n t zonesare presen betlvee il uorescent Ilor,r,ish-green -v-c n a o l h e z o n e s i s i l i b i n i n n d t a . x i i o i iin t h e c h r o m a t o g r a m r t o b t a i n e d' v i t hh e t e s Is o l u t i o n .

Top of the plate

[*,;;i;,;;;*
I' pirppLrs, Silllbunt mLtridnLIltt cti l . c e n t o i s i l l ' n r a r i nc x p r c s s e ta s I t ) ( . i r i e dd r l l g ) .

nl I i l u , r r c s c cz o r t c

fiiu.rr,"r.',.,,...,il,-,;;;- I
( zorresilibirrin) ] z r l t r o r e s c e n to t r c i):;;jif;

l ' r r r t o l i n : l n o r a n g ci l u o r c s c e n l

I
i

l A . T h e a c h e n ei s s l r t l n g l l 'c o m p r e s s e , e.l o n g a t e ' o b o v a t e ' a b o L r i r - Sn ' r ml o n g . 3 m m b r o a d a r l c l1 . 5 m m t h i c l < :l h e t 1 l o i , r t e s r - i r i r c e ss m o o t h . r n c s h i n l u i t l - r . ri r e v o r p a l e r .lai.k brorvn sLreaked r.'ariabiv color,rr brorvn gror-tncl l l o n g i t L r d i n a l ltro' g i v e a n c , r , e r a pl a i e i r e r " i s ho l ' b r o r v n . c o l o u r : t h e i r L r i ti s t a p e r i n ga t t h e b r s e a n d c r o r ' ; n e d . i t . a t h e a p e x r . r ' i t h g l i s t e n i n g p a l e 1 ' e i l o ue x l e n s i o nf b r m i n g ' th a c o l l a f a b o u i 1 m m h i g h s L r r r o L l l l t i i n g e r e m a i n so i t h e the transversel)'. iruit shotvs.i n;irrotv,brolvn slvle. Cr,rt r o u l e r a r e e a n c l2 i a r g e .d e n s e . v h i t eo i h "c o t r ' l e d o n s . t B . R e t l r r c eo a p o r v d e rl 3 5 r j 1 2 . 9 . 1 2 )T i i e p o u ' d e rt s ve b i . o , , v n i s h l l o u 'l i i t h d a r k e r s p e c k s .E r : i L r i n eu n . i e r i l R. microscopeusing c/z/oralhyilrote soltLtiott The po"r'cler s ihe ioilorvingCiagnosticcharacters: ii-agmenls shr.r.,', o s o i l h e e p i c a r pc o m p o s e e l f c o l o L r r l e sc e l l s .p o i v g o n a l i ' a i n s u r t a c er , i e u .t h e i r - r m e n p p e a r i n g a i r l 1 l a r g eo r a s o r d a s m a l l s 1 i t . e p e n c i i n qt n t h e o r i e n t i l i o n : g r o L r p s l p a r e n c h y , n r a t o c e l l sf r o m t h e p i g m e n tl a 1 e r .s o m e ' i i us b c i t h e m c o n t . r i n i n $ o l o u r i n gm a l t e r l ' h i c h a p p e a r s n g h l t s r e c , r ' e r l a b u n d a n lg i ' o L r po i l a r l e s c l e r e i d si ' o m t h e l a b t e s l a r , ' , i t h r i g i r t l e i l o u p i l t e d r v a l l s n c la n a r r o t ' l t i m e n : Lledparencl'r1'ma iragmenls of s mall-ce ocr-:isioneil)' ' , v i i hp i t t e c la n d b e a c l e d' a i l s : a b u n d a n tI h i n - r , r ' a l l e d l s p a r e n c h l ' m a t o uc e l l si r o m t h e c o t - v l e d o nc o n t a i n i n go i l s I o m g l o b u l e sa n d s c a t t e r e c ll u s l e rc r ) , s t a l s i c a L c i r - ro x a l a t e c a ie,,vlai.ger.prismaticcry'siaiscricalciittno.<alate.

