Nghin cu thnh phn ha hc cy c mc (Eclipta Prostrata L.

, Asteraceae)
Nguyn Th Thi
Trng i hc Khoa hc T nhin Khoa Ha hc Lun vn Thc s ngnh: Ha hu c; M s: 60 44 27 Ngi hng dn: PGS.TS. Phan Minh Giang Nm bo v: 2011
Abstract. Tng quan v Eclipta prostrata L; thnh phn ha hc ca Eclipta prostrata cng nh hot tnh sinh hc ca Eclipta prostrata v cc hp cht thnh phn. Nghin cu quy trnh chit cc hp cht hu c t phn trn mt t cy C mc (Eclipta prostrata L., Asteraceae). Phn tch sc k lp mng cc phn chit nh tnh cc phn chit v xc nh cc h dung mi sc k iu ch thch hp vi cht hp ph silica gel. Phn tch sc k v phn lp cc hp cht thnh phn chnh c bit l cc hp cht phn cc bng cc phng php sc k iu ch. Xc nh cu trc ha hc ca cc hp cht c phn lp bng phng php ph. a ra kt qu thc nghim: iu ch cc phn chit t cy C mc; phn tch sc k lp mng cc phn chit EA; phn tch sc k lp mng cc phn chit EP; phn tch sc k cc phn chit EA; hng s vt l v d kin ph ca cc hp cht c phn lp t EA; phn tch sc k cc phn chit EP; hng s vt l v d kin ph ca cc hp cht c phn lp t EP Keywords. Ha hu c; Ha hc; Cy c mc; Thnh phn ha hc Content Trong cc nghin cu y dc hin i cc hp cht thin nhin t cc cy thuc bao gm cc flavonoit, tecpenoit v ancoloit tip tc l ngun cung cp cc hp cht c tim nng cho cc th nghim hot tnh sinh hc. Eclipta prostrata L., syn. Eclipta alba L. (Asteraceae) l cy thuc dng ph bin trong Y hc c truyn Vit Nam v mt s nc trn th gii. Cc nghin cu ha hc cc loi Eclipta prostrata ca Trung Quc, Nht Bn v n xc nh c cc nhm hp cht tritecpen (oleanan v taraxastan) glycozit, cc flavonoit, cc coumarin, cc ancaloit-steroit v cc thiophen polyacetylen trong cc b phn ca cy Eclipta prostrata. Da trn cc kin thc dc l hc dn tc v cc hot tnh sinh hc ca cc phn chit nhiu hot cht c tc dng chng ung th, khng vim v chng HIV ca cc cht c phn lp t Eclipta prostrata c pht hin.

S a dng v cc nhm cu trc l th ca Eclipta prostrata, s tng quan ca cc cu trc ny vi nhiu hot tnh sinh hc quan trng v ngha khoa hc v thc tin ca vic t c s khoa hc cho vic s dng cy thuc C mc (Eclipta prostrata L., Asteraceae) ca Vit Nam thc y chng ti tip tc nghin cu mt cch h thng v ha thc vt ca cy C mc. Lun vn ny t mc tiu nghin cu phn lp sc k v xc nh cu trc cc thnh phn ha hc c bit l cc hp cht phn cc t phn trn mt t ca cy C mc ca Vit Nam. Cc kt qu nghin cu theo hng ny s l cc bc u tin trong cc chng trnh hin i ha Y hc c truyn Vit Nam. Cc hp cht phn cc cha c xc nh trong mt s nghin cu trc c phn lp trong nghin cu ny s dng cc phng php sc k hin i. H Cc (danh php khoa hc: Asteraceae hay Compositae), cn gi l h Hng dng, h Cc ty, l mt h thc vt c hoa hai