Tap ch Khoa hoc Trng ai hoc C n Th Phn A: Khoa hc T nhin, Cng ngh v Mi trng: 45 (2016): 28-32

DOI:10.22144/ctu.jvn.2016.508

KHO ST THNH PHN HA HC V HOT TNH KHNG VI SINH VT CA

TINH DU L TRU KHNG (Piper betel L.), H H TIU (PIPERACEAE)

Nguyn Thin Ch1, Nguyn Th Ngc Chm1, Phm Khnh Ngc1, Duy Phc1,
Dng Tng Kha1 v Nguyn Th Thu Thy2
1
Khoa Khoa hc T nhin, Trng i hc Cn Th
2
Khoa S phm, Trng i hc Cn Th

ABSTRACT
Thng tin chung:
Ngy nhn: 19/12/2015 Chemical composition and anti-microbial activity of essential oil from
Ngy chp nhn: 29/08/2016 leaves of Piper betel L. collected from Hau Giang province has been
investigated. Highest yield of oil (0.63 %) of P. betel L. leaves was
Title: successfully extracted by steam distillation. By using gas chromatography
Chemical composition and - mass spectrometry analysis (GC-MS), we have confirmed that the main
anti-microbial activity of component of P. betel L. essential oil is 4-Allyl-1,2-diacetoxybenzene, with
essential oil from leaves of content up to 34.55%. The in vitro anti-microbial activity was evaluated by
Piper betel L. Minimum Inhibitory Concentration (MIC) method. The results showed that
the essential oil from betel leaves inhibited the growth of 3
T kha: microorganisms species: B. subtillis, F. oxysporum and A. niger with the
Tinh du Tru khng, Piper MIC values of100, 200 and 200 g/mL, respectively.
betel L., hot tnh khng vi
sinh vt TM TT
Ln u tin, thnh phn ha hc v hot tnh khng vi sinh vt ca tinh
Keywords: du l Tru khng (Piper betel L.) thu hi ti tnh Hu Giang c kho
Betel oil, Piper betel L., anti- st. Tinh du l tru khng c ly trch thnh cng bng phng php
microbial activity chng ct li cun hi nc t hiu sut 0,63%. Bng phng php phn
tch sc k kh ghp khi ph (GC-MS), thnh phn ha hc chnh trong
tinh du l Tru khng c xc nh l hp cht 4-Allyl-1,2-
diacetoxybenzene vi hm lng 34,55%. Hot tnh khng vi sinh vt ca
tinh du c nh gi bng phng php pha long a nng xc
nh nng c ch ti thiu (MIC). Kt qu th hot tnh khng vi sinh
vt cho thy tinh du Tru khng c kh nng c ch s tng trng ca 3
chng vi sinh vt: B. subtillis, F. oxysporum v A. niger vi gi tr MIC
ln lt l 100, 200 v 200 g/mL.

Trch dn: Nguyn Thin Ch, Nguyn Th Ngc Chm, Phm Khnh Ngc, Duy Phc, Dng Tng
Kha v Nguyn Th Thu Thy, 2016. Kho st thnh phn ha hc v hot tnh khng vi sinh vt
ca tinh du l tru khng (Piper betel L.), h h tiu (Piperaceae). Tp ch Khoa hc Trng i
hc Cn Th. 45a: 28-32.
1 T VN ri n , Sri Lanka, Malaysia, Thi Lan, i
Loan v cc quc gia ng Nam khc (Guha,
Cy Tru khng c tn khoa hc l Piper betle
2006). nc ta, trong y hc c truyn Vit Nam,
L. hay Piper sriboa L., thuc h H tiu
l tru c s dng cho nhiu mc ch khc nhau
(Piperaceae), l mt trong nhng loi thc vt nhit
nh nhai tru chc rng, cha vim m chn
i quan trng khu vc chu , c trng rng
rng, nc sc l tru ra hoc p tr vt

