B Y T

I HC DC H NI

L HU TN

NGHIN CU C IM THC VT,

THNH PHN HA HC CA

CY NGI TIN

(Hedychium coronarium Koenig),

H GNG (Zingiberaceae) TRNG

TRT TI HUYN T LIM, H NI

KHA LUN TT NGHIP DC S

H NI - 2014

B Y T

I HC DC H NI

L HU TN

NGHIN CU C IM THC VT,

THNH PHN HA HC CA

CY NGI TIN

(Hedychium coronarium Koenig),

H GNG (Zingiberaceae) TRNG

TRT TI HUYN T LIM, H NI

KHA LUN TT NGHIP DC S

Ngi hng dn:

PGS.TS. Nguyn Vit Thn.

Ni thc hin:

B mn Dc Liu

Trng i Hc Dc H Ni

H NI-2014

MC LC T VN ............................................................................................................. 1

CHNG 1. TNG QUAN ....................................................................................... 2

1.1. TNG QUAN V H GNG (ZINGIBERACEAE) ..................................... 2

1.1.1. V tr phn loi h GNG (ZINGIBERACEAE) ........................................ 2

1.1.2. c im thc vt h Gng (ZINGIBERACEAE) ..................................... 2

1.1.3. Phn loi thc vt h Gng ....................................................................... 3

1.2. TNG QUAN V CHI HEDYCHIUM ............................................................ 4

1.2.1. c im thc vt v phn b chi Hedychium ........................................... 4

1.2.1.1. c im thc vt chi Hedychium .......................................................... 4

1.2.1.2. Phn loi thc vt chi Hedychium ........................................................... 4

1.2.1.3. Nhng tc dng c ch ca chi Hedychium ............................................. 4

1.2.2. c im mt s loi thuc chi Hedychium c Vit Nam ...................... 5

1.2.2.1. Hedychium coronarium Koenig .............................................................. 5

1.2.2.2. Hedychium bousigonianum Pierre ....................................................... 10

1.2.2.3. Hedychium coccineum Hamilt .............................................................. 11

1.2.2.4. Hedychium ellipticum (Smith) Kuntze .................................................. 11

1.2.2.5. Hedychium gardnerianum Roscoe ........................................................ 12

1.2.2.6. Hedychium forresti Diels....................................................................... 12

1.2.2.7. Hedychium villosum Wall ..................................................................... 12

CHNG 2. I TNG V PHNG PHP NGHIN CU ........................ 14

2.1. NGUYN LIU, PHNG TIN NGHIN CU ...................................... 14

2.1.1. Nguyn liu nghin cu ........................................................................... 14

2.1.2. Phng tin nghin cu ............................................................................ 14

2.1.2.1. Ha cht v dng c .............................................................................. 14

2.1.2.2. Thit b dng trong nghin cu ............................................................. 14

2.2. PHNG PHP NGHIN CU .................................................................. 15

2.2.1. Nghin cu thc vt bng phng php nghin cu hin vi ................... 15

2.2.2. Nghin cu bng phng php ha hc ................................................... 15

2.2.3. Nghin cu v tinh du thn r Ngi tin ................................................. 15

CHNG 3. THC NGHIM V KT QU ....................................................... 17

3.1. Nghin cu v thc vt ................................................................................... 17

3.1.1. M t hnh thi cy v gim nh tn khoa hc ....................................... 17

3.1.2. Nghin cu vi hc vi phu dc liu ....................................................... 18

3.1.3. Nghin cu vi hc bt dc liu .............................................................. 21

3.2. Nghin cu v ha hc ................................................................................... 23

3.2.1. Ct tinh du ............................................................................................... 23

3.2.1.1. Xc nh hm m dc liu ti ........................................................... 23

3.2.1.2. Xc nh hm lng tinh du cc b phn Ngi tin ............................ 23

3.2.2. nh tnh cc nhm cht trong dc liu bng phn ng ha hc. .......... 23

3.2.2.1. nh tnh flavonoid. .............................................................................. 23

3.2.2.2. nh tnh coumarin. ............................................................................... 24

3.2.2.3. nh tnh saponin. ................................................................................. 25

3.2.2.4. nh tnh alcaloid. ................................................................................. 26

3.2.2.5. nh tnh tanin ....................................................................................... 26

3.2.2.6. nh tnh anthranoid. ............................................................................. 27

3.2.2.7. nh tnh glycosid tim. .......................................................................... 27

3.2.2.8. nh tnh acid hu c ............................................................................ 28

3.2.2.9. nh tnh ng kh ............................................................................. 28

3.2.2.10. nh tnh acid amin ............................................................................. 29

3.2.2.11. nh tnh polysaccharid....................................................................... 29

3.2.2.12. nh tnh cht bo ............................................................................... 29

3.2.2.13. nh tnh caroten ................................................................................. 29

3.2.2.14. nh tnh sterol .................................................................................... 30

3.2.2.15. nh tnh iridoid .................................................................................. 30

3.2.3. nh tnh dch chit MeOH ton phn bng sc k lp mng .................. 32

3.2.4. nh tnh dch chit sau khi thy phn bng sc k lp mng ................. 32

3.3. Nghin cu v tinh du ................................................................................... 36

3.3.1. Sc k lp mng tinh du thn r Ngi tin ............................................ 36

3.3.2. Sc k kh-khi ph (GC-MS) tinh du thn r Ngi tin ....................... 38

BN LUN .............................................................................................................. 41

KT LUN V KIN NGH................................................................................... 43

LI CM N

Trong thi gian qua, em nhn c rt nhiu s gip , ng vin

ca thy c, gia nh v bn b. Nhng iu ht sc qu bu gip em n

lc hon thnh kha lun ny.

Vi lng knh trng v bit n su sc, em xin gi li cm n chn

thnh ti PGS.TS. Nguyn Vit Thn, ging vin b mn Dc liu trng

i hc Dc H Ni, ngi thy tn tnh hng dn, ch bo em trong

sut qu trnh thc hin kha lun.

Em xin chn thnh cm n cc thy c gio, cc anh ch k thut vin ti

b mn Dc liu gip v to iu kin thun li cho em thc hin

kha lun ti b mn.

Em xin trn trng cm n ban Gim hiu, phng o to, cc thy c

trng i hc Dc h Ni truyn cho em kin thc cng nhit huyt ca

cc thy c trong sut nhng nm hc tp ti trng.

V cui cng, em xin chn thnh cm n gia nh, bn b, ngi thn

bn ng vin, gip em thc hin kha lun.

H Ni, thng 5 nm 2014

Sinh vin

L Hu Tn

DANH MC CC K T VIT TT

K t vit tt Tn y

SKLM Sc k lp mng

Rf H s lu

UV Ultraviolet

TT Thuc th

GC-MS Gas Chromatography-Mass Spectrometry

Var. Variate

IC50 Inhibition Concentration at 50%

DANH MC CC BNG

Bng 3.1 Kt qu nh tnh s b cc nhm cht trong thn r Ngi tin Trang 31

Bng 3.2 Chng trnh nhit h thng GC-MS Trang 38

Bng 3.3 Thnh phn v hm lng cc cht trong tinh du thn r

Ngi tin

Trang 39

DANH MC CC HNH

Hnh 1.1 Cu trc ha hc mt s cht c hot tnh sinh hc c

phn lp t thn r Ngi tin Trang 7

Hnh 2.1 Cu to b dng c ct tinh du ci tin Trang 16

Hnh 3.1 nh c im mt s b phn cy Ngi tin Trang 17

Hnh 3.2 Vi phu l Ngi tin Trang 20

Hnh 3.3 Vi phu thn r Ngi tin Trang 20

Hnh 3.4 Mt s c im bt l Ngi tin Trang 22

Hnh 3.5 Mt s c im bt thn r Ngi tin Trang 22

Hnh 3.6

Sc k dch chit MeOH ton phn thn r Ngi tin

khi trin khai trong h dung mi Toluen-EtOAc-A.formic

(5,5:4,5:0,5)

Trang 33

Hnh 3.7 Sc k dch chit sau khi thy phn ca thn r Ngi tin

khi trin khai h Toluen-EtOAc-A.formic (7:3:0,1) Trang 33

Hnh 3.8

Kt qu lng gi sc k dch chit MeOH thn r

Ngi tin di n 254nm h dung mi Toluen-EtOAc-

A.formic (5,5:4,5:0,5)

Trang 34

Hnh 3.9

Kt qu lng gi sc k dch chit MeOH di n

366nm h dung mi Toluen-EtOAc-A.formic

(5,5:4,5:0,5)

Trang 35

Hnh 3.10

Kt qu lng gi sc k dch chit MeOH sau khi

phun TT Vanilin/H2SO4 c h Toluen-EtOAc-

A.formic (5,5:4,5:0,5)

Trang 35

Hnh 3.11 Sc k Tinh du thn r Ngi tin trong n-Hexan khi

trin khai h dung mi Toluen-Ethyl acetat (98:2) Trang 36

Hnh 3.12 Kt qu lng gi sc k tinh du thn r Ngi tin

di n 254nm h dung mi Toluen-EtOAc (98:2) Trang 37

Hnh 3.13

Kt qu lng gi sc k tinh du thn r Ngi tin

sau khi phun TT Vanilin/H2SO4 h dung mi Toluen-

EtOAc (98:2)

Trang 37

1

T VN

Vit Nam nm trong vng kh hu nhit i m gi ma nn c thin

nhin u i cho h thc vt phong ph v a dng, trong c bit phi k n

ti nguyn cy thuc.

Theo kt qu iu tra ca Vin Dc liu nm 2006, Vit Nam c 3948 loi

thc vt bc cao, bc thp v nm ln c dng lm thuc. Tuy nhin, phn ln

cc cy thuc c s dng theo kinh nghim dn gian hoc theo Y hc c truyn

m cha c nghin cu mt cch chuyn su v y .

Ngi tin l loi cy bi thn r thng mc ven b hoc c ngi dn

trng lm cnh, lm ba ngi trong tn ngng mt s dn tc thiu s. Thn r c

tc dng cha au d dy, y bng trng hi, cm mo, phong thp nhc mi,

rn cnv cha mt s cht c hot tnh chng ung th ang c th gii quan

tm nghin cu.

