B GIÁO D C & B YT O ==oo0oo== BIDENS PILOSA L., - 2014 - Bidens minh -72]; i: Bidens pilosa - - ; alcaloid, saponin, , -phenyl-hepta-1,3,5 triyn (74,46%) + Có là: Natri 3,5-dimethoxy-4-O- -L-rhamnosyl-benzoat (BP19) (R,R)-4- (hydroxymethyl)-5- -methoxy- -hydroxyphenyl]-2,2-dimethyl-1,3-dioxolan Pilosolane) (BP20); + Có Bidens: (1 R, R)-guaiacyl glycerol (BP1); Scopolin (BP2); Benzyl-O- -D-glucopyranosid (BP3); D-threo-guaiacylglycerol-8-O- -D-glucopyranosid (BP4); Maltol 3-O- -D-glucopyranosid (BP5); Ficuscarpanosid B (BP6); 4-allyl-2,6-dimethoxy phenol-1-O- -Dglucopyranosid (BP13); 3-[(4'-hydoxy-3'-methoxyphenyl) propan-1,2 diol-2-O- -D-glucopyranosid (BP15); (3S,6S)-6,7-dihydroxy-6,7-dihydrolinalool (BP17); 3-(4'-hydroxy-3'-methoxyphenyl) propan-1,2-diol (BP18); Daucosterol (BP22); + Có Bidens pilosa L.: Quercetin-3- -L-arabinofuranosid (BP7); Quercetin-3- -D-galactopyranosid (BP8); + Có Bidens pilosa L.: 7-phenyl-2- hepten-4,6-diyn-1-ol (BP11); Stigmasterol (BP23) (Bidens pilosa L.) Bidens pilosa L - 50 - - in vitro (HL-60) (IC50: 18,5 - ó9 85,7 µg/ml); c 50 BP7, BP8 BP23 36,5 µg/ml, 68,1 µg/ml 183,6 µg/ml -60) 50 BP20, BP2 BP23 có tác 50 BP19 BP20 - , có kim - - chi Bidens, loài Bidens pilosa L Bidens ng the gian (10 16 g/ngày) an toàn viêm gan, ng, 29 hình, 138 40 trang) (13 trang) A Bidens nói chung lồi Bidens pilosa L., nói riêng 2.1 Nguyên tháng - Ban W 20g - - (methanol, ethanol, n-butanol, ethylacetat, cloroform, n-hexan ) 2.2 - :T 9] - -MS -0,063mm (240 430 mesh, -50 m, Fujisilisa Chemical Ltd), Sephadex LH20 -Alufolien 60 F254 (Merck - rham W.B., Turner A theo quy - o vòng kháng , kháng - carbon - in vitro - dùng Y - S Microsoft Excel SD) - có 11 carotenoid, acid amin, tanin, phytosterol, - alcaloid, saponin, f - - 3.1.2 X - 127] 3.1.3 3.1.3.1 X tính theo tt/kl 0,18 ± 0.01%, cịn tính theo kl/kl 0,11 ± 3.1.3.2 P 1-phenyl-hepta-1,3,5-triyn (74,46%), -isoindol-1,3(2H)-dion (3,07%) 3.1.4 C hình 3.2: - BP1, BP2, BP3, BP4, BP5, BP6, BP7, BP8, BP11, BP13, BP15 - BP17, BP18, BP19, BP20, BP22, BP23 3.1.5 Xác BP1 - 260 (c= 0,3, MeOH) , BP1 3.6: BP1 # a,b C C (ppm) a, c H DEPT (ppm) HMBC (H (ppm) C) mult (J in Hz) 135.4 134.3 C - 111.7 111.1 CH 6.89 br s 