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LI CM N
Em xin trn trng cm n qu Thy C b mn Dc Liu Khoa Dc Trng i Hc Y Dc Cn Th tn tnh hng dn gip em hon thnh ni dung quyn bo co ht mn ny. Em xin trn trng cm n n Thy PGS.TS Trn Hng v TS. Nguyn Vit Knh b mn Dc Liu Khoa Dc Trng i Hc Y Dc TP. H Ch Minh truyn t nhiu kin thc qu bu gip em hiu v lm tt quyn bo co ny. Em cng chn thnh cc bn trong lp Dc K32 phi hp vi em thc hin tt cc ni dung thc nghim ca dc liu Ch rng ca.

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T VN Theo T chc Y t Th gii, 80% dn s ton cu s dng cc loi tho dc truyn thng bo v sc khe. Vin Thc vt hc Trung Quc khng nh, cng vi Trung Quc v Lo, Vit Nam l mt trong nhng nc c ti nguyn cy thuc phong ph nht vi s lng trn 3.800 loi cy lm thuc trn tng s hn 10.600 lai thc vt. Th trng dc liu v thuc c ngun gc t dc liu ang pht trin mnh m. Chnh v th, Vit Nam cn tn dng tim nng v ngun dc liu to ln ny pht trin cng nghip chit xut dc liu v y mnh nghin cu cc ch phm thuc mi t cy thuc. Tuy nhin, theo thng k ca Cc Qun L Dc Vit Nam, thuc t dc liu ch chim khong 30% tng s thuc ng k trong nc, hn 90% nguyn liu sn xut thuc trong nc vn phi nhp khu. V vy, pht huy tim nng ca ngun dc liu p ng yu cu chm sc sc khe nhn dn i hi cc c quan chc nng cn c gii php hiu qu. Mt trong s l vn xy dng cc tiu chun cho dc liu. y cng l mt yu cu cp bch i vi cc nh nghin cu dc liu. c th s dng dc liu lm nguyn liu thuc, thuc th i hi ngi ta phi xy dng cc tiu chun cht lng, ng thi xy dng cc phng php th nh gi cc tiu chun . Ch rng ca, mt dc liu qu c tc dng cha tr rt nhiu bnh cng cn phi c mt tiu chun cht lng c th trc khi c dng lm thuc. V th, vn Xy dng tiu chun cho dc liu Ch rng ca l mt vn cp thit. Quyn bo co ny s cung cp nhng vn c lin quan n dc liu Ch rng ca nh tng quan v thc vt hc, thnh phn ha hc, tc dng dc l...v ngh mt tiu chun kim nghim.

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NI DUNG
1. TNG QUAN TI LIU: 1.1. Thc vt hc: 1.1.1. Tn gi: [1],[2],[3] Cy ch rng ca

Tn khoa hc: Phyllanthus urinaria L., h thu du (Euphorbiaceae).

Tn khc: vng cao nguyn Ty Bc gi l trn chu tho (c ht chu),

vng h ng bng sng Cu Long (C Mau, Kin Giang) gi l me t ng. Ngoi ra, ch rng ca cn c cc tn gi khc nh dip ho thi, lo nha chu, cam kim, rt t, khao ham (Ty), prakphle (Campuchia)
Theo y s Tu Tnh trong ng dc thn tho tng th c tn l ch

v cc vng nng thn, ch ci sau khi con xong thng ra vn tm cy dip h chu nhai nut lin tc 2, 3 ngy, va cm mu, va bo v sinh mng, mt bn nng t v do to ha ban cho loi vt. V vy, cy ch phi l tho dc hu ch. Cn dn min ng Nam b v cc nh ch bin Nam dc gi l dip h chu v di k l c tri trn, bng lng xu thnh chui cho tr chi bn hng. 1.1.2. Phn loi: Gii Plantae Plants Phn gii Tracheobionta Thc vt c mch Nhm Spermatophyta Thc vt c ht Ngnh Magnoliophyta Thc vt ht kn Lp Magnoliopsida Hai l mm B Malpighiales H Euphorbiaceae Tng Phyllantheae Phn tng Flueggeinae Chi Phyllanthus Loi Phyllanthus urinaria 1.1.3. M t: [1],[2],[3] Cy tho sng hng nm hay sng dai, cao 20-30cm, c th cao n 60-70cm. Thn nhn thng c mu hng . L mc so le, hnh bu dc, xp xt nhau thnh 2 dy nh mt l kp hnh lng chim, mt trn xanh lc nht, mt di xanh xm nht, di 1-1,5cm, rng 3-4mm; cung l rt ngn.

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Hoa c mc k l, c cung ngn, n tnh cng gc; hoa c u cnh

c 3 l i, 3 nh, ch nh ngn;hoa ci cui cnh, 6 l i, hnh bu trng. Qu nang hnh cu, hi dt, mc r xung di l, c kha m v c gai; ht hnh 3 cnh.

Ma hoa: thng 4-6; ma qu khong thng 7-9.

1.1.4. Phn b, sinh thi: [1],[3] Phn b ch yu vng nhit i v cn nhit i. Vit Nam mc ri rc khp ni, tr vng ni cao lnh. Trn th gii cng phn b mt s nc nhit i chu khc nh n , Malaysia, Thi Lan, Campuchia, Lo v Trung Quc.
Ch l cy a m v a sng hoc c th hi chu bng, thng mc ln

trong cc bi c, rung cao, nng ry, vn nh v i khi vng i. Cy con mc t ht vo cui xun, sinh trng nhanh trong ma h v tn li vo gia ma thu. Do kh nng ra hoa kt qu nhiu, ht ging pht tn gn nn cy thng mc thnh m dy c, i khi ln t c c di v cy trng khc.

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phng Ty, ch rng ca v mt s loi thc vt cng h

Euphorbiaceae c gi tn l Chanca Piedra. Tn Chanca Piedra c xut x t th ng ca 1 b lc ngi da Peru vi ngha Break Stone, tm dch l cy tn si. (T Chanca c ngha to break lm b hoc b gy; t Piedra c ngha stone sn, si). Ch rng ca l 1 cy thuc ph bin c s dng t lu i Nam M iu tr nhiu loi bnh khc nhau nh ph thng, bnh gout, st rt, thng hn, cm cm, kit l, ung nht, l lot, tiu ng. 1.1.5. B phn dng, thu hi ch bin: [1],[3]
B phn dng: ton cy trn mt t.

