Tiiu ban: cdc chdt co hogt tinh sinh hqc ISBN: 978-604-913-012-0
TIEM NANG DANH GIA CHAT LlTONG DlTOC LIEU VIET NAIVI BANG
PHLTOTVGPHAP DAU VAN TAY SAC K Y '
THE POTENTIAL OF CHROIMATOGRAPHIC FINGERPRINTS FOR QUALITY EVALUATION OF VIETNAMESE HERBAL MEDICINES
Nguvin Hoai Nam, Chau Van Minh, Phan Van Kiem, Nguyen Xuan Cuotig, Nguyen Tien Dat, Pham Van Cuong, Nguyen Van Hung, Doan Thi Mai Huotig,
Nguyin Hai Dang, Hoang Le Tuan Anh.
Vien Hda sinh biin, Vien Khoa hoc va Cdng nghe Viet Nam 18 - Hoang Quoc Viet, Cdu Gidy, Ha Ndi
Email: naminpc(a),gmaiI.com
Tom tat:
Tu xa xua dugc lieu cd truyin Viet Nam vd cdc sdn phdm tu dugc lieu dd dugc su dung rdng rdi trong chdm sdc sue khde cdng ddng. Tuy nhien vlic ddnh gid dinh linh vd dinh lugng vi muc do an todn vd hiiu luc cua y hgc truyin thdng cdn rdt thieu hut vd chua dgt dugc nhdng tiiu chi cdn thiit di pup cho viec su dung y hgc cd truyin Viet Nam rdng khdp tren phgm vi todn the gi&i. Trong nhirng kit qud gdn ddy vi ddnh gid chdt lugng dugc lieu Viet Nam, phuong phdp ddu vdn tay sdc ky dp dung trong viec ddnh gid chdt lugng dugc lieu Mallotus vd nghien ciru kit hgp giira ky thudt ddu vdn tay sdc kyvdky thudt xu ly sd lieu nhdm chi thi nhirng peak tiim ndng liin quan den hogt tinh chdng d xy hda cua mdt sd mdu Mallotus cUng da dugc cdng bd. Bdo cdo ndy di cap din tiim ndng ddnh gid chdt lirgng dugc lieu Viet Nam bdng phuong phdp ddu vdn tay sdc ky.
Abstract:
The traditional Vietnamese herbal medicines (VHMs) and their preparations have been widely used for thousands of years throughout Vietnam. However, the quantity and quality of their safety and efficacy data are far from sufficient to meet the criteria needed to support Us use worldwide. In our recent studies on quality evaluation of VHMs, the development of HPLC fingerprints for Mallotus extracts and the combination of fingerprint technology with data-handling techniques, which allows us to indicate the peaks potentially responsible for the antioxidant activity of several Mallotus species, were investigated. Based on these results, this paper reviewed the potential of chromatographic fingerprints for the quality evaluation of Vietnamese herbal medicines.
I. GlOfI THIEU CHUNG
Thao dugc cd mat d nhieu noi tren thi gidi, dac biet la cac khu vuc nhiet ddi. Thao dugc ddng vai trd quan trong trong cac lieu phap chira benh bang y hoc cd truyen tai cac nude phuong Ddng. Nhieu qudc gia cd tiem nang vl dugc lieu nhu Trung Qudc, An Do da va dang cd nhieu chinh sach thuc ddy viec sir dung cac bai thudc trong y hgc cd truyen nhdm mang lai nhung ngudn Igi to Idn trong viec cham sdc siic khde cdng ddng va xuat khau dugc lieu. Cho den thdi diem nay Trung Qudc la qudc gia cd nhilu thanh cdng trong nghien ciin va iing dung dugc lieu cd truyin. Hien nay cd rdt nhilu san pham dugc lieu xuat xii tir Trung Qudc, vi du
Hoi nghi Khoa hqc ky niem 35 ndm Vien Khoa hqc vd Cong nghe Viet Nam - Hd Noil0/2010
nhu san pham tir cay Bach qua {Ginkgo biloba) dudi dang udng, vien nen va thudc tiem dugc diing de dieu tri giai doan sdm ciia benh Alzheimer, sa sut tri tue, ii tai do mach gay nen; san pham tir nhan sam {Panax Ginseng) dudi dang vien diing lam thudc bd dudng, lam cho cudng trang co the...Nhiing san phdm nay da va dang cd mat tren thi trudng the gidi va dugc rat nhieu ngudi quan tam va sir dung. Dk cd dugc thanh cdng dd, cac nha khoa hoc Trung Qudc da cd rat nhieu nd lire nghien ciiu va cdng bd cac nghien cim ve danh gia chat Iugng cac loai dugc lieu phd bien cua bo tren cac tap chi cd uy tin tren the gidi [1]. Trong khi dd d cac qudc gia thudc khu vuc Ddng Nam A va An do, ngudn dugc lieu hoan toan chua cd sir chuan hda.