Mc d c nhiu tc dng nh vy nhng cc nghin cu hin nay v cy

Ngi tin cn t v cha thc s y . V vy, ti thc hin kha lun NGHIN

CU C IM THC VT, THNH PHN HA HC CA CY NGI

TIN (Hedychium coronarium Koenig), H GNG (ZINGIBERACEAE)

TRNG TRT TI HUYN T LIM, H NI l 1 phn ca ti cp

trng NGHIN CU BO TN V PHT TRIN MT S NGUN GEN

CY THUC QU HIM C TIM NNG VIT NAM nhm mc tiu

gp phn xy dng c s d liu s dng cy mt cch c hiu qu.

thc hin c mc tiu trn, ti c tin hnh vi cc ni dung sau:

1. Nghin cu cc c im thc vt ca mu cy: m t c im hnh thi,

c im vi phu, c im bt l, thn r Ngi tin.

2. Nghin cu thnh phn ha hc ca b phn dng: nh tnh cc nhm cht

bng phn ng ha hc, sc k lp mng.

3. Nghin cu v tinh du: xc nh thnh phn, hm lng tinh du, d tm h

sc k lp mng tinh du ca thn r Ngi tin.

2

CHNG 1. TNG QUAN

1.1. TNG QUAN V H GNG (ZINGIBERACEAE)

1.1.1. V tr phn loi h GNG (ZINGIBERACEAE)

Theo Thc vt ch ng Dng [34] v h thng phn loi ca Takhtajan

2009 [17], v tr ca h Gng (Zingiberaceae) trong gii thc vt nh sau:

Gii thc vt (Planta)

Ngnh Ngc lan (Magnoliophyta)

Lp Hnh (Liliopsida)

Phn lp Loa kn (Liliidae)

B Gng (Zingiberales)

H Gng (Zingiberaceae)

1.1.2. c im thc vt h Gng (ZINGIBERACEAE)

Theo sch Thc vt ch ng dng [34], h Gng c cc c im sau:

Cy c, sng lu nm. Thn r khe, c khi phng ln nh c. Thn kh sinh

khng c hoc mc rt cao, do cc b l m nhau to thnh. L n, nguyn, xp

thnh 2 dy song song. B l ko di to thnh li nh. Phin l c gn song song.

Cm hoa dng bng, chm, mc gc (t thn r) hay trn ngn (trn thn kh

sinh). Hoa c mu, ln, d nhu nt, i xng 2 bn, lng tnh. i 3, dnh nhau

to thnh ng, trn chia 3 thy, thy gia ln hn 2 thy bn [6]. Nh 1, bao phn 2

, ch nh nc, hnh lng mng. 3 nh thoi ha dnh nhau to thnh cnh mi ln,

mu sc sc s, 2 nh cn li tiu gim mc khc nhau, c khi ln hn cnh

hoa, hay thnh dng ru 2 bn gc ch nh hu th, c khi tiu gim hon ton. B

nhy 3 l non, dnh nhau to thnh bu di, 3 , nh non trung tr, mi nhiu

non, c khi ch cn 1 .Vi nhy th ra ngoi, 2 vi cn li khng sinh sn, tiu

gim gc vi hu th. Qu nang, t khi l qu mng. Ht c ni nh v ngoi nh

[3]. C cc t bo tit tinh du nm ri rc m mm. Do cc b phn ca cy

u c mi c bit [3].

3

1.1.3. Phn loi thc vt h Gng

Theo T in Cy Thuc ca V Vn Chi, H Gng gm 45 chi vi khong

1300 loi phn b vng nhit i v nhit i [6]. Vit Nam c 21 chi, 100

loi, phn ln mc hoang, mt s loi c trng lm thuc, hng liu, gia v v

c trng trt vi din tch ln nh Gng, Ring, a lin, Tho qu c trn 20

loi thng c dng lm thuc vi cc tn u khu, a lin, Ring, ch tr, Sa

nhn, Tho qu, Ngh, Gng, phn ln c dng trong cng nghip dc. Cc loi

khc dng trong dn gian [3].

Theo Thc vt ch ng Dng [34] v Cy C Vit Nam ca Phm Hong

H [9], H Gng gm 2 phn h:

- Costoideae (Phn h ma d): L mc theo hnh xon c, khng c mi

thm, b khng ch.

- Zingiberoideae (Phn h Gng): L mc song nh, c mi thm, b ch.

Phn h Gng chia lm 3 tng: Zingibereae, Hedychieae, Globbeae. Gm

cc chi:

Chi Hedychium Chi Elettariopsis

Chi Alpinia Chi Geostachys

Chi Cenolophon Chi amomum

Chi Catimbium Chi Curcuma

Chi Languas Chi Zingiber

Chi Achasma Chi Stahlianthus

Chi Nicolaia Chi Kaemfperia

Chi Elettaria Chi Siliquamomum

Chi Boesenbergia

4

1.2. TNG QUAN V CHI HEDYCHIUM

1.2.1. c im thc vt v phn b chi Hedychium

1.2.1.1. c im thc vt chi Hedychium

Cy c, mc di mt t hoc sng ph sinh trn cy thn g. Thn r pht

trin phnh ln thnh c, thn gi mc cao do cc b l m nhau to thnh. B l ko

di to thnh li nh, d thy; phin l hnh ch nht hoc mi mc. Cm hoa

dng bng chm mc t gc, nhiu hoa, hoa nhiu mu; l bc xp lp ln nhau

hoc xp lng lo, bao quanh 1 hay nhiu hoa [19], [34]. i hoa hnh ng, c

ch ra 1 bn, hnh chp ct hoc c 3 rng. ng trng di, mnh, thy hoa n to

hoc dng mnh hp. Nh lp hnh cnh hoa, ln hn thy trng hoa. Cnh hoa gia

ln, nh x i. Ch nh thng di, him khi tiu bin, bao phn nh lng, y

chia ra [34]. B nhy 3 l non, nh nhau to thnh bu di. Qu nang hnh cu, 3

mnh v, c nhiu ht bn trong [9].

1.2.1.2. Phn loi thc vt chi Hedychium

Chi Hedychium c khong 50 loi, phn b ch yu vng Himalaya v cc

vng cn nhit i khc ca chu . Ngoi ra cn c chu Phi (Madagasca) [5],

[19]. Trung Quc c 28 loi (18 loi c hu) [19]. Hin nay, cy c trng

nhiu ni vi mc ch lm cnh hoc ly tinh du hoa, thn r dng lm nc

hoa, cha bnh [5]. Theo Sch Thc vt ch ng Dng v Cy C Vit Nam ca

Phm Hong H, nc ta c 11 loi v th thuc chi Hedychium [9], [34].

1.2.1.3. Nhng tc dng c ch ca chi Hedychium

Cc loi trong chi Hedychium c hoa rt thm v cc -lacton diterpen ,

khng no c nhiu hot tnh sinh hoc hu dng. c bit l cc -lacton , khng

no thuc dy labdan. Theo Kumakara Abe, chng c tc dng c ch s to thnh

v s hot ng ca tc nhn NF-kB gy ng th v nhiu bnh nguy him khc.

Do nhng c tnh ny m cc -lacton , khng no thuc dy labdan diterpen

ang l i tng c cc nh khoa hc trn th gii c bit quan tm nghin

cu [12].

5

Tinh du thn r cc loi Ngi tin c nghin cu ch yu cha cc

monoterpen v sesquiterpen, c bit l 1,8-cineol (0,1% -42%), linalool (0,1%-

56%), -pinen (3%-17%), -pinen (4%-31%) v (E)-nerolidol (0,1%-20%). Chng

u c tc dng dit nm, vi khun gy hi nh: Colletotrichum fragariae,

Stephanitis pyrioides, Aedes aegypti, Solenopsis invicta [18]. Mt s loi tinh du

c mi d chu c dng lm nc hoa.

1.2.2. c im mt s loi thuc chi Hedychium c Vit Nam

1.2.2.1. Hedychium coronarium Koenig

Tn gi khc: Ngi tin, Bch yn, Bch ip, Garlannt flower, White hedychium.

M t: Thn r mp, t phn nhnh, thn nhn, ging cy ging, gng. l mc sole,

khng cung, hnh di hp- mi mc, di 40-50cm, rng 5-10cm [5], [34], gc tht

li, u thun nhn, mt trn nhn mu lc sm bng, mt di nht c lng d

rng [32]. B l to, c kha mng, li nh kch thc 2-3cm [5]. Cm hoa hnh

trng mc ngn thn, di 5-7cm, gm nhiu l bc lp ln nhau [9]. L bc v l

bc con c mu lc u, hoa to, i dng ng, rng khng r, trng c ng di hn

i [6], nhn, c 3 cnh, hai cnh bn hnh mc, cnh gia rng, lm u, nh c

ch nh di, bao phn c cc ko di thnh ca, trung i dng bn. C nh lp

dnh nhau na di, bu c lng [5], [9], [34]. Qu khi chn mu vng, ht mu

[9]. Ma hoa qu t thng 7 n thng 10 [9], [34].

Theo thc vt ch ng Dng [34], loi Hedychium coronarium chia lm 5

th da theo kch thc, mu cnh mi v ch nh:

A. Cnh mi rng

a. Cnh mi mu trng

a1. Ch nh mu trng...Hedeychium coronarium

a2. Ch nh mu ..Hedychium maximum

b. Cnh mi mu vng...Hedychium flavum

B. Cnh mi di, hp

a. Cnh mi mu vng..Hedychium flavescens

b. Cnh mi trng..Hedychium chrysoleucum

6

Phn b: Ch yu mt s vng ni cao thuc cc tnh Lo Cai ( xung quanh

Hong Lin Sn, Bt Xt), Lai Chu (Phong Th, Sn H, Ta Cha), H Giang

(Qung B, ng Vn) c trng ri rc Sn La, Lai Chu, Lo Cai, Tuyn

Giang, Yn Bi, Ph Th v H Ni (Gia Lm, T Lim) [34]. Chng mc t nhin

xen ln mt s cy g nh, cy bi, cy tho ven rng kn thng xanh, ven i,

cao 1400-1800m [5]. Ngi tin l cy pht trin tng i mnh, ph hp vi th

nhng v kh hu nhit i. mt s nc vng chu M latin thm ch cn xp

n vo loi cy xm thc. Bi vy, khi c trng trt quy m ln th tim nng s

dng v gi tr cy mang li rt ln bi ngoi ly hoa lm cnh, lm nc hoa th

thn r Hedychium coronarium cng c nhiu tc dng hu ch.

