Nghien c&u khoa hgc cdng nglij
NGHIEN CLFU THANH P H A N HOA HOC CClA CAY TAM GU"!
TAXILLUS CHINENSIS (DC) Dans. S 6 N G TREN CAY GAO
NGUYEN X U A N T H O Y ' " , TRLTONG NGOC H C J N G '", NGUYEN TH! N G A « . VO T I E N C H I N H ™, LLTU V A N CHlNH '"
I. M d DAU
Cay ttei giii than go Taxillus chinensis (DC) Dans, thudc ho Tdm giii (Lorandiaceae) la cay ban ky sinh, hat mdc cd the sdng tren nhieu cay than gd khac nhau [1]. Trong y hoc cd tiuyin, cay tSm girl duac su dung rgng rai tiong cac don thudc cua ngudi Trung Qudc de dilu tii nhung bieu hien cua benh ve xuong, khop nhu dau lung, dau diu gdi, ehung viem khdp, benh say thai, cao huyet ap, dau that nguc, dot quy, chung loan nhip tim, beo phi, nhiem khuan va dieu tri ung thu [2, 3, 4, 5]. 6 Viet Nam, cay tdm gui Taxillus chinensis (DC) Dans, dugc tim thay tren nhieu cay chu than go khac nhau nhu cay gao, cay dau, cay mit, cay lim va thanh phan hda hgc ciia cay tam giri phu thugc vao cdy chu, do vay boat tinh sinh hgc cua loai nay tren cac cay chu khac nhau ciing khdng gidng nhau [6], dan den sir da dang vl thanh phan hda hpc va hoat tinh sinh hgc ciia loai nay. Trong sd cac cay tam giri than g5 Taxillus chinensis (DC) Dans., tSm giii sdng tien cay gao dugc sir dung nhieu hon ca, ngudi dan d mgt sd dja phuong nhu Phu Thg, Vinh Phiic sii dung tam giri cay gao de giai nhiet, giai rugu, dieu trj viem cau thdn, tang cudng sire khde cho phu nu" sau khi sinh... Nghien cim ve hda hgc cho thay trong cay tam giri gao cd chira cac flavon nhu quercetin-3-O-a-l-rhamnoside, kaemferon-3-O-a-L-rharanoside, quercetin-3-O-jff-D-glucuronide, 3,3',4',5,7-pentahydroxyflavan va tocopherolquinon [7, 8, 9]. Trong chuong tiinh nghien ciiu ve mdt sd Igai tdm gui than go d Viet Nam, chung tdi tiep tuc nghien cim them ve tdm giti sdng tren cay gao va mdt sd cay khac.
Trong khudn khd bai bao nay, ehimg tdi gidi thieu mdt sterol ldn ddu tien dugc phan lap tir tdm gui sdng tien cdy gao.
II. THlTC NGHIEM
2.1. Nguyen lieu: La va than cua tdm giri cdy gao (Taxillus chinensis (DC) Dans.) dugc thu hai vao tiiang 6 nam 2013 d Tam Ndng, Phii Tho. Mdu cay dugc dinh ten khoa hpc bdi ThS. Vu Tien Chinh - Vien Sinh fliai va Tai nguyen sinh vdt, man thuc vjt duoc gia lai tieu ban tai Trucmg Cao ding Y ti Phu Thg.
2.2. Thiet bi: Phd NMR dupc ghi ten may Brucker AM500 FT-NMR, sit dung TMS la chdt ndi chudn. Sdc ky ldp mdng thuc hien tren ban mdng trang sin silicagel tren dl nhdm 60 F254 (Merck) va phat Men bdng den tir ngoai d budc sdng 254 nm hodc dimg tiiudc thii la dung dich H2SO4 10%. Sdc ky cdt dugc tiln hanh vdi chdt hap phu silicagel (Merck), co hat 0,040 - 0,063 mm va 0,015 - 0,040 mm.
^ 2.3. Chief xuat va phan l|ip cac ch^t: Bgt cay tam gui gao (2,5 kg) dugc chiet bang MeOH cdng nghiep. Dich chilt dugc cd quay dudi dp suat giam tiiu dugc 30 g cao chiet tdng. Cao chilt MeOH dugc phdn tan tiong nudc sau dd chiit ldn lupt vdi n-hexan, etylaxetiit ya «-butanol, tiip theo la loai bd cac dung mdi dudi dp sudt giam de thu cac cao chiet n-hexan (12 g), etylaxetat (4 g) va n-butanol (9 g).
