin hnh thy phn hai mu Na v chit vi CHCl3 tin hnh
sc k lp mng.
Tin hnh:
Chun b dch chm sc k: Ngm 10g bt c Na trong 20 ml methanol trong
vng 24h. Lc ly dch trong. Thm ng lng HCl 10%. Thy phn trong vng 3
gi nhit 100C. ngui, chuyn dch lc vo bnh gn 50ml. Lc vi CHCl3
3 ln, mi ln 5ml. Gp cc dch chit CHCl3 vo mt cc c m sch. C ti cn
sau ha tan cn trong methanol ly dch chm sc k.
Chm ng thi dch chit hai mu trn cng mt bn mng bng my chm
sc k CAMAG LINOMAT 5.
Trin khai sc k trn cc h dung mi:
H I: Toluen-ethyl acetat-acid formic (6:2:0,5).
H II: Ete du ha-ethyl acetat-aceton-acid formic (3:1:0,5:0,05).
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36
H III: Cloroform-MeOH (9:1).
H IV: Cloroform-aceton (9:1).
H V: Ethyl acetat-acid formic (4:1).
Sau khi trin khai sc k, h I, h II, h IV cho kt qu tch tt. Sc k ca
chng nh sau:
Kt qu h I: Toluen-ethyl acetat-acid formic (6:2:0,5).
Ia Ib Ic
Vi dch chit thy phn ca hai mu Na khi trin khai h dung mi: toluen-
ethyl acetat-acid formic (6:2:0,5), kt qu phn tch cho thy:
Trc khi phun thuc th di nh sng t ngoi (UV 254nm):
Mu N2 quan st c 2 vt, vt m nht c Rf=0,24.
Mu N6 quan st c 3 vt, vt m nht c Rf=0,24.
Trc khi phun thuc th di nh sng t ngoi (UV 366nm):
Mu N2 quan st c 6 vt, vt m nht c Rf=0,16.
Mu N6 quan st c 8 vt, vt m nht c Rf=0,15.
Sau khi phun thuc th di nh sng thng:
Hnh 3.7. Sc k dch chit
thy phn ca hai mu Na vi
h dung mi toluen-ethyl
acetat-acid formic (6:2:0,5) :
Ia. =254nm
Ib. = 366nm
Ic. Sau khi phun TT Vanilin
1%/H2SO4.
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37
Mu N2 quan st c 5 vt, vt m nht c Rf=0,80.
Mu N6 quan st c 10 vt, vt m nht c Rf=0,79.
Nhn xt:
- bc sng 254nm: Hai mu dch thy phn ca hai loi Na khi trin khai trn
sc k c nhng vt tng ng nhau. Trn sc k ca dch chit mu N6 c
nhiu vt hn trn sc k ca dch chit mu N2 v vt c Rf=0,82 d dng nhn
bit s khc bit so vi mu N2.
- Khi hin mu bng thuc th nh sng thng ta thy chng c mt s vt
tng i ging nhau. Tuy nhin trn sc k mu N6 trong h dung mi ny tch
r hn v trin khai vi nhiu vt hn. Trn sc k ny dch chit mu N6 c
vt c Rf=0,60 d dng nhn bit s khc bit vi dch chit mu N2.
V vy h toluen-ethyl acetat-acid formic (6:2:0,5) c th dng phn bit
hai loi Na trn.
Kt qu h II: Ether du ha-ethyl acetat-aceton-acid formic (3:1:0,5:0,05).
Vi dch chit thy phn ca hai loi Na khi trin khai h dung mi: Ether du
ha-ethyl acetat-aceton-acid formic (3:1:0,5:0,05) kt qu phn tch cho thy:
Trc khi phun thuc th di nh sng t ngoi (UV 254nm):
Mu N2 quan st c 2 vt, vt m nht c Rf=0,30.
Mu N6 quan st c 6 vt, vt m nht c Rf=0,31.
Trc khi phun thuc th di nh sng t ngoi (UV 366nm):
Mu N2 quan st c 6 vt, vt m nht c Rf=0,19.
Mu N6 quan st c 9 vt, vt m nht c Rf=0,19.
Sau khi phun thuc th di nh sng thng:
Mu N2 quan st c 5 vt, vt m nht c Rf=0,84.
Mu N6 quan st c 12 vt, vt m nht c Rf=0,85.
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38
IIa IIb IIc
Nhn xt:
- bc sng 254nm: Dch chit ca hai loi Na khi khai trin trn h dung mi
trn c nhng vt tng i ging nhau. Tuy nhin dch chit mu N6 trin khai
nhiu vt hn so vi mu N2, cc vt c Rf tng ng l 0,28; 0,74; 0,90.