TEsTs

d 8 L o s so n d r y i n gt 2 . 2 . J 2 1m e x i m u m . ( )p e r c e n t . e t e r m i n e d : eu b i o n i . ( t ( t (g o i t h e p o r ' r ' c l e rd rc lg ( 5 0 0 )1 2 . 9 . 1 2 )y 'c l r fi n g i n a n o v e na t 1 0 5' C f o r 2 h . m 8 T o t a la s hQ . , 1 . 1 6 1 a x i m u m . 0 p e rc e n t . ASSAY Liqr-ricl chromatographv 2.2 9i. 12. '[est d 5 s o l t t t i o n P l a c e . 0 0g o i t h e p o r v d e r ed r u g ( 5 ( l 0 ) . a i pa rd i) 1 2 . 9 . 1 2n a c o n t i n r - r o u s - e r t r a c t po n r a t r -As .d 1 0 0m l ior R or light petroleLtm and heatin a r.valer-bath B h. I. : \ l l o l vt h e t l e i a t t e d r u g i o d r l a l r o o ml e m p e r a t n r e n a e app c o n t i n u o u s - e x t r a c t i o n a r a l u s ,x t r a c ith e l a t t e rr v i t h for ml 1tl{) oi nethanol l? in a r',.'aler-bath 5 h. Evaporale e o c t h e m e t h a n o l ie s t r a c itn t ' t t c L rto a r r o l t r mo i a b o u t3 0 m l . t c F i l t e ri n t o a 5 0 m l v o l u m e t r i i l a s k .r i n s i n g h e e r t r a c t i o n methanol[?. ml iiaskand fhe iiiter.and dilutingto 5lt.() r'r'ith m l m n l l i l u t e5 . ( )m i o l t h i s s o l L i t i o t o 5 ( l . t l l r , " i t h e t h a n oR . a solutton.Dissolve quanlitl'of milk thistle Relbrence to drg CRSequii'aient 5.()m$ ot sitntlttrtlisecl e,ttract 1 o L s i l i b i n i nn , g ) i n m e t h a n oR a n d c l i l L r tte 5 0 . 1m l r v i t hl h e i same olvent. s Column: - - s t . - c - ( 1 . 1 2m , O = 4 m m : 5 /: - sttttionorE phase: oc:terlecgi:sityl gel for silica R chrr:nutogrttpAy (5 pm).