Thnh phn ha hc: Thn r Ngi tin ch yu cha tinh bt, glucose, acid amin,

cht bo, nha, tinh du, acid hu c [30]. Ngoi ra, ngi ta cn phn lp c

(E)-labda-8(17), 12-dien-15,16-dial, coronarin B, 7-hydroxycoronarin B, coronarin

D [10], [12], coronarin-D ethyl ether, coronarin-D-methyl ether, Labda-8 (17),

11,13-trien-15 (16)-olid, mt ester ca acid labda-8 (17,11,13)-trien-15-al-16-oic,

isocoronarin D, coronarin E, coronarin F [27], (+)-14-hydroxylabda-8(17), 12-

dien-16,15-lacton (isocoronarin D), 14,15,16-trinorlabda-8 (17), 11-E-dien-13-al

[10], [12], [24]. Phan Vn Kim v cc cng s (2011) cng b nghin cu mi

nht v cc hp cht c phn lp t thn r Ngi tin Sapa. Trong bo co ny,

h phn lp c ba labdan diterpen mi, t tn l coronarin G, H, I [28]. Cc

cht ny c tc dng c ch sn xut cc cytokine ca t bo ui gai c ngun gc

t ty xng l cc tc nhn gy vim hoc tin gy vim c gii phng ra khi b

kch thch bi LPS (lypopolysacchrid). Cc cht coronarin A, coronarin B,

coronarin C, coronarin D v (E)-labda-8(17),12-dien-15,16-dial l cc cht c i

vi t bo [5], [9].

Thnh phn tinh du thn r kh Ngi tin ch yu gm: 1,8-cineol

(37.44%), -pinen (17.4%), -pinen (6.73%), -terpineol (6.7%) [19], [20], trong

khi tinh du thn r ti gm: 1,8-cineol (41.42%), -pinen (10.39%), -terpineol

(8.8%), cymen (4.08%), -pinen (4.06%), terpinen-4-ol (3.55%) [5], [20]. c

7

nhiu nghin cu cho thy tc dng dit khun ca tinh du thn r Ngi tin trn

nhiu loi vi khun nh: Trichoderma, Candida albicans, Pseudomonas

aeruginosa, Bacillus subtilis [20], [31].

Tinh du hoa Ngi tin rt thm, cha ti 175 thnh phn, trong c

linalol, methyl-benzoate, cis-jasmon, methyl-jasmon, methyl epi-jasmonat, indol,

nhiu cht nitril v oxim. Cc cht: trans-ocimen, 2- v 3-methylbutanal oxim,

2-exohydroxy-1,8-cineol, methyl epi-jatmoat, cis-jasmen lacton to ra mi hoa Ngi

tin [5]. Tinh du l c eucalyptol (Trung dc t hi, 1993) [22].

Cu trc ha hc mt s diterpen c hot tnh sinh hc nh sau:

Coronarin A (C20H28O2) Coronarin B (C20H30O4) Coronarin D (C20H30O3)

Isoconarin D (C20H30O3) Coronarin E (C20H28O) Villosin C (C20H24O6)

Hnh 1.1. Cu trc ha hc mt s cht c hot tnh sinh hc c phn lp t

thn r Ngi tin [27], [28], [36].

8

Tc dng dc l

- Tc dng khng vi sinh vt: Tinh du hoa Ngi tin ti c tc dng c ch

s pht trin ca nm men bia (l mm sinh bnh cho cy v ng vt),

nhng tc dng yu trn mt s vi khun khc. Tinh du ca cnh non v

chi cy c ch s pht trin ca si nm Aspergillus flavus nng 100

ppm v gy c cho nm nng 3000 ppm [5], [18].

- Tc dng trn ng vt thn mm: Tinh du thn r Ngi tin c tc dng

trn sn ln Taenia solium mnh hn piperazin phosphat, nhng li km hn

hexylresorcinol trn Bunostomum trigonocephalum v Oesophagostomum

columbianum [5], [20]. Cao chit t ht Ngi tin c tc dng dit ng vt

thn mm Lymnaea cubensis v L.columella. Cao cng c tc dng dit cn

trng trn loi rp Macrosiphumrosae [5].

- Tc dng li niu v chng tng huyt p: Cao Ngi tin chit bng cn 50%

t l nhit thp, sau lm bay hi cn, c tc dng li niu v chng

tng huyt p chut cng trng bnh thng v chut cng trng c gy

tng huyt p [5], [20].

- Tc dng h st, chng vim, khng khun: Do tinh du l Ngi tin c cha

cht Eucalyptol. Ngoi ra cn lm gin ph qun v gim au [5], [20].

- Tc dng chng ung th: Do cc labdan diterpen lacton c phn lp t

thn r Ngi tin gy c ch s to thnh v c ch hot tnh ca NF-kB

(cha kha trung gian ca vim nhim, nguyn nhn nhiu loi bnh ung

th). Mt s nghin cu kho st hot tnh gy c t bo invitro cho thy

Coronarin D c tc dng c ch mnh s pht trin 3 dng ung th ngi:

ung th gan Hep-G2 (IC50 0,78 g/ml), ung th phi Lu (IC50 0,96 g/ml)

[8], ung th c vn tim RD (IC50 0,67 g/ml) [14] v tc dng gy c cc

dng t bo ung th dy thn kinh (SK-N-SH), ung th v (MCF-7) [9], ung

th c t cung (Hela). Villosin (mt labdan diterpen -lacton , khng no)

c tc dng c ch mnh dng t bo ung th phi ngi NCI-H187 mnh

(IC50 0.4 g/ml) [12], [24], [27].

9

Tnh v, cng nng

Thn r v qu Ngi tin c v cay, mi thm, tnh m, c tc dng khu

phong tr thp, n trung, tn hn. Tinh du gy trung tin [5], [6], [13].

Cng dng

Thn r v qu dng cha au bng lnh, bng y chng, tiu ha km.

Thn r sc ly nc sc ming cha hi ming, ung cha cm st, au mnh

my, phong thp, nhc mi gn xng. Dng ngoi th ly thn r ti gi nt,

nc ung, b p cha rn cn, n ng tn thng [6]. Thn cy thi nh, sc,

ngm cha vim li, vim amidan. Hoa c th dng n. Tinh du hoa l mt loi

hng liu cao cp [5], [13]. Trn th gii, nhiu nc cng s dng thn r ngi

tin cha 1 s bnh thng thng nh: Hawaii dng lm thuc tr thi mi [6],

n dng r cy tn bt lm thuc h nhit v tr t thp [6], [29], [30],

Mluyc, Indonexia dng nu nc sc ming, tinh du thn r tr giun [6], Vn

Nam, Trung Quc dng cha n ng tn thng, phong thp, cm mo, bch i

nng lnh. Qu dng tr y trng bng, n ung khng tiu [6], [22].

Hedychium coronarium var. flavescens l mt loi tho dc c s dng

lu i trong cc bi thuc c truyn ca n , Nepal, Ty Tng [16], [23]. Thn

r c dng tr cc chng ri lon tiu ha, nn, bun nn, tiu chy, bnh v gan,

gim au, h stBt thn r rang vng dng tr hen suyn [30], sc vi r cy

Bch hng dng tr bnh lao [24].

Cch dng, liu lng

Ngy dng 6-12g thn r kh sc hoc tn bt ung, dng ring hay phi hp

vi cc v thuc khc [13].

Bi thuc c Ngi tin

- Cha st: Thn r Ngi tin, hnh, tha l (liu lng bng nhau) dng ti,

gi nt, p [5], [29], [30].

- Cha au bng lnh, y trng, tiu ha km: Thn r kh hoc qu (6-

12g) sc ly nc hoc tn bt ung [13].

10

- Cha au mnh my, phong thp. nhc mi xng: Thn r (6-12g) sc ly

nc hoc tn bt ung [13], [22].

- Cha hi ming, vim li, vim Amidan: Thn r thi nh, sc ly nc sc

ming hng ngy [13].

- Cha n ng tn thng, rn cn: Thn r ti gi ly nc ung, b p

ln vt thng, p ln ch rn cn sau khi s cu [13].

1.2.2.2. Hedychium bousigonianum Pierre

Tn gi khc: Ngi tin Bousigon

M t: C ph sinh, thn r pht trin mnh, thn gi cao 1-1,2m. L c phin thon

hp, nhn hnh mi mc, di 30-50cm, rng 7cm, c 2 mt u nhn, khng lng.

Li nh hnh trng, di 1,1-2cm. Cm hoa tha, di 20cm, l bc c lng, di

2,5cm. Hoa to, mu vng. Tiu nhy lp hp, di 4cm, mi xoan, ch n . Nh

dng si, di 5cm. Non so c lng. Qu nang, nt 3 mnh, cha nhiu ht, ht

hnh elip. Ma hoa vo thng 5, 6 hng nm [9], [34].

Phn b: Cy c ngun gc t vng Himalayas ca Nepal v n sau pht

tn ti khu vc Nam chu Phi v Nam M. Loi cn phn b Nam Trung Quc,

Malaixia, c v Vit Nam. Cy mc nhng vng c kh hu mt lnh [8]. Vit

Nam, cy c tm thy trong rng kn thng xanh, Lt [9].

Thnh phn ha hc: Khi chit tch cc phn on ca dch chit ton phn thn

r Ngi tin Bousigon kt hp dng khi ph xc nh c 3 cht c hot tnh sinh

hc l: villosin, epimer ca ete metyl coronarin D v geniposid [8]. Cc thnh phn

chnh ca tinh du thn r Ngi tin Bousigon c thu hi Vnh Phc gm: 1,8-

cineol, linalool, -terpineol, -tumeron, 37,94-zerubon, borneol, myrtenal

bicycle[8].

Tc dng: Th nghim invitro cho thy cn chit t thn r cy ngi tin Bousigon

c hot tnh chng 2 loi vi khun gy bnh ng rut l E.coli gy bnh tiu chy

v B.subtullis gy bnh vim i trng [8].

