Nghien CLFU - Ky thuat<br />
<br />
Nghien ciiu thanh phan hoa hoc va<br />
hoat tinh khang khuan cua tinh dau ran ma<br />
{Centella asiatica (L.) Urban)<br />
Nguyen Ngoc Hanh, Le Ngoc Thach<br />
Bo mon Hoa hgc Huv co- - Khoa Hod hoc<br />
Dgi h()c Khoa hoc Ty nhien,<br />
Dgi hoc Quoc gia Tp. Ho Chi Minh<br />
<br />
Dat van de Chiit tach<br />
Rau md cd tdn khoa hoc Id Centella asiatica Tinh diu du-gc chiit tach bing phu-ong phap<br />
(L.) Urban, thudc ho Apiaceae, cdn cd cdc tdn goi chu-ng cit hoi nu-dc (hydrodistillation, HD) dun<br />
khac la tich tuylt thao, lidn tiln thao. Ho Apiaceae ndng co diln (conventional heating<br />
cd 10 loai rau md khac nhau, trong dd Centella hydrodistillation, CHHD), hoac chilu xg vi sdng<br />
asiatica du-gc trdng chuydn canh rdng rdi tgi nhilu (microwave irradiation hydrodistillation) cd thdm<br />
noi d mien Nam Viet Nam'^' nu-dc vdo nguydn lieu (with water, MIHD) vd<br />
Nhilu qudc gia tren thi gidi dd cd nghidn ciPU khdng thdm nu-dc vdo nguyen lieu (without water,<br />
v l tinh diu ndy nhu- d Trung Qudc '^', Malaysia '^', MIHD) SLP dung Id vi sdng gia dung d i tiln Sanyo<br />
Banglasdesh '"', Sri Lanka '^', Tdy-Himalaya '^'' An EM-D553N, 2.450 MHz, 750 W, vdi bd Clevenger<br />
Od '^, Nam Phi '^' theo SciPinder (9/2010). 2000 ml (dng ggn tinh diu nhe) '^^' Mdi lln chiit<br />
Tinh dau rau md cd nhilu hogt tinh sinh hoc tach SLP dung 400 g nguydn lieu xay nho vd 200<br />
ddng lu-u y nhu-: hogt tinh khang khuIn (gram ml nu-dc. Lip hd thong chu-ng d t hoi nu-dc vd<br />
du-ong: Bacillus subtilis, Staphylococcus aureus; tiln hanh dun trong nhu-ng khoang thdi gian nhit<br />
gram am: Escherichia coli, Pseudomonas dinh (tinh tip giot ngu-ng tu diu tidn). De ngudi,<br />
aeruginosa. Shigella sonnei); hogt tinh khang chiit tach phin tinh diu trong dng ggn bing<br />
nim: Aspergillus niger, Rhizopus oryzae, Candida diethyl ether. Lam khan nu-dc dung djch ether<br />
albicans, Fusarium solani, Colletotrichum musae; nay bing Na2S04 khan. Sau dd loc vd thu hdi<br />
hogt tinh du-gc hoc trong mdt sd bdi thudc ddn dung mdi bing cd quay. Xdc djnh khdi lu-gng tinh<br />
gian'^^'^'^'^"'; hogt tinh xua duoi bai mudi diu cao nhit thu du-gc trdn khdi lu-gng nguydn<br />
Anopheles stephensi gdy bjnh sot rdt'^°'; thdnh lieu tu-oi tu-ong ipng, ti Id ndy cd thi xem Id ham<br />
phin cua my phim du-dng da'^^' lu-gng tinh diu (%) trong nguydn lidu (hoac hidu<br />
Din nay, rit it nghidn ciPU cd gid trj cung clp suit tinh diu). Do dd, hdm lu-gng nay cdn tiiy<br />
diy du thdng tin vd tinh diu rau md d Viet Nam, thudc vdo phu-ong phap chiit tach'^^'<br />
vi ly do dd chiing tdi tiln hdnh nghien ciPU tinh Thanh phin hoa hoc<br />
dau rau md d l ddng gdp vdo danh muc cac cay Thanh phin hda hoc cua tinh diu du-gc phdn<br />
tinh diu cd hogt tinh sinh hoc d Viet Nam. tich bing phu-ong phap sic ky khi ghep khoi<br />
Thyc nghiem pho (GC/MS), du-gc tiln hdnh trdn mdy Agilent,<br />
Nguyen lieu GC 7890A MS 5975C. Cot TR-5MS (30 m,<br />
Phin trdn mat dit cua rau md du-gc thu hai 0.25 mm, 0.25 |im film). Su- dung helium Idm khi<br />
tgi xa Than Cu-u NghTa, huydn Chdu Thanh, tinh mang d ap suit 19.248 psi. Nhidt dd budng tidm<br />