c ' C . T h i n l a 1 ' e r h r o m ; i t o g n p h r( 2 2 . 2 7 \ { \ cl T o 1 . 0 g o i p r r r v c l e r e dr L r g5 ( ) 0 ) 2 9 . 1 2 ' i T e s t. s o [ t t i i o n . . .tlohilephase acld iil r'ti r-tintethanol 1?.Heat unJer rcih-rr in a - ntrthile R R. plt,tse phosphoric ttcidR: methanol Lrater .1: 'C tor'5 min. Crrril nd iilter.Evaporate a rvaler-balh i 7{) a i i ) . 5 : 3 5 : iIr J1 . . ' l l : n t h e i i i t i . r t et o t l r l t l e s sa n d t l i s s l l v e [ h e r e s i d r - ri e 1 . 0 m l - nohile phaseB: pltospltoric R. ttcitlR: ntetltanol u'aterR o i n t e t h r t n o ll l . f" [ { 1 ) . 5 : 5 t ) : 5 ( '}, I l : fl I [!clcrcnce .:olutton. Dis-.ol'"'e rrl3 ol'-qi1i'!irirn ancl ) l o b i l e p h a s eB J [ng oi ttt,tili[in f? in iit rnl oi rncllranol R ]-ime ) l o b i l e p h a s e. \ P[ute:'l'L(.' si!it'o qel pltite [1. ucld fl. ocatone R .ttr.thi/e,lhr1.sc: atthgdtous l.orn'tic I / n t e t ' h l b n c c l ' t k t r i t l e? ( . J . 5 . l t r . . r : i . l l ' I l . r ir l l { ) p l o i t h e t ' : s l s r l l r . t t . i or l . i L 0 p l ' l i t n e )ltltltc'atiort: ,"fl'crairL-r' rttirln, ls b:rrlds. sol o D c t . e / o p r r t c r i t :l e i ' e tp a t h r r i 1 { tc m [ ) r r t i r t r ]r l l r ) l ) - l { ) 5 ' ( - . [ ) e t e t ' ! i r t rs :i . r l v t h e u . i r n t p l : i t e ' v l [ ] r l tl t ) { I s o l r ' r I t r l t l t r-'.tlrr in ntctltttttttlll 1i ttl'diPltcrttrll'tttir: t.tt-itl ilr|litt(ti'!litrl l t t t ds i r h s t r l t i c t r tsy ,r ; r ir t ' i t h ; r : . l tr : I s , r l r . r l - i o tr l lp ni{larL)q()l lli0 R trt rttt'litiinttiIi. .\li,t"',ttl Lii-fior i}() niin ;:r. lrttcicr.itl i tte ilt ulti:iviolei li lir t ;rL.ltr'r t'i 11,:.srr1Ls: llclO',1' :;t.lt:tt', ..'ll rr)rlts piisent in th.' Ihc sec iir'-'itec s0l u ti 0tl ob c it r,riltJ io'J.r;.rnls Ili rre.i' r'i l ii l.;lc''': fr a l r r li h e t e s t s o l r - r t i o n . i i t ' l ! t i i t t t o r ur.r l h c l r r r . i t l i ea t l d l ; l o u ' r o l e. 0 . S m l , m i n . r l ) a t c c t i o n : s p c c t r o p h o i o n l c t ea t l . 9 E t r t - r l Ittj:t:tir,tn: 1()pi .l0 min. Ii tlecessarv. /it lcrrlron li;rre: silibinin Ll - .r!-.r-rr-rL l t d . i r i slih c t i m e p e r i o t l so i t l r c g r . i t l i e r r t : . ,S1:l.ilc/n i I o lti I i t a : retelence solutiotl : .;i t - re solti!itttt: ntininti-int1.Ebettvc':t't il,c ilcllis ciue lo . s i l i b i n i n . \a n t l s i l i b i n i ul - l :
l ___~~( l~Ll,__~~ ln " ( ( 9 o l| ") l)) (( ^IL~__ Jent ( )l 0 l0 ~ l()() lO0- ()

COc-tf= rJCJs ftpl/J/,/t:FF p

09;r cps .rntl ttther tett-; (

2425

Uilk thistle dt1' exlrac:t,refined and -ctandarcliscd

EtlR PEAN PHARMACOP

EIA60 |

r r 90pcr ceJ1t to110per cent ofthc non1in l colltent t^t, ``l824), of silymarin,expressed as sihbinin(CJ5H Ol() statcd on thc Iabcl ' hc nolinal contcnt of s )llllarin is :(dried !iu1in thc ranfc30per cent to65per cent l A l , o c a t e s i l i c l i s t i r t .s i l i cil; r r i rr . s r li l . r i n i n . s i l ib i r r i r r ' ) . aCt} CXLl i i s o s i l i b i n i r A l n c l i s o s i l i b i r l i rjli b v t o t l l p l t r i s t t tu ' i t h t h t in he u'itlr rrlrll IhisIIc sIantlartl isedcirg' 1 contcnt of synlaC0rrcsponds to suppiie.1 clrronratrrgrant sL/P,7o'/ co ll's/`fc'/~x/f cl sf`f (bot d / l t l n t h c c h r o t t t : r t l r g r r ro tb l a i l l e J r t ' i t l rl i r e t c s i s /C c extntcl L1i.\. lt 20pcr cent to45pcr cent, t 1P=C)l ' 482 ) s o l u t i r . r nh e p e a k c l u ct o s i l i . l i ; r t t i n t l l ' r ' a n ' j t l s i z c .b e a b s c i l i C5 l o r b e p l e s e t t ta s t h e p r i n c i l l ; r p c a i i . D e t c r t n i l c t l l r : i r t . r u 1 calculated with rclf,rCnCe to total s ymarin . ri t l r e p c a l i s d u e t o s i i r c r i : , t i rs . l i d i : r n i r ls i l i b i n i nA , s i l i b i n i l tI J , stt 2or 2cc C r9f f`?H ds fbf`7f,?B rs s f l i s o s i l i b i n i r A i r n d i s o s i l i i - ' i l i rB . t (bth C H?lO 4824 40pcr cent to65pcr ccnt, e c a :ference to total silyl arin C a l c u l a t el h e p e r c c r t L a gc o n t e l l to 1 ' s i l l ' m ; r r i n ,a i c u l e l e d s caIculated :ith r( s l s r l r h r r n .u s i t r g I r r t i , l l t r r ; i rtr \: l:\ 1 1 r ' 1 1 1 , ' ii b ` ` rfo'fsl,s /bf ? ^rls `?f,7 c st 2or/ cr, 1 iOlJ-482) per ccnt to20per cent, (botl)C ith calculated re1erencc to total s rin yn)( 15+ `1 /l l()()() l 2+= J ~"|
d o t h e c h r o n r e t o g r a t n b i a i t i e i s s r r n i l a rt t i t h e ppl a oSram sLr ieil rt'jth nr i/k t h i s I le -st n ii t rri i sc d . ; rr1v1113f dr11e.rlrLtclCll5. i . . i 7 + . - l l t ! . 7 / r. ) x i i ( l r l u l 1