11

1.2.2.3. Hedychium coccineum Hamilt

Tn gi khc: Ngi tin

M t: Cy thn r pht trin, thn gi cao 1,5-2m. L khng cung, c phin di

25-50cm, rng 3-5cm, hnh mi mc, c khi hnh tim. Li nh kch thc 1,2-

2,5cm, bao ly thn gi. Cm hoa hnh tr, cao 15-25cm, rng 6-7 cm. L bc pht

trin, di 3-4cm, khng lp ln nhau, c t lng u l bc, gn nh vng xon 3.

Hoa mu , i di 3cm, cnh hoa di 3cm, cnh mi 2 thy, ng trng hoa mnh

mai, nh hoa di 4,5cm, non so c lng [9], [34]. Phn b vng m ly, vng

ni cao [9], [34].

Thnh phn ha hc: Tinh du thn r ch yu cha: ,-pinen (13,5-7,5%),

camphen (2,3%), linalool (26,7%), bornyl acetat (8,4%), curcumen (4,1%), (E)-

nerolidol (4,6%), spathulenol (3,1%) [20]

1.2.2.4. Hedychium ellipticum (Smith) Kuntze

Tn gi khc: Ngi tin bu dc, Gandasulium elipticum (Smith) Kuntze.

M t: Cy thn tho cao 1m, l c phin bu dc, kch thc 20-4010-15cm, mp

l nu, cao 1cm, b khng lng. Cm hoa nghing, mc dy. Kch thc 123-4cm.

L bc khng lng di 3cm, hoa trng, di 8cm, vnh c lng nhung, non so c

lng [9]. Phn b rng kn thng xanh [9].

Thnh phn ha hc: Sineenard Songsri v cc cng s (i hc Chiang Mai,

Thi Lan) nghin cu thn r cy Hedychium ellipticum cho thy cha mt s cht

c hot tnh sinh hc ging cc loi Hedychium khc nh: coronarin D, coronarin E,

16-hydroxylabda-8 (17),11,13-trien-15,16-olid, 15-methoxylabda-8 (17),11,13-

trien-15,16-olid, (E)-Labda-8 (17),12-dien-15,16-dial, Villosin, (E)-14,15,16-

trinorlabda-8 (17),11-dien-13-oic acid, (E)-15,16-bisnorlabda-8 (17), 11-dien-13-on.

Ngoi ra, cn c 1 s cht khc nh zerumin A, zerumin B, ellipticin, doxorubicin.

Th nghim invitro cho thy dch chit thn r H.ellipticum c tc dng khng

Mycobacterial tuberculosis, c tc dng c ch t bo ung th nim mc ming

(KB), ung th v (BC), ung th phi t bo nh (NCI-H187). Tinh du thn r

Hedychium ellipticum ch yu cha: 1,8-cineol (83,90%), -pinen (1,20%),

12

terpinen-4-ol (1,37%), -terpineol (1,26%), -pinen-10-ol (0,64%), -pinen

(0,83%), pinocarveol (0,55%)[20]

1.2.2.5. Hedychium gardnerianum Roscoe

Tn gi khc: Ngi tin Gardner, Kanili ginger.

M t: Cy thn tho, thn gi cao n 1m, dng ging gng. Phin l bu dc,

khng cung, mp cao. Cm hoa cao, l bc xanh, 21cm; cnh hoa hp, di 3cm,

rng 3-4mm. Nh lp dp mu vng ti, mi vng chanh c 2 thy, tiu nhy mu

cam m, ch ta. Non so khng c lng [9]. Phn b lt [14].

Thnh phn ha hc: Tinh du l Hedychium gardnerianum cha: Sesquiterpen

hydrocarbon (50,9%), Oxygen sesquiterpen (15,9%), -Calacoren (16,2%), ()

Cedreanol (15,9%), 3,4-Dimethyl-3-cyclohexen-1-carboxaldehyd (10,5%), trans--

Bisabolen (4,0%), 8,9-Dehydroisolongifolen (4,8%), 9,10-Dehydroisolongifolen

(4,8%), Cadalin (5,0%), -Cadinen (3,4%)[32]

Tc dng: Nghin cu cho thy tinh du l H.gardnerianum c tc dng c ch

acetylcholine esterase gip chng trm cm; c ch oxy ha lipid trong c th gip

chng oxy ha v c tc dng kh quan khi h tr iu tr bnh Alzheimer [32].

1.2.2.6. Hedychium forresti Diels

Tn gi khc: Ngi tin l hoa rng

M t: Cy c, cao 1,2m, c mi thm. Phin l thon nhn, 4010cm, mp cao 2,5-

3cm, c 2 mt nhn. Cm hoa cao 15cm, l bc di 3-4cm, hoa mu vng ti, mi

tam gic, c 2 thy thon [9]. Phn b Sapa, cao 1500m [9].

Thnh phn ha hc: Tinh du thn r cha: -pinen (14%), 1,8-cineol (10,1%),

linalool (56%), (E)-nerolidol (1,1%), -terpineol (1,1%), terpinen-4-ol (2,1%), -

terpinen (3,4%), p-cymen (1,4%), limonen (0,9%)[20]

1.2.2.7. Hedychium villosum Wall

Tn gi khc: Ngi tin lng.

M t: Cy thn tho, cao 1,5-2 m [33]. L c phin di, thon, hp, di 30cm, nhn

2 mt, mp cao 2-3cm. Cung l di 1-2cm, i khi tiu bin. Li nh di 3-4 cm,

rng 1cm. Cm hoa dy, di 10-15cm, hoa mc dy c. L bc c lng nhung,

13

mu nu nht, di 2-2,5cm, rng 1cm, khng lp ln nhau. Cnh hoa ph ngn hn,

hoa trng, ng i 2cm. Cnh hoa v tiu nhy lp di 3cm, mi hp, di 2,5cm, x

2 thy su. Vi nhy di 4,5cm, bao phn rt ngn, ch nh di 5cm. Non so c

lng. Ma hoa vo thng 2,3 hng nm [9], [34]. Phn b Bc b [9], [34].

Thnh phn ha hc: Trong thn r cha 14,15,16-trinorlabda-8 (17), 11-(E) dien-

13-carboxylic acid, 13, 14, 15, 16-tetranorlabda-8 (17)-en-12-carboxylic acid, (E)-

15,16-dinorlabda-8 (17), 11-dien-13-on, villosin, coronarin E v -sitosterol [33].

14

CHNG 2. I TNG V PHNG PHP NGHIN CU

2.1. NGUYN LIU, PHNG TIN NGHIN CU

2.1.1. Nguyn liu nghin cu

Mu nghin cu cy Ngi tin c thu hi cnh ng trng hoa x Ty

Tu-Qun Bc T Lim- H Ni vo thng 1/2014. Bao gm:

+ Mu cy ti mang l, thn r, qu nghin cu v mt thc vt.

+ Mu thn r ti, ra sch, thi nh, sy trong t sy 60 n kh,

nghin bng thuyn tn, bo qun trong ti nilon kn, ch mt lm nguyn liu

nghin cu v mt ha hc.

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.

- Ha cht: Javen, A.acetic, xanh methylen, son phn, nc ct, Na2SO4 khan,

- Dung mi hu c: chloroform, ethylacetat, acid formic, ether du ha, toluene,

- Thuc th: cc thuc th dng trong phn ng nh tnh v sc k.

- Bn mng trng sn Silica gel 60 F254 ca Merck.

- Dng c: dng c thy tinh v cc dng c khc dng trong phng th nghim

(cc c m, bt s, thuyn tn, a thy tinh, lam knh, bnh nn)

- B dng c: B dng c ct tinh du ci tin.

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 o m Sartorious.

- My chm sc k CAMAG LINOMAT 5.

- My chp nh sc k CAMAG REPROSTAR 3.

- My vi tnh vi phn mm winCATS v VideoScan.

15

- My nh Canon ISUS 115.

- My sc k kh khi ph (GC-MS) Agilent Technologies.

2.2. PHNG PHP NGHIN CU

2.2.1. Nghin cu thc vt bng phng php nghin cu hin vi

- c im vi phu: Mu thn r, l Ngi tin c ct bng dng c ct cm tay,

chn lt ct mng, 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 nhm gp phn xy dng tiu chun nguyn liu [2], [3].

- Soi bt: L, thn r Ngi tin c sy kh, nghin nh thnh bt bng thuyn tn

v ci s, ry qua ry 180m ly bt mn, ln tiu bn v quan st, chp nh di

knh hin vi xc nh cc c im bt theo ti liu [2], [3].

2.2.2. Nghin cu bng phng php ha hc

- nh tnh cc nhm cht hu c c trong mu bt thn r Ngi tin bng phn ng

ha hc [7].

- Sc k lp mng: S dng bn mng Silicagel trng sn ca Merck. Chm sc k

bng my. Sau khi trin khai, hin vt bng n cc bc sng 254nm v 366nm,

phun thuc th hin mu Vanilin/H2SO4 c. Chp nh sc k, phn tch sc k

bng phn mm WinCATS v Video Scan.

2.2.3. Nghin cu v tinh du thn r Ngi tin

- Ct tinh du thn r Ngi tin theo phng php ct ko hi nc bng b dng c

ct tinh du ci tin (Hnh 2.1, trang 16) [2].

16

Hnh 2.1: Cu to b

dng c ct tinh du ci

tin [2]

A. Bnh cha dc liu

B. B phn treo ng

hng tinh du

C. ng sinh hn

- Xc nh m thn r ti bng my o m Sartorious, nguyn tc o theo

cng thc:

a = 100% (mti -mkh)/mti

Trong : a: hm m dc liu ti

mti: Khi lng thn r ti

mkh: Khi lng thn r kh tuyt i

- Xc nh hm lng tinh du thn r Ngi tin theo dc liu kh tuyt i.

Cng thc tnh: XTd% = Vtd/ (mti- mti a) 100%

Trong : XTd%: hm lng phn trm tinh du thn r

Mti: khi lng dc liu ti (g)

a: m dc liu ti.

- Sc k bn mng tinh du.

- Tin hnh sc k kh khi ph (GC-MS) xc nh thnh phn ha hc tinh du

thn r Ngi tin.