Tiln Giang, la viing chuydn canh rau md noi 250 °C. Tl Id chia ddng 1/20, t h i tich tidm 1 ^il.<br />
tiing d miln Nam Viet Nam. Tdn khoa hoc cua Chu-ong trinh nhidt: Nhiet do diu 60 °C, tang 3<br />
rau md du-gc Bd mdn Thgc vdt hoc, Khoa Sinh °C/phut din 240 °C. Ghi nhan khdi pho m/z<br />
hoc, Dai hoc Khoa hoc Tg nhidn Tp. Hd Chi trong khoang 30-500, nang lu-gng ion hda 70<br />
Minh xdc nhan. eV. Thu- vidn pho NIST 2008.<br />
<br />
TAP CHI DUQC H Q C - 12/2011 (SO 428 NAM 51) 27 c<br />
Nghien CLFU - Ky thuat<br />
Chi s d v§t ly va hda hgc nhit cua phu-ong phap chieu xg vi sdng (cd thdm<br />
xac dinh ty trong b i n g ty trong k l thuy tinh. nu-dc) n g i n hon 5,2 l l n so vdi phu-ong phap dun<br />
Su- dgng khiie xg k l WYA SABBE d l xac djnh ndng e6 d i l n nhu-ng ham lu-gng tinh d i u thu<br />
chi so khuc xg. Gdc quay cg-e du-gc xdc djnh trdn du-gc tip phu-ong phap c h i l u xg vi sdng (cd thdm<br />
tnen quang k l A, KRUSS OPTRONIC P8000. nu-dc) t h i p hon 2,5 l l n so vdi phu-ong phdp dun<br />
cac ehi s l acid (lA), ester (IE), savon hda (IS) ndng c l d i l n . D i l u ndy cd t h i giai thich la tinh<br />
du'O'c xac dinh theo tidu chuIn Phap '''*' d i u rau ma cd t h i chipa nhieu d u phan khdng<br />
Hoat tinh khang khuIn phdn cgc, thu-dng Id cac logi hgp chit<br />
Thgc hidn tgi phdng thi nghidm Vi khuIn, hydrocarbon hon la cac hgp c h i t chipa oxygen.<br />
khog Vi sinh midn djch, Vidn Pasteur Tp. H i Chl Vi vdy tinh d i u rau ma thu du-gc theo phu-ong<br />
Minh. Hoat tinh khdng vi sinh vdt du-gc thii' phap chilu xg vi sdng cd t h i khdng phan anh du-gc<br />
nghidm theo phu-ong phdp khuleh tdn trdn tg nhidn, cho ndn trong cdc khao sat phia sau chi<br />
thgeh. trai vi sinh vdt vdi mdt s6 lu-gng n h i t dinh su- dgng logi tinh d i u thu du-gc theo phu-ong phap<br />
va deu Idn trdn m$t thgeh. Dat dia g i l y (d = 6 chu-ng d t hoi nu-dc dun ndng d diln.<br />
mm) da t i m tinh d i u , theo cac n i n g dd tu- Thanh p h i n hoa hoc<br />
nguydn chat d i n pha lodng d i n , Idn b l mdt sang 2: Thdnh phin hda hoc cua tmh diu<br />
thgeh Tmh d i u se khuleh tdn vao trong thgeh Rau Md<br />
va wc che sg tdng tru-dng cua vi sinh vat tgo ra STT Cau phin % (GC/MS)<br />
vdng khdng vi sinh vat trdn deu chung quanh 1 p-Cimen 0,22<br />
dia gily. Du-dng kinh cua vdng trdn ndy cho biet d-Limonen 0,2<br />
2<br />
kha nang khang vi sinh vdt cua tinh d i u ''^'<br />
3 Linalol 1.39<br />
Ket qua va t h a o luan 4 3-Nonen-2-on 0,41<br />
C h i i t tach 5 G-Terpineol 0.42<br />
Chu-ng d t hoi nu-dc dun ndng co d i l n du-gc 6 a-Cubeben 0,89<br />
t i l n hdnh tu' 2 gid den 8 gid. Chu-ng c i t hoi 7 P-Elemen 4,28<br />
nu-dc chieu xg vi sdng (cd thdm nu-dc, vd khdng 8 -(3-Cariophilen 4,56<br />
them nu-dc) t i l n hdnh tu- 42 phiit tdi 52 phiit. 9 Tujopsen 0,55<br />
K i t qua du-gc ghi trong bang sau. 10 P-Farnesen 56.09<br />
Bang 1 : Hdm Iwang tinh diu thu dwac theo 11 Allo-aromadendren 1,52<br />
phwang phdp ly trich 12 Acoradien 1,11<br />
Chwng cat hai nwoc Thai gian Hiim Iwqng 13 8-Cedren 0.37<br />
Dun nong co dien 6 gia 0,0374 14 P-Patchoulen 0,46<br />