41

i a l e ao f t h e p e a kd u et o s i l j c r i s t i nn t h e i^ijthil.ietestsolul,ion; obtarned chromatogran n l ; i r e ao f t h e p e a l ic l u e o s i l i C l a n iin t h e r c h r o m a t o g r a o b t a i n e dv i t ht h e t e s ts o l u t i o r r : n

PRODUCTlON The extract is produced f1 onl the herbal drug by an appropriate procedure,usi11 one or nnore of the folloxvin solvents _ eth\,l acetate _ acetonc or mixture of acetone and water ,ater; _ ethano1or n1ixture of ethanol and _methano1ormixtureofmethanolandwater CHARACTERS `' :yellowish-brown`amorphous po HpPc 9der

00

A o a r . e a f t h e p e a kd u et o s i l i b i n r n i n t h e t ob c h r o m a t o { i ' a n r t a i n e d' i t h t h e t e s ts o l u t i o n l n a r e ao i t h e p e a l id u et o . s i l i b i n iB i n t h e u c h r o m a t o g r a o b l a i n e d ' i t ht h e t e s ts o l u t i o l . t ; m n a r e ao f t h e p e a kd u et o i s o s j l i b i n iA i n i h e d c h r o m a t o g r a o b t a r n ew j t h t h e t e s ts o l u t i o n ; m n a r " e o f t h e p e a kd u et o i s o s i l i b i n iB i n t h e a u c h r o m a t o g r a o b t a i n e d ' i t ht h e t e s ts o l u t i o n : m

45

A d a r e ao f t h e p e a l < u et o s i l i b i n i n i n t h e u:ithtfre relerence chronatoSranoblained solution; l a r e ao f t h e p ea k d u et o s i l i b i n i n J i n t h e n'ith the reference obtained chron.ratograrl soluiion: dry massof nrilk thisllestdndardised exlractCRS t e r - i s etd p r e p a r eh e r e f e r e n cs o l u t i o ni.n g r a n s ; o j I i r a s so f t h e d r u ! t o b e e x a ml r e d i.n g r a m s r co t c o n r b i n e p e r c e n t a g e t t l e no f s i l i b i n i n \ d drg and silibininB in mill; thistlcslandardised e.rlractCll,S: p e r c e n t a gl e s so n d n ' i n go f t h e d r u g . o