17

CHNG 3. THC NGHIM V KT QU

3.1. Nghin cu v mt thc vt

3.1.1. M t hnh thi cy

Hnh 3.1: c im mt s b phn cy Ngi tin

1.nh cy

2.Cm l bc

3.Qu

4.Ht

5.Thn r

1

2

3

4

5

18

3.1.1. c im hnh thi cy v gim nh tn khoa hc

c im hnh thi mu thu hi: Cy thn tho cao 1-2m ging cy gng (1). L

khng cung (1), hnh di mi mc, nhn mt trn, c lng d rng mt di,

di ti 20-45cm, rng 12cm, thon di. Li nh m ly thn gi. Cm hoa hnh

trng, dng nn, l bc hp, lp ln nhau (2). Qu nang (3), nt 3 mnh, bn trong

c nhiu ht mu (4). Thn r pht trin, dng c ging gng (5).

Gim nh tn khoa hc: Tin hnh i chiu cc c im mu nghin cu vi cc

thng tin c ghi chp trong Cy c Vit Nam [9], Thc vt ch ng dng [34],

Thc vt ch Trung Quc [19], cy c TS. Nguyn Quc Huy (B mn Thc

Vt, i hc Dc H Ni) gim nh tn khoa hc l Hedychium coronarium

Koenig (Ph lc 1).

3.1.2. Nghin cu vi hc vi phu dc liu

* Tin hnh:

- Tin hnh lm ring vi phu gn l, thn r ring, ct bng dng c ct vi phu

cm tay, chn cc lt ct mng. Ty lt ct dc liu bng cloramin B bo ha ti

khi lt ct trng hon ton ty sch cc cht trong t bo, ch gi li mng t bo

nhm quan st tiu bn d hn. Ra sch bng nc ct nhiu ln. Ngm trong dung

dch acid acetic 5% ty clorid ca cloramin B. Nhum xanh methylen ( pha

long theo t l 1:4) trong vng 15 pht. Ra sch nhiu ln bng nc ct. Nhum

son phn trong 5 pht. Ra sch nhiu ln bng nc ct. t vi phu vo mt

git glycerin trn phin knh,y lamen, soi trn knh hin vi. Chp nh cc vi phu

bng my Canon IXUS 115 vt knh 10x ca knh hin vi.

* Kt qu

- c im vi phu ca l Ngi tin:

Quan st di knh hin vi (Hnh 3.2), ta thy cc c im sau:

Phn gn l: Mt trn hi lm, mt di li. Biu b trn v di gm cc t

bo trn, nh, xp u n (1). St lp biu b trn v di l m mm gm cc t

bo hnh trn hoc a gic c thnh t bo mng (2). C nhiu b libe-g xp u

n thnh nhiu vng cung hng v pha gn chnh, g trn (3), libe di (4).

19

Vng m cng m ly libe (6). B libe-g gia, hng cui t trn xung ln nht

v nh dn v 2 pha. Xen k cc b libe-g hng th 3 l cc m khuyt (5). Ri

rc trong m mm c cc t bo tit tinh du (8).

Phn phin l: Biu b trn v di gm 1 lp t bo hnh ch nht, nh, xp

u n (7). C 2 mt u c l kh. Gia 2 biu b trn, di l m mm gm cc

t bo hnh trn hoc a gic khng u nhau (2). Ri rc trong m mm c cc b

libe-g, g trn, libe di, vng m cng di libe. Ngoi ra cn c cc t bo

tit tinh du (8).

- c im vi phu ca thn r Ngi tin:

Quan st di knh hin vi (Hnh 3.3), ta thy cc c im sau:

Vi phu ct ngang thn r Ngi tin thng c hnh trn. Ngoi cng l lp

bn gm 10-12 lp t bo. 6 lp t bo bn ngoi l cc t bo hnh a gic, 6 lp

bn trong l cc t bo hnh ch nht xp thnh cc dy xuyn tm (1). St lp bn

l m mm v, gm cc t bo hnh trn hoc a gic, thnh t bo mng (2). Ri

rc trong m mm v c cc t bo tit tinh du (3). Pha trong xut hin nhiu m

khuyt (4). Tip theo l ni b gm 1 lp t bo hnh ch nht xp u n (5). St

lp ni b c nhiu b libe-g nm trong m mm rut (6, 7). M mm rut cu to

t cc t bo hnh trn hoc a gic (8), to hn t bo m mm v, xen ln c cc t

bo tit tinh du, cc mch (9).

20

Hnh 3.2: Vi phu l Ngi tin

1.Biu b gn l 5.M khuyt

2.M mm 6.Vng m cng

3.G 7.Biu b phin l

4.Libe 8.T bo tit tinh du

Hnh 3.3. Vi phu thn r Ngi tin

1.Bn 6.Libe

2.M mm v 7.G

3.T bo tit 8.M mm rut

4.M khuyt 9.Mch

5.Ni b

21

3.1.3. Nghin cu vi hc bt dc liu

* Tin hnh

Sy kh l, thn r Ngi tin trong t sy 600C sau dng thuyn tn

nghin nh. Ry ly bt mn, dng kim mi mc ly bt dc liu cho ln phin

knh nh sn nc ct, y lamen ln v quan st di knh hin vi. Xc nh

nhng c im vi hc ca bt l, bt thn r Ngi tin v chp li bng my nh.

* Kt qu:

- c im vi hc bt l Ngi tin:

Bt c mu xanh, mi thm, v cay. Quan st di knh hin vi (Hnh 3.4,

trang 22) thy c mt s c im: C cc mnh m mm gm cc t bo hnh trn

hoc a gic thnh mng xp st nhau (1), c nhiu mnh biu b mang l kh (2),

cc mnh phin l (3). Ri rc c cc mnh mch (4,5), b si (6).

- c im vi hc bt thn r Ngi tin:

Bt c mu vng nht, mi thm, v cay. Quan st di knh hin vi (Hnh

3.5, trang 22) thy c mt s c im: Ri rc c cc mnh mang mu (1). Mnh

bn vi cc t bo xp thnh cc dy xuyn tm (2). Ht tinh bt hnh trng hoc

hnh chung, c cc vn ng tm, khng r. ng ring r (6) hoc thnh m (3),

ng knh 0,04-0,07mm. Ngoi ra cn c cc mnh mch (4), (5).

Nhn xt: Bt l Ngi tin khng c c im c trng nhn bit so vi cc bt

l khc. Trong khi , bt thn r c mt s c im c trng nh c nhiu mnh

bn gm nhiu lp t bo hnh a gic, xp thnh cc hng xuyn tm. C cc ht

tinh bt hnh trng hoc hnh chung, kch thc 0,04-0,07mm, vn ng tm

khng r nt.

22

Hnh 3.4: Mt s c im bt l Ngi tin

1. Mnh m mm 2. Biu b mang l kh 3. Mnh phin l

4,5. Mnh mch 6. B si

Hnh 3.5: Mt s c im bt thn r Ngi tin

1. Mnh mang mu 2. Mnh bn 3. m tinh bt

4,5. Mnh mch 6. Ht tinh bt

23

3.2. Nghin cu v ha hc

3.2.1. Ct tinh du

3.2.1.1. Xc nh m thn r ti

Ct nh thn r Ngi tin ti loi b tp, ra sch, lau kh. Sau cn

chnh xc 1g dc liu ti v cho vo ci s nghin nh. a vo my xc nh

m dc liu.

Kt qu: m thn r NGait tin ti: a= 82,48% .

3.2.1.2. Xc nh hm lng tinh du cc b phn Ngi tin

Phng php ct tinh du: Ct ko bng hi nc.

Dng c: S dng b dng c ct tinh du ci tin, B mn dc liu, i hc

Dc H Ni (Hnh 2.1) [2].

Tin hnh:

- Thi nh thn r ti lm sch tp. Sau nghin nt bng thuyn tn.

- Cn dc liu thn r ti mti = 469,82g.

- Cho dc liu vo bnh ct tinh du, nc va ngp mt dc liu. Lp b

dng c ng quy nh.

- Ct trong vng 4 ting (tc ct 2-3ml/pht). Sau ngng ct, sau 15 pht c

th tch tinh du phn chia vch ca dng c hng tinh du.

- Rt ly tinh du vo l thy tinh c np Silicon.

Kt qu: Vtinh du = 0,3ml

p dng cng thc (Trang 16): XTd% = 0,365%

Trong : XTd%: hm lng phn trm tinh du trong thn r

mti=469,82 g

a=82,48%

3.2.2. nh tnh cc nhm cht trong dc liu bng phn ng ha hc.

3.2.2.1. nh tnh flavonoid.

Cn khong 10g bt thn r Ngi tin cho vo bnh nn 100ml, thm 50ml

cn 90. un cch thy 10 pht, lc nng ly dch lc lm cc phn ng sau:

24

a. Phn ng Cyanidin (Phn ng Shinoda)

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, nhn

thy dung dch chuyn t mu vng sang mu hng Phn ng dng tnh.

b. Phn ng vi kim

Phn ng vi hi amoniac: Nh 1 git dch chit ln t giy lc, sy kh, ri

h trn ming l c cha amoniac c m nt, i chiu vi t giy nh git

dch chit i chng thy mu vng ca vt m ln r rt Phn ng dng tnh.

Phn ng vi dd NaOH 10%: Cho vo ng nghim 1ml dch chit, thm vi

git dung dch NaOH 10%, thy dch chit chuyn t vng sang vng m, khi un

nng thy xut hin mu Phn ng dng tnh.

c. Phn ng vi FeCl3

Cho vo ng nghim nh 1ml dch chit. Thm vi git dung dch FeCl3

5%.Phn ng dng tnh khi xut hin kt ta xanh en Phn ng dng tnh.

d. Phn ng diazo ha

Cho 1ml dch chit vo ng nghim, kim ha bng dung dch kim dung

dch NaOH, thm vi git thuc th diazo mi pha, lc u, un nng trn ni cch

thy trong vi pht thy xut hin mu Phn ng dng tnh.

Kt lun s b: Thn r Ngi tin c cha flavonoid.

3.2.2.2. nh tnh coumarin.

Ly khong 5g bt thn r Ngi tin cho vo bnh nn 100ml, thm 50ml cn

90. un cch thy 5 pht, lc nng qua bng. Dch lc thu c dng lm cc

phn ng sau:

a. Phn ng m ng vng lacton :

- 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 khng thy hin tng g

Phn ng m tnh.