Chieu xa vi song 15 P-Chamigren 0,51<br />
52 phiit 0,0150<br />
(co them nu-oc) 16 p-Selinen 0,34<br />
Chieu xa vi song 17 P-Panasinsen 0,85<br />
48 phiit 0,0146<br />
(khSng them nu-dc) 18 y-Murolen 0,26<br />
Ham lu-gng tinh dau tu-ong doi thap phii hgp 19 6-Cadinen 0.72<br />
vdi k i t qua giai p h l u hgc t u y i n tinh d i u rau ma 20 Spatulenol 0.59<br />
vi kich thu-dc tiii tilt nho va mat dd r i t t h i p . Khi 21 Oxid cariophilen 3,64<br />
toan than rau ma cdn nguydn, khd ngCpi t h i y miii 3,4-Dimetil-3-ciclohexen-1 -<br />
thorn, khi vd ndt thi mdi ngu-i t h i y miii thom 22 3,55<br />
carboxaldehid<br />
thoang thoang. 2-lsopropil-5-metil-9-metilen-<br />
23 0,82<br />
Sg so sanh ham lu-gng tinh d i u thu du-gc biciclo[4,4,0]decen<br />
theo ba phu-ong phdp cho t h i y phu-ong phap 24 a-Murolen 0.51<br />
chilu xa vi sdng (cd thdm nu-dc) cho k i t qua cao 25 tau-iVlufolol 1.76<br />
hon phu-ong phdp chilu xa vi sdng (khdng thdm 6-lsopropeniJ-4,8a-dimetil-<br />
nu-dc). Do phu-ong phdp c h i l u xg vi sdng (khdng 26 1,2,3,5,6,7,8,8a-octahidro-2- 0,27<br />
them nu-dc) SLP dung lu-gng nu-dc t g nhidn cd s i n naptalenol<br />
trong nguydn li^u d l Idi cuon tinh d i u nhu-ng Hgp chat hidrocarbon 73,71<br />
lu-gng nu-dc dd khdng du. Thdi gian chiit tdeh tot T i n g cpng 86,56<br />
<br />
<br />
28 TAP CHI DUQC HQC - 12/2011 (SO 428 NAM 51)<br />
Nghien CLFU - Ky thuat<br />
Bang 3: So sdnh thdnh phin hda hoc cua tinh diu thwc nghiem vdi tdi lidu tham khdo<br />
Ham IwQ-ng ciu phin chinh (% GC/MS)<br />
Sri Tay An Nam<br />
ciu phan chinh Thwc Malaysia Bangladesh<br />
[3] Lanka Himalaya Do Phi<br />
nghiem 1*1 isi m [7b] [8]<br />
<br />
Apiol 4,49<br />
Bicidogermacren 11,22<br />
Oxid cariophilen 3,64 1,9 2,51 3,3 15,1 0,27<br />
P-Cariophilen 4,56 26,8 20,3 12 23,2 10,3 19,8<br />
a-Copaen 14<br />
P-Elemen 4,28 4,71 1,58<br />
Mircen 0,4 0,85 6,55<br />
p-Farnesen 56,09 Vet 4,45 5 6,3<br />
Germacren D 10,0 6,02 5,4 4,6 6,29<br />
a-Humulen 33,7 23,9 9 23,1 11,3 21,06<br />
<br />
Phu-ong phap GC/MS cho thiy tinh diu rau ThiP nghiem hoat tinh khang vi sinh vat<br />
ma thgc nghidm cd 27 chit, chiem khoang Bang 5: Kit qud khdng vi sinh vat cua tinh<br />
86,56%. Ngodi ra cdn cd mdt so hgp chit khac diu rau md<br />
khdng the nhan bilt du-gc do hdm lu-gng qua Dwang kinh vong khang<br />
thip hoac chl d dgng vdt. Trong dd thdnh phan Vi sinh vat vi sinh vat (mm)<br />
chinh la p-farnesen (56,09%), p-eariophilen 10° 10' Iff^ 10'^<br />
(4,56%), p-elemen (4,28%), oxid cariophilen Staphylococcus aureus 35 10 8 6<br />
(3,64%).<br />
Escherichia coli 28 7 6 6<br />
So vdi cac d u phin chinh trong tinh diu rau<br />
Bacillus subtilis 29 10 7 6<br />
ma thu du-gc tgi Malaysia '^', Bangladesh '"', Sri<br />
Proteus vulgaris 23 7 6 6<br />
Lanka '^', Tay Himalaya '^', An Do '^''', vd Nam<br />
Candida albicans 19 10 6 6<br />
Phi '®' tu-ong ddi gidng nhau, nhu-ng vdi hdm<br />
lu-gng p-farnesen cao, cd the rau md d Viet Tgi ndng do tinh ddu rau ma nguydn chdt va<br />
Nam thudc vd mdt chemotype khdc hin vdi cac 10"\ thiy vdng khdng vi sinh vat ddu xuit hidn<br />
nu-dc chung quanh. trdn tit ca cac chung vi sinh vat thu- nghidm:<br />
