Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tha Phdn A: Khoa hgc Tv nhien, Cong nghf vd Mdi irudng: 35 (2014): 131-136
Tap chi Khoa hpc Tri/dng Dai hoc Can Thtf website: sj. ctu. edu. vn
K B B A | ^
KHAO SAT THANH PHAN HOA HQC VA HOAT TINH SINH HQC CUA TINH DAU NGAI SAY {ZINGIBER MONTANUM)
Trin Thi Thuy Duong', Nguydn Trpng Diic', H6 Nhu Quynh', Dang Kieu Nhung', Tuong Le My Tu' va Bui Thi Buu Hue'
' Khoa Khoa hgc Tu nhien, Truong Dgi hgc Cdn Tho Thong lin chung:
Ngdy nhgn: 16/05/2014 Ngdy chtip nhgn: 29/12/2014 Title:
Research the chemical components and biological activities of Ngai say's essential oil (Zingiber montanum) Tirkhda:
Ngdi sgy, chimg cdt loi cuon hoi nuoc, chung cdt loi cuon hai nuoc co su ho trg cua vi song, dgc tinh tren ti bdo ung thu Keywords:
Ngai say, water distillation, steam dislillation, microwave-assisted water distillation, cytotoxic
ABSTRACT
Three different methods of extraction of essential oil from rhizome of Ngai say collected from Bay Nui, An Giang have been studied including water dislillation (NSC-1), steam distillation (NSC-2) and microwave-assisted water distillation (NSC-3). The overall yields of the obtained essential oils were found to be almost the same in the cases of NSC-2 and NSC-3 methods (1.439% and 1.423%, respectively) and higher than that of NSC-1 (1.261%). Negligible amount of essential oil was found for trunks and leaves based on the water distillation method (6.00x10"^ % and 4.33x10"^ %, respectively). GC-MS analysis showed that the essential oil from rhizome obtained under studied methods comprised mainly of 4-terpinenol (27 - 35%), sabinene (15 - 26%) and 1,4- bis(methoxy)-triquinacene (7 - 28%). Evaluation of bioaclivity showed that the essential oils of Ngai say's rhizome had no anti-mtcrobial and anti-fungal properties: negligible anti-oxidation activity compared to that ofvitamine C but more significant compared with other essential oils such as Curcuma longa or Cinnamomum camphor a Ness; and possessed good cytotoxic acitivites toward MCF-7, MCF7/ADR, MDA-MB-231 andHela species.
TOM TAT
Ba phuang phdp trich ly linh ddu tie cii Ngdi sgy thu hdi tgi Bdy NHI tinh An Giang dd dugc nghien cuu bao gdm phuong phdp chung cdt loi cudn hai nude true tiep (NSC-1), phuong phdp chung cdt ldi cuon hai nude gidn liip (NSC-2)^, vd phuang phdp chung cdt loi cudn hai nude co su hd trg ciia vi song (NSC-3).
Hieu sudt tinh ddu thu dugc Irong hai phuang phdp NSC-2 vd NSC-3 Id khd giong nhau (ldn lugt Id 1,439% vd 1,423%) vd cao han so vdi phuang phdp NSC-1 (1,261%). Ben cgnh do, tinh ddu thdn vd Id Ngdi sgy cUng dugc trich ly bdng phuang phdp chung cdt loi cudn hai nude true tiep nhung higu sudt khong ddng ki (tuang ung la 6x1 (T^% vd 4,33x10'^%). Kit qud phdn tich bdng phuang phdp GC-MS cho thdy thdnh phdn tinh ddu cu thu dugc trong cd ba phuang phdp chuyeu bao gom 4-terpinenol (27 - 35%), sabinene (15 - 26%) vd 1,4-bis (methoxy)-triquinacene (7 - 28%). Kit qud ddnh gid hogt linh sinh hgc cho thdy linh ddu cU Ngdi say thu dugc cd hogt tinh khdng oxi hoa khong ddng ki khi so vol vitamin C nhung tdi han tinh ddu Nghe nhd vd Long ndo; khong cd hogt linh khdng khudn vd khdng ndm tren cdc chung vi sinh vgt thu nghidm nhung thi hiin tdt hogt tinh gay dgc tren 4 ddng ti bdo ung thu: ung thu vi (MCF-7), ung thu vu da khdng thudc (MCF7/ADR), ung thu biiu mo tuyin (MDA-MB- 231) vd ung thit CO tit cung (Hela).
Tgp chiKhoa hgc TrudngDgi hgc Cdn Tho Phdn A: Khoa hgc Tv nhien, Cong nghi va Mdi trudng: 35 (2014): 131-138
1 G i d l THIEU
Hg Giing a Vidt Nam ed td 17 din 20 chi vd tten 100 lodi. Cdc ciy hg Gdng da dugc sd dimg tu Idu ddi 6k Idm thudc tri bdnh nhu Rilng nep giup tidu hda; Nghe frj dau da day, ldm mau ldnh vet thuong; Gdng gidp tieu hda, ttj ho, dau bimg,...