28
Tap ch Khoa hoc Trng ai hoc C n Th
thng, bng, l lot, mn nht, chm, l tru
ngm trong nc si dng nh mt cha bnh
vim kt mc... ( Tt Li, 2003).
Hin nay, c mt vi cng trnh nghin cu v
cy Tru khng cc tnh min Bc, min Trung
Vit Nam v ch dng li bc u nh nghin
cu v thnh phn ho hc ca tinh du l Tru
Hi Dng (Phm Th Chnh v ctv., 2009) Hc
Mn (Nguyn Nho Dng, 2011), cng nh phn
lp mt s hp cht t cc cao chit l tru (Phm
Th Chnh v ctv., 2009) m cha ch trng vo
kho st cc hot tnh sinh hc ca tinh du. Bi
vit ny trnh by kt qu nghin cu v thnh
phn ho hc v kh nng khng vi sinh vt ca
tinh du l tru trng Hu Giang, mt a phng
thuc vng ng bng sng Cu Long nhm ng
gp c s khoa hc cho tc dng iu tr cc bnh
do nhim khun v nm ca tinh du Tru khng,
ng thi lm phong ph thm ngun dc liu c
vng ng bng sng Cu Long.
2 PHNG PHP NGHIN CU
2.1 Nguyn liu
L Tru khng c thu mua trc tip ti vn
tru huyn V Thy, tnh Hu Giang v c nh
danh bng cch tham chiu vi ti liu tham kho
(Shukla, R., 2015). Nguyn liu cn ti c x
l s b, loi tp cht, ra sch v xay nhuyn
trc khi tin hnh ly trch tinh du.
2.2 Phng php ly trch tinh du
Tinh du l Tru khng c ly trch bng
phng php chng ct li cun hi nc trc tip,
vi b chng ct tinh du Clevenger, kt hp vic
s dng mui NaCl h tr s khuch tn ca
tinh du trong qu trnh li cun tinh du theo hi
nc. ng thi kho st s nh hng ca thi
gian chng ct, th tch v nng dung dch mui
c s dng trong qu trnh chng ct n hm
lng tinh du thu c.
Bng 1: Mt s tnh cht vt l ca tinh du Tru khng
Tnh cht Mu, mi v
Khng tan trong nc, tan trong cc
Dung dch du mu vng
Kt qu dung mi hu c: methanol,
nht, mi c trng, v gt.
chloroform, diethyl ether.
3.2 Kho st cc yu t nh hng n qu
trnh ly trch tinh du
3.2.1 nh hng ca thi gian ly trch
Tin hnh kho st nh hng ca thi gian ly
Phn A: Khoa hc T nhin, Cng ngh v Mi trng: 45 (2016): 28-32
2.3 Phn tch thnh phn ha hc ca tinh du
Thnh phn v hm lng cc cu t c trong
tinh du c phn tch bng phng php sc k
kh ghp khi ph (GC MS), thc hin trn thit
b GC MS ca THERMO SCIENTIFIC Trace
GC Ultra ISQ, loi ct s dng l ct TG SQC
(15 m x 0,25 mm x 0,25 m), kh mang helium.
Chng trnh nhit cho my sc k kh c
thit lp nh sau:
Nhit u: 70oC, gi 2 pht 150oC, gi
2 pht 175oC, gi 2 pht 230oC, gi
2 pht.
2.4 Th nghim hot tnh khng vi sinh vt
kim nh
Th nghim hot tnh khng vi sinh vt kim
nh ca tinh du Tru khng c thc hin trn
phin vi lng 96 ging (96-well microtiter plate)
theo phng php ca Vander Bergher v
Vlietlinck (1991). Th nghim c thc hin ti
phng Sinh hc thc nghim, Vin Ho hc cc
hp cht thin nhin, Vin Hn Lm Khoa hc v
Cng ngh Vit Nam. Mu tinh du c pha
long theo cc thang nng thp dn, tnh
nng c ch ti thiu (MIC)- l nng m
vi sinh vt b c ch gn nh hon ton. Mu c
MIC 200 g/mL l c hot tnh.
3 CC KT QU V THO LUN
3.1 Tnh cht vt l ca tinh du
Cc tnh cht vt l ca tinh du Tru khng
c ly trch bng phng php li cun hi nc
u ph hp vi cc ti liu tham kho
(Sugumaranet al., 2011, Adeltrudes and Marina,
2010).
T trng quay cc
tan
(d20)
( )
0,94 +4,29
trch n hm lng tinh du thu c trong 150 g
l tru ti, c nh cc yu t nh nhit , nng
v th tch dung dch NaCl. Kt qu c trnh
by trong Bng 2.
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Tap ch Khoa hoc Trng ai hoc C n Th Phn A: Khoa hc T nhin, Cng ngh v Mi trng: 45 (2016): 28-32