148.5 147.0 C - 147.3 145.3 C - 116.1 114.7 CH 6.69 d (8.0) 120.5 119.1 CH 6.69 br d (8.0) 74.9 72.8 CH 4.38 d (6.0) 77.9 75.9 CH 3.44 m 64.3 3-OCH3 62.6 CH2 4, 3,30 dd (6.0, 11.5) 3,14 dd (4.0, 11.5) 55.8 55.6 3.73 s 4-OH - - 8.69 s 3, 4, -OH - - 4.86 s -OH a b c 125MHz, 500MHz, 4.43 s # 13) [61] C BP1 13 R, R)-guaiacyl BP1 R, - Bidens Pimpinella anisum L [61] Hình 3.3 BP1 BP2 BP2 BP2 a,b * (ppm) DMSO 160.5 113.3 114.2 109.8 146.0 150.0 103.0 148.9 112.3 99.7 73.1 76.7 69.6 77.1 60.7 C (ppm) DMSO 160.4 113.2 114.1 109.3 146.6 149.9 103.0 148.9 112.2 99.7 73.3 76.7 69.6 77.1 60.6 V trí C 10 1' 2' 3' 4' 5' 6' 6-OCH3 56.0 H HMBC (ppm, J, Hz) 6.31 d (9.5) 7.95 d (9.5) 7.29 s 7.14 s 5.06 d (7.5) 3.28 3.29 3.15 3.43 3.43 dd (2.0, 12.0) 3.67 dd (5.5, 12.0) 3.81 s 56.1 5], a *C a,c C 2, 10 2, 5, 4, 6, 7, 6, 10 1', 2', 4' 3', 6' 4', 5' b 125 MHz, c500 MHz BP2 BP2 scopolin Hình 3.5 BP2 59,20 (c= 0,67, MeOH) BP3 H - 7,28 H 3,28 H 4,38 (1H, d, J = 7,5 Hz) H 3,93 (1H, d, J = 12,0 Hz) H 3,72 (1H, d, J = BP3 * V trí C C a,b (ppm) C a,c (ppm) H (ppm, J, Hz) 139.0 139.1 - 128.6 129.2 7.44 m 1, 3, 129.2 129.3 7.36 m 2, 129.1 128.7 7.30 m 3, 5, 2, 129.2 129.3 7.44 m 2, 128.6 129.2 7.36 m 1, 3, 71.6 71.7 4.96 d (12.0) 1, 2, 3, 1' 4.70 d (12.0) 1' 103.2 103.3 4.38 d (7.5) 7, 2', 3', 5' 2' 75.0 75.2 3.28 m 3' 78.0 78.1 3.29 m 4' 71.6 71.8 3.32 m 5' 77.9 78.0 3.36 m 6' 62.8 62.8 3.93 dd (2.0, 12,0) 4', 5' 3.72 dd (5.5, 12,0) a 3OD [43], *C 13 b 125 MHz, c500 MHz C-NMR, DEPT, -O- -D- BP3 3OD, BP3 benzyl-O- - - Cedronella canariensis [43] Hình 3.6 BP3 48,10 (c= 0,5, MeOH) BP4 H (1H, dd , J = 2,0, 8,0 Hz), 6,78 (1H, d, J = 8, J H J 3,89 (3H, s) C- BP4 C C C C 111,7 4,44 (1H, d, H 13 7,03 (1H, d, J = 1,5 Hz), 6,83 71,4 149,0 ppm -threo- guaiacylglycerol-8-O- -D-glucopyranosid L-threo-guaiacylglycerol-8-O- -D-glucopyranosid BP4 BP4 D-threo-guaiacylglycerol-8-O- -D-glucopyranosid Hình 3.8 BP4 BP5 BP5 a,b C1 C2 (ppm) (ppm) (ppm) mult (J = Hz) (ppm) 1' 2' 3' 4' 5' 161.8 143.3 175.9 116.9 155.3 15.4 106.1 75.6 78.4 71.1 79.0 164.6 143.3 177.0 117.2 157.1 15.9 105.0 74.9 77.9 71.2 77.4 164.5 143.6 177.2 117.3 157.1 15.8 105.5 75.4 78.5 71.1 78.0 6' 62.6 68.5 62.5 HMBC (H C) 6.46 d (5.5) 8.02 