Ngai Say ciing thugc hg Giing, cd tdn khoa hgc l i Zingiber montanum. Cac kdt qud nghien cuu trdn thi gidi eho thdy tinh diu Ngdi Sdy cd rdt nhilu hogt ti'nh sinh hgc quy nhu khdng nim (Md. Nazrul Islam Bhuiyan et al, 2008; Ibrahim bin Jantan el ai, 2003), khing oxy hda (Saowaluck Bua-in el al, 2009) vi cd ddc tinh vdi tl bio ung thu (Trdn Cdng Luan, 2010). CJ Viet Nam, cic nghidn cdu vl boat ti'nh sinh hgc eiia tinh ddu Ngii Say edn rdt ban che. Dang chd -^ li cdng ttinh n^idn cuu eua Trin Cdng Lugn va ctv di dgt dugc nhung thinh cdng budc diu trong vide nghien cdu khd ning khing ung thu v i khdng nim cda tinh diu Ngdi say. Tu do cho thiy vide nghien cvai quy frinh trich ly, xdc djnh thinh phin hoi hgc vd hoat tinh sinh hgc cda tinh ddu Ngdi Say ed ^ nghia guan frgng c i vd mit khoa hgc ldn thyc tien, gop phin vio hudng nghidn ciiu t|in dyng nhftng ngudn nguyen lieu sin ed de tgo ra cie sin phim tinh ddu thien nhidn chat lugng cao, gid tiianh thdp, nhim phyc vu tdt hon nhu eiu cu$c sdng. Tii cdng trinh nghidn cdu cda nhdm tic gii nay, 6k tii phdt frien hodn thien them viec nghien ciiu tinh diu Ngii siy bdng cich md rdng nghien ciiu frdn than vi li Ngii say. Thdm vao do la xdy dung quy frinh ttich ly tinh dau Ngdi say theo ba phuong phip li chung cit Idi cudn hoi nude true tilp, giin tidp vi cd su hd frg ciia vi sdng d quy md phdng thi nghiem. Ve khdo sat hogt ti'nh sinh hgc cua tinh dau, de tai nghien cdu them hogt tinh khing oxy hda, k h a n g ^ u i n ^ die bidt Jigat _ tt'nh khang ung thu frdn 4 'ddng td bao img thu: ung thu vu (MCF-7), ung thu vii da khing thudc (MCF7/ADR), ung thu bieu md hiyin (MDA-MB- 231) vi ung thu ed tii cung (Hela).
2 PHlTONG PHAP NGHIEN CiTU VA THtrC NGHI£M
2.1 Nguyen lieu
Nguydn li|u duge sd dung Id than, la va eu Ngdi siy {Zingiber montanum) duge thu mua true tidp tai vudn d Biy Nui, An Giang vd dugc dinh danh tgi Bd mdn Sinh, Khoa Khoa bge Tu nhien, Trudng Dgi hgc Cdn Tha va tham chieu vdi tii lidu tiiam khio (Nguyen Thi Kim Hud, 2009). Todn bd cay Ngdi s§y sau khi thu mua vl duge nia sgch bun
dat, hui bin va logi bd phin hu, dip. Cic boa chit su dung frong nghien ciiu cd ngudn gde Trung Qudc.
2.2 Thuc nghiem
2.2.1 Trich ly tinh ddu tir cu Ngdi say 6 mdi phuong phdp trich ly, sau khi qui trinh chung cat kdt thuc thu lay hoin toin cdt nude vd tmh diu vio binh tam giic, &i ngudi. Tiln hinh chiet tach tinh dau bang diethyl ether, lim khan bdng NajSO^, Igc, cd dull dung mdi dl thu tinh diu sin pham. Cdn, xdc dinh hidu suit.
Mdi thi nghiem dugc tien hanh ba lan, ldy gid trj trung binh.
a. Chung cat loi cuon hoi nude true tiip (NSC-1)
Cd Ngdi say cd do im 7,53% dugc xay nhuyln.
Can 200 g mlu, eho vdo buih cdu cun^ vdi 500 mL nude cat hai lin, lip hd thing chung cit hoin ehinh vi tien hinh chung cdt tinh dau trong 2,5 gid. Hidu suit tinh diu thu duoc theo phuong phdp ndy Id 1,261%.
b. Chimg cdt loi cudn hoi nude gidn liep (NSC-2)
Cu Ngii siy ed do am 7,53% duge xay nbuyen.
Can 200 g mdu, cho vio binh ciu cd day dudi duge chin ludi thep vi gdn vdi binh chda 500 mL nude cdt. Nguydn lidu khdng tidp xiic true tidp vdi nude cit va ngudn nhidt. Tien hdnh chung cat tinb ddu frong 12 gid. Hidu suit tinh diu thu duge theo phuang phdp nay Id 1,423%.
c. Chimg cdt loi cuon hai nuac co sif ho trg cda vi song (NSC-3)
Cu Ngai say cd dd im 7,53% dugc xay nhuyln.