- bc sng 366nm: Trn sc k ca dch chit mu N6 khai trin nhiu vt
hn so vi dch chit mu N2 v trn sc k ca dch chit mu N6 c cc vt
hunh quang mu . C th s b d on trong dch chit mu N6 c
chlorophyl.
- Thuc th hin mu nh sng thng: Dch chit mu N6 trin khai trn sc k
vi nhiu vt hn v vt c Rf=0,69 d dng nhn thy s khc bit so vi mu
N2.
Sc k ca hai loi Na ca h dung mi ether du ha-ethyl acetat-aceton-
acid formic (3:1:0,5:0,05) bc sng 366nm l tng i khc nhau. V vy h
thch hp dng so snh hai loi.
Hnh 3.8. Hnh nh sc k dch
chit thy phn hai mu Na vi h
dung mi ether du ha-ethyl acetat-
aceton-acid formic (3:1:0,5:0,05) :
IIa. =254nm
IIb. =366nm
IIc. sau khi phun TT Vanilin
1%/H2SO4
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39
Kt qu h IV: Cloroform: aceton (9:1).
IIIa IIIb IIIc
Vi dch chit thy phn ca c hai loi Na khi khai trin h dung mi:
Cloroform-aceton (9:1) kt qu phn tch cho thy:
- Trc khi phun thuc th di nh sng t ngoi (UV 254nm):
Mu N2 quan st c 4 vt, vt m nht c Rf=0,26.
Mu N6 quan st c 5 vt, vt m nht c Rf=0,27.
- Trc khi phun thuc th di nh sng t ngoi (UV 366nm):
Mu N2 quan st c 4 vt, vt m nht c Rf=0,03.
Mu N6 quan st c 8 vt, vt m nht c Rf=0,05.
- Sau khi phun thuc th di nh sng thng:
Mu N2 quan st c 2 vt, vt m nht c Rf=0,84.
Mu N6 quan st c 11 vt, vt m nht c Rf=0,83.
Nhn xt:
- bc sng 254nm: Sc k trin khai trn h dung mi trn ca hai mu Na
t c s khc bit. Chng u tch c 3 vt r.
Hnh 3.9 Hnh nh sc k dch
chit thy phn hai mu Na vi h
dung mi Cloroform-aceton (9:1)
:
IIIa. =254nm
IIIb. =366nm
IIIc. sau khi phun TT Vanilin
1%/H2SO4
-
40
- bc sng 366nm: Sc k ca mu N6 c nhiu vt mu tch r rng. S
b xc nh mu ny c chlorophyl.
- Khi hin mu bng thuc th: Mu N6 trin khai c nhiu vt hn mu N2.
Nh vy h dung mi chloroform: aceton (9:1) thch hp phn bit hai loi.
Kt lun: Vi dch chit thy phn ca hai loi Na trn, la chn c 3 h
dung mi tch tt v cho kt qu phn bit r rng hai loi Na.
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41
3.4. BN LUN
mt s a phng Vit Nam, tn gi Na rt quen thuc vi mi
ngi dn. Tuy cng tn gi l Na nhng mi loi li c nhng c im hnh
thi v gi tr s dng khc nhau. Qua nghin cu thc a, tham kho cc ti liu
thc vt hc nh thc vt ch Trung Quc v di s t vn ca TS. Nguyn Quc
Huy hai mu Na thu hi ti Nho Quan-Ninh Bnh v vn Quc Gia Ba V c
tin hnh gim nh tn khoa hc v xc nh tn khoa hc ca 2 mu nghin
cu nh sau:
Mu N2: Amorphophallus paeoniifolius (Dennst.) Nicolson, h Ry (Araceae).
Mu N6: Amorphophallus coaetaneus S. Y. Liu & S. J. Wei, h Ry (Araceae).
Cy Na c ngi dn s dng trong dn gian lm lng thc cng nh
cha bnh trong y hc c truyn. Gn y, nhiu nghin cu cho rng glucomannan
trong cy Na c nhiu ng dng tuyt vi. Tuy nhin nghin cu mi ch gii hn
trn mu Na Tha Thin-Hu.
ti nghin cu phn bit hai loi Na ti Ninh Bnh v vn Quc Gia Ba
V l mt trong nhng nghin cu u tin Vit Nam v phn bit c im thc
vt v thnh phn ha hc. N l mt nghin cu rt c tim nng gp phn to
ngun gen qu phc v cng tc nghin cu bo tn ging cy trng qu. Trong
khun kh ca ti phn bit c c im v thc vt cng nh thnh phn
ha hc ca hai loi.