Thu hi, s ch: - Thu hi quanh nm, tt nht vo ma h. Thng dng ti. C th dng cy phi hoc sy kh. - Khi 3/4 s cy ra hoa th thu hoch la 1. Ct cy cha gc 20cm ( cc cnh ng mau pht trin). Ct xong, ti bn cho cy 1 ln thu hoch la 2. - Ra sch, thi ngn, b li phi trong rm, hong gi hoc phi trc tip ngoi nng. Khi b thn thy cy kh rn l c. Thu ht, l rng cho vo ti kh, sch kn bo qun. L v cnh kh lm thuc. - Thng ngi ta thu hi cy ch rng ca vo ma h - bi thi im ny cc hot cht cha bnh ca cy t ngng cao nht. 1.1.6. Phn bit vi mt s loi cng h Euphorbiaceae: - Trn thc t nc ta c nhiu loi ch nhng thng gp nht l 3 loi ch rng ca, ch qu trn, ch thn xanh nn cn phn bit ch rng ca vi ch qu trn v ch thn xanh.
Cy Ch qu trn (Phyllanthus niruri Linn): cy tho mc hng nm,

nhn. Thn mu hng nht, cc cnh c gc. L thun, t c gc ln u. L km hnh di trong sut. Cm hoa nch gm 1 hoa c v 1 hoa ci hoc ch c hoa ci. Hoa c c cung rt ngn, i 5-6 hnh bu dc mi mc, a mt gm nhng tuyn rt b, nh 3. Hoa ci cng c cung ngn, i 5-6 ging hoa c nhng rng hn mt t, a mt hnh u c 5 thy su, cc

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vi nhy rt ngn, ri nhau ch i u, bu hnh cu. Qu nang hnh cu. Ra hoa t thng 1-10. Cy mc di trong vn, gp khp ni trong nc ta. Kinh nghim nhn dn lm thuc thng tiu, thng sa.
Ch thn xanh (Phyllanthus amarus Schum. Et Thonn.): cy cao 40cm

n 80cm, thn trn, bng, mu xanh, phn nhnh u, nhiu. L mc so le xp thnh 2 dy xt nhau trng nh l kp hnh lng chim. Phin l hnh bu dc, di t 5mm n 10mm, rng 3mm n 6mm, mu xanh sm mt trn, mu xanh nht mt di. Hoa c v hoa ci mc thnh cm. Hoa c c cun ngn 1mm n 2mm, i 5, c tuyn mt, nh 3, ch nh dnh nhau. Hoa ci c cun di hn hoa c. Qu nang, nhn, hnh cu, ng knh 1,8mm n 2mm, c i tn ti. Cha 6 ht hnh tam gic, ng knh 1mm, ht c

Phyllanthus niruri L. sc dc lng. 1.2. Thnh phn ha hc: [ 1],[3],[4],[6],[9],[11]

Phyllanthus amarus S.

1.2.1. Cc nhm hp cht ho hc c trong Phyllanthus urinaria L.: Lignan: phylanthin, phytetralin, hypophylanthin, urinatetralin, dextrobuscherlin, 5-demethyoxynirathin, urinaligran Acid: acid hexacosanoic Alkanol: triacontanol. Benzenoid: acid gallic
Coumarin:

trimethylester

dehydrochebulic

acid,

methylbrevifolin

carboxylate, ellagic acid

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Ester: montanoic acid ethyl ester Flavonoid: quercetin, quercitrin, isoquercitrin, rutin, kaempferol
Phytallate: phyllester. Sterol: daucosterol, -sitosterol. Tannin: geraniin Triterpene: lupeol acetate, amyrin

1.2.2. Mt s hot cht c hot tnh sinh hc cao trong cy Phyllanthus urinaria L. 1.2.2.1. Phyllanthin (C24H34O6) - Khi lng phn t: M = 418,523 vC. - C dng tinh th hnh kim ngn, khng mu. - Nhit nng chy: 960C. - max (EtOH) = 230; 280nm. - []30D = + 15.50C. 1.2.2.2. Hypophyllanthin (C24H30O7) - Khi lng phn t: M = 430,491 vC. - Kt tinh trong ete du ha dng tinh th hnh kim di, khng mu. - Nhit nng chy: 1280C. - []30D = + 3.80C.

1.2.2.3. Quercetin - C dng tinh th hnh kim mu vng (EtOH) long. - Nhit nng chy: 3140C. - max (EtOH) = 258nm.

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1.2.3. Mt s cng thc ca cc cht khc

Gallocatechin 1.2.4. Mt s nghin cu ngoi nc: [9] Hp cht t nhin Alkaloids Flavonoids

Catechin

Tn hp cht Securinine, Norsecurinine, Epibubbialine, Isobubbialine (Houghton et al., 1996) Catechin, Gallocatechin, Quercetin, Quercitoside v Rutin (Morton, 1981; Foo, 1993) Amariin, Amariinic, Amarulone, Corilagin, Glucopyranoside (Foo and Wong, 1992; Foo, 1993, Foo, 1995) Phyllanthin v Hypophyllanthin (Morton, 1981; Chevallier, 2000) Acid Gallic (Foo, 1993) Acid Ellagic, Phenazine v dn xut Phenazine (Foo, 1993)