Cdn 200 g mdu, cho vdo binh cdu cung vdi 125 mL nude cit, lap hd thdng chung cat hoin ehinh, didu ehinh cdng suat Id vi song d 750 W vd tien hanh chung cit frong 1,5 gid. Hidu suit tinh diu thu dugc theo phuang phap niy li 1,439%.
2.2.2 Phdn lich thdnhphdn tinh ddu bdng ky Ihugl GC/MS
Thanh phdn hda hgc cda tinh ddu cu Ngdi say dugc phdn ti'ch bdng phuong phap sdc ky khi ghep khdi phd frdn may GC-MS eua hang Thermo Scientific tai Bg mdn Hda, Khoa Khoa bge Ty nhien. Cgt su dung trong phdn tich Id cgt TG-SQC (15 m X 0,25 mm x 0,25 pm), khi mang la Heli, vdi cdc didu kidn phin tich nhu sau:
Tgp chi Khoa hgc Trudng Dgi hgc Cdn Tha Phdn A: Khoa hgc Tv nhiin, Cdng nghf vd Mdi Irudng: 35 (2014): 131-138
fiieu ki^n sac ky
Chuong frinh nhiet do Injector Khi mang Dieu kien khdi pho
Nhidt dg dau d dO^C, gid 2 Nhiet do budng tidm:
phut. 240X.
Ram I: Tdng len I50°C vdi Chl dd tiem mdu chia tdc dd 5°C/phut, gift 2 phut. ddng: 50 mL/phdt.
Ram 2: Ting Ien 250°C vdi Ti Id chia dong: 42.
toe dp 5°C/phdt, gift 3 phut
Logi khi: Heli. Nhi?t do dudng fruyen Che dd ding ddng, khdi phd: 275°C.
tdc do 1,2 Nhiet do ngudn cap niL/phdt. ion: 230*'C.
Khoi quet: 4 0 - 5 0 0 amu.
2.2.3 Phdn lich chi so vdt ly vd hoa hgc cua tinh ddu
Tinh ddu cu Ngdi say trich ly tu 3 phuong phdp sau khi dugc xac dinh thanh phdn hda hgc, tiep tuc dugc xdc djnh eic chi sd vat 11 vi hda hgc nhu ty trgng (d), gdc quay cue (a'^), chi sd acid (L^), chi so savon hda (IS), chi sd ester (IE).
2.2.4 Xdc dinh khd ndng khdng oxy hoa Nguyen tdc cua phuang phdp Id su dyng 1,1- diphenyl-2-picrylhydra2yl (DPPH). Hoat tinh khdng oxy hda cua mau thu duge dinh gia dua tren khd ndng logi bd gdc tu do thdng qua vide lam giim miu eua DPPH, dugc xac dinh bdng do hip thu quang cua dung dich sau phdn dng do tai budc sdngX = 517nm.
Cdch tidn hinh:
DPPH gde: 1 mg hda tan frong 2 mL ethanol 99,7%.
- DPPH: 200 pL DPPH gdc pha loang tiianh 2 mL trong ethanol vdi ndng dg li 0,006%.
- Mau thu dugc pha loing den ndng do phu hgp trong ethanol 99,7%. Sau dd tien hdnh pha cdc dung djch phan ung theo Bdng 1.
Bing 1: Pha dung dich thir nghidm hogt tinh khing oxy hda
STT 1 2 3 4 5 6
Dung dich mau thu ()iL) 0 50 100 150 200 250
Ethanol (|iL) 250 200 150 100 50 0
DPPH
(nD
250 250 250 250 250 250 l. = 517nmDe thdi gian phin dng 30 phut d 37°C, frong bdng tdi, sau do do dg hip thu quang cda cdc dung djch d budc sdng 517 nm. Phin ttdm quet gdc ty do (Scavenging effect) DPPH cua miu tiiu dugc linh theo cdng thdc sau:
S C % = [(Atring-Amiu)/A,tang] X 100 Thi nghiem duge lap Iai ba ldn, tinh ket qua trung binh.
Lap dd thi bilu hien mdi tuong quan gifta SC va thi ti'ch miu thu da dimg, td dd tinh dugc gii trj ECso ciia tinh dau.
2.2.5 Xdc dinh hogt tinh khdng ndm, khdngkhudn Hogt tinh khing nim, khing khudn dugc kiem djnh theo phuong phap khuech tdn frong bdn thgch, thgc hidn d Phdng kiem nghiem Hda - Ly - Vi sinh, Vidn Pasteur, Thanh pho Hd Chi Minh vd phuang phdp pha loing da ndng do A& xic djnh ICso, thyc hien d Phdng Hda sinh dng dung, Vien Hda hgc thugc Vien Han Idm Khoa bge vi Cdng ngh? Viet Nam.