Tap chl Khaa hoc vo Cong nghe nhiet d&i, S6O6, 03 - 201A
Nghien c&u khaa hoc cong nghe
Phan lap cac chat: Cao chiet n-hexan dugc chay cdt sac ky dimg he dung mdi rira giai «-hexan : axeton tii 19 : 1 din 1 : 1 thu dugc 5 phan doan (HI - H5), tiip theo phdn doan H3 (1,1 g) dugc chay sac ky cdt vdi he dung mdi n-hexan : etylaxetat 10 : 1 tim dupc 40 mg chit 1 (TGH3). Phdn doan H4 (900 mg dupc chay tiep sac ky cot - sihcagel vdi he dung mdi riia giai n-hexan : etylaxetat 9 ; 1 sau dd kit tinh lai trong n-hexan thu dupc 75 mg chdt 2 (TGH4). S8c ky cot thudng - silicagel cao chiet etylaxetat (1,8 g) vdi he dung mdi etylaxetat: metanol 1 9 : 1 , sau do kit tinh lai ttong etylacetat thu dupc 70 mg hgp chdt 3 (TGE2). Tir cao chiit n- butanol, sau khi chay sac ky cdt - silicagel vdi he dung mdi rua giai cloroform : metanol = 4 : 1 , tiip theo la 1 : 1 thu dupc 3 phan doan (Bl - B3). Phdn doan B3 (380 mg) tiep tuc dugc chay sac ki cdt - silicagel vdi he dung mdi cloroform : metanol: nudc 1 : 1 : 0,1 di nhan dugc 50 mg chdt 4 (TGB).
Hop chdt 1 (TGH3): Chdt ddu khdng mau. ESI-MS [M+H]*: 297. 'H-NMR (CDCI3, 500 MHz, ppm) 6 5,41 (m, IH, H-2), 4,15 (d,J=7 Hz, 2H, H-1), 2,03 (m, 2H, H-4), 1,67 (s, 3H, H-20), 1,53 (m, IH, H-15), 1,43 (m, 4H, H-5, H-7, H-11), 1,39-1,02 (m, 14H, H-6, H-8, H-9, H-10, H-12, H-13, H-14), 0,86 (m, 12H, H-16, H-17, H-18, H-19). "C-NMR (CDCI3, 500 MHz, ppm) 6 140,3 (C-3), 123,1 (C-2), 59,4 (C-1), 39,9 (C-4), 39,4 (C-14), 37,4 (C-10, C-12), 37,3 (C-8), 36,7 (C-6), 32,8 (C-7), 32,7 (C-11), 28,0 (C-15), 25,1 (C-5), 24,8 (C-13), 24,5 (C-9), 22,7 (C-17), 22,6 (C-16), 19,7 (C-18, C-19), 16,2 (C-20).
Hop chat 2 (TGE2): Chdt rdn, mau vang Mp: 182-183°C. ESI-MS [M-1]":
447. ' H - N M R (CDCI3, 500 MHz, ppm) 6 7,36 (d, J = 2 Hz, IH, H-2'), 7,33 (dd, J, = 2 Hz, 72 = 8 Hz, H-6'), 6,93 (d, J= 8 Hz, H-5'), 6,39 (d, J = 2 Hz, H-8), 6,22 (d, 7 = 2 Hz, H-6), 5,37 (d, J = 1,5 Hz, H-1"), 4,24 (dd, Ji = 1,5 Hz, J2 = 3,5 Hz, H-2"), 3,77 (dd, J, =3,5 Hz, J2 = 9,5 Hz, H-3"), 3,44 (q, J, = 6,5 Hz, 7. = 10 Hz, H-4"), 3,36 (d, J= 9,5 Hz, H-5"), 0,96 (d,J=6 Hz, 3H, H-6"). '^C-NMR (CDCI3, 500 MHz, ppm) 6 179,6 (C-4), 165,9 (C-7), 163,2 (C-5), 159,3 (C-9), 158,5 (C-2), 149,8 (C-4'), 146.4 (C-3'), 136,2 (C-3), 123,0 (C-6'), 122,9 (C-1'), 116,9 (C-5'), 116,4 (C-2'), 105,9 (C-10), 103,5 (C-1"), 99,8 (C-6), 94,7 (C-8), 73,3 (C-4"), 72,1 (C-3"), 72,0 (C- 2"), 71,9 (C-5"), 16,7 (C-6").