d (6.0) 2.48 s 4.82 d (8.0) 3.40 dd (8.0, 9,0) 3.28 dd (8.0, 9.0) 3.38 dd (9.0, 9.0) 3.42 m 3.68 dd (5.0, 12.0) 3.84 dd (2.0, 12.0) CD3OD at 500 MHz, 125 MHz [78], a 5D5N [117], C2 C1 a,c H C 3OD, 3, 2, 4, 2, 3 1', 5' 4' 5' 2', 4' b 125, c 500 MHz BP5, 3-O- -DBP5 - -O- -D-apiofuranosyl- -D-glucopyranosid [78], -O- -D-apiofuranosyl- -D- - C BP5 Maltol 3-O- -D-glucopyranosid O CH3 O 2' 3' HO OH 4' OH O O 1' O OH CH3 5' HO 6' OH O O OH OH O Hình 3.9 BP5 +16,10 (c= 0,3, MeOH) BP6 H 3,87 (s H 6,73 (s H 3,18 C 56,9 (x2); nhóm hydoxy-3'-methoxyphenyl) propan-1,2 diol-2-O- -D- (6''-O-galloyl)- 5'' C- BP15 BP15 3-[(4'-hydoxy-3'-methoxyphenyl) propan-1,2 diol-2-O- -DBidens H3CO CH2OH 3' OH 2'' 1' HO 5'' 1'' HO OH O 3'' O 5' Hình 3.17 4'' 6'' OH BP15 -15,10 (c= 0,3, MeOH) BP17 BP17 BP17 a,b # C C (ppm) (ppm) a, c H DEPT (ppm) mult (J in Hz) 5.12 dd (2.0, 17.5) HMBC (H C) 111.1 110.7 CH2 147.4 146.4 CH 72.6 71.7 C 41.1 40.0 CH2 27.0 25.5 CH2 1.57 m/ 1.52 m 79.6 78.1 CH 3.00 d (10.0) 72.7 71.6 C 25.8 24.7 CH3 0.97 s 6, 7, 9 26.1 26.1 CH3 1.02 s 6, 7, 28.7 27.9 CH3 1.13 s 2, 3, 10 a b c 125MHz, 500MHz, # 2,3 4.92 dd (2.0, 11.0) 5.83 dd (11.0, 17.5) 1, 1.72 ddd (4.5, 12.5, 17.5) 1.32 ddd (4.5, 12.5, 17.5) 4, 5, 7, - S, 6S) -6,7-dihydroxy-dihydrolinalool [62] C BP17 10 OH OH Hình 3.19 -d5 (3S, 6S)-6,7-dihydroxy- dihydrolinalool HO 3, BP17 BP1, BP4, h BP18 khung phenylpropanoid 3-(4'-hydroxy-3'- methoxy-phenyl) propan-1,2-diol CH2OH H3CO 2' 3' 1' 6' 4' OH 5' HO Hình 3.20 BP18 BP19 H- BP19 H7,34 ppm (2H, s 5,30 ppm (d, J = 1,5 Hz), H = 3,0, 1,5 Hz), H 3,93 ppm (dd, J = 9,0, 3,0 Hz), H (d, J = 6, 3,45 ppm (t, J = 9,0 Hz), H H 4,15 ppm (dd, J 4,31ppm (m), H 1,24 ppm J= H 13 o BP19 methyl ( C C 17,8), nhóm metoxy ( 107,7 ppm (2CH), 103,5, 72,1, 72,3, 73,8, 71,2 ppm), nhóm C C ( H , DEPT 135o C 175,0, 153,9 (2C), 107,7 C 4-O- -L- C -1 C- BP19 -4-O- -L- -8 ppm nhóm COO - BP19 O H3CO O6 O 6' HC HO3 4' Hình 3.21 3' HO 1' O 5' OCH3 2' OH BP19 * a,b C Aglycon Rham BP19 (ppm) a,b (ppm) C a,c H (ppm)mult (J in Hz) 126.9 106.4 152.8 137.7 152.8 106.4 167.1 134.00 107.7 153.9 137.7 153.9 107.7 175.0 102.1 70.3 103.5 72.1 5.30 d (1.5) 4.15 dd (3.0, 1.5) 70.5 71.6 72.3 73.8 3.39 dd (9.0, 