2.2.6 Xdc dinh dpc tinh tren ti bdo ung thu Thu nghidm ddc tinh tten td bdo cua tinh dau cu Ngdi say ly trich tii 3 phuang phdp dugc thuc hien tren 4 ddng tl bdo ung thu: ung thu vu (MCF-7), ung thu vu da khdng thuoc (MCF7/ADR), ung thu xuat phdt td bieu md tuyln (MDA-MB-231), ung thu cd td cung (Hela) bdng phuong phap MTT.
Thii nghidm dugc tiln hdnh d Phdng thi nghidm Dugc lidu, Khoa Dugc, Trudng Dgi hgc Quoc Gia Seoul, Hin Qudc.
Phuong phip MTT (3-(4,5-dimetiiyl-2- thia2olyl)-2,5-diphenyl-2i/-tefrazolium bromide) dugc md ta nbu sau: eic ddng tl bio ung thu dugc nudi cay ttong dTa 96 gieng ed mit do 5000 tl bao/gidng, vdi mdi trudng nudi cdy Id DMEM h i ^ glucose, 10% FBS, 1% A/A, u d nhidt dg 37''C, 5%
CO2. Sau 24 gid nudi ciy, dTa 96 giing duge chuyin ddi sang mdi trudng DMEM khdng cd FBS. T I bdo dugc xii \-$ vdi tinh diu Ngdi siy pha loing bdng dung mdi DMSO d cic ndng do khic nhau vi dim bdo n6ng dg cudi cda DMSO frong mdi ttudng nudi cay khdng vugt qui 0,05% (v/v) dl ti-inh gay dgc dung mdi. Sau 48 gid, 20 pL dung djch MTT ed ndng dg 2 mg/mL dugc cho vio m6i giing vd tilp tyc u frong 4 gid. Enzyme cellular
Tgp chiKhoa hgc TrudngDgi hgc Cdn Tito Phdn A: Khoa hgc Tv nhien, Cdng ngh$ vd Mdi truong- 35 (2014): 131-138 cua nhimg t l bdo sdng sot sd chuyin hda MTT
tiidnh fonnazan khdng hi hda tan trong mdi trudi^
nudi ciy. Formazan se dugc hda tan bdng DMSO vd do dd hip diu d budc sdng 550 nm. Thinh phan phdn trim ciia tl bao sdng sdt bidu thj ddc tinh cua tinh diu Ngdi sdy, khi d^e ti'nh cdng cao thi sd lugng te bao sdng sdt cdng thap. Thinh phan phdn trim dgc thih dugc xdc djnh bdng cdch so sdnh do hip thu d giing thd nghidm vdi do hip thy d gieng ddi ehiing. Mdi thu nghidm ed do lap lai ba lin. Sd dyng pMn mem Sigma PlotlO.O dl tinh toan gia tti IC50.
3 KET QUA VA THAO LUAN 3.1 Trich ly tinh dau tu- cu Ngai sgy Ba phuang phip trich ly tinh diu tix cu Ngai say d quy mo phdng thi nghiem duge tiln hinh khio sit bao gdm: trieh ly tbeo phuong phap chung cit ldi cuon hoi nude true tiep vdi bg Clevenger (NSC-I); ttich ly theo phuang phap chung eit ldi cuin hoi nude gidn tilp (NSC-2) vd ttich ly bing phuong phdp chung cit Idi cudn hoi nude cd su hd frg cua vi sdng (NSC-3).
O phuang phdp chung cat ldi cudn hoi nude tryrc tilp (NSC-1): Tiln hdnh khdo sit ba ylu td inh hudng tdi hieu suit tinh diu ttich ly bao gdm tiie tich nude eit (mL), thdi gian chung eit (gid) vd eich xu ly miu (xay nhuydn va xdt nhd) bing each cd dinh hai ylu td vd thay ddi yeu td khdo sdt dl xdc djnh gid tti tot nhit. Kdt qui tim duge didu
Bing 2: Dilu kien tot nhat cho qua trinh ly trich tinh dau cii Ngai siy
kidn chung cit tdt nhat nhu sau: iing vdi lugng mau la 200 g (xay nhuydn), Iugng nude cit su dung li 500 mL vi frong flidi gian chung cit la 2,5 gjd. Trong dilu kien nay, hieu suit tinh ddu thu dugc tuong ung Id 1,261 %.
D I xdy dyng phuang phap chung cat ldi cudn hoi nude giin tiep (NSC-2), dl tii tiln hinh cd djnh hai ylu td da duge xac djnh la tdt nhit td quy trinh frich ly (NSC-l) la till tich nude cit (500 mL) v i cdch xu ly mdu (xay nhuydn), thay ddi thdi gian chung cit lan lugt d cae gii tti: 6, 8, 10, 12 vi 14 gid. Ket qua eho thay, thdi gian 12 gid Id khodng thdi gian tdt nhdt dng vdi hidu suit tinh diu thu duge Id 1,423%.