Hgp ch^t 3 (TGB): Chdt rdn man vang, Mp: 175-176°C. ESI-MS [M-H]":
477. ' H - N M R (CDCI3, 500 MHz, ppm) 5 8,00 (d, J = 2 Hz, IH, H-2'), 7,50 (dd, J, = 2 Hz, J2 = 8,5 Hz, IH, H-6'), 6,88 (d, 7 = 8,5 Hz, IH, H-5'), 6,41 (d, J = 2 Hz, IH, H-6), 6,22 (d, / = 2 Hz, IH, H-8), 5,36"(d, 7 = 7 Hz, IH, H-1"), 3,57 (m, 4H, H-2", H-3", H-4", H-5"). "C-NMR (CDCI3, 500 MHz, ppm) 5 179,4 (C-4), 176,0 (C-6"), 166,1 (C-7), 162,9 (C-5), 159,1 (C-9), 158,4 (C-2), 149,8 (C-4'), 145,9 (C-3'), 135,8 (C-3), 122,7 (C-6', C-1'), 118,3 (C-5'), 116,4 (C-2'), 105,6 (C-10), 104,3 (C-1"), 100,0 (C-6), 94,8 (C-8), 78,1 (C-3"), 77,4 (C-5"), 75,5 (C-2"), 73,4 (C-4").
Tap chi Khaa hac va Cong nghe nhiet d&i. So 06, 03 - 2014
Nghien c&u khaa hgc cong ng/i|
Hgp chdt 4 (TGH4): Chdt rdn man tidng, Mp: 151-152°C. ESI-MS [M+1]*:
411. 'H-NMR (CDCI3, 500 MHz, ppm) 6 5,35 (m, IH, H-6), 5,24 (dd, Ji = 8 Hz, J2
= 15 Hz, IH, H-23), 5,17 (dd, J, = 7,5 Hz, J2 = 15 Hz, IH, H-22), 4,72 (d, 7 = 1,5 Hz, IH, H-27b), 4,69 (d, J = 1,5 Hz, IH, H-27a), 3,52 (m, IH, H-3), 2,42 (m, IH, H- 24), 2,27 (m, 2H, H-4), 1,95-2,06 (m, 3H, H-7a, H-12b, H-20), 1,84 (m, 2H, H-7b, H-lb), 1,69 (m, 2H, H-16), 1,65 (s, 3H, H-26), 1,32-1,58 (m, 7H, H-1 la, H-I5b, H- 28, H-2, H-8), 1,04-1,31 (m, 5H, H-la, H-15a, H-12a, H-16a, H-17), 1,02 (d, J = 5 Hz, 3H, H-21), 1,01 (m, 2H, H-14, H-1 lb), 1,00 (s, 3H, H-19), 0,94 (m, IH, H-9), 0,85 (t, J = 3,5 Hz, 3H, H-29), 0,69 (s, 3H, H-18). '^C-NMR (CDCI3, 125 MHz, ppm) 5 148,6 (C-25), 140,7 (C-5), 137,2 (C-22), 130,0 (C-23), 121,7 (C-6), 109,5 (C-27), 71,8 (C-3), 56,8 (C-14), 55,8 (C-17), 52,0 (C-24), 50,1 (C-9), 42,3 (C-4), 42,2 (C-13), 41,0 (C-20), 39,7 (C-12), 37,2 (C-1), 36,5 (C-10), 31,9 (C-7, C-8), 31,6 (C-2), 28,7 (C-16), 25,7 (C-28), 24,3 (C-15), 21,1 (C-11), 20,8 (C-19), 20,2 (C-21), 19,4 (C-26), 12,1 (C-29), 12,0 (C-18).