Trong tinh dau thgc nghidm, nhdm hgp chit Staphylococcus aureus, Escherichia coli, Bacillus<br />
hydrocarbon chilm 73,71%. Kit qua ndy phii subtilis, Proteus vulgaris, Candida albicans.<br />
hgp vdi kit qua thgc nghidm vd chiit tach. Vi Tgi nong do 10"^, thiy vdng khdng vi sinh vat<br />
hdm lu-gng cac hgp chit cd chipa oxygen it hon tiep tuc xuit hidn trdn cdc chung<br />
rit nhilu so vdi ham lu-gng cac hydrocarbon ndn Staphylococcus aureus, Bacillus subtilis.<br />
sg chu-ng d t hoi nu-dc du-di sg chieu xg vi sdng Tw ndng do 10"^ khdng thay xuit hidn vdng<br />
trd ndn khdng thich hgp cho logi tinh diu nay. khang vi sinh vat trdn tit ca cdc chung thCp nghidm.<br />
Chi sd vat ly va hoa hoc Kit luan<br />
Tinh dau cd mau vang nhgt, miii thom dd chju. Sg khao sdt tinh diu rau ma, Centella asiatica<br />
Bang 4: Chi sd vat ly vd hda hoc cua tinh (L.) Urban trong tgi Tiln Giang cho thiy:<br />
diu rau ma Ham lu-gng tinh diu thip (0,015-0,037%).<br />
Ty trong (30 °C) 0,8484 Tuy nhidn rau iTid dang du-gc chuydn canh rdng<br />
Chi so khuc xa (23 °C) 1,4798° rai, nlu muon du-a vdo san xuit Idn cd t h i tdn<br />
Goc quay cgc (20 °C) -6,761° dung Igi t h i nay va ap dung cac he thong chiit<br />
tach tinh diu trong cdng nghiep.<br />
lA 8,29<br />
NhQ-ng thdng tin ve thdnh phin hda hoe,<br />
IE 25,76<br />
cdc chl sd hda ly, hogt tinh khdng khuan cua<br />
IS 34,05<br />
tinh diu rau ma thgc nghidm gdp phin lam giau<br />
<br />
<br />
TAP CHi DUOC H Q C - 12/2011 (SO 428 NAM 51) 29<br />
Nghien CLFU - Ky thuat<br />
bd su-u t$p cac cay tinh d i u cd hogt tinh sinh asiatica aerial pahs. Indian Perfumer 51(1), 57-58<br />
hoe d Vidt Nam (2007).<br />
8. O. A. Oyedeji, A. J. Afolayan, Chemical<br />
Summary<br />
composition and antibacterial activity of the essential<br />
Essential oils of Centella asiatica (L.) Urban oil of Centella asiatica growing in South Africa,<br />
grown in Tien Giang province were extracted by Phanvaceutical Biology (2005), 43(3), 249-252 .<br />
strnnhdistillation with conventional heating or 9. (a) J. Minija, J. E. Thoppil, Antimicrobial activity<br />
microwave irradiation Twenty six compounds of Centella asiatica (L.) Urb essential oil, India<br />
were indentified by GC/MS Of them, the main Perfumer 47(2), 179-181 (2003): (b) J. S. Jangwan,<br />
constituent was (i-farnesene (56 09%) The A M Painuly, Phytoconsituents of essential oil of<br />
Centella asiatica and their antibacterial activity. Acta<br />
chemical and physical properties of the so<br />
Ciencia Indica 34(1), (2008),137-140<br />
obtained oils were observed and measured.<br />
10. S. Rajkuma, A. Jebanesan, Repellent activity<br />
Some factors influencing the oil yield and its<br />
of selected plant essential oils againt the malarial<br />
antimicrobial properties were studied fever mosquito Anophles stephensi. Tropical<br />
Key words: Centella asiatica. chemical Biomedicine (2007) 24(2), 71-75<br />
composition, (i-farnesene. antimicrobial activity. 11 Armaud Ruel. Skin-care cosmetic composition<br />
Tai lieu t h a m k h a o comprising rice starch. Oleum cocois, and Shea butter, Fr.<br />