Ddi vdi phuong phdp chung cit Idi cudn hoi nude cd su hd ttg eua vi sdng (NSC-3), dl tii tiln hanh kliio sdt cdc ylu td inh hudng tdi bi?u suit tinh dau cu Ngii say bao gdm cdng suit Id vi sdng (W), the ti'ch nude cat (mL), thdi gian chung cit (gid) bdng cdch cd djnh hai ydu td vi thay ddi mdt yeu td d nhilu gii frj, dong thdi giu nguyen gii trj cda yeu td da dugc xic djnh li tdt nhdt tu quy trinh ly frich tinh diu bdng phuong phip (NSC-I) Ii each xu Iy mau (xay nhuydn). Ket qua tim dugc dieu kien frich Iy tdt nhat nhu sau: cdng sudt ehilu cua Id vi sdng li 750W, lugng nude cdt sd dung li 125 mL va thai gian chung eit la 2,5 gid.
Didu kien tdt nhat tim dugc eho ba quy frinh frich Iy tinh diu cu Ngii sgy vd hidu sudt tinh dau thu dugc tuong ung dugc tdm tit frong Bing 2.
Phinmg phap NSC-1 NSC-2 NSC-3
Lirgng nude chung cat (mL) 500 500 125
Thdi gian chung cat (gid) 2,5 12 1,5
Cach xu* ly nguyen Ij^u Xay nhuydn Xay nbuyen Xay nhuydn
Cdng suat Id vi song (W) 750
Hieu suit tinh diu (%) 1,261 1,439 Tii Bdng 2 cho thdy hidu suat tinh dau eu Ngdi
say thu dugc bang phuong phdp chung cit ldi cudn hoi nude giin tilp (NSC-2) tuy cao hon so vdi phuong phdp chung cdt ldi cudn hoi nude true tiep (NSC-1) nhung ddi hdi thdi gian chung cit rit dii (12 gid). Trong khi dd, phucmg phap chung cit Idi cudn hoi nude ed sy hS frg cua vi sdng (NSC-3) cdn thdi gian chung cdt ngdn nhit ddng thdi cho hidu sudt tinh ddu cao nhit frong ca ba phuong phdp.
3.2 Trich ly tinh diu tiir than vi la Ngai say Phuong phdp chung cit ldi cudn hoi nude true tilp dugc ip dung dl fri'eh Iy tinh ddu td thdn va Id Ngii say. Thi n ^ d m dugc Idp lai ba lin, moi lin vdi 300 g than ho§c Id Ngdi say da dugc xii ly so
bd (rda sgch bui bin, logi bd nhdng phdn bj hu, dap, dd khd ty nhidn frong bdng mit, xay nhuydn).
Lugng nude eat sd dyng cho mdi man chung eit li 500 mL vi thdi gian chung cit Id 5 gid. Kit qua cho thdy: hieu suat tinh dau thdn vi l i thu dugc theo phuong phip niy lin lugt la 6x10"^ vi 4,33x10'' %. Td ddy eho thdy, thin vi Ii Ngdi say CO hdm lugng tinh ddu rat thdp, khdng ding kl so vdi cu Ngii siy. Chinh vl viy, dl tdi ehi tiln hinh phin tieh thanh phdn hda hgc cung nhu hoat tinh sinh hoc cua tinh diu td cu Ngdi say.
3.3 Phan tich thanh phan hda hgc cua tinh dau cu Ngai siy
Thanh phin hda hgc cua tinh diu cu Ngai say duge tiln hanh phan tich bang phuong phip GC-
Tgp chi Khoa hgc Trucmg Dgi hgc CSn Tha Phdn A: Khoa hgc Tv nhien, Cdng nghf va Mdi Irudng: 35 (2014): 131-1^8 MS. Kdt qui dugc tdm tdt ttong Bang 3. Nhin Ngii say thu dugc bing cd ba phuong phdp la chung cd su khdc biet ve thdnh phin cda tinh diu gidng nhau bao gdm 4-terpinenol (27 - 35%), frich Iy dugc d ba phuong phip nghien eiiu. Tuy sabinene (15 - 26%) vi l,4-bis(methoxy)- nhien, thinh phin nhilu nhit frong tmh diu cu triquinacene (7 - 28%).