Q0 ^

?1

`72,

IDENTIFICATION Thin layer chromatoraphy(2~92 esr sc/`/fo Diss01ve0250g ofthe extractto be examined BoFP in5mlof` f and5mg o J+P, ` aactoso/ r Dissolve2mgofsf`fD r d P of/a flo in]0m1of 7 oF P c ' o :r PFJ/ C sf`fc r/,r (5 um)10rT/~C sf /P / R(210um)) '` ccr0 , Cfd, sc / dro sr?fc ^`afr /D `(8 1675 `o`'j ?c c '/I^ll : H/,'zfccz/fo10ul I0r8ull ofthe test solubon and10ul10r lce efere solution,as bands 2uIl of the1 `?/oVer a path of10cl z, rcJoP Ior6cm1 D/ f/,z`at100-105 C //, z/`al,'fc C/d c a10g//l soIution or sprayvitl c/fr, Dc/ / P //ll// ?f7z0 s/c9 in o`fP and subsequenuy spray Cl //ro with a50 /l solution of 2czc`'r, fl9 irl 2L iIle in Allovv the plate to dry in air for about30n,in and exal ultravioletlightat365nm w Pcsztr see bel the sequel1ce of zones prescntin the chronlatogras obtailled with the reference solution and reen the test soIution Furdlermore.other yellov ish- ~een fluorescent zOnes may be present bet the zOncs due to ralu0btained xsith sihbinin and taxifo1ill in the chromato the test solution
Top of the platc Silibjnir : a lcllon'ish-green z fluorescerL ole

0I/2008:207I ILK THIsTLE DRY EXTRACT, REFINED AND STANDARDIsED silybimarianiex actumsiccum
L L

z r l A l , e l i o u s h ' g r e efn u o r e s c e n to n e (silibinin l

'l'axiiolin : i l n o r a l f e 1l u o r e s c e t l t z0lr(

t A n o r a n g t f l u o r e s c e r zto n e r ( t a rj f u l i n ) l i \ v c l l o u ' i s S r e c rf i u o r c s c e nz o t t e h

raffinatun)et nOr1natu1 INITION DEI Dl`extract,refh ed and standardised r/ J //Plst,/,&fr r 8

produced fl'on

Rcfercnce soluliott

2426

(cctt:er pages) See the inforntatiott section on general moncsgraphs

POEIA60 EUROPEAN PHARMAC

dihydrate llinocyclinehydrochloride

TESTs Lossondrying(28 AsS Y I.iquicl chrolnt1togrdphy(~ PP~t,) 7Vsf st/ffrl l)isso|ve60()l11g ofthe cxtractto bC exan1hlCd in/,,Or/ tz,lo anddiluteto1000nllwiththesamesolvent r / 2f` rs/`c rro Dissolve a quantity of e `LP/0 CC sl,/t /'l/L9 r/tlcrC/x Scorrespondingto1o0mg rdfsLp srld o and ditute to l()()O ml with 21g)of silibi11in in llld (` the samc solvcnt Co : ~s`zc r=()125m,0=4mm sro 2C 2sc rfo yp/2 oc/ 'J sf/fcc/Fr,r 0 C d/o~t/`0P (5um) y Fx D mt1ximum50perce!lt

n t e C a l c u l a t t h e p e r c e n t a l c o n t e n o f t h e s u mo i i s o s i l i b i n iA e r-rsing the to B, lvith reierence total silymarin, and isosilibinin : f o l l o r v i ne r p r e s s i o n g ( +rl;) H)() fil_ F) +F;+Fl +/1+F F1 `) ' F, area of the peak due to s icristin in the chromatogran1obta"led xsith the test solution; idianin in the area ofthC pcak due to s chron1atogranl obtah)ed xA/ith the test solution; ibinin k in the area ofthe peak due to s ith l ostttined the test s0lution chromatoEra area of the pet1k due tO s ibinirl B in the chromatogran obtained vith the test s01ution k in the area ofthe peak due to isos"ibinin ith l obtai11ed the test so1ution chromatogra area ofthe peak due to isOsdibinin B h1the chrolatogran1obtained N.,ith the test solution area ofthe peak due tO s"ibinir1A in the chromatogram obtained with the re rence solution ibinin B in the area ofthe peak due to s chromatogram obtained with the re rence solution; mass of silibinin in the reference solution,in s gra lnass of the extract to be exalllined.in grtims
01/2008:1030 corrected6.o MINOCYCLINE HYDROCHLORIDE DIHYDRATE lvIinOcychni hydrOchlOridu 1dihydricum

ttsL, i1/llb`FP p Ei AD cllz f,r ~ 2obtD H r,sz, c clc`d`,`2/ o`P,tt tzrB J tlsc ' O 351;51 1 I 5 F~ , f`2 2o/ ,Lt. /2rP o/fc `2l z, scB `c,sP czC 2 l 7 / 50 In (0 5() = Time _ mv_____ms (|-28 2S-35 35-36 36-51 08m F o /O "->l(lO 1(" min 0 MobiIe phase 1"0->() lI ___ 0 () l()() l()0 0 bile phase B ~ 100

i "