Bng 2: Kt qu kho st thi gian ly trch tinh du Tru khng bng phng php chng ct li
cun hi nc
Thi gian ly trch (pht) 60 120 180 240 300
Hm lng tinh du (% v/w) 0,17 0,21 0,37 0,63 0,64
C nh cc yu t: khi lng nguyn liu (150 g), nhit chng ct (100oC), nng dung dch NaCl (15%) v th
tch dung dch (500 mL)
Bng 2 cho thy hm lng tinh du thu c v c c nh tip tc kho st cc yu t
ph thuc vo thi gian chng ct, hm lng tinh khc.
du tng khi thi gian chng ct tng v vi thi 3.2.2 nh hng ca nng dung dch NaCl
gian chng ct l 240 pht, hm lng gn nh t
Vi thi gian ly trch ti u c xc nh
cao nht. Kt qu ny ph hp vi nhng nghin
nh trn, nhm nghin cu tip tc kho st s nh
cu tng t, thi gian chng ct ti u c cc
hng ca nng dung dch NaCl n hm lng
nhm nghin cu chn u khng vt qu 240
tinh du thu c t 150 g l tru ti. Chng ct
pht (Rawatet al., 1989; B. Prakashet al., 2010).
thu tinh du ln lt nhng nng dung dch
Mt khc, khi ly trch vi thi gian lu hn th
NaCl khc nhau, c nh th tch dung dch l 500
lng tinh du tuy thu c nhiu hn nhng
mL. Kt qu c th hin trong Bng 3.
khng ng k. V vy, tit kim thi gian, thi
gian ly trch tinh du ti u c chn l 240 pht
Bng 3: Kt qu kho st s nh hng ca nng dung dch NaCl n hm lng tinh du ly trch
bng phng php chng ct li cun hi nc
Nng dung dch NaCl (%) 0 5 10 15 20
Hm lng tinh du (% v/w) 0,16 0,23 0,30 0,63 0,43
C nh cc yu t: thi gian chng ct (240 pht), khi lng nguyn liu (150 g), nhit chng ct (100oC) v th
tch dung dch (500 mL)