Hydrolysabletanins

Lignan chnh Phenolic Polyphenols

1.3. TC DNG CNG DNG: [1],[3],[4],[5],[10],[12],[13],[14] 1.3.1. Tc dng dc l

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iu tr vim gan: Ti Vit Nam, kh nhiu cng trnh nghin cu v tc dng iu tr vim gan ca Dip h chu c tin hnh, chng hn: nhm nghin cu ca L V nh Tng (Hc Vin Qun Y - 1990 - 1996) thnh cng vi ch phm Hepamarin t Phyllanthus amarus; nhm nghin cu ca Trn Danh Vit, Nguyn Thng Dong (Vin Dc Liu) vi bt Phyllanthin (2001). Tc dng trn h thng min dch: - Vo nm 1992, cc nh khoa hc Nht Bn cng khm ph tc dng c ch s pht trin HIV-1 ca cao lng Phyllanthus niruri thng qua s km hm qu trnh nhn ln ca virus HIV. - Nm 1996, Vin nghin cu Dc hc Bristol Myezs Squibb cng chit xut t Dip h chu c mt hot cht c tc dng ny v t tn l Nuruside. Tc dng gii c: - Ngi Vit Nam, n , Trung Quc dng Dip h chu tr cc chng mn nht, l lot, inh ru, rn cn, giun. Nhn dn Java, n dng cha bnh lu. Theo kinh nghim dn gian Malaysia, Dip h chu c th dng tr cc chng vim da, vim ng tit niu, giang mai, vim m o,... - Cng trnh nghin cu ti Vin Dc liu - Vit Nam (1987 - 2000) cho thy khi dng liu 10 - 50g/kg, Dip h chu c tc dng chng vim cp trn chut th nghim. iu tr cc bnh ng tiu ha: Cy thuc c kh nng kch thch n ngon, kch thch trung tin. Ngi n dng cha cc bnh vim gan, vng da, kit l, to bn, thng hn, vim i trng. Nhn dn vng Haiti, Java dng cy thuc ny tr chng au d dy, ri lon tiu ha,.. Bnh ng h hp: Ngi n s dng Dip h chu tr ho, vim ph qun, hen ph qun, lao,. .. Tc dng gim au:

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Kenneth Jones v cc nh nghin cu Brazil khm ph tc dng gim au mnh v bn vng ca mt vi loi Phyllanthus, trong c cy Dip h chu. Tc dng gim au ca Dip h chu mnh hn indomethacin gp 4 ln v mnh hn 3 ln so vi morphin. Tc dng ny c chng minh l do s hin din ca acid gallic, ester ethyl v hn hp steroid (beta sitosterol v stigmasterol) c trong Dip h chu. Tc dng li tiu: Y hc c truyn mt s nc s dng Dip h chu lm thuc li tiu, tr ph thng. Mt nghin cu ca trng i hc Dc Santa Catarina (Brazil-1984) pht hin mt alkaloid ca Dip h chu (phyllan thoside) c tc dng chng co tht c vn v c trn, cc nh khoa hc nh vo iu ny gii thch hiu qu iu tr si thn, si mt ca cy thuc. iu tr tiu ng: Tc dng gim ng huyt ca Dip h chu (Phyllanthus niruri) c kt lun vo nm 1995, ng huyt gim mt cch ng k trn nhng bnh nhn tiu ng khi cho ung thuc ny trong 10 ngy. 1.3.2. Mt s cng trnh nghin cu v tc dng dc l ca cy Dip h chu:
Cc nh khoa hc chng minh dch chit ca Phyllantus c tc dng c

ch mnh HBV, thng qua c ch c ch enzym ADNp (DNA polymerase) ca HBV, lm gim hot HbsAg v Anti- HBs. Theo cc nghin cu, cy dip h chu cha mt s enzyme v hot cht c tc dng cha vim gan nh phyllanthine, hypophyllanthine, alkaloids v flavonoids... Ngi ta cho rng ch rng ca c tc dng c ch mnh HBV- DNA (virut vim gan B trn h m di truyn) v lm cho virut b o thi, khng bm vo c ADN ca ngi. Nhng bnh nhn vim gan do HBV sau khi s dng thuc c ch rng ca, c phc hi enzym transaminase t 50-97%, bilirubin ton phn tr v bnh thng. Mt nghin cu cho thy, 50% yu t ly truyn ca virus vim gan B trong mu mt i sau 30 ngy s dng loi cy ny (vi liu 900 mg/ngy).

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Trong thi gian nghin cu, khng c bt k s tng tc no gia Dip h chu vi cc thuc khc. Theo mt nghin cu tin hnh nm 1995, cy thuc ny c tc dng li tiu, gim huyt p tm thu ngi khng b tiu ng v gim ng k ng huyt bnh nhn tiu ng. Th nghim trn chut cng trng cho thy: Phyllanthin v hypophyllanthin chng li tnh c hi t bo gy ra bi carbontetraclorid v galactosami; cht triterpen triacontanol c tc dng bo v gan chng li tnh c hi t bo gy ra bi galactosamin. Trn in vitro: cy ch c tc dng khng virus vim gan B. Mt lot nhng acid phenolic phn lp t cc lo Phyllanthus c tc dng c ch DNA polymerase ca virus vim gan B. Cy ch c tc dng khng khun i vi t cu vng, trc khun m xanh, trc khun coli, Shigella dysenteriae, S.flexneri, S.shigae, Moraxella, v khng nm i vi Aspergillus fumigatus. Acid galic cha trong cy c tc dng khng khun yu. Mt dn cht phenolic v mt flavonoid phn lp t cy ch c tc dng khng khun mnh hn v khng nm r rt. Cao chit vi cn-nc t cy ch c tc dng gim au chut nht trng v cao cn methylic c tc dng h ng huyt trn chut cng trng i tho ng. Nm 1998, trn th gii c nc cng b nghin cu thnh cng iu tr vim gan do virus B bng Dip h chu ng. Trn th trng Vit Nam cng c nhiu ch phm tr vim gan do HBV, trong thnh phn c ch rng ca. Ngoi ra, cn dng cha l lot, mn nht khng lin ming: L ch rng ca, l thm lm n tai, lng bng nhau, inh hng 1 n, gi nt, p vo ch au.
Bnh vin Qun khu IV th nghim lm sng iu tr vim gan B mn

tnh vi hepaphyl c cha bt Dip h chu ng ca x nghip dc phm Trung ng 25 trn 54 bnh nhn. Sau 4 thng theo di, kt qu cho thy

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cc bnh nhn gim hoc mt cc triu chng lm sng ca vim gan B, phc hi nhanh chc nng gan; Trung tm nghin cu trng v ch bin cy thuc thuc Vin Dc liu (B Y t) cng nghin cu thnh cng v a vo sn xut i tr "Tr dip h chu. Loi tr ny c tc dng gii nhit, tr gip tiu ha, gii c do ru v bia. 1.3.3. CNG DNG [7],[8] T 2.000 nm nay, y hc c truyn ca nhiu dn tc s dng dip h chu cha vng da, lu, tiu ng, u x tuyn tin lit, hen, st, khi u, au n ko di, to bn, vim ph qun, ho, vim m o, kh tiu, vim i trng... N cn c p ti ch cha cc bnh ngoi da nh l lot, sng n, nga,... Cha au hng, inh ru, mn nht, vim da, l nga, sn h huyt au bng, tr em ta li, chm m. Ngoi ra cn dng cha bnh gan, st, rn rt cn v dng cy ti gi p, hoc dch p cy ti bi ngoi, liu lng khng hn ch. King k: ph n c thai khng nn dng. Trong y hc dn gian n : cy ch c tc dng li tiu, nc p l cho vo sa da dng cho tr em lm n ngon ming, h st, st trng, tr ri lon tiu ho, ph, bnh lu v bnh ng niu-sinh dc. nc sc l lm mt da u. M cy bi cha mn nht v lot kh lnh. Nhng chi non tr l, vng da. gy sy thai. Malaysia: lm sch li tr em v kch thch n ngon ming. Papua Niu Ghin dng h st. Brunei thuc p t l cng vi sa da tr bnh u ma. Campuchia tr st rt. Thi Bnh Dng dng iu kinh v gy sy thai. Guaham tr l, qun o Solomon th dng l cha au ngc.
Ngi Peru tin rng dip h chu c tc dng kch thch bi tit nc mt,