Mot sd loai dugc lieu dugc thu thap khdng chinh xac va nay sinh sir nham Ian tir sir gidng nhau ve binh thai hoc cua cay trong cimg noi thu mau, tir sir nhan dang cay hoae tir nhiing sai sdt tir cac mau thuc vat nhap khau. Mot vi du dien hinh tir mot loai dugc lieu phd bien cd ten Ddc boat. Doe boat thudng la ten goi cho cac re ciia loai thudc chi Angelica (ho Triic Dao Apiaceae) nhu Xuyen doe boat {A. laxiflora Diels va A. megaphylla Diels), huong doe boat {A. pubescens Maxim.). Loai dugc lieu nay ciing cd the thudc mot chi kbac nhu Nguu vi ddc boat {Heracleum hemsleyanum Michx.), ddc boat Idng mem {Heracleum lanalum Michx.).
Hay tham chi thudc mot ho kbac nhu Cim nhdn ddc boat {Aralia cordata, ho Nhan Sam, Araliaceae) hoae nhu ciing mot loai dugc lieu cd ten Thd tam that nhung cd den vai loai kbac nhau cd cimg ten nhu vay, dd la Gynura pseudochlna, Stahlianthus thorelll Gagn, thudng dugc sir dung thay the cho Panax nologinseng ma khdng cd sir quan tam den nhirng thay ddi ve hda hoc va boat tinh sinh bgc ciia cay. Ngay ca cay Che vang {Jasminum subtrlpllnerve BL.) diing de chGa cac benh viem nhiem, da day, se rat de bi nham Ian vdi cay La ngdn {Gelsemlum elegans Benth.), mot cay cd doe tinh rat cao, cd the gay chet ngudi. Ngoai ra cdn cd rat nhieu loai dugc lieu kem chat lugng, dugc lieu bi lam gia va dugc lieu bi phdi trdn. Van de nay hien nay khdng chi xay ra d Viet Nam ma cdn d cac nude A Ddng, noi sir dung rat nhieu ngudn thao dugc lam thudc chiia benh. Van de nay ciing dugc nhac den trong "Hudng dan cho phuong phap nghien cuu, thir nghiem y hoe cd truyen" cua td chiic Y te the gidi (WHO) nam 2000. "Mac dii da tdn tai va sir dung qua hang the ky, mac dii da dugc diing phd bien trong sudt vai thap ky qua, y hoc cd truyen van chua dugc chinh thiic cdng nhan tai hau bet cac qudc gia [2]. Nhiing nguyen nhan tren khien eho chat lugng dugc lieu d nude ta hien nay khdng dam bao, gay anh hudng true tiep den siic khoe cdng ddng [3]. Chinh vi the, viec kiem soat chat Iugng dugc lieu can dugc thuc hien chat che qua dd se gdp phan tang hieu qua sir dung va khai thac dugc tiem nang ciia dugc lieu cd truyen. Kinh nghiem ciia qudc gia cd nhieu thanh cdng trong quan ly va danh gia chat lugng dugc lieu cho tbay viec danh gia chat lugng dugc lieu can thuc hien bai ban va quy md. Nam 2000, co quan quan Iy dugc Trung Qudc thira nhan va phat trien phuong phap dau van tay sac ky trong danh gia dugc lieu. Tiep dd rat nhieu cac nghien cuu danh gia chat lugng bao gdm cac nghien cuu ve hda hoc, boat tinh va co sd dii lieu dau van tay sac ky dugc cdng bd tren cac dugc dien, tren cac tap chi cd uy tin tren the gidi [4-6]. Cac nghien ciiu bai ban ciia Trung Qudc da gdp phan dua cac san pham tir dugc lieu ciia ho dat chuan hda va dugc thira nhan tai cac thi trudng khd tinh nhat nhu chau Au va My [7].