T bng kt qu trn, c th thy hm lng du ra ngoi. Mt khc, do lm tng nhit si

tinh du thu c ph thuc vo nng dung dch
NaCl thm vo, hm lng tinh du tng khi nng
mui tng v t cao nht nng mui l
15%. Vic thm mui NaCl vo qu trnh chng
ct nhm mc ch tng hiu sut ly trch do NaCl
ha tan trong nc s to thnh dung dch in ly
dn n lm tng nhit si (hay lm tng p sut
hi) ca nc, gim s bay hi ca nc trong hn
hp; do , tinh du ly trch c nhiu hn (W. F.
Furter and R. A. Cook, 1967). Tuy nhin, khi
chng ct vi nng ln hn, lng tinh du thu
c gim, iu ny c th gii thch l do khi s
dng mui nng cao th cc lp biu b ngoi
cha tinh du b co li, ngn cn s khuch tn tinh
Bng 4: Kt qu kho st s nh hng ca th tch nc ct thm vo n hm lng tinh du ly
trch bng phng php chng ct li cun hi nc
Th tch nc ct (mL)
Hm lng tinh du (% v/w)
C nh cc yu t: thi gian chng ct (240 pht), khi lng nguyn liu (150 g), nhit chng ct (100oC) v nng
dung dch mui (15%)
Kt qu cho thy hm lng tinh du tng khi
th tch nc ct cho vo bnh cu tng, hm lng
t cao nht khi lng nc thm vo l 500 mL.
Ngoi ra, do iu kin tin hnh th nghim khng
cho php tng th tch dung dch v nguyn liu
cho vo bnh cu vt qu 2/3 th tch bnh nn
khng th kho st nhng th tch ln hn. V
ca nc nn vic thm mui NaCl vo qu trnh
chng ct khng th p dng cho nhng tinh du
c nhit si thp hoc km bn vi nhit.
3.2.3 nh hng ca th tch nc ct thm vo
Kho st s nh hng ca th tch nc ct
thm vo n hm lng tinh du ly trch t 150 g
l tru ti bng cch tin hnh chng ct tinh du
ln lt vi nhng th tch nc khc nhau, c
nh nng dung dch NaCl l 15%. Mc ch
nhm xc nh th tch nc ti u cho qu trnh
chng ct, trnh s dng lng nc qu d,
khng c li cho vic ly trch tinh du do trong tinh
du cha nhiu hp cht d tan trong nc. Kt
qu c th hin Bng 4.
200 300 400 500
0,10 0,19 0,38 0,63
vy, chn 500 mL l th tch ti u trong phm vi
nghin cu ny.
Phn tch v tng hp nhng kt qu trn, iu
kin ti u ly trch tinh du Tru khng bng
phng php chng ct li cun hi nc quy
m phng th nghim c xut nh trong
Bng 5.

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Tap ch Khoa hoc Trng ai hoc C n Th Phn A: Khoa hc T nhin, Cng ngh v Mi trng: 45 (2016): 28-32

Bng 5: Kt qu iu kin ti u cho qu trnh ly trch tinh du l Tru khng bng phng php
chng ct li cun hi nc
Khi lng Thi gian ly Th tch nc Nng dung Nhit Hm lng cao
nguyn liu (g) trch (pht) (mL) dch NaCl (%) (0C) nht (%)
150 240 500 15 100 0,63
3.3 Thnh phn ha hc ca tinh du Bng 6: Kt qu th nghim hot tnh khng vi
Bng 5: Thnh phn ha hc chnh tinh du sinh vt
Tru khng thu hi ti Hu Giang Nng c ch
Thi gian Hm Vi sinh vt ti thiu (MIC)
STT Thnh phn (g/mL)
lu (pht) lng (%)
1 5,52 4-Terpineol 2,08 Escherichia coli (-)
Vi khun
Phenol, 2-methoxy-3- Pseudomonas
2 13,45 19,79 Gr (-) (-)
(2-propenyl) aeruginosa
3 18,42 -Muurolene 1,33 Bacillus subtillis 100
Vi khun
4 20,97 -Cadinene 2,22 Staphylococcus
Gr (+) (-)
5 22,06 Acetyleugenol 20,14 aureus
6 27,12 (+)-tau-Muurolol 3,16 Aspergillus niger 200
Nm si
7 27,79 -Cadinol 5,51 Fusarium oxysporum 200
4-Allyl-1,2- Candida albicans (-)
8 28,41 34,55 Nm men Saccharomyces
diacetoxybenzene (-)
9 Thnh phn khc 11,22 cerevisiae
T kt qu GC-MS cho thy thnh phn ha T kt qu Bng 5 cho thy tinh du l Tru
hc chnh ca tinh du Tru khng khu vc Hu khng th hin hot tnh c ch 3 chng vi sinh vt
Giang l cc hp cht thuc nhm terpene (4- kim nh: vi khun Gram (+) Bacillus subtillis,
terpineol, -muurolene, -cadinene,(+)-tau- nm mc Aspergillus niger v Fusarium
muurolol, -cadinol) v cc hp cht l dn xut oxysporum vi gi tr MIC ln lt l 100, 200 v
ca phenol nh phenol, 2-methoxy-3-(2-propenyl); 200 g/mL. Trong , cc loi nm mc
acetyleugenol; 4-allyl-1,2-diacetoxybenzene. Tng Aspergillus khng ch gy bnh thc vt m cn
hm lng ca tt c cc cu t chim gn 90% gy ra cc bnh nghim trng ngi nh cc
lng tinh du ly trch c. Trong , hp cht 4- bnh nm Aspergillosis (Thomas, 2015); nm
allyl-1,2-diacetoxybenzene chim hm lng cao F.oxysporum gy ra bnh ho vng nhiu loi
nht (34,55%), kt qu ny ph hp vi ti liu thc vt, loi nm ny gy hi nhiu loi cy trng
tham kho v thnh phn ca tinh du Tru khng trn tt c cc b phn, c bit b phn gc v r
vng Nam B (Nguyn Th L v Trn Th Hng ca cy (Rosado-lvarez, 2014). Kt qu ny cho
Vn, 2010). C th thy cc cu t chim hm thy tim nng khng khun v khng nm ca tinh
lng cao nht trong tinh du u l nhng dn du Tru khng c th ng dng c vo thc t.
xut ca phenol nn tinh du ny c nhit si, t 4 KT LUN
trng v chit sut cao. Cc dn xut phenol c mt
trong tinh du c nhng tc dng sinh hc tt nh Nghin cu qu trnh chit xut tinh du t l
tc dng khng khun, khng nm, chng oxy ha, Tru khng bng phng php li cun hi nc
c tnh t bo (Roy and Vijayalaxmi, 2013, quy m phng th nghim xut iu kin ti
Bhanu P.et al., 2010). u v thi gian (240 pht), nng dung dch
NaCl (15%) v th tch nc ct thm vo bnh cu
3.4 Hot tnh khng vi sinh vt
(500 mL) vi hiu sut t c cao nht l 0,63%.
Vi nng ban u l 400 g/mL, tinh du p dng phng php phn tch hin i GC-MS
c pha long cc thang nng thp dn, t cho thy tinh du ly trch c t l Tru khng
xc nh gi tr MIC. Kt qu th nghim c trng tnh Hu Giang thuc ng bng sng Cu
trnh by trong Bng 6. Long c cha cc hp cht ha hc nh 4-allyl-