Bang 3: Thanh pban hda hgc cua tinb dau cu Ngai sgy
STT Thinh phan hda hgc NSC-1 NSC-2 NSC-3
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25
4-Terpinenol Sabinene
1,4-bis(Methoxy)-triquinacene y-Terpinene
P-Sesquiphellandrene P-Pinene a-Terpinene p-Cymene a-Pinene p-Myrcene 4-Thujanol a-Thujene
2-Allyl-l,4-diinethoxy-6-methy!benzene a- Terpinolen
a-Humulene (-)-Zingiberene I-Terpinenoi Trans-Piperitol a>Piperitol a-Tei-pineol acetate (Z)-Sabinene hydrate Camphene
5,6-bis(Hydroxymethyl)-3a,4,5,6,7,7a-hexahydro-l//-inden-l-one P-Bisabolene
a-Terpineol
27,05 15,21 7,45 6,03 4,65 4,4 4.26 3,62 3,57 3,3 3,11 2,36 2,04 2 1,51
-
1,36 1,22 1,03
-
0,97
-
0,7 0,62
-
34,62 25.92 15,86 6,48 1.17 3
-
1,24 1,18 1,26 0,44
-
2,03 1,12 0,36
-
1,33 0,37 0,26 0,99 0,27 0,54
35,31 20,37 28,38 4,07 0,72 0,91 0,58 0,66 2,77 0,68 1,77 0,19 0,35 1,47 0,13
3.4 Kit qua phin tich mdt sd ehi tidu hda ly ciia tinh diu cii Ngai say
Mdt so chi tieu hda ly quan frgng ctia tinh diu cu Ngii sliy nhu tj frgng^ gdc quay cyc. chi sd acid, chl so savon hda vi"ehP'so"ester'dugc tien hinh phdn tich. Ket qud dugc trinh biy ttong Bing 4.
Nliin chung, tinh diu cu Ngii say frich ly td ba phuong phdp khae nhau ed sy khdc nhau vl chi so vgt ly, hda hgc. Kdt qui nay cung phu hgp vdi kit qui phdn tich thdnh phin hda hgc cda ba miu tinh diu thu dugc: thdnh phan hda hgc khac nhau din ddn cac tinh chit hda ly nhu gde quay c\fc, chi sd acid, ehi so savon vd chi si ester ciing kliac nhau.
Bang 4: Chi so vgt ly, hda hgc cua tinh dau cu Ngii siy
Dac diem NSC-1 NSC-2 NSC-3
Ty trpng Goc quay circ Chi so acid Chi s6 savon h6a Chi so ester
0,8360 -10,06 3,02 47,31 44,29
0,8714 -8,51 2,58 55,04 52,46
0,8564 -5,64 4,59 37,67 33,08 3.5 Kit qua dinh gid boat ti'nh sinh hgc cua tinh dau cii Ngai say
3.5.1 Hogt tinh khdng oxy hoa bdng phuang phdp DPPH
Hoat tinh khing oxy hda cua tinh ddu cd Ngii siy dugc xic dinh bing phuang phdp DPPH. Kit
r<p chi KhoG hoc TnaitgDgl htcCAiTlur Phin A: Khoa hfc Tr nhien. Cmg nghe vo USi Inmng: 35 (2014): I3I-I3S qua tinh diroc gia trj ECso cua tinh dJu cii Ngai say
va so sinh vol gia tri ECso cila chit doi chiing la vitamin C va mgt s6 loai tinh diu khic (Sara Albino Antunes et a!., 2012) duoc tnnh bay trong Bing 5.
Bdng 5; Gii tri ECso ciia cdc lo?i tinh diu va vitamin C
Ten NSC-1 NSC-2 NSC-3
Cinnamomum camphora Ness (long nSo) Curcuma longa (nghe nha) Vitamin C
ECsi 1 (na/mL) 545 470 315
2094,172 4,870 Td ddy cho thiy eic loai tinh diu khio sit deu cd hogt tinh khing oxy hda khdng ddng ke so vdi vitamin C, mgt hgp chat dugc biet cd boat tinh khing oxi hda rat tot va cd nhieu iing dung thuong
Bang 6: Gia tri dirdng kinh vdng vd khuin cua tinh dau cii Ngai siy
mgi. Tuy nhien, so vdi hai loai tinh diu khio sdt thi tinh ddu cu N^ii say cd hogt tfnh khdng oxi hda tdt hon va tdt nhit la tinh diu eii Ngii say trich ly tu phuang phdp chung cit ldi cudn hoi nude cd su ho ttg ciia vi sdng (NSC-3).
3.5.2 Hogt tinh khdng ndm vd khdng khudn a. Thu nghiem hogt tinh khang ndm vd khdng khudn bdng phuang phdp khuich tdn tren dia thgch vd do dudng kinh vong uc chi
Do thinh phin hda hgc ciia cd ba loai tinh diu khd gidng nhau ndn dl tai tien hanh khdo sdt boat tinh khdng nim, khang khuin frdn mdu tinh ddu NSC-1 bang phuang phip khudch tin fren dia thgch va do dudng kmh viing uc chd 6t sang Igc so bd trudc.
Tinh ddu thu nghidm bao gdm tinh ddu nguyen chit Co vd tinh ddu pha lodng frong 1 the tich dimethyl sulfoxide, theo thd ty lin lugt la Ci, C2, C3, Ci. Lugng dung thii nghidm l i 25 pL.