II. C d s d PHU^OTVG PHAP DAU VAN TAY SAC KY VA THIET BI NGHIEN ClTU Theo dinh nghia, dau van tay sac ky (chromatographic fingerprint) ciia mot mau thao dugc la mdt sac ky dd the hien dugc dugc tinh va tinh chdt hoa hoc dien hinh cua mot dich chiet mau thao dugc dd. Mau sac ky dd nay phai dam bao cac yeu cau ve sir xuat hien "rd rang" hay "md nhat", sir "gidng" hay "khae nhau" ciia cac ylu td hoa hoc cd mat trong dugc lieu [8]. Bude dau tien ddng vai trd quan trong trong quy trinh tao mau ddu van tay sdc ky dd la viec chuan boa qua trinh thu thap mau, xac dinh ten phan loai dugc lieu va ehilt xudt mau nghien cim [2, 7]. Bude tiep theo la viec khao sat boat tinh sinh hoc ciia mau nghien cim. Qua
Tiiu ban: cdc chdt co hogt linh sinh hqc ISBN: 978-604-913-012-0
trinh nay dugc thuc hien theo muc dich danh gia chat lugng dugc lieu ciia timg phdng nghien cim. Cac boat tinh sinh hoc thudng dugc xem xet la: boat tinh khang sinh, boat tinh chdng dxy boa, hoat tinh gay doe te bao, boat tinh chdng virus.. .Viec kilm soat chdt Iugng ciing bao gdm ca viec xac dinh hoat tinh tu nhien vdn cd ciing nhu ca doe tinh ciia chiing. Xac dinh rd rang boat tinh hoae doe tinh ciia thao dugc se lam cho cac san phdm cd gia tri cao bon trong nhiing iing dung dugc pham va thuc pham [7]. Tiep dd viec lira ebon phuong phap phan tich ddng vai trd quan trong vi viec lira chpn, sir dung cac phuong tien phii hgp vdi ddi tugng nghien ciiu se mang lai hieu qua cao trong qua trinh danh gia chat lugng dugc lieu. Mdi thiet bi va cdng cu nghien cim sir dung trong viec danh gia chat Iugng dugc lieu deu cd tinh nang uu viet va nhugc dilm, vi du nhu sdc ky Idp mdng cd uu viet la sir dung don gian, Iinh boat, tdc do phan tich nhanh, de thuc hien trong moi dieu kien. Tuy nbien, mot van de gap phai khi sir dung sdc ky ldp mdng dk danh gia dugc lieu dd la trong mau thao dugc thudng cd nhieu thanh phdn, cac thanh phdn nay cd mirc do phirc tap cao, dl phan buy va bay hoi, cd nhieu thanh phdn chdng cheo va d Iugng rdt nhd thi phuong phap sac ky Idp mdng gap nhieu khd khan. Hon thi, dir lieu ciia sdc ky ldp mdng la dir lieu mot chieu khdng thoa man dugc yeu cdu ciia phan tich dii lieu trong dilu kien hien nay. Them vao dd viec chuyen dii lieu de cd the ling dung cac phuong phap boa thdng ke de tien hanh xir Iy sd lieu gap nhieu khd khan, ddi khi khdng thi thuc hien dugc viec chuyen ddi de xir Iy sd lieu thu dugc [9]; Sac ky khi cd do nhay cao va thich hgp ddi vdi cac mau cd thanh phan de bay hoi, phuong phap nay gap khd khan khi cac boat chat ciia mau thao dugc cd tinh phan cue cao va khdng phai la thanh phan dl boa hoi [10]; Sac ky ldng cao ap ket hgp vdi cac dau dd chuyen dung DAD, FLD, tan xa anh sang ELSD, va ket hgp vdi khdi phd (hinh 1) rat hieu dung trong xay dung cac dau van tay sac ky [6]. Cudi ciing la viec xir ly sd lieu thu dugc bang cac phuong phap hda thdng ke (chemometrics) nhu phuong phap phan tich thanh phan chinh (PCA), phan tich nhdm (HCA), phuong phap binh phuong tdi thieu rieng phan (PLS). Bude nghien ciiru