Pham ThOy Linh vd Dtg Tap chl KHOA H(JC & CONG NGHE
N G H I E N c i r u B A O C H E C A O T H U 6 C C H I E T X U A T T f C A Y D O N L A D O (EXCOECARIA COCHINCHINENSIS L O U R . ) T H U H A I T A I T H A I N G U Y E N
Phatii Thity Linh , Nguyen Quoc Thjnh, Pham Thi Tuyet Nhung Tnrdng Dai hoc Y Duac - DH Thdi Nguyen T O M T A T
Muc tieu: Bao chg dupe cao d$c chiet xuit tCr cSy Don la dd thu hai tai Thai Nguyen; khao sat va xSy dyng tigu chuin co sd cua cao dac chiSt xuit ttr c§y Don Id do thu hai tgi Thai Nguyen.
Phuxmg p h i p nghien ciin: Kiem tra tinh diing cua duoc liSu theo phuang phap ghi yong Dug-c dien Viel Nam IV [3]; Tien hanh khao sat digu kien chiSt xuit: Dung moi, nhiet dp, ty le dung moi/dupc ligu; khao sat vk xay dyng mpt s6 tigu chuin co sd cua cao dSc. Ket qua: Cao dgc co the chit mim, ddng nhit, mau nau sam, mCii thorn, vj ding, chat; co chua flavonoid, mat khoi lupng do iam khd: 18,60 ± 0,50 (%) (a=0,05); pH = 3,97 ± 0,20 (a=0,05). Ket luan: Xay dung dupe quy trinh bio chl cao dac chiet xuat ttr cty Don la do thu hii tgi Thai Nguyen bing phuong phap ngam d nhi$t dp ph6ng vdi cdn 90%, tj IS dung moi tren dupe ligu la 12/1; bao che dupe cao d5c vdi khoi lupng la: 33,63 ± 0,37 (g) (a=0,05); xay dyng dupe du thao tieu chuan co sd ciia cao dac la _Don Id do gdm cdc chi tlSu: Cam quan, djnh tinh, mat khdi lupng do \km kho, do ddng nhit va pH.
Til'kh6a: Dan la do, flavonoid, dj ting, cao dgc, Thdi Nguyen
D A T V A N D E
Dj ling la mpt benh kha phd bien d Vigt Nam, Bgnh anh hudng nhieu den tinh than va chat lugng cupc song cua con ngudi, V a o nhQ'ng nam 1970 cua the ky trudc, t>' le cac bgnh dj ling (dj irng thuoc, viem mui di iing, hen phe q u a n . , , ) cd xu the tang nhanh [7], Dac bipt trong nhung nam gan day, khi mdi trydng ngay cang 6 nhigm, khf h | u thudng xuyen thay doi dpt ngpt, cac tac nhan gay dj dng cQng x u i t hign nhigu han, lam anh hudng den c h i t lugng cugc song ciia con ngudi.
Cac thuoc tay y digu trj dj iing bao gom thu6c khang histamin vh corticoid. Hai nhdm thuoc nay deu cd nhigu tac dyng phy. Trong khi dd, nude ta cd ngudn d u p e lieu phong phii, nhan dan ta cd nhidu kinh nghigm su dyng cac vj thudc chua dj irng vdi ft tac dyng phy hon. Do dd vigc nghien ciiu vj thuoc chong dj irng c6 nguon goc d u a c lipu la can thiet.
Don la dd d u p e bidt den vdi tac dyng chita dj ling [ I ] . T r u d c day da c6 mpt so nghien ciiu ve thyc vat, hda hpc va tac dyng sinh hpc ciia Don la dd tgi Ha Npi, Tuyen CJuang, Thai Binh [18], [9], Cay thuoc dupe trdng nhigu d Thai Nguygn. N h u n g chua co nghien ciiu
Tel: 0977 404151, Email-phamlinhl702@gmaiic<
dieu chg cao va chuan hda cao cua vj thudc thu hai tgi Thai Nguygn vdi myc dich hudng tdi tao ra san pham ho tra dieu tri benh di li'ng. Vi vgy, chiing tdi thyc hign de tai
"Nghign cii'u bao chg cao thu6c chigt xuat tii eay Dan la do (Excoecaria cochinchinensis Lour.j thu hai tai Thai Nguyen" nhim gdp phan nang cao gia trj sir dung ciia dupe ligu nay va tign tdi phat trien mpt san pham t u d u p e ligu dg hd trp digu t n chiing benh dj ling. Dg tai dugc thuc hien vdi hai muc tieu:
Bao che duac cao dgc chiet xuat tii cay Don la dd thu hai tgi Thai Nguyen; khao sat va xay dyng tieu chuan c a sd ciia cao dac chiet xuat tir cay Don la dd thu hai tgi Thai Nguygn.