3.0) 3.45 t (9.0) 70.1 17.8 56.1 71.2 17.8 57.0 4.31 m 1.24 d (6.5) 3.88 s 3,5-OCH3 a HMBC (H C) b MeOD; 125 MHz; c 7.34 s 7.34 s - 1, 3, 4, 1, 2, 4, 5, * 3, syringic acid-4-O-a-L-rhamnosid C [132] + -ESI MS Qua phân tích, m/z, 379.0810 m/z, 401.0627 m/z - + - m = [M-Na ] =343 m/z m12 - .H Cl, m1- = [M-Na++HCl]-= 379 m/z (không Na Cl, m1-= [M-H++NaCl]- [137] O H3CO H3CO C O Na H O R O Na - CO2, - H+ (-) Na+ + + C C H+ O O H+ OCH3 R OCH3 [C14H18O7Na]- = m3-= 321.211 Hình 3.23 -ESI- BP19 Natri 3,5- dimethoxy-4-O- -L-rhamnosylBP20 B pilosa, + 18,1 (c = 0,3, MeOH) BP20 Hình 3.24 BP20 BP20 a,c a,b * C H (ppm) C 109.1 85.3 79.8 110.1 85.4 80.5 61.5 61.7 27.43 27.4 27.47 27.4 131.4 130.6 114.5 111.4 145.7 149.2 145.9 147.9 115.9 116.2 119.2 120.7 -OCH3 56.4 a b c MeOD; 125 MHz; 500 MHz; * C 2,2-dimethyl-1,3-dioxolan -d6 [64] HMBC (H C) (ppm) mult (J in Hz) a,b (ppm) 3.84 m 4.75 d (8.5) (6a) 3.76 dd (12.0, 3.0) (6b) 3.61 dd (12.0, 5.0) 1.50 s 1.55 s 6.99 d (1.5) 6.80 d (8.5) 6.87 dd (8.5, 1.5) 3.88 s (R,R)-4-(hydroxymethyl)-5- 4, 2b, 1', 2' 6a, 6b, 5', 8, 7, 1', 3', 6', 1', 3' 1', 2', 4', 3' -dihydroxyphenyl] - BP20 NMR - BP20 -ESI-MS BP20 C13H18O5 1, m2, m3, m4) BP20 R,R)-4-(hydroxymethyl)-5- - hydroxyphenyl]-2,2-dimethyl-1,3Pilosolane BP22 Qua phân tíc D BP22 , BP22 Daucosterol 26 22 18 19 HO HO O 24 27 29 20 17 28 O HO OH Hình 3.28 BP22 - o BP23 BP22, qua phân tích C BP23 [45] tigmasterol BP23 Stigmasterol Hình 3.29 BP23 - 160 g - Ch 50 3.2.2 Th Bacillus pumilus, Escherichia coli Salmonella typhi - 25: 25 1) Lô 2: Diclofenac ( 10 mg/ kg) 10,0 g/kg 20,0 g/kg p phù (%) 44,39 ± 4,16 35,36 ± 20,35 3,35 43,20 ± 2,68 4,92 38,88 ± 12,40 4,67 p2-1,4-1 < 0,05, p3-1>0,05 p2-3,2-4 < 0,05 Sau 2) phù phù so (%) 34,66 ± 4,09 20,78 ± 40,05 4,19 28,07 ± 19,03 5,94 30,92 ± 10,08 3,26 p2-1,3-1, 4-1 < 0,05 p2-3,2-4 < 0,05 3) 4) (%) (%) 25,46 ± 2,15 15,76 ± 38,08 2,14 15,34 ± 39,72 2,67 20,40 ± 19,85 3,21 p2-1,3-1, 4-1 < 0,05 p2-4 < 0,05, p3-2>0,05 8,53 1,58 11,47 2,00 6,45 2,15 8,10 1,95 ± ± ± ± 24,00 5,00 p2-1,3-1, 4-1 > 0,05 carrageenin 6, 3.27, 3.28: Lô p (ml) 2,53 ± 0,15 p1-2,3,4,5,6< 0,05 p2-3,4,5,6>0,05 18,58 p3-4> 0,05 23,72 p5-6>0,05 1,94 ± 0,11 23,32 p3-5>0,05 1,88 ± 0,12 