Vi k h u i n Gram duong
Vi khuan Gram am V i n a m
T e n
Listeria monocytogens Staphylococcia aureus Enterococcus faecalis Bacillus cereus Bacillus sublillis Klebsiella pnemoniae Salmonella typhi Escherichia coli Pseudomonas aeruginosa Candida albicans
Dir&ug k i n h C . 11 8 10 6 6 6 6 3 6 17
v o n g vo k h u a n Cl 10 6 10 6 6 6 6 6 6 14
( m m ) C i
9 6 9 6 6 6 6 6 6 13
Cs 8 6 7 6 6 6 6 6 6 10
C4 7 6 7 6 6 6 6 6 6 8 Budng kinh 16 thgch: 6 mm
Dudng ldnh vong vd khudn = 6 mm: khdng co ddu hiiu diet khudn Duong kinh vdng vd khudn > 6 mm: xudt hien ddu hieu diet khudn
b. Thil nghi?m hoat tinh khdng ndm, khdng khudn bdng phucmg phdp pha lodng da nong dg xdc dinh ICso
Qua kdt qui khio sat khi ndng khdng nim, khing kbuin bing phuong phap khulch tin fren dia thgch v i do dudi^ kinh vung uc chl cd thi
nhin thay tinh ddu cu Ngii say cd dau hieu diet mgt sd ehiing vi khuin va nam kiem dinh. Chi'nh vi vay, de tdi tidp tyc xac dinh gid ttj IC50 cua tinh dau cu Ngdi siy frich ly dugc tu ba phuong phap dd cd kit luin chi'nh xac. Ket qud dugc frinh biy frong Bing 7.
Tgp ehi Khoa hoc Tnrdng Dgi hgc Cdn Tha Phdn A. Khoa hgc Tv nhiin, Cong nghe vd Mdi Irudng- 35 (2014): 131-138
Bang 7: Gia trj ICso Vi sinh vgt Vi khuin Gram dm Vi khudn Gram duong Vi nam
cua tinh diu cu Ngai siy Ten
Stiq)hylococcus aureus Bacillus subtilis Lactobacillus fermentum Salmonella enterica Escherichia coli Pseudomonas aeruginosa Candida albicans
I C ^ (i^/mL) NSC-1 NSC-2
>128 >128
>128 >128
>128 >128
>128 >128
> 128 >I28
>128 >128
> 128 >128
.
_ N S C : 3 _
00 00 00
AAA
>128
>128
>128
>128 ICSO < 128pg/mL' eo hogt tinh khdng vi sinh vol kiim dmh
1C50 > 128 pg/mL: khdng eo hogt linh khdng vi sinh vgt kiem dinh Bang 7 cho thiy tmh diu cuNgii say frich ly td ba phuang phip khdng cd hoat tinh kliang cdc chung vi khuin vd vi nam thd nghiem.
3.5.3 Xdc dinh doc linh vai te bdo ung Ihu Ba mdu tinh dau cu Ngii sdy dugc tiln hanh Bang 8: Dgc ti'nh vdi te bao ung thu ciia tinh dau c
ddnh gid hogt ti'nh giy ddc frdn 4 ddng td bio ung thu: ung thu vu (MCF-7), ung tiiu vu da khang tiiudc (MCF7/ADR) ung thu bilu md tuyln (MDA- MB-231) va ung tiiu cd til cung (Hela). Kdt qui dugc trinh biy ttong Bang 8.
I Ngai siy
Hop chat Ddng te bao/ICso (ug/mL)°
MCF-7 MDA-MB-231 Hela
NSC-1 NSC-2 NSC-3
4-HydrQxytamoxifen^
9,75±0,49 6,3±0,20 5,91+0,60 4,21 ±0,29
13,06+0,28 5,96+0,68 5,1+0,11 3,54±0,29
14,30+0,53 11,06±0,74 4,29+0,80 6,29±0,26 4,S5±0,42 6,29±0,31 3.51±0,62 4,27±0.82 a: Cdc gid tri dugc the hiin theo gia tri trung binh ± SD lap lai 3 ldn
b: Nhom doi chung duang
Theo Vien nghien cdu ung thu qudc gia Hoa KJ, mdt chit cd IC50 < 20 pg/niL duge xem la cd ddc tinh ddi vdi td bao. Kdt qua IC50 cdn giup so sdnh mdt cdch day du hoat tinh gay ddc td bio gida cie mSu vdi nhau. Ndu miu tinh dau nao cd gia frj ICso thdp hon, tiic ndng dd ue chd 50% thdp hon Ihi tinh diu dd cd hogt tinh gay ddc td bio manh hon.
CJ diy, ci ba mlu tinh diu deu cd IC50 < 20 [ig/mL cho thdy cdc mdu tinh diu nay ddu cd tilm nang khing ung thu. Tuy nhien, hai mdu tinli dau cu Ngdi say NSC-2 vd NSC-3 ed hoat tinh gay dgc ddi vdi td bao cao hon so vdi mau tinh dau NSC-1.