DOI TU'ONG VA PHU'ONG PHAP N G H I E N c i r u
Doi tirong nghien cihi
La d u p e phoi va say khd ciia cay D a n la do (Excoecaria cochinchinensis Lour.> thu hai tai Thai Nguygn.
P h u w n g p h a p n g h i l n ciru
Phiamg phdp xdc dinh ti'nh dung cda dime lieu - Cam quan: Quan sat d anh sang thudng. Md ta mau sac, hinh dang, kich thudc va miii vj ctia d u p e lieu.
Phgm Thiiy Linh vo Dtg Tap chi KHOA HQC & CONG NGHE 177(01): 33-38 - Soi bpt: Thai nhd dupe ligu, say khd, tan
thanh dgng bpt thd, ray lay bgt mjn. Lam tieu ban bpt bang p h u a n g phap gipt ep. Quan sat dac diem dudi kfnh hien vi. Chup anh cac dac diem quan sat duoc,
- Djnh tinh: Tien hanh dinh tfnh 2 nhdm hgp chat chinh la flavonoid va c h i t mau theo phuong phap dupe ghi trong chuyen luan Dupe ligu va thudc tii dugc ligu (Dan la do) - Du(xc diin Viet Nam IV [3].
Phirffng phdp xdy dung quy trinh bdo che cao dgc chiit xudt tit Id cdy Bon Id do Tien hanh khao sat cac dieu kien chiet x u i t gom: Dung m6i, nhigt dp, ty Ig dung mdi/dupe ligu. Lya chpn dieu kign chiet xuat cd it tap va cd hieu suat chiet xuat cao hon.
Djch chiet dugc logi tgp bang phuang phap thi'ch hpp. Djch chigt da logi tap duac cat thu hdi dung mdi dudi ap suat giam va cd each thiiy tdi khi thu dugc cao dac.
Phiamg ph^ khdo sdt vd xdy dimg tieu chuan Cff sd cOa cao dac chiet tir Id c^ Don Id do
• Cam quan: Quan sat cao d anh sang thudng.
Md ta mau sac, the chat va miii vi ciia cao.
- Djnh tinh: Djnh tfnh flavonoid trong cao dgc bang cac phan iing hoa hpc theo phuang phap ghi trong tai lieu [2], [3] va [5].
M i t khoi lugng do lam khd: Xac djnh b i n g phuang phap quy dinh tgi Dugrc diin Viel Nam IV{Ph}^\\tc 9.6) [3],
pH: x a c djnh theo phuong phap ghi trong Duac diin Viit Nam IV{Phu luc 6.2) [3], - Dp dong nhit; Cho cao Ign lam kfnh, dgt lamen len ep sat, quan sat dudi kfnh hign vi dg kiem tra dp dong nhat ciia cao, Phuffng phdp xu ly so liiu
Ket qua thyc nghigm luu trii va ti'nh toan b i n g phin mgm M I C R O S O F T EXCEL. Xii 1^ so ligu thyc nghiem b i n g p h u a n g phap th6ng kg vdi khoang tin cgy 9 5 % ( a = 0,05).
KET QUA NGHIEN C U U Xac dinh tinh diing c u a ducrc l i | u - C a m q u a n : La hinh b i u due hai dau thudn nhpn, dai 5 cm den 11,5 cm, rdng 2,5 cm den 34
4 cm. Cudng la dai 0,5 cm den I cm. Phien la nguygn, mep la cd rang cua nhd, mat tren mau xanh la dam, mgt dudi mau dd tia Cd 10 dgn 12 cap gan Idng chim noi ro d mat dudi cua la, lorn d mat tren la. Hinh anh dupe lieu dugc trinh bay d hinh 1.