III. KET QUA VA THAO LUAN
Can tnic ciia cac hpp chdt TGH3, TGE2 va TGB dupc xac dinh bang cac phucmg phap phd NMR va MS. So sanh vdi cac ket qua do nhdm Vu Xudn Giang va cac cdng su cdng bd cd the di den ket ludn THG3 la 3,7,11,15-tetra metyl-2- hexadecen-1-ol, TGE2 la quercetin-3-O-a-L-rhamnoside va TGB la quercetin-3-0-
^-D-glucuronide. Tuy nhien, hgp chat TGH4 lai la mdt chdt chua timg dupc phdn lap tir loai Taxillus chinensis (DC) Dans
Hinh 1. Mdt sd hgp chdt phdn lap tit loai Taxillus chinensis (DC) Dans.
Tgp chi Khoa hgc va Cong nghe nhiet d&i. So 06, 03 - 2014
Nghien c&u khoa hgc cong nghe
Pho ' C-NMR cua TGH4 cho thay hgp chat nay co 29 cacbon gdm 5 nhdm metyl, 10 nhom metylen, 10 nhom metin va 4 nhom cacbon bac 4 (pho DEPT).
Trong sd 10 nhdm metin co mat tiong phan tii thi chi cd 1 nhom metin thupc kieu hydroxymetin dugc d|c trung bdi 6c 71,8 ppm. Ngoai ra, 3 nhom metin khac xudt hien d vimg tiudng thdp cd 5c xu4t hien lin lugt d 137,2; 130,0 va 121,7 ppm chiing to day la cac nhom metin thugc 2 noi doi biet lap, cac dii kien ciia ph6 '^C-NMR va su CO mat ciia pic ion phan tu tai m/z 410 tien pho khoi lugng EISM cho phep xac dinh cong thiic phan td ciia chat nay la C29H46O va cho biet cau tnic cua chSt nay co chiia 4 vong, mgt nhom hydroxy. Nhu vay TGH4 la mgt sterol co b6 khung gi6ng nhu cholesterol.
Tren ph6 H-NMR xuat hien tin hieu ciia mgt nhom metyl d dang tiiplet co 5H 0,85 ppm (J= 3,5 Hz) thugc nhom etyl. Trong 3 nhom metyl co dang tin hieu singlet thi mgt nhdm xuat hien d vj tri cd 5H 1,65 ppm chiing td nhom nay dugc gan vdi nguyen tir cacbon mang noi doi, tin hieu cac proton ciia 2 nhdm con lai xuat hien lan lugt tai 5H 0,69 va 1,00 ppm thupc \k 3H-18 va 3H-19. Nhom metyl cu6i cimg c6 dang tin hieu doublet d 5H 1,02 (J= 5 Hz) dugc quy cho 3H-21. Q vimg trudng thip hem CO tin hieu d 6H 3.52 ciia nhom metin gan vdi -OH, dua vao dang phan tach tin hieu cho biet proton nay co cau hinh J3. Cac nh6m metin cua cac n6i doi cQng dugc n h ^ dang g6m H-27a (5H 4,69 ppm) va H-27b (5H 4,70 ppm), H-22 (5H 5,17 ppm, dd, J; = 7,5 Hz, J2 ^ 15 Hz), H-23 (8H 5,24 ppm, dd, J/ = 8 Hz, J^ = 15 Hz) vdi hieu ling mai nha cua dang trans, cuoi cimg la proton metin cua H-6 vdi tin hieu chan rgng tai 6H 5,35 ppm. Su quy ket cac tin hieu ciia hgp chat nay cdn dugc khing dinh bdi cac phd 2 chiiu HSQC va HMBC.
Hinh 2. Pho HMBC ciia hgp chat TGH4
Tgp chi Khoa hgc va Cdng nghi nhiit ddi. So 06, 03 - 2014
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Tir phan tich cac dil lieu phd d ti-en va so sanh vdi cac dil lieu ciia hpp chdt sterol tach ra tir cac loai Clerodendrum (VERBENACEAE) da dugc Akihisa cdng bd trong tai heu [10] cho phep kit ludn TGH4 la 22-dehydroxclerosterol hay (22E)- poriferasta-5,22,25-ttien-3^-ol mdi dupc phat Men trong cay tam gui gao.
IV. KET LUAN
- Tir cdy tdm giri gao (Taxillus chinensis (DC) Dans.) da phan lap dugc 4 chdt gdm phytol (3,7,11,15-Tetra metyl-2-hexadecen-l-ol), (22£)-poriferasta-5,22,25- ttien-3^-ol, quercetin-3-O-a-L-rhamnoside va quercetin-3-O-glucuromde
- Cdu tnic ciia cac chdt dugc xac dinh bdng cac phuong phap phd nhu H- NMR, '^C-NMR HSQC, HMBC va teen co sd so sanh vdi cac tai lieu tham khao.