nay cd vai trd vd ciing quan trong, ket qua nghien cim se la cau tra ldi cudi ciing cho viee img dung phuong phap de giai quyet van de gi. Cd nhieu each danh gia dii lieu nhu dua tren cac bieu dd hoae cac dd thi, hoae dua tren sir kiem tra cac gia tri ket qua (score) va gia tri nap (loading). Thdng qua dd cac mdi lien he giiia cac dau van tay sac ky (hinh 2) vdi cac thanh phan trong dugc lieu nhu: cac hgp chat chi thi, boat tinh sinh hoc, ddc td...dugc giai quyet va hien thi. Dua vao cac tieu chi cu the nhu so sanh cac sd lieu dau van tay theo ly thuyet thdng tin (information theory), danh gia sir gidng nhau giiia cac mau (similarity estimation), nhan dang hda hoc (chemical pattern recognition), sir tuong ddng giiia cac sac dd (spectral correlative chromatogram)... ta thu dugc cac ket qua phan tich, cac ket qua cd the ehi don gian la ve sir gidng nhau hoae khae nhau giira cac dau van tay sac ky, cac mdi lien be, cac gia tri du doan ve hoat tinh [6,11].
Hinh 1. Thiit biphdn lich LC-MS Hinh 2. Dir lieu ddu vdn lay sdc ky
Hoi nghi Khoa hqc ky niem 35 ndm Vien Khoa hqc vd Cong nghe Viet Nam - Hd Noil 0/2010
III. MOT SO KET QUA NGHIEN CUtJ B U 6 c DAU
^ Chi Mallotus d Viet Nam cd rat nhieu tiem nang trong Iinh vuc y dugc, vdi su phat tnen cua san pham tir Mallotus (binh 3) thi van de danh gia dau vao cho nguyen lieu Mallotus la mot yeu cau rdt cdn thilt.
Hinh 3. Sdn phdm thuc phdm chuc ndng tir Mallotus apelta
^ De giai quyet van dl tren, phuong phap ddu van tay sdc ky dugc ap dung dk danh gia chat lugn^ eua nguyen lieu Mallotus vdi muc tieu trudc mdt la phat triln phuong phap ddu van tay sac ky ddi vdi ddi tugng Mallotus, muc tieu cao hon la khao sat va xay dung md hinh du doan boat tinh chdng d xy hda. Viec phat triln phuong phap ddu van tay sdc ky tren ddi tugng Mallotus dugc xay dung tudn tu tir viec lira chon thilt bi nghien ciiu, tdi uu cac dilu kien phii hgp eho chiit mau, cac dilu kien cho sdc ky, thilt kl thi nghiem, tiln xir Iy sd lieu (hinh 4) va danh gia dti: lieu bdng cac phuong phap hda thdng ke (hinh 5) [ 12].
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Hinh 4. Kit qua ddnh gid trin PCA dd Hinh 5. Gidn dd PC1-PC2 cua 26 mdu Mallotus, lieu sau khi tiin hdnh tiin xu ly su dung bdn fingerprint sdc ky 60 phut
^ Viec khao sat md hinh du doan boat tinh chdng dxy boa cac ddu van tay sdc ky cua cac mdu Mallotus cung dugc xay dung. Muc tieu la xay dung dugc md hinh du doan boat tinh chdng d xy hda cua cac mau dua tren ddu van tay sdc ky. Md hinh dugc xay dung tren co sd ldy cac sd lieu thuc nghiem thir boat tinh chdng d xy hda lam sd lieu co sd. Tilp dd la viec ap dung cac phuong phap xii ly sd lieu hda thdng ke nhilu bien dl khao sat boat tinh chdng dxy hda cua cac mau thuc thdng qua viec so sanh giu:a cac mau cd gia tri hoat tinh chdng dxy hda thuc nghiem vdi sd lieu ddu van tay sdc ky cua cac mau cdn khao sat. Cac kit qua thu dugc bude ddu cho thay kha nang du doan boat tinh chdng dxy hoa thdng qua cac mdu ddu van tay sdc ky hoan toan cd the thuc hien dugc [13].