Hlnh 1. Duac liiu Dem Id do - Bot: Cd mau xanh nau, miii hac nhe. Quan sat dudi kinh hien vi thay: Manh mach mgng (3), mgch xoin (6) dii-ng rieng le hay trong cac mo;
sgi (4); manh md mem (2); tinh thg caici oxalat hinh cau gal ( I ) ; manh bieu bi cd nhieu tg bao lo khf kieu song bao (5). Anh chyp cac dac diem bpt dupe lieu Don la dd dudi kinh hiln vi dupe trinh bay d hinh 2.
Hinh 2. Anh chup cdc ddc diem bgt duac li$u Dan Id do du&i kinh hien vi Ghi chij; I: Tinh thg caIci oxalat hinh ciu gal, 2:
Manh md mgm; 3: Manh mgch mgng; 4: Manh mach xoin; 5 Manh bieu bi mang lo khi kieu song
bao; 6: Spi - D i n h t i n h ;
+ Flavonoid: Phan iing cyanidin va phan iing vdi kiem duong tfnh. N h u vay trong dugc lieu cd flavonoid.
+ C h i t mau: Dich chigt vdi con 50% cd mau dd tfa, N h u vay, trong dugc ligu cd chit mau.
Kit lugn: D u g c lieu dat tieu c h u i n Duac Dien Viit Nam IV [3] v e cac chi tieu: Cam quan, bpt, dinh tfnh.
Xay d y n g q u y t r i n h b a o c h i cao d a c - K h a o sat n h i l t dp v a d u n g moi chiet xual:
Phgm Thily Linh vd Dtg Tgp chi KHOA HOC & CONG NGH$ 177(01): 3 3 - 3 8
Nghign Cliu da sii dyng phuang phap ngam
Ignh tai nhiet do phdng vi cd the tiln hanh d quy md nhd (quy md phdng thf nghigm) cflng nhu khdng ddi hdi may mdc, trang thiet bj dit tien. Mat khac, phucmg phap chigt ndng tuy thdi gian chiet nhanh hon nhung Igi cd the gay pha hiiy hoat chit.
Tien hanh khao sat dung mdi ethanol d cac ndng do 70% va 90% do tham khao 2 tai lieu trudc do la [3] va [6] da sii dyng 2 dung mdi nay dg chiet xuit flavonoid toan phin la thanh phan cd tac dyng chira dj iing.
Bang I. Anh huang cua ly Ii dung
Kit qud nghien ciru cho thdy: Dich chiet ethanol 70% cd mau dd tia, ethanol 90% cd mau vang dim.
Nhgn xet: Dich chiet ethanol 70% cd nhieu tap hon ethanol 90%. Do dd, chpn dung mdi ethanol 90% dg khao sat cac digu kien tigp theo.
- Khao sat ty le dirp^c lieu va dung moi:
Tien hanh khao sat cac ty Ie dung m6i/dupc ligu la: 20/1, 12/1, lO/I sau khi chgn dugc cac dieu kien vg dung m6i va nhiet dd chiet xuat, Anh hudng ciia ly le dung mdi/duac ligu den hieu suat chiet dupe trinh bay trong bang 1.
mdi/duac hiu din hiiu sudt chiit Khoi lirpng duo'c
ligu kho kigt (g)
STT Ty Ig dung mdi/duo'c ligu Khoi iu'png cao
khd kift (g)
Higu suat
chiit (%) 20/1
12/1
10/1182,22 183,11 181,34
36,45 33,61 22,55
20,00 18,36
12,44Nhdn xet: Ket qua khac sat cho thay ty le 20/1 va 12/1/ cho hieu suat chiet cao hon so vdi ty le
10/1. Tuy nhien, sy chenh lech giua 2 ty Ip 20/1 va 12/1 la khdng qua Idn. Vi vgy chiing t6i lya chpn ty Ig dung mdi/dugc ligu la 12/1 la bJ' le phCi hpp dg chilt xuit, tilt kiem dugc dupe ligu dung mdi va boa chat.