tng cng chc nng gan v dng n iu tr si mt, si thn. H x vn cy thuc, un si (nh sc thuc), cho thm cht nc chanh, chia ung 4 ln trong ngy. N cng c dng cha vim bng quang, vng da ph, ri lon tiu ha, au bng kinh. Ngi Brazil, Haiti cng dng cy thuc ny

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cha cc bnh tng t. Ti cc vng khc Nam M, Dip h chu c s dng rng ri tr vim gan B, vim ti mt, mt s bnh l thn, thng phong, st rt, thng hn, cm, cm lnh, kit l, au d dy, mn nht, l lot, ung c. N cn c s dng nh mt thuc gim au, kch thch ngon ming, kch thch trung tin, ty giun, li tiu, iu ha kinh nguyt ph n... Ti nhiu nc chu , ngi dn cng dng Dip h chu cha vim gan, vng da, hen ph qun, lao, kit l, lu, vim ph qun, vim da, vim ng tit niu, giang mai. Bi thuc c dc liu dip h chu: Cha suy gan do nghin ru, mt - Dip h chu : 10g, Cam tho t : 20g - Cch dng : Sc ung thay nc hng ngy. Cha vim gan do virus B - Dip h chu ng: 100g Ngh vng : 5g. - Cch dng : Sc nc 3 ln. Ln u 3 chn, sc cn 1 chn. Ln 2 v 3 vo 2 chn nc vi 50g ng, sc cn na chn. Chia lm 4 ln, ung trong ngy. Cha suy gan do ru, st rt, nhim c do mi trng hoc cc trng hp hay ni mn, ni mn do huyt nhit. - Dip h chu ng 12g, Cam tho t 12g. - Sc nc ung hng ngy thay tr. Cha sn mt, sn thn. - Dip h chu ng 24g. Sc ung, sc lm 2 nc va tn dng c hot cht va ung thm nhiu nc. Nu y bng, n km gia thm Gng sng hoc Hu phc. ngn chn si ti pht, thnh thong nn dng dip h chu di hnh thc hm ung thay tr, liu khong 8 n 10g mi ngy. Cha st rt. - Dip h chu 16g, Tho qu 12g, Thng sn 16g, H kh tho 12g, Binh lang 8g, inh lng 12g.

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- Sc ung. Cha nht c sng au: - Cy ch mt nm vi mt t mui gi nh, ch nc chin vo, vt ly nc ct ung, b p ch au. Cha b thng t, chy mu: - Cy ch vi vi gi nh, p vo v thng. Cha l lot, thi tht khng lin ming: - L cy ch , l thm lm, bng nhau, inh hng 1 n, gi nh p. Cha bnh chm mn tnh: - Cy ch v, xt nhiu ln, lm lin tc, hng ngy s khi. Cha vim gan vng da, vim thn i , hoc vim rut tiu chy, hoc mt au sng : - Cy ch 40g, m 20g, dnh dnh 12g. - Sc ung. Mt s ch phm c ngun gc t dc liu ch thn xanh c trn th trng hin nay Ch phm EQIDO ca cng ty TNHH Mai Hn THNH PHN: Tnh cht cy ch rng ca 100% thin nhin CNG DNG: Tng cng sc khng c th H tr iu tr cc chng bnh vim gan, h men gan trong bnh gan, bo v gan trc tc ng ca ru v thuc THNH PHN: H tr iu tr bnh inh ru, thn kinh, sn Dip h chu ..... 2 g hu, vim rut, tiu chy, cam tch,....c phn Ch phm Livbilnic ca cng ty T dc...v... 1 vin Traphaco CNG DNG: Vim gan do virus, c bit vim gan siu vi B mn tnh. D phng v iu tr vim gan do ru, vim gan do thuc. Ri lon chc nng gan

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Ch phm VG-5 ca cng ty c phn Dc Danapha THNH PHN Cao Dip H Chu...100mg Cao C Nh Ni .......................50mg Cao Nhn Trn ........................130mg Cao Ru Bp ............................ 50mg T dc va 1 vin CNG DNG: - H men gan, tng cng chc nng gan, phc hi t bo gan. - H tr iu tr vim gan cp v mn tnh. 1.4. TIU CHUN CHT LNG V PHNG PHP TH 1.4.1. Cc ch tiu kim nghim Ch Rng Ca theo Dc in Vit Nam IV (xem chuyn lun dc liu) Yu cu kim nghim dc liu ch rng ca c trnh by cc ch tiu sau: c im cm quan, c im vi hc, nh tnh, th tinh khit ( m, t l vn nt, cht chit c trong dc liu), mt s vn khc (ch bin, bo qun, cng nng, tnh v) v ch tiu nh lng l xc nh hm lng cht chit c trong dc liu. 1.4.1.1. nh tnh A. Ly 5 g dc liu kh, tn nh, thm 50 ml ethanol 90% (TT), lc u ri un hi lu trong cch thy 30 pht. Lc, c cch thu cn 3 n 4 ml. Chia i dch lc vo 2 ng nghim lm cc phn ng sau y: ng 1: Thm 4 - 5 git acid hydrochloric (TT), ri thm vo mt t bt Magnesi (TT), xut hin mu .