Tiiu ban: cdc chdt co hogt tinh sinh hqc JSBN: 978-604-913-012-0
IV. KET LUAN CHUNG
Kit qua ling dung phuong phap dau van tay sac ky ap trong danh gia chat lugng dugc lieu Mallotus 6 Viet Nam dl cap trong bao cao nay chi la giai doan bdt ddu ciia qua trinh danh gia chdt lugng dugc lieu cd truyen Viet Nam trong thdi gian tilp theo. Trong dieu kien quan ly chdt lugng dugc lieu hien nay, dl nghi sir dung phuong phap ddu van tay sdc ky trong danh gia chdt lugng dugc lieu cd truyin Viet Nam la hoan toan can thiet. Trong thuc te, nghien cim danh gia chdt lugng dugc lieu cd truyin Viet Nam la mot ITnh vuc nghien cim lien nganh, can sir tham gia lien nganh cua nganh hda hoc, dugc hoc, y hoc, sinh thai hoc va tham chi ca sd lieu thdng ke dk ciing nhau tap trung giai quyet van de nay sinh trong kiem soat chat lugng dugc lieu Viet Nam.
Loi cam on:
Tac gia xin cam on kinh phi ho trg ciia dl tai hgp tac qudc tl theo Nghi dinh thu Viet Nam-Bi giai doan 2009-2012.
TAI LIEU THAM KHAO
1. R-S. Xie and T.A. van Beek, Joumal of Chromatography A, 1216(11), 2009,1931- 2210.
2. WHO, Guidelines for the Assessment of Herbal Medicine, World Health Organization, Munich, Germany, 1991, p 2, accessed 25 January 2006.
3. http://cpv.org.vn/cpv/Modules/News/NewsDetail.aspx?co_id=30085&cn_id=388636#
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4. Pei-Shan Xie, Albert Y. Leung, Journal of Chromatography A, 1216(11), 2009, 1933- 1940.
5. Xiao-Hui Fan, Yi-Yu Cheng, Zheng-Liang Ye, Rui-Cbao Lin, Zhong-Zhi Qian., Analytica Chimica Acta, 555, 2006, 217-224.
6. Yi-Zeng Liang, Peisban Xie, Kelvin Chan, Joumal of Chromatography B, 812(1-2), 2004, 53-70.
7. Drug Administration Bureau of China, "Requirements for studying fingerprints of traditional Chinese Medicine Injection", Bejing, China (2002).
8. P. Valentao, P.B. Andrade, F. Areias, F. Ferreres, R.M. Seabra, J. Agric.Food Chem. 47, 4579(1999).
9. Shufen Cui, Boqiang Fu, Frank Sen-Chun Lee, Xiaoru Wang, Journal of Chromatography B, 828(1-2), 2005, 33-40.
10. M. Adahchour, J. Beens, R.J.J. Vreuls, U.A.Th. Brinkman, TrAC Trends in Analytical Chemistry, 25(8), 2006, 821-840.
11. Yi Chen, Shang-Bin Zhu, Ming-Yong Xie, Shao-Ping Nie, Wei Liu, Chang Li, Xiao- Feng Gong, Yuan-Xing Wang, Analytica Chimica Acta, 623(2), 2008, 146-156.
12. N. Nguyen Hoai, B. Dejaegher, C. Tistaert, V. Nguyen Tbi Hong, C. Riviere, G.
Chataigne, K. Phan Van, M. Chau Van, J. Quetin-Leclercq, Y Vander Heyden, Journal of Pharmaceutical and Biomedical Analysis, 50(5), 2009, 753-763.
13. C. Tistaert, B. Dejaegher, N. Nguyen Hoai, G. Chataigne, C. Riviere, V. Nguyen Thi Hong, M. Chau Van, J. Quetin-Leclercq, Y Vander Heyden, Analytica Chimica Acta, 649(1), 2009, 24-32.