- Quy trinh bao che cao:
Sau khi khao sat mpt so dieu kign chigt xuat, dg tai da dua ra quy trinh bao che cao dgc nhu sau:
La Don la dd dugc nghien thanh bdt. Xac djnh mat khdi lupng do lam khd bang phuang phap dugc ghi trong Dui^-c dien Viit Nam /F'(phy luc 9.6) [3] cho ket qua mat kh6i lupng do lam kh6 ciia dugc ligu la 11,11%. Tien hanh d6ng thdi 6 lan nhu tren, m6i lan 200 g b6t dugc lieu thu dugc 6 miu cao, ky hieu lin lugt la CI, C2, C3, C4, C5, C6,
Quy trinh chiet xuat va bao che cao dgc chiet tii dupe lieu la Don la d6 (sau day gpi tat la cao dgc Don la dd) dupe tdm tat d sa do hinh 3.
The tfch cdn de chiet, khdi lugng cao va higu suat chiet xuat thu dugc cua 6 mau cao dac dugc trinh bay trong bang 2.
H
I) [biiy tinhdjm miuco ax_L
.Tli!in2400mI.M-njtm2400mLtt'
[ « T «
_^[^
—*| Lgclo:
Hinh 3. Sadd torn tdt quy Irinh chiet xudt vd bdo chi cao dgc Dan Id do
Phgm Thiiy Linh vd Dtg Tap chf KHOA HQC & CONG NGH$ 177(01): 3 3 - 3 8 Bang 2. Thi tich cdn di chiet vd hiiu sudl chiit xudt thu dugc cua 6 mdu cao
Hi^u su^t chi^t Khoi lu'ffng cao
d | c khfi kigl (a) Tong lu'png con
chiet (ml)
Khii lu'png ducrc ligu
khfi kigt (g)
(%)
1 2 3 4 5 6
4800 4800 4800 4800 4800 4800
177,78 177,78 177,78 177,78 177,78 177,78
33,50 33,72 33,64 34,01 33,01 33,89
18,84 18,97 18,92 19,13 18,57 19,06 Thong ke; Khoi lupng cao d^z kho kigl 33,63 ± 0,37 (g) (a-0,05)
Higu suit chilt xuit: 18,92 ±0,21 (%) (a=0,05)
- K h a o sat va xay d u n g m o t so chi tieu kiem n g h i l m cao d^c D o n la d o
+ C a m q u a n : Cao dac-cd the c h i t mem, dong nhit, mau nau sam, miii t h a m , vi dang, chat + M a t khoi luffng d o l a m khfi: Ket qua xac djnh m i t khoi lugng do lam khd ciia cao dac dupe trinh bay d bang 3 .
Bang 3. Kit qud xdc dinh mdt khdi tug'ng do ldm kho (%) cua cao ddc Dan Id do Trang binh
Thiing kg
18,03 19,01 18.60 J: 0,50 (%)(a-Q,05)
18,45 19,11 Nhgn xet- Ck 6 mau cao deu cd mat khoi lugng do lam khd khdng qua 2 0 % , thda man tigu chuan v e m i t khdi lupng do lam kho cua cao dac theo Dirac dien Viel Nam IV (Pha luc I . I ) [3], De nghj: Mat khoi lugng do lam kho cua cao dgc D a n la dd khdng qua 20,0%.
+ Dinh t i n h : Bang cac phan iing hda hgc thudng quy theo phucmg phap ghi trong tai ligu [2], [4]
va [5]. K i t qua thi nghigm cho thay 6 mau cao dgc Don la dd dgu cd c h u a flavonoid.
De nghj: Cao dac phai chiia flavonoid
+ p H : Kgt qua do pH ciia 6 mau cao dugc trinh bay d bang 4, Mdi mau cao dac tiln hanh do 6 lin, liy ket qua trung binh. K i t qua xac dinh pH cua cao dgc la Don la dd dugc trinh bay trong bang 4 dudi day.
Bang 4. Ket qud xdc dinh pH ciia cao ddc Dan Id do
Trung binh Th6ng ke
3,92 3,80 3,97±Q,10 (a=0,05)
4,02 Nhdn xet: Cao dgc Don la dd cd tfnh acid y l u . pH cua dung djch cao dgc Don la do 1% trong nudc(kh6i lugng/thl tich) la 3,97 ± 0,10 ( a = 0,05).
De nghi: Dung djch cao dac Don la do 1%
trong nude (kh6i lupng/the tich) phai cd pH t u 3 , 8 0 den 4,20.