l mm. H Asteraceae phn b rng khp th gii, nhng ph bin nht ti cc khu vc n i v min ni nhit i. Cc chi trong h ny c chia thnh 13 tng. Ch c mt trong s 13 tng ny l Lactuceae, c th l c khc bit c th coi l mt phn h (phn h Cichorioideae), cc tng cn li, phn ln l chng ghp ln nhau, c a vo phn h Asteroideae. Cc loi thuc v h Cc c cc c trng sau: cm hoa dng u, bao phn hu tnh, tc l vi cc nh hoa kt hp li vi nhau ti cc g ca chng bi cc bao phn, to thnh ng, bu nhy vi s phn b c bn ca cc non hoa, cc non hoa trn mt bu nhy, mo lng (chm lng trn qu), qu l loi qu b (to thnh t mt l non v khng n ra khi chn). Eclipta prostrata L. (syn. Eclipta alba L.), h Cc (Asteraceae), c tn thng dng Vit Nam l C mc, C nh ni, Nh ni, hn lin tho, mc hn lin, kim lng tho. C mc thuc loi cy thn tho hng nin, cao t 10-60 cm, mc b hoc c khi gn nh thng ng, c lng trng, cng, tha. Thn mu lc hay mu nu nht hay hi ta. L mc i, phin l di v hp c 2,5 1,2 cm. Mp l nguyn hay c rng ca cn, hai mt u c lng. Hoa trng tp hp thnh u nch l hay u cnh, cc hoa ci hnh li ngoi, cc hoa lng tnh hnh ng gia. Qa b dt c 3 cnh, c cnh di 3 mm. Vng phn b ca cy C mc trn th gii kh rng v n l loi cy nhit i, mc hoang ch m mt. C mc c dng iu tr cc bnh nh: nn ra mu t d dy, chy mu cam, i ra mu, xut huyt t cung, vim gan mn tnh, vim rut, l, tr em suy dinh dng, tai, rng tc do non, suy nhc thn kinh, nm da, ezecma, vt lot, b thng, chy mu, vim da. Ngoi ra C mc cn c dng lm thuc st trng trong bnh ho lao, vim c hng, ban chn, l nga, au mt, sng rng, au d dy, bnh nm ngoi da gy rng tc. Cch dng: dng ti hay gi ly nc ung, hoc sao chy en vi liu 15-30 g sc ung. Dng ring hoc phi hp vi ng sen, l trc b. Trong trng hp st trng cng dng sc ung hoc gi ti ly nc ung, b p. C th dng ti xoa tay cha rt do vi, cha nm ngoi da v nhum tc c mu tm en. Vin chng lao Trung ng v Bnh vin lao K71 pha ch thnh thuc tim cm mu, tim bp tht, mi ngy 1-3 ng (2 ml). C ni sn xut thnh cng dng cao nn thnh vin dng cm mu. -Sitosterol (cht I)

Hp cht I c phn lp di dng tinh th hnh kim khng mu, Rf = 0,38 (TLC, silica gel, n-hexan-axeton 6:1, v/v), hin mu tm vi thuc th vanilin/H2SO4 c 1%. Cht I c nhn dng l -sitosterol trn c s so snh TLC v co-TLC vi cht chun -sitosterol. Metyl gallat (cht II) Hp cht II c phn lp di dng tinh th hnh kim mu trng, Rf = 0,70 (TLC, silica gel, n-hexan-etyl axetat-axit fomic 10:10:1, v/v/v), vt cht hin mu vng vi thuc th vanillin/H2SO4 c 1%, hin mu xanh thm vi thuc th FeCl3 5%. Ph 1H-NMR (CD3OD) ca II cho thy s c mt ca hai nhm tn hiu cng hng t proton, trong tn hiu H 3,83 (3H, s) ca 3 proton ca mt nhm metoxy v tn hiu H 7,06 (2H, s) ca 2 proton vng thm tng ng. Ph 13C-NMR v DEPT (CD3OD) ca II ch ra s c mt ca cc nhm sau: mt nhm cacbonyl este C 169,1 (s), 4 cacbon sp2 (4C) ca mt vng benzen th 4 ln trong c c 3 cacbon lin kt vi oxi [C 146,5 (s), 139,8 (2 s) v 121,5 (s)] v 2 nhm metin sp2 (2CH) ca vng benzen th [C 110,1 (2 d)] v mt nhm metoxy C 52,3 (q). Cc d kin ph NMR cho thy II c cu trc ca metyl gallat ph hp vi cc d kin ph ca cht chun metyl gallat. Eclalbasaponin I (cht III) Hp cht III c phn lp di dng bt v nh hnh mu trng, Rf = 0,66 (TLC, silica gel, CH2Cl2-MeOH 2:1, v/v), vt cht hin mu en vi thuc th vanillin/H2SO4 c 1%. Ph ESI-MS (+) ca III cho pic gi ion phn t m/z 819,2 ([M + Na]+) cho gi thit v mt cng thc phn t C42H68O14 ca cht III. Ph 1H-NMR (CD3OD) ca III cho thy s c mt ca 7 nhm metyl bc ba di dng singlet (7s) H 0,80, 0,86, 0,89, 0,97, 0,98, 1,07 v 1,38; mt nhm oxymetin H 3,0 (1H, dd, J = 14,5 Hz, 4,0 Hz), mt proton olefinic ca mt ni i th ba ln H 5,33 (1H, t, J = 3,0 Hz) v hai proton anomeric ca 2 nhm glucopyranosyl H 4,34 (1H, d, J = 7,5 Hz) v 5,36 (1H, d, J = 8,0 Hz). Ph 13C-NMR v DEPT (CD3OD) ca III bao gm 42 cacbon trong s c 30 cacbon ca mt khung tritecpenoit v 12 cacbon ca 2 nhm glucopyranosyl. Tritecpenoit sapogenin cha 7 nhm metyl (7q) C 33,3, 28,6, 27,3, 25,1, 17,8, 17,0 v 16,1; 9 nhm metylen (9t) C 47,8, 40,2, 36,4, 36,3, 34,2, 31,6, 27,1, 24,5 v 19,3; 5 nhm metin (5d) C 90,9, 74,0, 57,2, 48,2 v 42,2 trong s c 2 nhm oxymetin (C 90,9 v 74,0); 6 C bc bn (6s) 50,1, 42,7, 40,9, 39,9, 37,9 v 31,3; mt ni i th 3 ln C 123,7 (d) v 144,6 (s) v mt nhm cacbonyl este C 177,3 (s). Cc tn hiu cng hng t cacbon 13 ca hai nhm glucopyranosyl xut hin C 106,7 (d), 78,7 (d), 78,2 (d), 75,7 (d), 71,7 (d) v 62,8 (t) v 95,8 (d), 78,3 (d), 77,7 (d), 74,9 (d), 71,2 (d) v 62,5 (t). Cc cu hnh ca 2 nhm glucopyranosyl c xc nh da trn hng s tng tc (J = 7,5 Hz v 8,0 Hz) ca cc proton anomeric. Cc d kin ph NMR ca cht III xc nh mt cu trc glycozit ca mt axit olean-12-en-28-oic. Cc nhm -D-glucopyranosyl c gi thit l lin kt vi C-3 v

C-28 da vo cc gi tr C-3 v C-28 v ha lp th H-3 c xc nh nh hng s tng tc J = 14,5 Hz ca H-3. Cc d kin ph NMR ca cht III hon ton ph hp vi cu trc ca saponin tritecpenoit eclalbasaponin I [12]. Hp cht ny c phn lp ln u tin t cy Eclipta alba ca Nht Bn. Eclalbasaponin II (cht IV) Hp cht IV c phn lp di dng bt v nh hnh mu trng, Rf = 0,65 (TLC, silica gel Merck, h dung mi EtOAc-H2O-HCOOH 85:15:10, v/v/v), vt cht hin mu tm vi thuc th vanillin/H2SO4 c Ph ESI-MS (+) ca IV cho cc pic gi ion phn t m/z 635,3 ([M + H]+) v 657,0 ([M + Na]+) cho gi thit v mt cng thc phn t C36H58O9 ca IV. Ph 1H-NMR (CD3OD) ca IV cho thy s c mt ca 7 nhm metyl bc ba di dng singlet (7s) H 0,80, 0,87, 0,90, 0,98, 0,99, 1,08 v 1,39; 2 nhm oxymetin H 3,03 (1H, d, J = 14,5 Hz, 4,0 Hz) v 4,56 (1H, s br); mt proton olefinic ca mt ni i th ba ln H 5,32 (1H, t, J = 3,5 Hz) v mt proton anomeric ca mt nhm glucopyranosyl H 4,34 (1H, d, J = 7,5 Hz). Ph 13C-NMR v DEPT (CD3OD) ca IV bao gm 36 cacbon trong s c 30 cacbon ca mt khung tritecpenoit v 6 cacbon ca mt nhm glucopyranosyl. Tritecpenoit sapogenin cha 7 nhm metyl (7q) C 33,4, 28,6, 27,3, 24,9, 17,8, 17,0 v 16,1; 9 nhm metylen (9t) C 47,7, 39,9, 36,6, 36,3, 34,3, 32,6, 27,1, 24,5 v 19,3; 5 nhm metin (5d) C 90,8, 75,3, 57,2, 48,2 v 42,1 trong s c 2 nhm oxymetin (C 90,8 v 75,3); 6 C bc bn (6s) C 49,8, 42,7, 40,7, 40,2, 37,9 v 31,4; mt ni i th 3 ln C 123,5 (d) v 145,1 (s) v mt nhm cacbonyl axit cacboxylic C 181,5 (s). Cc tn hiu cng hng t cacbon 13 ca nhm glucopyranosyl xut hin C 106,7 (d), 78,3 (d), 77,7 (d), 75,7 (d), 71,7 (d) v 62,9 (t). Cu hnh ca nhm D-glucopyranosyl c xc nh da trn hng s tng tc (J = 7,5 Hz) ca proton anomeric. S ph hp ca cc d kin ph NMR thuc khung tritecpenoit ca cht IV vi ca eclalbasaponin I (III) v s xut hin ca mt nhm axit cacboxylic (C 181,5) cho thy nhm glucopyranosyl duy nht ca IV phi v tr C-3. Trn c s cc phn tch chuyn dch ha hc v hng s tng tc (J = 14,5 Hz v 4,0 Hz) ha lp th C-3 v C-16 ca hp cht ny ph hp vi ca cht eclalbasaponin I. Cc d kin ph ny hon ton ph hp vi cu trc ca saponin tritecpenoit eclalbasaponin II [12]. Hp cht ny c phn lp ln u tin t cy Eclipta alba ca Nht Bn. Cht ny cng c thng bo l c trong cy C mc trng min Nam [2]. Norwedelolactone (cht V) Hp cht V nhn c di dng bt v nh hnh mu trng, Rf = 0,21 (TLC, silica gel Merck, h dung mi EtOAc-H2O-HCOOH 85:15:10, v/v/v), vt cht hin mu vng vi thuc th vanillin/H2SO4 c 1%. Ph 1H-NMR (CD3OD) ca V cho thy s xut hin cc tn hiu cng hng ca 2 proton vng thm tng tc octo (J = 2,0 Hz) H 6,88 (1H, d, J = 2,0 Hz) v 6,97 (1H, d, J = 2,0 Hz) v 2 proton vng thm v tr para H 7,18 (1H, s) v 7,36 (1H, s). Ph 13C-NMR v DEPT (CD3OD) ca V cho thy c 15 tn hiu cacbon sp2 ca mt cht coumestan vi cc tn hiu ca bn nhm metin (4d) C 99,6, 101,8, 105,2 v 106,1 ph hp vi cc d kin ph 1H-NMR; 12 tn hiu cng hng cn li l ca cc tn hiu