V thc vt:
- c im hnh thi: C bn phn bit c chng qua c im c, c im cung l.
- c im vi phu: Hai bt Na u c cc c im c bn nh mnh m mm
thnh mng mang ht tinh bt, tinh th canxi oxalat, mnh mch, cc ht tinh bt
n v tinh bt kp. ti so snh, phn bit c im bt c hai loi Na qua
cc c im v canxi oxalat v kch thc tinh bt.
V thnh phn ha hc:
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42
- tin hnh xc nh s b s c mt ca cc nhm cht hu c trong dc liu bng
cc phn ng ha hc c trng v la chn c h dung mi thch hp tin hnh sc
k lp mng nhm so snh s khc bit v thnh phn ha hc gia hai loi.
- Kt qu nh tnh cho thy: Thnh phn ha hc ca hai loi Na tng i ging
nhau chng u cha saponin, tanin, ng kh, polysaccharid, acid amin, acid
hu c, steroid.
- Sc k lp mng: Dch chit methanol th vi nhiu h dung mi cho kh
nng tch km nn tin hnh thy phn v chit bng cloroform chm sc
k. Vi dch chit thy phn, la chn ra h dung mi tch r c c bn phn
bit c hai loi Na trn. T c th phn bit c thnh phn ha hc ca
hai loi ny.
Vic phn tch, so snh v ch ra cc c im hnh thi, c im bt c to c s
bc u cho vic nh hng, nhn bit, phn bit cc loi trong chi Amorphophalus, t
to c s d liu, l lch mu trnh nhm ln trong vic thu hi, s dng dc liu.
Nh c th a vo tiu chun kim nghim dc liu. T nhng c im hnh thi
gip la chn ging tt c tim nng v sn xut pht trin vng trng mang li gi tr
kinh t cao cho mi a phng.
Vic so snh, nh tnh s b cc nhm cht v thnh phn ha hc trong hai
mu Na to nhng bc m u tin cho vic la chn mu nghin cu phn
lp nhng hp cht c ngha v mt dc l nh glucomannan.
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43
KT LUN V XUT
1. KT LUN
V c im thc vt
- m t, so snh c c im hnh thi, vi hc bt, vi phu c ca hai loi
Na.
V thnh phn ha hc
- Bng cc phn ng nh tnh s b xc nh c trong c ca hai loi Na
nghin cu c cha saponin, ng kh, polysaccharid, acid amin, acid hu c,
steroid.
- Phn tch dch thy phn ca hai loi Na bng sc k lp mng trn nhiu h
dung mi cho kt qu so snh, phn bit tt cc h:
H I: Toluen - ethyl acetat - acid formic (6:2:0,5).
H II: Ether du ha - ethyl acetat aceton - acid formic (3:1:0,5:0,05).
H IV: Cloroform - aceton (9:1).
2. XUT
T nhng kt qu t c, ti c mt s xut sau:
- Nghin cu iu kin sinh trng v nng sut mu Na ti Ninh Bnh a cy
Na l mt cy trng mang li ngun li kinh t cho a phng.
- Nghin cu hnh thc s dng ca hai mu Na c bit l mu Na ti Ba V.
- Tip tc nghin cu thnh phn ha hc ca hai loi Na, ng thi nghin cu
hm lng glucomannan trong mi loi.
- Xy dng tiu chun kim nghim cho mi mu Na khi a vo sn xut.
-
TI LIU THAM KHO
Ti liu ting vit
1. Huy Bch v cng s, Cy thuc v ng vt lm thuc Vit Nam tp II,
Nxb Khoa hc K thut, tr. 78.
2. B mn Dc liu (2007), Thc tp dc liu phn kim nghim bng phng
php hin vi, trng i hc Dc H Ni, H Ni.
3. B mn Dc liu (2007), Thc tp dc liu phn kim nghim bng phng
php ha hc, trng i hc Dc H Ni, H Ni.
4. B mn Thc vt (2004), Thc tp Thc vt v nhn bit cy thuc, trng i
hc Dc H Ni, H Ni.
5. B Y t (2007), Dc liu hc, Nxb Y hc, tp I.
6. B Y t (2007), Dc liu hc, Nxb Y hc, tp II.
7. B Y t (2007), Thc vt hc, Nxb Y hc, tr. 358-359.
8. B Y t , Dc in Vit Nam IV. Ph lc 9.8.
9. V Vn Chi (1997), T in cy thuc Vit Nam, Nxb Y hc, tr. 617.
10. V Vn Chi (2003), T in thc vt thng dng, Nxb Khoa hc v K thut,
tp I, tr. 272.