+ Dp 6hng n h a t : Can 1 g cao dac D a n la do cho len lam kinh, dgt lamen len ep sat, quan sat dudi kfnh hiln vi d l k i l m tra dp dong nhit ciia cao. Hinh anh cac mau cao dgc quan sat dudi kinh hiln vi dugc trinh bay d hinh 4.
Nhdn xet: Cac mau cao dgc cd mau vang, ddng nhat, khdng cd vang moc, ba dugc lieu hoac vgt la.
36
Hlnh 4. Anh chup cdc mdu cao ddc quan sdt dudi . kinh hiin vi vgt kinh 10 ''-•
Di nghi: Quan sat dudi kinh h i l n vi, cao dgc Don la dd phai d6ng nhit, khdng dupe co vang moc, ba d u g c lieu hoac vgt la.
BAN LUAN
Qua vigc xac dinh tinh ddng ciia dupe ligu da dua ra dugc mpt so tieu chuan de lya chpn
Pham Thuy Linh va Dtg Tgp chi KHOA HQC & CONG NGHE 177(01): 33 - 38 nguyen ligu dau dl bao che cao dac tii' la cay
Dan la do, Dl xay dyng quy trinh bao chl cao dac Dan la dd, dl tai da tien hanh khao sat mdt so dieu kien chiet xuat dl thu dugc cao dgc cd chOra flavonoid va cd higu suat chiet cao han. Tuy nhign, neu cd chit chuin dl djnh lugng dupe hogt chit, rdi tir do lya chpn ra quy trinh cd ham lugng hoat chat cao han se giup thu dugc cao dac cd tac dung toi uu.
Tuy nhign do thieu chat chuin nen chua thi tien hanh khao sat. Nhu da bilt flavonoid la nhu'ng chat cd dp phan cue trung binh. Tuy nhien, neu sii dung cac dung mdi huu co it phan cue dk chiet xuit nhu chloroform, ethyl acetat, n-hexan... se gay dgc hai vdi siic khde con ngudi va mdi trudng, khdng thd ap dyng d quy md Idn. Hdn hgp ethanol va nude dugc lua chpn vi day ciing la dung mdi cd the chiet dugc nhilu hoat chat tii phan cyc din it phan cue (gom cd cac flavonoid trong Don la dd la thanh phan tac dyng ch6ng di iing [1]) re tiln, kh6ng dgc hai, de kiem, it dgc hgi va pho bien. Cac tai lieu chii yeu diing cdn 70% va 90% [3] va [6] de chiet xuit djch chilt flavonoid toan phan (la chat cd tac dyng chong di iing) nen de tai lya chgn 2 dung mdi nay de khao sat, Lya chpn bao che cao long se riit ngan thdi gian cat thu hdi dung mdi, nhung lai khdng the logi het c6n ngn nhdm nghien cii'u da chpn hudng xay dung quy trinh bao che cao dac.
Do lugng dugc lieu thu hai dugc khdng nhieu va de tiet kigm dung mdi hda chat nen de tai khdng tiln hanh khao sat cac ty le 7/1 va 5/1.
Vi vay, hien nay di d6i vdi vipc su dyng dugc lieu thi ngn cd hudng bao tdn va phat trien dugc ligu Dan la dd.
De tai da tien hanh xay dyng dugc dy thao tieu chuan ca sd ciia cao dac la Don la dd gom cd cac chi tieu: Cam quan, dp dong nhat, djnh tinh, mat khdi lupng do lam khd va pH.
Qua chi tieu djnh tfnh thiy rang trong qua trinh bao che cao dgc khdng lam mit di thanh phin flavonoid (la thanh phin cd tac dung chong dj ling trong la cay Dan la do). Do pH
cho thay cao cd tfnh acid trung binh. Tinh chit dd la do trong Dan la do cd chiia cac thanh phan cd tinh acid va trung tinh la:
Flavonoid 1,5%, saponin, coumarin, anthranoid, tanin, dudng khii [6]
Day la lan dau tign cd nghien ciiu xay dung quy trinh bao che cao dac va tigu chuan hda cao dac Don la do thu hai tgi Thai Nguygn KET LUAN
Xay dyng dugc quy trinh bao chl cao dac chiet xuat tir cay Dan la dd thu hai tgi Thai Nguyen bang phuang phap ngam d nhiet dp phdng vdi cdn 90%, ty le dung mdi tren dugc ligu la 12/1. Bao che dugc cao dgc vdi khoi lupng la: 33,63 ± 0,37 (g) (a=0,05); xay dung dupe dy thao tieu chuan ca sd ciia cao dgc la Don la dd gdm cac chi tieu: Cam quan, do dong nhat, djnh tfnh, mat kh6i lupng do iam kh6 va pH.