cacbon singlet (12s) C 100,9, 104,6, 115,6, 145,5, 147,0, 151,4, 155,8, 155,9, 156,9 v 160,6 v nhm cacbonyl lacton C 160,7 (s). Cc d kin ph NMR khi c so snh vi cc d kin ph ca mt s cht benzofuran [14] xc nh c cu trc 1,3,8,9-tetrahydroxycoumestan (norwedelolacton) ca cht V. Cc d kin ph NMR ca V hon ton ph hp vi ti liu cng b cho norwedelolacton [15]. Hesperidin (Hespentin 7-O-rutinosid) (cht VI) Hesperidin c phn lp di dng bt v nh hnh mu trng Rf = 0,33 (TLC, silica gel Merck, h dung mi EtOAc-H2O-HCOOH 85:15:10), vt cht hin mu en vi thuc th vanillin/H2SO4 c 1%. Ph 1H-NMR (DMSO-d6) ca cht VI cho cc tn hiu ca 2 proton tng tc octo ca vng thm H 6,13 (1H, d, J = 2,0 Hz) v 6,14 (1H, d, J = 2,0 Hz), mt vng benzen th ba ln vi 3 tn hiu proton cng hng H 6,90-6,96 (3H, m), v mt nhm oxymetin H 5,50 (1H, d, J = 12,0 Hz, 3,0 Hz) lin kt vi mt nhm metylen v tr ca mt nhm cacbonyl H 2,78 (1H, dd, J = 17,0 Hz, 3,0 Hz) v 3,12-3,83 (1H, m). Cc tn hiu cng hng t proton ny xc nh mt cu trc flavanon cho cht VI. Cc tn hiu khc thuc v hai gc ng vi cc proton anomeric H 4,53 (1H, s) v 4,97 (1H, d, J = 7,5 Hz) v mt nhm metoxy H 3,78 (3H, s). Ph 13C-NMR (DMSO-d6) ca cht VI khng nh cu trc flavanon vi cc tn hiu cng hng t cacbon 13 ng ch nht ca vng A th ioxy 5,7 [C 95,6 (d, C-8), 96,5 (d, C-6), 103,4 (d, C-10), 162,6 (s, C-9), 163,1 (s, C-5) v 165,2 (s, C-7)], vng B th ioxi 3,4 [C 112,2 (d, C-5), 114,2 (d, C-2), 117,9 (d, C-6), 131,0 (s, C-1), 146,5 (s, C-3) v 148,0 (s, C-4)], v vng C ca mt flavanon vi nhm CH2 CHO [C 78,4 (d, C-2) v 42,1 (t, C-3)] v nhm cacbonyl C-4 [C 196,9 (s, C-4)]. chuyn dch ha hc (C 196,9) ca nhm cacbonyl cho thy nhm ny lin kt vi nhm 5-OH qua mt lin kt hiro, trong trng hp khng c lin kt hiro C-4 < 180. Cc tn hiu cacbon 13 cn li ph hp vi ca mt nhm glucopyranosyl [C 66,1 (t, C-6), 68,3 (d, C-5), 70,8 (d, C-4), 75,6 (d, C-3), 76,3 (d, C-5), 99,6 (d, C-1)] v mt nhm rhamnopyranosyl [C 17,8 (q, C-6), 70,3 (d, C2), 70,7 (d, C-3), 72,1 (d, C-4) v 100,6 (d, C-1)]. Nhm rhamnopyranosyl lin kt vi nhm glucopyranosyl v tr C-6 gy ra s chuyn dch v pha trng thp ca C-6 (C 66,1). Cc cu hnh cho nhm rhamnopyranosyl v cho glucopyranosyl c xc nh t hng s tng tc ca cc proton anomeric (J tng ng l br s v 7,5 Hz). V tr ca cc nhm rutinozit ( -L-rhmanopyranosyl(16)- -D-glucopyranosyl) v metoxi [C 55,8 (q)] c xc nh l C-7 v C-4 qua s so snh cc d kin ph NMR vi cc hp cht flavanon glycozit. Do , cu trc ca cht c xc nh l hesperetin 7-O-rutinozit (hesperidin) [9]. Cc d kin ph 13C-NMR cn chng minh s tn ti ca ng phn epime 2R ca hesperidin (epime 2S). Cc epime ny trong thc t khng khc bit nhiu v cc d kin ph 1 H-NMR [9], tuy nhin s phn tch k lng cc tn hiu ph cacbon 13 cho thy mt s tn hiu xut hin dng kp (doublet) cho php nhn dng s tn ti ca epime 2R c bit l cc tn hiu cng hng vng B [C 112,1 (d, C-5), 114,1 (d, C-2), 117,8 (d, C-6), 130,96 (s, C-1), 146,48 (C-3) v 147,98 (s, C-4)], cc tn hiu vng A [C 162,5 (s, C-9), 163,0 (s, C-5) v 165,1 (s, C-7)] v mt s tn hiu ca cc nhm ng [C 66,07 (t, C-6), 69,7 (d,