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ng 2: Thm 3 - 4 git dung dch st (III) clorid 9% (TT), xut hin mu xanh tm. B. Ly 1 g bt dc liu, thm 5 ml nc, un si, lc. Ly 2 - 3 ml dch lc ngui, thm 1 - 2 git dung dch gelatin 2% (TT), xut hin ta bng. C. Phng php sc k lp mng (Ph lc 5.4) Bn mng: Silica gel GF254 Dung mi khai trin: Cloroform - methanol ( 9:1 ). Dung dch th: Ly 5 g bt dc liu vo bnh nn nt mi, thm m bt dc liu bng dung dch ammoniac m c (TT) , yn 30 pht. Thm 50ml ethanol 96% (TT), un cch thy 30 pht. Ly dch cht ethanol c n cn, ha tan cn bng dung dch acid sulfuric 3% (TT) ( 2 ln, mi ln 30ml ), lc. Kim ha dch lc bng amoniac m c (TT) n pH 9-10 , lc dung dch ny vi cloroform (TT), ( 2 ln, mi ln 20 ml ). Gp dch chit cloroform, lc, bay hi trn cch thu n cn 2 ml, dng lm dung dch th. Dung dch i chiu: Ly 5g bt Dip H Chu (mu chun), tin hnh chit tng t nh i vi dung dch th. Cch tin hnh: Chm ring bit ln bn mng 20 l mi dung dch trn. Sau khi trin khai sc k, ly bn mng ra, kh nhit phng, phun thuc th Dragendorff (TT). Sc k ca dung dch th phi c cc vt cng gi tr Rf v mu sc vi cc vt trn sc k ca dung dch i chiu. 1.4.1.2. m: khng qu 12% (Ph lc 9.6 , 1g, 100oC, 4 gi). Cch tin hnh Dng dng c sy bng thy tinh ming rng y bng c np mi lm b ng mu th; lm kh b trong thi gian 30 pht theo phng php v iu kin qui nh trong chuyn lun ri cn xc nh khi lng b. Cn ngay vo b ny mt lng chnh xc mu th bng khi lng qui nh trong chuyn lun vi sai s 10%. Nu khng c ch dn g c bit th lng mu th c dn mng thnh lp c dy khng qu 5mm. Nu mu th c kch thc ln th phi nghin nhanh ti kch thc di 2mm trc khi cn. Tin hnh lm kh trong iu kin qui nh ca chuyn lun. Nu dng phng php sy th nhit thc cho php ch chnh lch 20C so vi nhit qui nh. Sau khi sy phi lm ngui ti nhit

Trang 17

phng cn trong bnh ht m c silicagel ri cn ngay. Nu chuyn lun khng qui nh thi gian lm kh c ngha l phi lm kh n khi lng khng i, tc l s chnh lch khi lng sau khi sy thm mt gi trong t sy hoc su gi trong bnh ht m so vi ln sy trc khng qu 0.5mg. Nu mu th b chy nhit thp hn nhit sy qui nh th trc khi a ln nhit , cn duy tr t mt n hai gi nhit thp hn nhit nng chy ca mu th t 50C n 100C. Nu mu th l dc liu, khi chuyn lun ring khng c ch dn g c bit th tin hnh sy trong t sy p sut thng. Dc liu phi c lm thnh mnh nh ng knh khng qu 3mm; lng em th t 2g n 5g; chiu dy lp mu th em sy l 5mm v khng qu 10mm i vi dc liu c cu to xp. Nhit v thi gian sy theo yu cu ca chuyn lun ring. 1.4.1.3. T l vn nt Qua ry c kch thc mt ry 3,150 mm : khng qu 8% (Ph lc 12.12). Cn mt lng dc liu nht nh ( p gam ) c loi tp cht. Ry qua ry c s quy nh theo chuyn lun ring. Cn ton b phn lt qua ry (a gam). Tnh t l vn nt (X%) (t kt qu trung bnh ca ba ln thc hin) theo cng thc:

X% =
Ghi ch:

a 100 p

Lng dc liu ly th (tu theo bn cht ca dc liu) t 100g n 200g. i vi dc liu mng manh th ch lc nh, trnh lm vn nt thm. Phn bi v bt vn khng phn bit c bng mt thng c tnh vo mc tp cht. 1.4.1.4. Cht chit c trong dc liu Khng t hn 7,0% tnh theo dc liu kh kit. Tin hnh theo phng php chit nng (Ph lc 12.10), dng ethanol 96% (TT) lm dung mi. Phng php chit nng vi dung mi l nc: Nu khng c ch dn c bit

Trang 18

trong chuyn lun ring, cn chnh xc khong 2,000g - 4,000g bt dc liu c c bt na th cho vo bnh nn 100ml hoc 250ml. Thm chnh xc 50,0 hoc 100,0 ml nc, y kn, cn xc nh khi lng, yn 1 gi, sau un si nh di hi lu 1 gi, ngui, ly bnh nn ra, y kn, cn xc nh li khi lng, dng nc b sung phn khi lng b gim, lc qua phu lc kh vo mt bnh hng kh thch hp. Ly chnh xc 25 ml dch lc vo cc thy tinh cn b trc, c trong cch thy n cn kh, cn thu c sy 105 0C trong 3 gi, ly ra ngui trong bnh ht m 30 pht, cn nhanh xc nh khi lng cn. Tnh phn trm lng cht chit c bng nc theo dc liu kh. Phng php xc nh cc cht chit c bng ethanol hoc methanol: dng phng php tng t nh phng php cc cht chit c bng nc. Ty theo ch dn trong chuyn lun ring m dng ethanol hoc methanol c nng thch hp thay nc lm dung mi chit. Vi phng php chit nng th nn un trong cch thy nu dung mi chit c si thp. 1.4.2. Cc ch tiu kim nghim theo Dc in n II (The Ayurvedic Pharmacopoeia of India, part I, volume I) [8] 1.4.2.1. Tp cht: khng c qu 2% (Ph lc 2.2.2) Ly mt phn i din t mt thng cha ln, hoc ly ht khi lng nu gi khngqu 100g, v tri thnh mt lp ,mng trn mt a hoc khai ph hp. Quan st di nh sng ban ngy bng mt thng. Nu c bt c nghi ng phn t no, chuyn vo a Petri, v quan st di th knh 10X di nh sng ban ngy.
X%= a x100 p

a: Khi lng tp cht tnh bng gam b: Khi lng mu th tnh bng gam 1.4.2.2. Tro ton phn: khng c qu 16% (Ph lc 2.2.3) Cn chnh xc khong 2-3g dc liu mn, t chy tronh mt chn s hoc Pt c cn b nht khng qu 4500C cho n khi khng cn cacbon, ngui v cn. Nu tro khng cn cacbon khng th thu c bng cch ny, ly ht khi tro cho vo nc nng, thu cc cn trn giy lc khng tro, t phn b v giy lc,