25

b. Phn ng diazo ha

Cho 1ml dch chit vo ng nghim, kim ha bng dung dch NaOH, thm

vi git thuc th diazo mi pha, lc u, un nng trn ni cch thy trong vi

pht thy xut hin mu Phn ng dng tnh.

c. Quan st hunh quang ca cc vt coumarin di nh sng t ngoi khi tc dng

vi dung dch kim.

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 ng xu ri chiu

tia t ngoi trong vi pht. B ng xu ra, quan st tip di n t ngoi thy phn

khng b che c hunh quang sng ging phn b che Phn ng m tnh.

d. Vi thng hoa

Cho mt t bt thn r Ngi tin vo np chai kim loi. t ln bp in c

li amian, cho bay ht hi nc trong dc liu. t trn ming np nhm mt

phin knh trn c t t bng thm nc lnh. un nh di np nhm, sau

5 pht ly lam knh ra, ngui, soi di knh hin vi khng thy tinh th hnh kim

Phn ng m tnh.

Kt lun s b: Thn r Ngi tin khng cha coumarin.

3.2.2.3. nh tnh saponin.

a. Quan st hin tng to bt

Cho vo ng nghim ln 0,1g bt thn r Ngi tin, thm 5ml nc. Lc

mnh trong 5 pht. yn v quan st thy hin tng to bt bn vng sau

15pht.

b. Hin tng ph huyt

Cho mt git mu b loi fibrin ln phin knh, nh thm mt git dch

chit nc ca thn r Ngi tin. Quan st hin tng ph huyt trn knh hin vi.

c. Phn ng phn bit Saponin steroid v Saponin triterpenoid

Ly 1g bt thn r Ngi tin cho thm 5ml cn un si cch thy trong 15

pht. 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

26

cn chit ri bt ng nghim, lc mnh c 2 ng trong 15s. yn nu ct bt trong

2 ng cao ngang 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: Ct bt trong 2 ng ngang nhau Dc liu cha saponin

triterpenoid

Kt lun s b: Thn r Ngi tin c cha saponin triterpenoid.

3.2.2.4. nh tnh alcaloid.

Cn 1g bt thn r Ngi tin cho vo bnh nn dung tch 50ml. Thm 15ml

dung dch acid sulfuric 1N. un n si. ngui. Lc dch lc vo bnh gn dung

tch 100ml. Kim ha dch lc bng dung dch amoniac 6N (khong 8ml) n pH =

9-10 (th bng giy qu hoc ch th mu vn nng). Chit alcaloid base bng

cloroform (chit 3 ln, mi ln 5ml). Gp cc dch chit cloroform, loi nc bng

natrisulfat khan, sau dng lm phn ng nh tnh.

Ly mt phn dch chit cloroform c chun b trn, em lc vi acid

sulfuric 1N hai ln, mi ln 5ml. Gp cc dch chit nc. Chia u vo cc ng

nghim nh, mi ng 1ml. Nh vo tng ng nghim 2-3 git ln lt cc thuc th

sau:

a. ng 1: thuc th Mayer, khng thy xut hin ta mu t trng n vng.

b. ng 2: thuc th Bouchardat, khng thy xut hin ta mu t nu n nu.

c. ng 3: thuc th Dragendorff, khng thy xut hin ta mu t vng cam n .

Kt lun s b: Thn r Ngi tin khng cha alcaloid.

3.2.2.5. nh tnh tanin

Ly khong 1,00g bt thn r Ngi tin cho vo bnh nn dung tch 50ml,

thm 20ml nc ct, un si trong 2 pht. ngui, lc. Dch lc dng nh tnh.

a. ng 1: ly 2ml dch lc, thm 2 git dung dch FeCl3 5% (TT) thy xut hin kt

ta mu xanh nu nht Phn ng dng tnh.

b. ng 2: ly 2ml dch lc, thm 2 git ch acetat 10% (TT) thy xut hin ta bng

Phn ng dng tnh.

27

c. ng 3: ly 2ml dch lc, thm 5 git dung dch gelatin 1% khng thy xut hin

ta bng trng Phn ng m tnh.

Kt qu: 2 phn ng a, b dng tnh. Phn ng c m tnh.

Kt lun s b: Thn r Ngi tin khng c tannin.

3.2.2.6. nh tnh anthranoid.

a. Phn ng Borntraeger

nh tnh anthranoid ton phn (dng glycosid v dng t do)

Cho vo ng nghim ln 1g thn r Ngi tin. Thm 5ml dung dch acid

sulfuric 1N. un trc tip trn ngun nhit n si. Lc dch chit cn nng qua

mt lp bng mng vo trong bnh gn dung tch 50ml. Lm ngui dch lc. Thm

5ml chloroform, lc nh, gn b lp nc. Gi lp cloroform lm phn ng.

Ly 1ml dch chit cloroform, thm 1ml dung dch amoniac. Lc nh. Khng

thy lp nc xut hin mu sim Phn ng m tnh.

Kt lun s b: Thn r Ngi tin khng c anthranoid.

3.2.2.7. nh tnh glycosid tim.

Ly 3g thn r Ngi tin, chit soxhlet vi n-hexan 1 gi. B dc liu sy

kh, cho vo bnh cu, un hi lu vi ethanol 40% trong 1 gi. Gn dch chit vo

cc c m, thm khong 3ml ch acetat 30%, khuy u. Lc loi ta, th dch lc

vn cn ta vi ch acetat, cho thm 1ml ch acetat na vo dch chit, khuy v lc

li. Tip tc th n khi dch chit khng cn ta vi ch acetat. Cho ton b dch

lc vo bnh gn v lc k vi hn hp chloroform:ethanol t l 4:1 (3 ln, mi ln

5ml), gn ly lp chloroform vo cc c m kh sch. Chia dch chit vo 6 ng

nghim nh, bc hi dung mi trn ni cch thu cho n kh. Cn cn li lm

cc phn ng nh tnh sau:

a. Phn ng ca khung steroid: Phn ng Liebermann

Cho vo ng nghim c cha cn trn 1ml anhydrid acetic, lc u cho tan

ht cn. Nghing ng 450. Cho t t theo thnh ng 0,5ml acid sulfuric c, trnh

xo trn cht lng trong ng. Nhn thy mt tip xc gia hai lp cht lng xut

28

hin mt vng mu tm . Lp cht lng pha di c mu hng, lp trn c mu

xanh l Phn ng dng tnh.

b. Phn ng ca vng lacton 5 cnh

- Phn ng Baljet

Cho vo ng nghim c cha cn trn 0,5ml ethanol 90%. Lc u cho tan

ht cn. Nh tng git thuc th Baljet (1 phn dung dch acid picric 1% v 9 phn

dung dch NaOH 10%) mi pha. Khng thy xut hin mu da cam

Phn ng m tnh.

- Phn ng Legal

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. Phn ng thy xut hin mu cam Phn ng m tnh.

c. Phn ng ca phn ng 2,6 - desoxy: Phn ng Keller - Kiliani

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 sulphuric c, trnh xo trn

cht lng trong ng. mt tip xc gia 2 lp cht lng xut hin 1 vng mu tm

. Lc nh, lp cht lng pha trn c mu xanh l Phn ng dng tnh.

Kt qu: Ch c phn ng a v c dng tnh.

Kt lun s b: Thn r Ngi tin khng c glycosid tim.

3.2.2.8. nh tnh acid hu c

Cn khong 3g bt thn r Ngi tin cho vo ng nghim to, thm 10ml nc

ct em un si trc tip 10 pht, ngui ri lc qua giy lc gp np. Cho vo

ng nghim nh khong 2ml dch lc, thm mt t tinh th Na2CO3. Thy c bt kh

thot ra Phn ng dng tnh.

Kt lun s b: Thn r Ngi tin c cha acid hu c.

3.2.2.9. nh tnh ng kh

Cn khong 3g thn r Ngi tin cho vo ng nghim to, thm 1ml nc ct,

un si. i vi cao s dng 1g v ha tan bng nc nng. Lc qua giy lc vo 1

29

ng nghim khc. Thm 1ml dung dch thuc th Felling A v 1ml dung dch

Felling B. un cch thy si vi pht. Thy xut hiu ta gch Phn ng

dng tnh.

Kt lun s b: Thn r Ngi tin cha ng kh.

3.2.2.10. nh tnh acid amin

Cn khong 3g thn r Ngi tin cho vo cc c m, thm 20ml nc ct,

un si vi pht. i vi cao s dng 1g v ha tan bng nc nng. Lc qua giy

lc vo 1 ng nghim khc. Thm vi git thuc th Ninhydrin 3%. un cch thy

si vi pht. Thy xut hiu mu tm Phn ng dng tnh.

Kt lun s b: Thn r Ngi tin c cha acid amin.

3.2.2.11. nh tnh polysaccharid

Chun b dch chit nh i vi phn ng nh tnh acid amin. Chia dch chit 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.

Thy mu ng 1 m hn ng 2 v ng 3 Phn ng dng tnh.

Kt lun s b: Thn r Ngi tin cha polysaccharid.

3.2.2.12. nh tnh cht bo

Cn khong 3g bt thn r Ngi tin vo cc c m 100ml, thm 10ml ether

du ha, bc kn, ngm 1 gi. Lc qua giy lc gp np ly dch lc. Nh 1 git

dch chit ln mnh giy trng, sy nh cho bay ht hi dung mi, khng thy vt

m trn giy Phn ng m tnh.

Kt lun s b: Thn r Ngi tin khng cha cht bo.

3.2.2.13. nh tnh caroten

Cho vo ng nghim nh khong 2ml dch chit ether du ha ca thn r

Ngi tin, c cch thy n cn, nh vi git H2SO4 c vo cn. Khng thy xut

hin mu xanh l Phn ng m tnh.

Kt lun s b: Thn r Ngi tin khng cha caroten.

30

3.2.2.14. nh tnh sterol

Cho vo ng nghim nh khong 2ml dch chit ether du ha ca thn r

Ngi tin, c cch thy n cn. Thm vo ng nghim khong 1ml anhydride

acetic, lc k cho tan ht cn. nghim ng nghim 450, thm t t H2SO4 c

theo thnh ng nghim. Quan st thy mt phn cch c vng tm , lp cht lng

pha trn c mu xanh l Phn ng dng tnh.