C-2), 73,0 (d, C-4), 99,5 (d, C-1)]. S xut hin ca cc cht chuyn ha th cp trong thin nhin thng c cho l theo cc con ng c th v lp th v s tham gia ca cc enzym trong cc bc ca con ng sinh tng hp. Tuy nhin cc ng phn lp th vn c th cng xut hin nh v d v cc ng phn epimeric 2R v 2S ca cc flavanon do l sn phm ph ca cc qua trnh gy bi cc enzym nht nh v cn c cc phng php nhn bit cc cht ny.Trong trng hp ca hesperidin (dng 2S) v epime 2R F. Maltese s dng ph 1H-NMR nhn bit gia 2 ng phn epime ny [9]. Tuy nhin, cc gi tr H ch phn bit nhau n v 0,01 ppm v khng phn gii tt trong trng hp cc cht ca Lun vn ny. Ph 13C-NMR c xc nh trong trng hp ca hesperidin (epime 2S) l phng php nhanh nhn bit cht ny v epime 2R ca n. Lun vn tt nghip Nghin cu thnh phn cy C mc (Eclipta prostrata L., Asteraceae) thc hin cc nhim v nghin cu phn lp v xc nh cu trc cc hp cht t cy C mc. Lun vn t c cc kt qu nghin cu sau : 1. xy dng c qui trnh chit v phn tch iu ch c cc phn chit n-hexan, iclometan, etyl axetat v phn chit nc cha cc hp cht phn cc t phn trn mt t cy C mc. 2. phn tch sc k lp mng (TLC) cc phn chit n-hexan, iclometan v etyl axetat xc nh iu kin sc k nh tnh v xc nh cc h dung mi thch hp cho phn tch sc k gradient cc phn chit ny. 3. Bng cc phng php sc k ct gradient phn lp c 4 hp cht sitosterol, metyl gallat, eclalbasaponin II v hesperidin t cc phn chit n-hexan, iclometan v etyl axetat v 3 cht eclalbasaponin I, norwedelolacton v hesperidin t phn chit nc. 4. Cu trc ca cc hp cht c phn lp c xc nh bng cch kt hp cc phng php ph hin i: ESI-MS, 1H-NMR, 13C-NMR v DEPT. 5. Nghin cu ca Lun vn dn n cc pht hin l th. -Sitosterol c xc nh l thnh phn chnh trong cc phn chit n-hexan v iclometan. Cc saponin tritecpenoit eclalbasaponin I v II cng vi cht coumarin norwedelolacton v flavanon rutinozit hesperidin c ch yu trong cc phn chit phn cc (etyl axetat v nc). Eclalbasaponin I v eclalbasaponin II l cc hot cht saponin tecpenoit quan trng ca cy C mc ; chng ln u tin c tm thy trong cy C mc trng min Bc Vit Nam. Metyl gallat v hesperidin c phn lp ln u tin t cy Eclipta prostrata L.. S xut hin ng thi ca hn hp epimeric 2R v 2S (hesperidin) ca cht 3,5,7trihydroxy-4-metoxyflavanon 7-O-rutinozit c pht hin v nghin cu chng t l c th xc nh nhanh hn hp ny bng ph 13C-NMR. References Ting Vit 1. V Vn Chi (1997), T in cy thuc Vit Nam, NXB Y hc, Thnh ph H Ch Minh.