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Tap ch Khoa hoc Trng ai hoc C n Th Phn A: Khoa hc T nhin, Cng ngh v Mi trng: 45 (2016): 28-32
1,2-diacetoxybenzene (34,55%); acetyleugenol
(20,14%); phenol, 2-methoxy-3-(2-propenyl)
(19,79%) v cc hp cht mono- v sesquiterpene
nh -cadinol (5,51%), (+)-tau-muurolo (3,16%),
-cadinene (2,22%), 4-terpineol (2,08), -
muurolene (1,33%). Kt qu nghin cu v thnh
phn ho hc trong tinh du Tru khng Hu Giang
c so snh vi kt qu ca mt nghin cu Phm Th Chnh, Dng Ngha Bang, Phan Thanh
tng t c thc hin n (Bhanu P. et al.,
2010) nhn thy c s khc nhau v cc hp cht
thuc nhm phenylpropene trong hai tinh du hai
vng nguyn liu khc nhau. Trong khi hp cht
ch yu trong tinh du Tru khng Hu Giang l 4-
allyl-1,2-diacetoxybenzene (34,55%) th eugenol l
thnh phn chim hm lng cao nht trong tinh
du Tru khng n (63,39%), hai hp cht
ny l ng phn v tr nhm chc ca nhau. C s Rawat, A.K.S., Tripathl, R.D. Khan, A.J. and
khc nhau trn c th l do kh hu, th nhng
hoc phng php ly trch tinh du.
Tinh du l Tru khng biu hin hot tnh
khng vi sinh vt tt vi kh nng khng vi khun
Gram (+) nh B.subtillis v cc loi nm gy hi
nh A.nigervF.oxysporum, y l c s gip ng
dng tinh du l tru nh mt loi khng sinh c
chit xut t thin nhin.
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