28,46 1,93 ± 0,16 5: 1,81 ± 0,17 2,06 ± 0,13 2: Diclofenac (10 mg/kg) 25,69 p4-6>0,05 Lô protein (mg/dl) 24,33 ± 1,35 p1-2,3,4,5,6 0,05; 41,67 ± 1,41 21,83 ± 1,88 p3-5>0,05 39,20 ± 1,74 p 53,50 ± 3,22 2: Diclofenac (10 mg/kg) albumin (mg/dl) 19,40 ± 1,81 p4-6>0,05 Lô p 9,45 ± 0,97 p2-3,4,5,6>0,05 p3-4> 0,05 8,71 ± 1,51 p5-6>0,05 8,15 ± 1,14 p3-5>0,05 8,49 ± 1,33 3.26, 3.27, 3.28 7,94 ± 1,08 8,33 ± 1,40 2: Diclofenac (10 mg/kg) p1-2,3,4,5,6< 0,05 p4-6>0,05 Lô p 70,87 ± 4,38 58,93 ± 3,34 p1-3< 0,05 55,20 ± 4,74 p1-4 0,05 58,11 ± 3,29 p2-3>0,05 54,09 ± 3,89 2: Diclofenac (10 mg/kg) p1-2< 0,05 p2-4>0,05 tic Lô NC 2: Aspirin 100 mg/kg 20 g/kg p - phút 23,10 ± 1,39 9,22 ± 1,62 14,30 ± 1,56 12,00 ± 1,87 p1-2,3,4< 0,05 p2-3,4 < 0,05 p3-4 >0,05 10 phút 21,50 ± 1,73 15,00 ± 1,15 14,40 ± 1,65 14,33 ± 1,00 p1-2,3,4< 0,05 p2-3,4 < 0,05 p3-4 >0,05 10 15 phút 14,00 ± 1,46 12,40 ± 0,86 12,60 ± 1,18 11,33 ± 0,90 p1-2,3,4< 0,05 p2-3,4 < 0,05 p3-4 >0,05 15 20 phút 11,50 ± 1,29 9,40 ± 0,96 10,20 ± 1,03 8,33 ± 0,87 p1-2,3,4< 0,05 p2-3,4 < 0,05 p3-4 >0,05 sau 10, 15, 20 25 phút (p>0,05) Lô NC p - sau 0) 21,88 ± 2,15 2: Morphin 10mg/kg 1) 22,33 ± 1,30 p>0,05 21,15 ± 1,86 30,81 ± 2,13 p0,05 22,08 ± 1,79 24,36 ± 2,35 p>0,05 p>0,05 20 25 phút 9,44 ± 0,96 6,60 ± 1,13 6,70 ± 0,63 5,78 ± 0,80 p1-2,3,4< 0,05 p2-3,4 < 0,05 p3-4 >0,05 ngày, 5) Lô n (IU/l) (2) (IU/l) (2) 420 ± 83 57 ± 17 1232 ± 283 1065 ± 129 861 ± 134 30,11 554 ± 175 47,98 1045 ± 220 15,18 922 ± 219 13,42 120 mg/kg 788 ± 155 36,06 497 ± 269 53,32 p p2,3,4,5-1 < 0,05; p3,4,5-2 < 0,05 ; p3-4 < 0,05, p3-5>0,05, p4-5 0,05, p3-5>0,05, p4-5>0,05 FLTP , s s t lô dùng silymarin không in vitro - BP20, BP2 BP23 - 50 185,9 µg/ml - C BP7, BP8 BP23 50 36,5 µg/ml, 68,1 µg/ml 183,6 µg/ml -C - 50 BP23 50 (IC50 BP19 BP20 50 25,3 µg/ml Bidens pilosa Bidens Bidens pilosa Bidens pilosa (Bidens pilosa var radiata Sch-Bip.) L 04% (kl/kl) GC- - phenylhepta-1,3,5-triyn (74,46%) - tính theo kl/kl) [20] 0,07% -phenylhepta-1,3,5-triyn Bidens pilosa L., Có Hai (hydroxymethyl)-5Pilosolane Bidens Bidens pilosa L Natri 3,5-dimethoxy-4-O- -L-rhamnosyl-benzoat (BP19) -methoxy- -hydroxyphenyl]-2,2-dimethyl-1,3-dioxolan (BP20) (R,R)-4- (Dictionary of Natural