Trong sd cac chit phdn lap dupc, hgp chit sterol 22-dehydroxclerosterol hay (22£)- poriferasta-5,22,25-trien-3^-ol ldn diu tien phan lap dugc tir loai thuc vat nay.
Lai cdm <m Cong trinh dugc hoan thdnh tit ngudn kinh phi ciia de Idi co sa truang Cao ding Y ti Phii Thg nam 2013 - 2014.
TAI LIEU THAM KHAO
1. Pham Hoang Hd, Cdy co Viet Nam, Nha xudt ban tre, 2000, 2, tt.37.
2. Ding B., Dai Y., Hon Y., Wu X., Chen X., Yao X., Four new hemilerpenoid derivatives from Taxillus chinensis, Fitoterapia,, 2013, Vol 86, p.1-5.
3. Zhang L., Koyyalamudi S., Jeong S., Reddy N., Bailey T., Longvah T., Immunomodulatory activities of polysaccharides isolated from Taxillus chinensis and Uncaria rhyncophylla. Carbohydrate Polymer, 2013, Vol 98, p.1458-1465.
4. Wang Y., Zhang S., Ma X., Tian W., Potent inhibition of fatty acid synthase by parasitic loranthus [Taxillus chinensis (DC.) DanserJ and its constituent avicularin. Journal of Enzym Inhibition and Medicinal Chemistry, 2006, 21(l):87-93.
5. Wang Y., Deng M., Zhang S., Zhou Z., Tian W., Parasitic loranthus from Loranlhaceae rather than Viscaceae potently inhibits fatty acid synthase and reduces body weight in mice. Journal of Ethnopharmacology, 2008, Vol 118, p.473-478.
6. Kefei Hu, Yonghua Li, Yukai Du, Benwei Su and Dong Lu, Analysis of 1- deoxynojirimycin component correlation between medicinal parasitic loranthus from loranlhaceae and their mulberry host trees. Journal of Medicinal Plants Research, 2011, 5(17):4326-4331.
7. Pham Thanh Ky, Vu Xuan Giang, Phan Van Kiem, Phdn lap vd xac dinh cdu true Trans-phytol va a-tocopherolquinon tit cdy lim giri - Taxillus chinensis (DC) Dans sing tren cdy ggo. Tap chi Dugc hgc, 7/2011,423, tr.36-39.
^ ^ Tgp chi Khoa hoc va Cong nghe nhiet d&i, So 06, 03 • 2014
Ngnien ciru khoa hgc cong nghi
8. Vli Xuan Giang, Phan Thanh Ky, Phan Van Kiem, Phdn lap va xac dinh cdu triic catechin vd quercituron tir cdy tdm gui - Taxillus chinensis (DC) Dans sdng tren cdy ggo. Tap chi Dugc hgc, 7/2011, 423, ti.29-35.
9. Vii Xuan Giang, Phan Thanh Ky, Phan Van Kiem, Phdn lap vd xde dinh cdu triic quercitrin vd afzelin tic cdy ldm gui - Taxillus chinensis (DC) Dans, song tren cdy ggo. Tap chi Dugc hpc, 3/2011, 419, ti.37-40
10. Hu K., Li Y., Du Y., Su B. and Lu D., Analysis of 1-deoxynojirimycin component correlation between medicinal parasitic loranthus from loranlhaceae and their mulberry host trees. Journal of Medicinal Plants Research, 2011, 5(17):4326-4331.
SUMMARY
STUDY ON CHEMICAL COMPOSITION OF TAXILLUS CHINENSIS (DC) Dans.
Four compoimds were isolated from whole plant of Taxillus chinensis (DC) Dans, including tocopherolquinon, quercetin-3-O-a-L-rhamnoside, quercetin-3-0-y9- D-glucuronide and (22£)-poriferasta-5,22,25-trien-3^-ol. Their structures were elucidated by spectroscopic methods such as NMR, MS.
Keywords: Taxillus chinensis, isolation, quercetin-3-O-a-L-rhamnoside.
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