Trn V Thin, Phng Vn Trung, Nguyn Ngc Hnh (2009) Phn lp echino cystic axit v eclabasapnin II t cy C mc Eclipta prostrata. H cc (Asteraceae), Tp ch Khoa hc, 11, 278-283. Ting Anh 3. Abdel-Kader M. S., Bahler B. D., Malone S., Werkhoven M. C. M., Troon F., David, Wisse J. H., Bursuker I., Neddermann K., Mamber S. W., Kington D. G. I. (1998), DNA-damaging steroidal alkaloids from Eclipta alba from the Suriname rainforest, J. Nat. Prod., 61, 1202-1208. 4. Govindachari T. R., Premila M. S. (1985), The benzofuran norwedelic acid from Wedelia calendulaceae, Phytochemistry, 24, 3068-3069. 5. Krishnaswamy N. R., Seehadri T. R., Sharma B. R. (1966), The structure of a new polythienyl from Eclipta alba, Tetrahedron Letters, 35, 4227-4230. 6. Lee M. K., Ha N. R., Yang H., Sung S. H., Kim G. H., Kim Y. C. (2008), Antiproliferative activity of triterpenoids from Eclipta prostrata on hepatic stellate cells, Phytomedicine, 15, 775-780. 7. Lee M. K., Ha N. R., Yang H., Sung S. H., Kim Y. C. (2009), Stimulatory constituent of Eclipta prostrata on mouse osteoblast differentiation, Phytother. Res., 23, 129-131. 8. Li C. C., Xie Z. X., Zhang Y. D., Chen J. H., Yang Z. (2003), Total synthesis of wedelolactone, J. Org. Chem., 68, 8500-8504. 9. Maltese F., Erkelans C., Vander Kooy F., Choi Y. H., Verpoorte R. (2009), Identification of natural epimeric flavanone glycosides by NMR spectroscopy, Food Chemistry, 116, 575-579. 10. Michels M. G., Bertolini L. C. T., Esteves A. F., Moreira P., S. Franca C. (2010), Anticoccidial effects of coumestans from Eclipta alba for sustainable control of Eimeria tenella parasitosis in poultry production, Veterinary Parasitology, 18732550. 11. Sing P., Sharma A. K., Joshi K. C., Bohlmann F. (1985), A futher dithienylacetylene from Eclipta erecta, Phytochemistry, 24, 615-616. 12. Sing P., Bhargava S. (1992), A dithienylacetylene ester from Eclipta erecta, Phytochemistry, 31, 2883-2884. 13. Tewtrakul S., Subhadhirasakul S., Cheenpracha S., Karalai C. (2007), HIV-1 protease and HIV-1 integrase inhibitory substances from Eclipta prostrata, Phytother. Res., 21, 1092-1095. 14. Tewtrakul S., Subhadhirasakul S., Tansakul P., Cheenpracha S., Karalai C. (2011), Antiinflammatory constituents from Eclipta prostrata using RAW264.7 macrophage cells, Phytother. Res.,25, 1313-1316. 15. Yahara S., Ding N., Nohara T. (1994), Oleanane glycosides from Eclipta alba, Chem. Pharm. Bull., 42, 1336-1338. 16. Yahara S., Ding N., Nohara T., Masuda K., Ageta H. (1997), Taraxastane glycosides from Eclipta alba, Phytochemistry, 44, 131-135. 2.