Kt lun s b: Thn r Ngi tin cha sterol.

3.2.2.15. nh tnh iridoid

Cn khong 3g bt thn r Ngi tin vo ng nghim to, thm 10 dung dch

HCl 1% ngm trong 5 gi. Lc qua giy lc gp np ly dch lc. Cho vo ng

nghim nh 1ml dch lc trn, thm 1ml thuc th Trim-Hill (10ml acid acetic +

1ml CuSO4 0,2% + 0,5ml HCl c). Khng thy xut hin mu xanh. Phn ng

m tnh.

Kt lun s b: Thn r Ngi tin khng cha iridoid.

Kt lun s b chung: Sau khi tin hnh nh tnh s b 15 nhm cht hu c v

ct tinh du th ta thy thn r Ngi tin cha Flavonoid, Saponin, Tinh du, Acid

hu c, ng kh, Acid amin, Sterol, Polysarcarid.

Kt qu phn ng nh tnh c tng kt bng sau y:

31

Bng 3.1: Kt qu s b nh tnh cc nhm cht trong thn r Ngi tin

Nhm cht Phn ng nh tnh Kt qu Kt lun

1

Phn ng vi hi NH3 +

Phn ng vi dd NaOH 10% ++

Phn ng vi dd FeCl3 5% +

Phn ng Cyanidin ++

Phn ng diazo ha +

2

Phn ng Libermann +

Phn ng Legal -

Phn ng Baljet -

Phn ng Keller-Kiliani +

3 Saponin Quan st hin tng to bt ++

C Quan st hin tng ph huyt +

4

Phn ng m, ng vng lacton -

Khng c Phn ng diazo ha -

Phn ng chuyn dng ng phn cis- trans -

Vi thng hoa -

5

Phn ng vi dd FeCl3 5% +

Phn ng vi dd gelatin 1% -

Phn ng dd ch acetat 10% +

6 Anthranoid Phn ng Borntraeger - Khng c

7

Phn ng vi TT Mayer -

Phn ng vi TT Dragendorff -

Phn ng vi TT Bouchardat -

8 Acid hu c Phn ng vi Na2CO3 + C

9 ng kh Phn ng vi TT Fehling A v Fehling B ++ C

10 Acid amin Phn ng vi TT Ninhydrin 3% ++ C

11 Cht bo To vt m trn giy - Khng c

12 Caroten Phn ng vi H2SO4 c - Khng c

13 Sterol Phn ng Liebermann + C

14 Polysaccharid Phn ng vi TT Lugol ++ C

15 Iridoid Phn ng vi thuc th Trim-Hill - Khng c

Khng c

Khng c

Flavonoid

Coumarin

Tanin

Alcaloid

C

Glycoside tim

Khng c

- Phn ng m tnh + Phn ng dng tnh ++ Phn ng dng tnh r

32

3.2.3. nh tnh dch chit MeOH ton phn bng sc k lp mng

Chun b dch chit chm sc k

Phng php ngm lnh: Cho 5g bt thn r Ngi tin vo bnh nn, thm

Methanol ngp mt dc liu. Ngm nhit phng trong 24h, gn ly dch chit,

lc qua giy lc thu c dch chm sc k.

Chm sc k: Chm bng my chm sc k CAMAG LINOMAT 5. Chp li nh

bc sng 254nm, 366nm v sau khi phun thuc th hin mu bng my CAMAG

REPROSTAR 3. S dng phn mm WinCATS, VideoScan x l hnh nh.

H dung mi

-H 1: Toluen-Ethyl acetat-Acid formic (5,5:4,5:0,5)

-H 2: Toluen-Ethyl acetat (93:7)

-H 3: n-Hexan-Ethyl acetat-Acid formic (6:4:0,5)

-H 4: Toluen-Ethyl acetat-Methanol-Acid Formic (4:5:0,5:1)

-H 5: Diclomethan-Methanol (6:4)

Thuc th hin mu: Vanilin/H2SO4 c

Kt qu: Sau khi tin hnh khai trin sc k dch chit MeOH ton phn trn cc h

dung mi khc nhau th nhn thy h 1 c kh nng tch vt tt nht.

3.2.4. nh tnh dch chit sau khi thy phn bng sc k lp mng

Chun b dch chit chm sc k

Cho 5g bt thn r Ngi tin vo bnh nn 100ml, thm dung dch H2SO4 1N

v nc t l bng nhau cho ngp qua mt thong dc liu. Lp ng sinh hn, un

hi lu trong 3 ting. ngui, lc qua giy lc, ly dch lc cho vo bnh gn.

Thm Cloroform 3 ln (mi ln 5ml), lc nh trnh to nh, gn v gp 3 dch

Cloroform mang i un n cn. Ha tan cn bng MeOH, lc qua giy lc ln na

v mang i chm sc k.

Chm sc k: Chm bng my chm sc k CAMAG LINOMAT 5. Chp li nh

bc sng 254nm, 366nm v sau khi phun thuc th hin mu bng my CAMAG

REPROSTAR 3. S dng phn mm WinCATS, VideoScan x l hnh nh.

33

H dung mi

-H 1: Toluen-Ethyl acetat-Acid formic (7:3:0,1)

-H 2: Toluen-Ethyl acetat (7:3)

-H 3: n-Hexan-Ethyl acetat-Acid formic (6:4:0,5)

-H 4: Toluen-Ethyl acetat-Methanol-Acid formic (4:5:0,5:1)

Thuc th hin mu: Vanilin/H2SO4 c

Kt qu: Sau khi tin hnh khai trin sc k dch chit sau khi thy phn trn cc h

dung mi khc nhau th nhn thy h 1 c kh nng tch cht tt nht.

a. Hnh nh sc k sau khi chp bng my CAMAG REPROSTAR 3

Hnh 3.6: Sc k dch chit MeOH

ton phn thn r Ngi tin khi trin khai

trong h dung mi Toluen-Ethyl acetat-

A.formic (5,5:4,5:0,5)

a.Sc k di n 254nm

b.Sc k di n 366nm

c.Sc k sau khi phun TT hin mu

Hnh 3.7: Sc k dch chit sau

khi thy phn ca thn r Ngi

tin khi trin khai h Toluen-

EtOAc-A.formic (7:3:0,1)

a.Sc k di n 254nm

b.Sc k di n 366nm

c.Sc k khi phun TT hin mu

.

34

Kt qu hnh 3.6: Sc k dch chit MeOH = 254 nm v khi phun thuc th

hin mu thy hin nhiu vt, cc vt tch tng i r, nhng = 366 nm thy

cc vt rt m nht, khng r.

Kt qu hnh 3.7: Sc k dch chit sau khi thy phn hin nhiu vt, cc vt r

hn dch chit MeOH ton phn.

b. Hnh nh sc k dch chit MeOH ton phn sau phn tch bng phn mm

winCATS v VideoScan, bao gm th v bng biu din cc gi tr.

Kt qu: S dng phn mm VideoScan (vi nhy 25pixels, hmin=100,

Smin=300) x l hnh nh lng gi c 8 vt. Cc vt tch nhau tng i r.

Trong , c vt th 6 (Rf=0,760) l m nht, hm lng tng i l 41,04%.

Hnh 3.8: Kt qu lng gi

sc k dch chit MeOH

thn r Ngi tin di n

254nm h dung mi Toluen-

EtOAc-A.formic (5,5:4,5:0,5)

35

Kt qu: Khc vi sc k di n 254nm, di n 366nm khi cng nhy

(25pixels, hmin=100, Smin=300) th phn mm Video Scan ch nhn bit c 3 vt

m.

Kt qu: Phn mm Video Scan ( nhy 25pixels, hmin=100, Smin=300) lng gi

c 8 vt trn sc k khi phun TT hin mu. Trong , vt th 9 (Rf=0,758)

m nht, c hm lng tng i ln nht. Khi i chiu vi kt qu phn tch

254nm ta thy vt ny c th cng chnh l vt Rf=0,760.

Hnh 3.9: Kt qu lng gi

sc k dch chit MeOH

di n 366nm h dung mi

Toluen-EtOAc-A.formic

(5,5:4,5:0,5)

Hnh 3.10: Kt qu lng gi

sc k dch chit MeOH sau

khi phun TT Vanilin/H2SO4

c h Toluen-EtOAc-

A.formic (5,5:4,5:0,5)

36

3.3. Nghin cu v tinh du

3.3.1. Sc k lp mng tinh du thn r Ngi tin

Chun b dch chit chm sc k

Pha long tinh du trong dung mi n-Hexan vi t l 1:15.

Chm sc k: Chm bng my chm sc k CAMAG LINOMAT 5. Chp li nh

bc sng 254nm, 366nm v sau khi phun TT hin mu bng my CAMAG

REPROSTAR 3. S dng phn mm WinCATS, VideoScan x l hnh nh.

H dung mi

-H 1: Toluen-Ethyl acetat (98:2)

-H 2: Cloroform

-H 3: n-Hexan-Ethyl acetat (7:3)

-H 4: Cloroform-Ethyl acetat (5:5)

TT hin mu: Vanilin/H2SO4 c

Kt qu: Sau khi tin hnh khai trin sc k TD/n-Hexan qua cc h dung mi khc

nhau th nhn thy h 1 tch vt r nht.

Khai trin h 1: Toluen-Ethyl acetat (98:2)

a. Hnh nh sc k sau khi chp bng my CAMAG REPROSTAR

Hnh 3.11: Sc k Tinh du thn r Ngi tin

trong n-Hexan khi trin khai h dung mi

Toluen-Ethyl acetat (98:2)

37

Kt qu: Tinh du khi chy sc k bn mng c tnh cht c bit. Di n c

bc sng = 366 nm th khng hin vt, di n = 254 nm th c t vt lm tt

hunh quang. Tuy nhin khi phun thuc th th s vt hin ln nhiu v mu tng

i m.

b. Kt qu lng gi sc k sau phn tch bng phn mm winCATS v

VideoScan

Nhn xt: Phn mm VideoScan ( nhy 25pixels, hmin=100, Smin=300) lng gi

sc k tinh du sau khi phun thuc th Vanilin/H2SO4 c 10 vt. Trong , c

vt th 6 (Rf=0,544) l m nht v chim hm lng tng i ln nht (51,80%).