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thm phn giy lc, bay hi n kh, v t nhit khng a 450 0C. Tnh ton t l phn trm tro so vi dc liu kh trong khng kh. 1.4.2.3. Tro khng tan trong acid: khng c qu 7% (Ph lc 2.2.4) Cho tro ton phn vo ni cha, thm 25ml HCl long. Thu phn khng tan trn giy lc khng tro (Whatman 41) v ra bng nc nng cho n khi nc lc trung tnh. Chuyn giy lc c cha cht khng tan vo ni nu kim loi ban u, lm kh trn mt bn nng v nung n khi lng khng i. cn ngui trong mt my sy kh trong 30 pht v cn ngay. Tnh ton hm lng ca tro khng tan trong acid so vi dc liu kh trong khng kh. 1.4.2.4. Xc nh hm lng cht chit c trong dch chit cn: khng t hn 3% (Ph lc 2.2.6). Ngm 5g bt dc liu th, c lm kh trong khng kh, vi 100ml ethanol vi nng ghi r trong mt bnh ngm kit khong 24 gi, lc thng xuyn trong 6 gi v yn trong 18 gi. Lc nhanh, phng mt mt dung mi, bc hi 25 ml dch lc n kh trong mt chn y phng nng c cn b, v sy kh 105 0 C, n khi lng khng i v cn. Tnh ton t l phn trm ca cc cht trong dch chit ethanol so vi dc liu kh trong khng kh. 1.4.2.5. Xc nh hm lng cht chit c trong dch chit nc: khng t hn 13% (Ph lc 2.2.7). Tin hnh theo ch dn xc nh cht trong dch chit ethanol, nhng s dng cloroform-nc thay v ethanol. 1.4.3. Tiu chun c s ca kim nghim nguyn liu Ch Rng Ca ca Cng Ty Xut Nhp Khu Y T Domesco: [7] 1.4.3.1. m: khng c qu 15%. 1.4.3.2. Tro ton phn: khng c qu 15%. 1.4.3.3. Lng tp cht: khng c qu 5%. 1.4.3.4. Kim tra d lng thuc tr su: s dng phng php nh lng bng GCMS - Chlorpyrifos: Khng c qu: 0.5 mg/kg

Trang 20

- Diazinon: Khng c qu: 0.5 mg/kg - Cypermethrin: Khng c qu: 0.1 mg/kg 1.4.4. Mt s ph nh tnh: [11] 1.4.4.1. Mt s dng ph ca Phyllanthin UV ABSORPTION SPECTRA UV - Absorption of 0.0192% w/v solution in methanol Cc thng s: Measuring Mode: Abs Scan speed: Medium Slit width: 1.0 Sample Interval : 0.2 No 1 2 3 wave length (nm) 280 229 206 Abs 0.354 0.965 2.19 4

Ph hng ngoi bin i (FTIR in KBr dispersion)

Isolat

Trang 21

ed phyllanthin exhibited the following peaks: 1517.9, 1481.2, 1463.9, 1446.5, 1419.5, 1313.4, 1269.1, 1247.9, 1236.3, 1178.4, 1159.1, 1139.9, 1109, 1055, 1041.5,1026.1, 964.3, 952.8, 858.3, 817.8, 788.8, 767.6 & 752.2. 1.4.4.2. Dng ph hng ngoi bin i ca Hypophyllanthin

Trang 22

1.4.5. Kt qu HPLC ca Phyllanthin v Hypophyllanthin Dng c: Mt h thng sc k Shimadzu bao gm my bm LC8A m hnh kp, Ai-t pht hin nh mng (SPD-M10A). Ct: E-Merck Nitrile Column (250mm x 4mm) Pha ng: Buffer (pH 2.8) : Acetonitrile (83:17) Tc dng: 1.9ml/min Detector pht hin bc sng 230nm.

Chormatogram of Phyllathin

PDA of Phyllathin

3D VIEW OF PHYLLANTHIN

3D VIEW OF HYPOPHYLLANTHIN

Chormatogram of Hypophyllathin

PDA of Hypophyllathin

Trang 23

2. KT QU THC NGHIM 2.1. Kt Qu Vi Phu: 2.1.1. Thn: Vi phu c thit din trn, c 2 n 3 gc li khng u nhau.

B g

Ht tinh bt M mm v Biu b

Trang 24

2.1.2. L: Gn gia mt di li nhiu, mt trn hi li

Phin l

L kh kiu song bo 2.1.3. R

Lng che ch a bo

Biu b trn

Trang 25

2.2. Kt Qu Soi Bt:

Mnh biu b

B si di

Mnh m mm

Mnh mch chm

Mch xon

Tinh th Calci oxalat

Trang 26 2.3. Kt Qu Phn Tch S B Thnh Phn Ha Thc Vt Nhm hp cht Thuc th Cch thc hin Nh dd ln giy Carr-Price H2SO4 Tinh du Triterpenoid t do Alkaloid Coumarin Atraglycosid Flavonoid Glycosid tim Bc hi ti cn LiebermannBurchard Thuc th chung alkaloid Pht quang trong kim KOH 10% Phn ng dng tnh Kt qu nh tnh trn cc dch chit Dch Dch chit cn Dch chit nc chit Khng Thy Khng Thy ether thy phn thy phn phn phn (-) () () (-) (+++) (-) () (-) (-) (+) (-) (-) (-) (-) (+) (-) () () (-) (-) (-) (++) (-) (-) (-) (-) (+++) (-) (-) (+) Kt qu nh tnh chung Khng Nghi ng Nghi ng Khng C nhiu C t Nghi ng Khng C Khng Khng Khng Khng C

Cht bo Carotenoid

Vt trong m Xanh chuyn sang Xanh dng hay xanh lc ng sang xanh dng C mi thm nu tm, lp trn c mu xanh lc Kt ta Pht quang mnh Dung dch kim c mu hng ti Dung dch c mu hng ti Tm mn Xanh

Mg/HCl Thuc th vng lacton Thuc th ng 2desoxy Anthocyanosid HCl KOH Proanthocyanidin HCl / t0

Trang 27 nhiu C nhiu C nhiu C it C C C C C

Tanin

Dd FeCl3 Dung dch Gelatin mui LiebermannBurchard TT Liebermann Lc mnh vi dd nc Na2CO3 TT Fehling Pha long vi cn 90%