Products CD-ROM (1982-2009)), website: ScienceDirect.com, Spinger.com, Pubmed.com - 127 Ge 18], [23], [26], [27] LD50 - LD50 52 Do 50 LD50 > 5g/kg [52] kim - dùng dùng dài ng L 50 16 g M m, oài B.pilosa 4.3.3 Trên k kim in có protein nói chung albumin nói riêng [11], [88] tính , viêm cho 4.3.4 20g/kg K 20g/kg 4.3.5 4, peroxid ho ALT vào máu, , LTP 133] 4.3.6 in vitro - -60) - hóm flavonoid BP7 BP8 50 50: 36,5 ± 5,5 68,1 ± 8,1 µg/ml BP20 50: 36,5 ± BP19 50 50: in vitro in vitro 4.3.7 , 73 có tác morin) 103 hoá acid arachidonic (phospholipase A2, cyclooxygenase, lipoxyge 73 kaempferol 3-(2,3-di-E-p-coumaroyl- -L-rhamnopyranosid); isorhamnetin 3-[O- -L-rhamnopyranosyl-(1-2)- -D-glucopyranosid] [20] L Quercetin-3- -L-arabinofuranosid, Quercetin-3- -D-galactopyranosid flavonoid có vai trị quan , Ngồi : Luteolin 20 hố nh có tác 73]; 102]; C - Bidens pilosa L.) có: alcaloid, saponin, có: alcaloid, saponin, flavonoid, acid amin, hép -phenyl-hepta-1,3,5 triyn (74,46%) + Có Natri 3,5-dimethoxy-4-O- -L-rhamnosyl-benzoat (BP19) (R,R)-4-(hydroxymethyl)-5- -methoxy- -hydroxyphenyl]-2,2-dimethyl-1,3-dioxolan + Có Pilosolane) (BP20); Bidens: (1 R, R)-guaiacyl glycerol (BP1); Scopolin (BP2); Benzyl-O- -D-glucopyranosid (BP3); D-threo-guaiacylglycerol-8-O- -D-glucopyranosid (BP4); Maltol 3O- -D-glucopyranosid (BP5); Ficuscarpanosid B (BP6); 4-allyl-2,6-dimethoxy phenol-1-O- -D- glucopyranosid (BP13); 3-[(4'-hydoxy-3'-methoxyphenyl) propan-1,2 diol-2-O- -D-glucopyranosid (BP15); (3S,6S)-6,7-dihydroxy-6,7-dihydrolinalool (BP17); 3-(4'-hydroxy-3'-methoxyphenyl) propan-1,2-diol (BP18); Daucosterol (BP22); + Có Bidens pilosa L.: Quercetin-3- -L-arabinofuranosid (BP7); Quercetin-3- -D-galactopyranosid (BP8); + Có Bidens pilosa L.: 7-phenyl-2-hepten- 4,6-diyn-1-ol (BP11); Stigmasterol (BP23) 50 - - nh gây 70mg/kg - 50: 18,5 Tá tràng ) Bidens pilosa L.) -37 sterol Bidens pilosa - trang 216-220 Bidens pilosa - - 289 Bidens pilosa 37, N 8, tháng 10/2012, trang 7-12 Bidens pilosa trang 34-37 Bidens pilosa trang 31-34 -O- -Dpilosa Bidens -44 Bidens pilosa -2013, trang 46-52 Bidens pilosa -2013, trang 55- 58 10 Pilosolane Sodium 3,5-dimethoxy 4-O- -L(Bidens pilosa L.) -2014, trang 59-63