Dgc biet, hoat tinh giy ddc t l bao ung thu cua hai mdu tinh diu cu Ngdi say NSC-2 va NSC-3 xip xi so vdi ddi chiing duong 4-hydroxytamoxifen, mgt md hinh thy the esttogen chgn lgc dugc ling dung frong dieu frj ung thu vu vd phiu thuit dilu frj ung thu vd hien nay.
i KET LUAN
Khao sat tinh diu Ngdi say frdng d Bay Nui, An Giang cho cdc kdt qui nhu sau:
- Hieu suit tinh ddu d thin v i l i Ngii say khdng dang kd so vdi phan cu.
- Ddi vdi cu Ngdi say, phuong phap chung cit ldi cudn cd su h6 trg cda vi sdng (NSC-3) cho hieu suit tinh dau cao nhdt trong thdi gian ngdn nhit frong cd ba phuong phap n g l i i e g n V i i ^ ^ ^ ^ ^
Tinh diu cu Ngai siy frich ly tii ba phuang phap kliac nhau ddu ed ba thanh phin ehinh la 4- terpinenol (27 - 35%), sabinene (15 - 26%i) va 1,4- bis(raetiioxy)-triquinacene (7 - 28%).
Tinh dau cu Ngdi say khdng ed boat ti'nh khdng nim, khdng kbuin ddi vdi cac chung vi khudn va ndm thd nghiem vd hoat tinh khdng oxy hda khdn^ dang kl so vdi Vitamin C, nhung tdt han tinh diu Long nao vd tinh diu Nghe nhi.
Tinh diu cu Ngii siy cd hoat tinh giy ddc frdn cdc ddr^ tl bio ung thu: img thu vd (MCF-7) ung thu vu da khang tiiudc (MCF7/ADR), ung tiiu bilu md tuyln (MDA-MB-231) vd ung thu cd tu cung (Hela)
T , . t r , . ™. , ^ •, ," - „ TTm- Phdn A: Khoa hoc Tv nhien, Cdng nghe vd Mdi Irudng:
Tgp chl Khoa hgc Trudng Dgi hgc Cantho ^ """
35 (2014): 131-138
LOn CAM TA
Nghidn cdu dugc tiiyc hi?n bdng ngudn kmh phi tii dl tii nghien cdu khoa hgc cua sinh vien (ma sd: TSV2013-03). Nhdm tdc gid xm chan tiiinh cdm on cd Hi Thj Kim Quy va Giao su Oh Won Keun (Khoa Dugc - Trudng Dgi hgc Qudc gia Seoul, Hin Qudc) da hd frg khao sat tiiu nghidm hogt tfnh gdy doc ttdn ede ddng tl bdo ung thu eua tinh ddu cii Ngdi say.
TAI LIEU THAM KHAO 1. Sara Albino Antunes, Weber da Silva
Robazza, Liziane Sehittier, Gilmar de Almeida Gomes, 2012. Synergistic and antimicrobial properties of commercial turmeric (Curcuma longa) essentijd oil against pathogenic bacteria. Ciencia e Teenologia de Alimentos. 32(3): 525-530.
2. Md. Nazrul Islam Bhuiyan, Jasim Uddin Chowdhury and Jaripa Begum, 2008.
Volatile constituents of essential oils isolated from leaf and rhizome of Zingiber cassumunar Roxb. Joumai of the Bangladesh Pharmacological Society. 3: 69-73.
3. Saowaluck Bua-in and Yingyong Paisooksantivatana, 2009. Essential Oil and Antioxidant Activity of Cassumunar Ginger (Zingiberaceae: Zingiber montanum (Koenig) Luik ex Dietr.) Collected from Various Parts of Thailand. Kasetsart Joumai: Natural Science. 43: 467-475.
4. Ibrahim bin Jantan, Mohd Salleh Mohd Yassin, Chen Bee Chin, Lau Lee Chen and Ng Lee Sim, 2003. Antifungal activity of the essential oils of nine Zingiberaceae spicies. Research Article of Pharmaceutical Biology. 41 (5): 392-397.
5. Nguydn Thi Kim Hue, 2009, Hg Gung (Zingibereceae) d Thit Son - An Giang:
Hinh thii, Gidi phdu, Phin loai vd Cdng dung. Luan dn Thac sT. Dai hpc c i n Tho.
c i n Tho, Viet Nam.
6. Trin Cdng L u ^ , Nguyin Thj Phuang Thdo vd Trin Thu Hoa, 2010. Thanh phdn hoi hgc vd mdt sd tac dyng sinh hoc ciia tinh dau ba loii ngdi sgy An Giang. Tap chi Y hgc Thanh phd Hd Chi Minh. Tap 14, Sd 2, Chuydn dl Y hgc Cd truyen, trang 151 - 156.