Hnh 3.12: Kt qu lng gi sc

k tinh du thn r Ngi tin

di n 254nm h dung mi

Toluen-Ethyl acetat (98:2)

Hnh 3.13: Kt qu lng gi sc

k tinh du thn r Ngi tin

sau khi phun TT Vanilin/H2SO4

h dung mi Toluen-Ethyl acetat

(98:2)

38

3.3.2. Sc k kh-khi ph (GC-MS) tinh du thn r Ngi tin

a. H thng GC-MS

- H thng GC: Agilent Technologies 7890A.

- H thng MS: Agilent Technologies 5975C.

- Ct sc k: HP-5MS

+ Chiu di ct: 30cm

+ ng knh ct: 0,25mm

b. Chng trnh khai trin

Tinh du thn r Ngi tin c pha long bng Cloroform (MECRK) n

nng 10-2 ri cho vo l thy tinh 2ml, y np chuyn dng, lc u, t vo

khay cha mu ca h thng tim mu t ng. Tim mu t ng. Th tch tim

mu: 1l, chia dng 1:50. S dng kh mang Heli, tc kh mang: 1ml/pht.

Chng trnh nhit :

Bng 3.2: Chng trnh nhit ca h thng GC-MS

Thi gian (pht) Nhit (C)

Ct

0-2 50

2-27 50-100

27-33,7 100-200

33,7-35,7 200

Nhit ha hi mu 250

Detector: Khi ph (MS) Aligent Technologies 5975C

c. Kt qu

Ph GC-MS c so snh vi th vin ph Willey, Flavor v Nist phn

tch kt qu. Hm lng phn trm tng i ca cc thnh phn c tnh d vo

din tch pic.

39

Bng 3.3: Thnh phn v hm lng cc cht trong tinh du thn r Ngi tin

STT Tn cht

Thi gian lu (pht)

Din tch Pic

Hm

lng (%)

1 Thujen 8,447 1947606 0,262

2 Pinen 8,717 45924867 6,184

3 Camphen 9,372 3069936 0,413

4 Sabinen 10,616 7138946 0,961

5 Pinen 10,725 168363778 22,671

6 Myrcen 11,601 6523434 0,878

7 Phellandren 12,213 9537142 1,248

8 3-Caren 12,523 2502720 0,337

9 Terpinen 12,900 1968601 0,265

10 o,p-Cymen 13,355 17044444 2,295

11 Limonen 13,577 31659304 4,263

12 1,8-Cineol 13,691 300701284 40,491

13 -Terpinen 15,382 7935689 1,069

14 Terpinolen 17,169 3299610 0,444

15 Linalool 18,018 3645854 0,491

16 Borneol 22,004 9757740 1,314

17 4-Terpineol 22,804 32607466 4,391

18 Terpineol 23,715 72369287 9,745

19 Terpinolen 30,428 5540300 0,746

20 Caryophyllen 31,588 2918656 0,393

21 Caryophyllen oxid 33,543 2851220 0,384

22 Spathulenol 33,948 1284150 0,173

23

4-Isobutylidene-2,2-

dimethyl-8-oxa bicycle

[3.2.1] oct-6-en-3-on

34,006 1545364 0,208

24 Longifolenaldehyd 34,459 2507968 0,338

Tng = 742645366 100%

40

Nhn xt: Kt qu GC-MS cho thy trong tinh du Thn r Ngi tin c

10 thnh phn chnh l: pinen (6,18%), sabinen (0,961%), pinen (22,671%),

cymen (2,295%), limonen (4,263%), 1,8-cineol (40,491%), borneol (1,314%),

myrcen (0,878%), 4-terpineol (4,391%), terpineol (9,745%). Trong , hm lng

1,8-cineol v -pinen rt cao. Ta i chiu vi sc k sau khi phun thuc hin

mu c 10 vt hin mu r rt, c phn tch bi phn mm VideoScan ( nhy

25pixels, hmin=30, Smin=300) tng ng vi 10 Pic. iu chng t SKLM kt

hp x l bng WinCat v VideoScan l phng php n gin nhng tng i

chnh xc.

41

BN LUN

Cy Ngi tin c ngi dn s dng cha 1 s bnh thng thng nh:

au bng lnh, kh tiu, phong thp, rn cn nc ta cng nh trn th gii,

ngoi nhng cng dng ngi ta cn i su vo nghin cu tc dng chng ung

th ca cc Diterpen phn lp t thn r cho nhiu kt qu kh quan. Cy pht trin

tng i tt khi c trng trt ly hoa lm cnh 1 s a phng. Tuy c

nhiu tc dng tt v tim nng pht trin ln nhng hin nay cy mi ch c

dng theo kinh nghim dn gian m cha pht huy c ht gi tr s dng. V vy,

vic tin hnh nghin cu c im thc vt, thnh phn ha hc ca cy l rt cn

thit, nhng kt qu t c s l c s d liu xy dng tiu chun kim

nghim cho cy Ngi tin khi c s dng trn quy m ln.

V phng php nghin cu: Ngoi nhng phng php thng qui nghin

cu v thc vt, ha hc nh vi hc, nh tnh phn ng ha hc, SKLM th vic kt

hp cc phng php hin i vo nghin cu dc liu ni chung v Ngi tin ni

ring l rt hu ch. Cc phn tch sc k ca phn mm WinCats v Video Scan

va chnh xc li va tin li khi cho ta lun kt qu s vt, Rf, Spic tng vt m

khng phi lm th cng. Phng php GC-MS li gip chng ta nh tnh v nh

lng cc thnh phn d bay hi, trong c tinh du. Vic ny gip chng ta

nghin cu chuyn su hn v c nh gi chnh xc hn.

V c im thc vt, tin hnh nghin cu c im hnh thi cy, c

im vi phu, c im vi hc bt l v thn r Ngi tin. y l ln u tin cc

c im thc vt ca cy c nghin cu 1 cch y , trong khi cc nghin cu

trc ch dng li m t hnh thi cy, phn tch hoa .

V ha hc, tin hnh kim nghim bt thn r ca cy bng cc phn ng

ha hc c trng. Tin hnh d tm h dung mi chy sc k cho dch chit

MeOH, dch chit thy phn, tinh du v kt hp x l bng phn mm chuyn

dng cho kt qu nhanh, chnh xc.

V nghin cu tinh du, tin hnh xc nh thnh phn ha hc, hm

lng tinh du trong thn r Ngi tin bng GC-MS cho kt qu rt chi tit. Trong

42

cc thnh phn chnh ca tinh du c 1,8-cineol c hm lng ln ti 40,491% c

nhiu tc dng rt qu v c gi tr kinh t, bi vy Ngi tin cng c th l 1 ngun

cung cp 1,8-cineol ng quan tm.

Nhng kt qu t c s l c s d liu khoa hc v cy Ngi tin phc

v ti NGHIN CU BO TN V PHT TRIN MT S NGUN

GEN CY THUC QU HIM C TIM NNG VIT NAM.

43

KT LUN V KIN NGH

Kt lun

V thc vt: Kha lun xc nh v chp nh c cc c im hnh thi cy:

l, qu, ht, thn r. Vi phu: L, thn r ca cy Ngi tin. c im vi hc: Bt l

(biu b cha l kh, m mm, mnh mch, mnh phin l), bt thn r (mnh

mch, b si, ht tinh bt, mnh mang mu,mnh bn, m mm mang tinh bt).

V ha hc: Qua cc th nghim nh tnh ha hc, kha lun xc nh c

trong thn r Ngi tin c cha: polysarcarid, flavonoid, saponin, tinh du, acid

amin, acid hu c, sterol, polysaccarid, ng kh. Ngoi ra cng xc nh c

h dung mi chy SKLM: Toluen-Ethyl acetat-A.formic (5,5:4,5:0,5) cho dch chit

MeOH ton phn, chp v x l hnh nh bng cc phn mm winCATS,

VideoScan gp phn lm c s d liu ha hc ca cy.

V nghin cu tinh du: ti xc nh c hm lng tinh du trong mu

thn r l 0,365% tnh theo dc liu kh tuyt i. tin hnh chy SKLM, GC-

MS xc nh c thnh phn ha hc, hm lng cc cht trong tinh du thn r.

Gm cc thnh phn chnh: pinen (6,18%), sabinen (0,961%), pinen

(22,671%), cymen (2,295%), limonen (4,263%), 1,8-cineol (40,491%), borneol

(1,314%), myrcen (0,878%), 4-terpineol (4,391%), terpineol (9,745%).

Kin ngh

Ngi tin l cy dc liu ang cn tng i mi, cha c nghin cu 1

cch y v chuyn su. Trn th gii cng nh Vit Nam, ngi ta quan tm

ch yu ti tc dng chng ung th ca cc Diterpen trong thn r. V vy, ngoi

kt qu nghin cu ca ti cng nh nhng nghin cu cng b trc ,

chng ti xut nghin cu thm thnh phn ha hc tinh du hoa, l ca cy

Ngi tin cng nh tc dng sinh hc ca chng v ca tinh du thn r. Nghin

cu a Ngi tin vo trng trt quy m ln khai thc ht tim nng ca cy.

44

TI LIU THAM KHO

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chuyn nghnh ha hu c, i hc Khoa hc T nhin.

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14. Nguyn Th Kim Thy (2013), Nghin cu thnh phn ha hc v hot tnh

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hc, Vin ha hc cc hp cht thin nhin.

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(Hedychium coronarium Koenig), Vin Sinh thi & Ti nguyn Sinh vt,

Vin KH& CNVN.

TI LIU TING ANH

16. Ajay Kumar Meena, et al (2010), Comparative Study on Family

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Vol.5, No.2, pp. 601-606.

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Hedychium coronarium J.Koenig, Journal of Agriculture and Biological

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26. Muhammad Lateef, et al (2013), An Account of Botanical Anthelmintics and

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2305-7211.

27. Nitirat Chimnoi, et al (2008), Labdane diterpenes from the rhizomes

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TI LIU TING PHP

34. M. H. Lecomte, Flore gnral de L'indo-chine VI, pp. 70-75.

PH LC

1. PHIU GIM NH TN KHOA HC

2. PH GC-MS TINH DU THN R NGI TIN