Denamde, Fr 2801505 Al 200110601 (2001).<br />
1 Pham Hodng Ho, C§y co Vi$t Nam, Nxb Trd.<br />
TpHCM. (1999), 949-951. 12. (a) L§ Ngoc Thgeh, Nghien cuu chuyen doi 16<br />
vi sdng gia dgng th^nh thiet bj chiet tach hgp chat<br />
2 Cheng-jian Zheng, Lu-ping Qin, Chemical<br />
thi6n nhien vd thue hi^n ting hgp huu ca, Tuyen t$p<br />
components of Centella asiatica and their<br />
Hpi th^o Sdng tao Khoa hpc v&i sw nghiep cong<br />
bioactivitive, Joumal of Chinese Integrative Medicine<br />
nghi$p hda. hien dai hda dat nwoc. Da Nang 19-<br />
(2007), 5(3). 348-351<br />
23/07/2006, 204-212 (2006), (b) Nguyen Duong<br />
3 K. C. Wong, G. L. Tan, Essential oil of Centella<br />
Thanh Thi, Tran Huu Anh, Le Ngoc Thach. The<br />
asiatica (L.) Urt^., Joumal of Essential Oil Research essential oil composition of Eryngium foetidum L. in<br />
(1994), 6(3), 307-309. South Vietnam extracted by hydrodistillation under<br />
4. Jasim Uddin Chowdhury, Nemai Chandra conventionnal heating and microwave irradiation,<br />
Nandi, Minhaj Uddin, Aromatic plants of Bangladesh: Joumal of Essential Oil Beanng Plants 11 (2), 154-161<br />
volatile oil constituents of Centella asiatica, Indian (2008) (c) Nicholas E. Leadbeater, Microwave<br />
Perfumer (2006), 50(2), 36-37 heating as tool for suitainable chemistry. CRC Press,<br />
5. S. C. Bhattacharyya, Constituents of Centella Boca Raton (2011).<br />
asiatica. I. Examination of the Ceylonese variety, 13. (a) L§ Ngoc Thgeh, Tinh d i u , Nxb D^i hQC<br />
Joumal of the Indian Chemical Society (1956), 33, Qudc gia. Tp HCM, 122-142 (2003); (b) Nguyen True<br />
579-86. Linh, Le Ngoc Thach, Study of the essential oil of<br />
6. Virenda P. Joshi, Neeral Kurnar, Bikram Singh, Limnophila rugosa (Roth.) Merr in the south of<br />
R. P Chamoli, Chemical composition of the essential Vietnam, Joumal of Essential Oil Bearing Plants<br />
oil of Centella asiatica (L.) Urb. from Western 14(3), 366-372 (2011).<br />
Himalaya, Natural Product Communications{2007), 14 (a) AFNOR, Huiles essentielles, Syndieat<br />
2(5), 587-590. National des Industries Aromatiques Alimentaires,<br />
7. (a) J. Minija, J. E. Thoppil, Volatile oil Paris 37-156 (1992); (b) K. Husnij Can Ba§er,<br />
constitution in two species of Apiaceace. Geobios Gerhard Buchbauer, Handbook of Essential Oils:<br />
(Jodhpur, India) 29(1), 59-60 (2002); (b) V. S. Rana, Science. Technology, and Applications, CRC Press,<br />
M A Balazquez, Volatile constituents of Centella Boca Raton, 83-120 (2010).<br />
<br />
<br />
<br />
BwcyC d a u . . . (Tlip theo trang 51)<br />
<br />
Tai lieu tham khao 2. Chinh phu: Lugt dau thau<br />
1. Bp Y /e - BO Tai Chinh (2007), Hwdng dan thwc 3. Chinh phu: Lu$t su-a doi so 38/2009/QH12<br />
hi$n dau thau cung wng thudc trong cdc ca sof y te 4. Chinh phu (2009), Hw&ng dan lu$t dau thau.<br />
cong Idp, Thong tu- lien tjch so 10/2007TTLT- BYT- Nghj djnh so 85/2009/NO-CP ngay 15/10/2009.<br />
BTC ngay 10/8/2007;<br />
<br />
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30 TAP CHi DUQC HQC - 12/2011 (SO 428 NAM 51)<br />
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