Xanh ru hay xanh en (Polyphenol) Ta bng trng (Tanin) nu tm, lp trn c mu xanh lc C vng mu tm Si bt Ta gch Ta bng trng vng nu

(-) (-) (+) (++) (++) (+++) (++)

(+++) (+++)

Triterpenoid thy phn Saponin Acid hu c Cht kh Hp cht polyuronic

(++)

nh gi theo cc mc sau: (-): Khng c (++++): C rt nhiu. Ghi ch :

(): Nghi ng,

(+): C t,

(++): C,

(+++): C nhiu,

C th c phn ng nhng khng thc hin

Khng c mt cc nhm hp cht trong dch chit

Trang 28

2.4. Kt Qu Sc K Lp Mng Bn mng: s dng bng mng trng sn Silica gel GF254 Merck Kch thc bng mng: 2,5 x 10 cm Dung mi khai trin: Cloroform - methanol ( 7:1 ). Dung dch th: Ly 5 g bt dc liu vo bnh nn nt mi, thm m bt dc liu bng dung dch ammoniac m c (TT) , yn 30 pht. Thm 50ml ethanol 96% (TT), un cch thy 30 pht. Ly dch cht ethanol c n cn, ha tan cn bng dung dch acid sulfuric 3% (TT) ( 2 ln, mi ln 30ml ), lc. Kim ha dch lc bng amoniac m c (TT) n pH 9 - 10 , lc dung dch ny vi cloroform (TT), ( 2 ln, mi ln 20 ml ). Gp dch chit cloroform, lc, bay hi trn cch thu n cn 2 ml, dng lm dung dch th. Cch tin hnh: Chm ln bn mng khong 20 l dung dch th. Sau khi trin khai sc k, ly bn mng ra, kh nhit phng, phun thuc th Dragendorff . Thu c sc k nh sau:

Bng mng c 5 vt: c cc gi tr Rf nh sau: 0.32 - 0.41 - 0.56 - 0.61 - 0.81 Nhn xt: bng mng c nhng vt tch tng i l xa v khng c hin tng trng vt, nhng do hm lng cc cht c chit trong cy qu thp nn cc vt khng c r. c bit, kt qu sc k c vt cng cua l kh c trng cho dc liu Ch rng ca. 2.5. o m: Kt qu o m ca dc liu Ch rng ca bng cn hng ngoi l 8.3%.

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3. XY DNG TIU CHUN DC LIU 3.1. Tiu chun ngh: Cn c vo cc ni dung trnh by trn, xin c ngh tiu chun kim nghim dc liu Ch Rng Ca nh sau: Ch rng ca Herba Phyllanthi urinariae B phn dng Ton cy ti hoc phi sy kh ca cy Ch rng ca (Phyllanthus urinaria L.), h Thu du (Euphorbiaceae). M t Cy cao khong 30 cm, thn gn nh nhn, mang nhiu cnh nh mu hi ta. L mc so le xp thnh hai dy xt nhau trng nh l kp lng chim. Phin l thun bu dc hay tri xoan ngc, di 5-15 mm, u nhn hay hi t, mu xanh sm mt trn, xanh nht mt di, khng cung hay c cung rt ngn. Hoa mu trng mc di l, n tnh, hoa c hoa ci cng gc, hoa c u cnh, hoa ci di. Hoa khng c cung hoc c cung rt ngn. Qu nang hnh cu, ng knh c th ti 2 mm, sn si, nm st di l. Qu c su ht. Ht hnh tam gic mu nu nht, lng ht c vn ngang. Vi phu Thn: Vi phu c thit din trn, c 2 - 3 gc li khng u nhau. Biu b gm 1 lp t bo hnh ch nht, dt, nm ngang khng u nhau; m mm v gm nhng t bo hnh trn hay hnh bu dc xp ngang, khng u, xp cha nhng khe nh, c cha t tinh bt. Mt vi tinh th calci oxalat hnh khi trong m mm tu. Gn l: Gn gia mt di li nhiu, mt trn hi li. Khng c m mm giu nh P. amarus. Tinh th calci oxalat hnh cu gai rt nhiu, tp trung vng m mm ngay di si. Phin l: Biu b trn l nhng t bo hnh ch nht dt. L kh kiu song bo. Lng che ch a bo (2 t bo), ngn, vch dy st mp l, khng c lng che ch. Bt

Trang 30

Bt mu xanh, c v hi ng. Soi knh hin vi thy: Mnh biu b gm nhng t bo thnh mng hnh ch nht. Lng che ch n bo hoc a bo. Mnh m mm gm nhng t bo a gic thnh mng. Mt vi m t bo m mm ang phn ho thnh m dy (thnh hi dy ln gc). B si di. Mnh mch chm v mch xon. Tinh th calci oxalat hnh cu gai.
nh tnh: nh phn 1.4.1.1 trn nhng ngh h dung mi khai trin sc k

lp mng l Cloroform - methanol (7:1 ).

m: khng qu 12% (phn 1.4.1.2 trn) T l vn nt:

Qua ry c kch thc mt ry 3,150 mm: Khng qu 8% (phn 1.4.1.3 trn).

Tp cht: Khng c qu 2% (phn 1.4.2.1 trn). Tro ton phn: Khng c qu 15% (phn 1.4.3.2 trn). Tro khng tan trong acid: Khng c qu 7% (phn 1.4.2.3 trn).

nh lng
Hm lng cht chit c trong dch chit cn

Khng c t hn 7% (phn 1.4.1.4 trn). Hm lng cht chit c trong dch chit nc Khng c t hn 13% (phn 1.4.2.5 trn).
nh lng d lng thuc tr su: (phn 1.4.3.4 trn).