11. Nguyn Vn n, Nguyn Vit Tu (1985), Phng php nghin cu ha hc
cy thuc, Nxb Y hc, Tp. H Ch Minh.
12. Phm Hong H (2000), Cy c Vit Nam, Nxb Tr, quyn III, tr. 359-362.
13. V Mai Hng, Trn V Ngc Thi, Nguyn Th Thu, Nghin cu sinh
trng ca cy Na Chung (Amorphophallus paeoniifolius) Tha Thin
Hu.
14. Tt Li (1999), Nhng cy thuc v v thuc Vit Nam, Nxb Y hc, tr. 136.
15. Nguyn Vn Mi (2001), Thc hnh sinh hc, Nxb Khoa hc k thut.
16. Nguyn Vit Thn (2013), Cy thuc Vit Nam v nhng bi thuc thng
dng, Nxb Th gii, tp 3, tr. 46,48.
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17. Nguyn Vit Thn (2003), Kim nghim dc liu bng phng php hin vi,
Nxb Khoa hc v K thut.
18. Ng Vn Thu (1990), Ha hc Saponin, khoa Dc, trng i hc Y Dc
thnh ph H Ch Minh.
Ti liu ting Anh
19. Dey YN, Ghosh AK (2009), Evaluation of anthelmintic activity of the
methanolic extract of Amorphophallus paeoniifolius tuber, Int J Pharm Sci Res, 1,
pp. 117121.
20. De S, Dey YN, Ghosh AK (2010), Anti-inflammatory activity of methanolic
extract of Amorphophallus paeoniifolius and its possible mechanism, Int J Pharma
Biosci, 1, pp. 18.
21. De S, Ghosh AK, Dey YN (2010), Evaluation of Analgesic activity of
methanolic extract of Amorphophallus paeoniifolius tuber by tail flick and acetic
acid-induced writhing response method, Int J Pharm Biosci, 1, pp. 662668.
22. Jayaraman Angayarkanni et al. (2010), Antioxidant potential of
Amorphophalus paeoniifolius in realation to their phenolic content, Bharathiar
University, 48, pp. 659 665.
23. Khan A, Rahman M, Islam MS (2008), Anibacterial, antifungal and cytotoxic
activities of amblyone isolated from Amorphophalus campanulatus, Indian J
Pharmarcol, 40, pp. 41- 44.
24. Li Heng, Zhu Guanghua, Peter C. Boyce, Niels Jacobsen (2010), Flora of
China, 23, pp. 1, 3, 23-33, 80.
25. Liu Pei-Ying, Wang Yulan, 1990. Physiological study on the storage of
elephant- foot-yam, In: Proceedings of the 60th Anniversary of the Chinese
Horticultural Society and the 6th
Annual Workshop: 29-32.
26. Malini Sen et al. (2009), Effects of petroleum ether extract of Amorphophalus
paeoniifolius tuber on central nervous system in mice, Indian J Pharm Sci, 71, pp.
651- 655.
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27. Shilpi JA, Ray PK, Sarder MM, Uddin SJ (2005), Analgesic activity
of Amorphophallus paeoniifolius tuber, Fitoterpia, 76, pp. 367369.
Ti liu Internet
28. http://www.botanyvn.com/cnt.asp?param=edir&v=Amorphophallus&list=genus.
29. http://ich.ac.vn/Tin-tuc/Hoat-dong-nghien-cuu-khoa-hoc/27556/tom-tat-ket-qua-
thuc-hien-de-tai-nghien-cuu-quy-trinh-tach-chiet-cau-truc-hoa-hoc-va-hoat-tinh-
sinh-hoc-cua-glucomannan-tu-cay-nua-amorphophallus-sp-ho-ray--araceae.
30. http://www.tchdkh.org.vn/khcn-dia-phuong/5266-thua-thien-hue-nghien-cuu-
ham-luong-chat-luong-tac-dung-duoc-ly-va-xay-dung-quy-trinh-san-xuat-
glucomannan-trong-cu-nua-amorphophallus-sp-ho-ray-araceae.html.
31. http://www.theplantlist.org.
32. http://www.vast.ac.vn/1.0/index.php?option=com_detai&view=detai&id=723%
3Anghien-cuu-trong-va-phat-trien-cay-nua-konjac-(amorphoph-allus)-va-mot-so-
loai--khac-trong-chi-nua-(ho-ray-%E2%80%93-araceae)-o-viet-nam-huong-toi-
viec-lay-cu-lam-nguyen-lieu-san-xuat-thuc-pham-chuc-nang-va-thuoc-dieu-tri-
benh-tieu-duong%2C-mo-mau-va-beo-phi&Itemid=54&lang=vi.
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PH LC