TAI LIEU THAM KHAO 1. Nguyen Thai An (2003), Nghien cuu dugc lieu dan Id do (Excoecaria cochinchinensis Lour var.
cochinchinensis), Lu^n an tiln sT trudng Dgi hpc Dupe Ha N6i,
2. Bp mdn Dupe lieu (2012), Phuang phdp nghien cuu dugc liiu, Tnidng Dai hpc Y Dupe th^nh pho H6 Chi Minh, thanh phd Ho Chi Minh, tr. 51 -104.
3. Bp Y tl (2009), Dugc diin Viil Nam 4, Nxb Y hpc, Ha N$i.
4. Pham Thanh KJ- (2007), Dugc Ii?u hgc, lap 2, Nxb Y hpc, Ha Noi.
5. Ngo van Thu, Trin Hung (2011), Dugc liiu hoc, tap I, Nxb Y hpc, Ha Npi
6. Lg Thu Trang (2013), Nghiin cuu diiu chi cao dgc vd thu mot s6 tdc dung sinh hgc cua bdi thudc Nuna, khda luSn t6t nghigp Dupe st, Trudng Dai hpc Dupe Ha Npi.
7.http://diungmiendich.comvn/diung/productView /354_636/html.d
8, An Thi Nguyen f2003), "Isolation of Flavonoids from Excoecaria cochinchinensis Lour, var cochinchinensis". Journal of Medicinal Materials - Hanoi. 8(4), pp. 103-107.
9. An Thai Nguyen (2001), "Preliminary investigation on some biological effects of leaves of excoecaria cochinchinensis Lour Euphorbiaceae", Pharmaceutical Journal, 298(2), pp. 20-21,
37
Phgm Thuy Linh vd Dtg Tgp chl KHOA HQC & CONG NGH$ 177(01): 33 - 38
SUMMARY
S T U D Y O F P R E P A R A T I O N E X T R A C T U M
F R O M EXCOECARIA COCHINCHINENSIS LOUR. H A R V E S T E D IN T H A I N G U Y E N
Pham Thuy Linh*, Nguyen Quoc Thinh, Pham Thi Tuyet Nhung TNU - Umversity of Medicine and Phamarcy Objectives: To prepare extractum from the leaves ai Excoecaria cochinchinensis Lour, havested in Thai Nguyen; to survey the conditions of prepare of extractum and build some indicators to test it from the leaves of Excoecaria cochinchinensis Lour, havested in Thai Nguyen. Methods;
Checking the right of pharmaceuticals under the Vietnam Pharmacopoeia IV [3]; conducting a survey of conditions to prepare extractum: Solvent (70% ethanol, 90% ethanol), temperature, the ratio of solvent/pharmaceutical; evaluating the quality of extractum determined in accordance with Vietnam Pharmacopoeia IV. Results: Have prepared exUactum whose characteristics are soft, uniform, dark brown, fragrant, bitter, sour, acrid; extractum contained flavonoids; humidity: 18,60
± 0.50 (%) (a=0.05); pH = 3.97 ± 0 20 (a-0,05). Conclusions: Have choose the conditions of prepare of extractum. Have examined the conditions and constructed process of extracting of extractum whose volume is 33.63 ± 0.37 (g) (a=0.05) from the leaves of Excoecaria cochinchinensis Lour.; connstructed drafting specific to test exfractum.
Keywords: Excoecaria cochinchinensis Lour., flavonoids, allergy, extractum. Thai Nguyen
Ngdy nh^n bdi: I I/I 0/2017; Ngdy phdn bi?n: 08/11/2017; Ngdy duyit ddng: 30/01/201S ' Tel: 0977 404151, Email: phamtinhl 702@gmail com
38