Ch bin Thu hi quanh nm nhng tt nht l vo ma h-thu, em v ra sch dng ti. C th ct tng on phi kh; hoc ra sch c cy, phi gn kh ri b li, phi m can tip n kh, khi dng loi b tp cht, ra qua nc, ct on 5 - 6 cm phi kh. C th ly l p li thnh bnh. Bo qun ni kh, trnh m, mc, mt. Tnh v, qui kinh Cam, kh, lng.Vo cc kinh can, ph. Cng nng, ch tr

Trang 31

Tiu c, hot huyt, li mt, thanh can sng mt, li thu. Dng khi vim gan hong n, vim hng, mn nht, vim da thn kinh, chm, vim thn, ph thng, si tit niu, vim rut, tiu chy. Cch dng, liu lng Ngy dng 8 20 g dc liu kh, dng thuc sc. King k Ph n c thai khng dng. 3.2. Gii thch tiu chun - Phn m t thc vt, vi phu, bt dc liu, nh tnh, m, t l vn nt, hm lng cht chit c trong dc liu, ch bin, bo qun, tnh v, quy kinh, cng nng, ch tr, cch dng, liu dng, tng k vn gi nguyn theo tiu chun ca Dc in Vit Nam IV. Ngoi ra, cn b sung thm mt s ch tiu nh sau:
Trong phn th tinh khit: ngoi cc ch tiu v m, t l vn nt, b

sung thm cc ch tiu: - Tp cht. - Tro ton phn. - Tro khng tan trong acid. Cc ch tiu trn l cn thit phi c b sung m bo cht lng dc liu, m bo tnh tinh khit ca dc liu, trnh cc sai s xy ra khi nh tnh, nh lng cc phn khng phi l dc liu.
Trong phn nh tnh:

- Trong phn sc k lp mng, khi tin hnh thc nghim trn dc liu thu hi th nhm thc tp chy vi h dung mi Cloroform - methanol (7:1 ) hi khc vi h dung mi DVN IV qui nh Cloroform - methanol ( (9:1 ). Thc nghim cho thy kt qu sc k lp mng h dung mi Cloroform - methanol (7:1 ) tch tt hn nn ngh tiu chun phn nh tnh sc k lp mng chy vi h dung mi Cloroform - methanol (7:1 ).

Trang 32

- Ngoi cc phng php nh tnh bng cc phn ng ha hc, nh tnh bng sc k lp mng, b sung thm cc ph hng ngoi bin i (ph FT-IR) ca phyllanthin v hypophyllanthin tng tnh c hiu ca cc phn ng nh tnh. Trong phn nh lng Dc in Vit Nam c ch tiu xc nh hm lng cht chit c trong dc liu, s dng dung mi ethanol 96%. V vy, khi xy dng tiu chun cho dc liu b sung thm ch tiu: - Xc nh hm lng cht chit c trong dch chit nc - C th nh lng phyllanthin v hypophyllanthin bng phng php HPLC da vo cc ph chun ca chng. Ngoi ra, vn d lng thuc tr su trong dc liu cng l vn ang c quan tm nn khi xy dng tiu chun cho dc liu b sung thm phn nh lng d lng thuc tr su trong dc liu. y cng l mt ch tiu kh quan trng.

Trang 33

KT LUN
m bo tnh an ton, hiu qu v cht lng ca thuc th yu cu cc

nguyn liu lm thuc phi t tiu chun nht nh, nht l cc nguyn liu lm thuc c ngun gc t dc liu. Xy dng mt tiu chun cho mt dc liu l yu cu rt quan trng, gip xc nh dc liu c t cc yu cu ca mt nguyn liu lm thuc hay khng.
Ch rng ca, mt dc liu rt thng gp v c nhiu tc dng cha

bnh cn c mt tiu chun c th kim tra cht lng dc liu trc khi dng lm thuc. Qua b tiu chun xy dng trn hi vng s gip cc sn phm thuc t dc liu ch rng ca s c cht lng cao hn, iu tr bnh hiu qu hn. B tiu chun trn c xy dng kh y ng vi nhiu ngun ti liu tham kho khc nhau. Tuy nhin, ty vo iu kin k thut mi ni m cc gi tr th nghim, cc phng php nh tnh, nh lng, s lun c cp nht, sa i ph hp vi s pht trin ca khoa hc k thut.

Trang 34

TI LIU THAM KHO Ti liu ting Vit 1. Cy thuc v ng vt lm thuc Vit Nam (2004), trang 438-441, NXB Khoa hc v k thut. 2. V Vn Chi (2003), T in thc vt thng dng, tp 2, trang 1943, NXB Khoa hc v k thut. 3. Tt Li (2009), Cy thuc v v thuc Vit Nam, NXB Y Hc. 4. Ng c Trng (2008), Nghin cu ha hc v nhn dng mt s nhm cht co trong cy ch rng ca Phyllanthus urinaria L., Euphorbiaceae, Thi Nguyn. Ti liu ting Anh 5. Bo Shen, Jun Yu, Shiyan Wang, Chu Eagle S. H., Wong V.W.S., Xin Zhou, Ge Lin, Sung Joseph, Chan Henry (2008), Phyllanthus Urinaria Ameliorates the Severity of Nutritional Steatohepatitis Both In Vitro and In Vivo, HEPATOLOGY, pp. 473-483 6. Chia-Chuan Chang, Yu-Chin Lien, Karin C. S. Chen Liu and Shoei-Sheng Lee (2003), Ligans from Phyllanthus urinaria. 7. Domesco Medical Import - Export JSC, Detailed information for Chamberbitter. 8. Government of India Ministry of Health and Family Welfare Deparment of Ayush, The Ayurvedic Pharmacopoeia of India, Part I, Volume I. 9. Joo B. Calixto, Adair R. S. Santos, Valdir Cechinel Filho, Rosendo A. Yunes (1998), A Review of the Plants of the GenusPhyllanthus: Their Chemistry, Pharmacology,and Therapeutic Potential. 10. Shin MS, Kang EH, Lee YI, A Flavonoid from Medicinal Plants Blocks Hepatitis B Virus-e Antigen Secretion in HBV-infected Hepatocytes. 11. Vivian Esther Fernan (2003), Initial characterization of crude extracts from Phyllanthus amarus schum. and thonn. quassia amara L. using normal phase thin layer chromatography.

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12. Yang CM, Cheng HY, Lin TC, Chiang LC, Lin CC. Taiwan (2005), Acetone, Ethanol & Methanol Extracts of Phyllanthus Urinaria Inhibit HSV-2 Infection in Vitro, Antiviral Res, pp. 24-30. 13. Zhonghua Shi, Yan He, Lin Chuang, The Inhibitory Effect of Chinese Herb Phyllanthus on Hepatitis B Virus in Vitro.
14. Zhou S, Xu C, Zhou N, Huang Y, Huang L, Chen X, Hu Y, Liao Y,

Mechanism of Protective Action of Phyllanthus Urinaria L. Against Injuries of Liver Cells, ZHONGGUO ZHONGYAO ZAZHI , pp. 109-111.