/t,c/fo:speCtrophotorneter at288nn1 D ,t/ccFfc, 10|l : /'o / sihbinin B=about3O mh1 PCr 2fZC adj11st the time periods ofthe gradient ution f Sys sf relererce soif necessary,

- rcsoF : minimum18between the peaks due to t, sihbininAandsilibininB - the chrOmatogram Obtained wit l the reference solution fs/r2 ar is sirn to the chro 1atograrn supphed with zfJ/tr sC/r ` xf,Jcr CPS C/ cl 7C/ s I'ocate the peaks duc tO s icristin,sdidianir ,sdibini11A, si ninB,isOsilibininAandiSOSdibininBush)gthe bi Jr tt/c/r~/ f/trfs/rc,rd c/ /rs chromato tal11Supphed with Jl% absent J Tl: lT F}}; z :

> | H3C'N H CH3 H3c

2H2
H

)( H N- CH3

C23H ClNOT,2H!C) 1136 4 8 |

i1 ' 5300

arin, Calculatc the pcrcentage contcnt of total s"yln vhlg e\presslon e\presscd as s"lbhlit1,usiIlg the lllDllo | t / [ L r ~| rl; P + | 7 it,
/' [- t.' ut
l

i,' "

s i c r C a l c t r l a i eh . l i r e c e n t J q e o n t c r l to l ' l l t c s t t t r o i s iIi c r i Ln t r g [rl trlf.tl sill'n'^arin u s i n t h e lr,ith reterence antl siiitliartiir. g f o l i o r v i n f ) . p i . s s i o l:t il )t) l /` h i i/ - ; l
, r o C a i c : - r 1 l;lrriiep e c c n t r { ec o n l c l l t i t h c s ' . L mr i ' s i l i b i n i n \ r t s i l , . ' u a r i nL r s i n g - h e , [t r e f e r e t t c er l , r t : r i a n c s i l i i r i n i nl , l i " v i t h: iollo,,', r.\'-ri-cssl0n ir,g

DEF'INITION 1 ./ta 7 (,ltS. S.5ai? 12aS)zr. Bi s(d i methvIami no )-'1 {l'12' 12a 5'r,6' 1I'tlioxo-1.4,14.5 i 1' I 2atetrah-v-drrrxy-1, rochioricle i tle e-2'carbttxanl h)'il tetr.icen octahvtlro ciihvcirlrte. prodlrct irom a iermelrt;ilion derivetl proclrict Semi-s1'ntlteti' p ) r C o n t e n t '!.r 6 . {p e c e r l tt o 1 0 2 . 5 e rc e n t{ a n h } " d r o L l s ). snbstunctt CHARAT]'IERS h ir n . l p p et t r ' r ce ' .) , e l l o r v 'v Q r o s c o p cc 'l s t a liln e p o t v c l e r ' e . ' i s p a r i l r g ls'o l u b l ei r r v a t e rs l i g f i t i ls r ' r l t r biln l Solubilitu: of r I e t h a n o il g i - p . i -c e r l t ) . t t t i s s o l v ci sl s o l L r t i o n s a l l t a l i l .lntl roricles c.trbonatcs. 1^r1'd I I)IJNTIITtC.Yi'I()N (L' .hrornstograph\" )' 2 7\' ,\, Thin-lai,'er
2427

{ j}

F 1+ I f +FJ|

1
1+ 1+ l;

and other text's (1) Cenerti! -\:i;i1i:a.s applg to alL mont)ltr'rtph.s

l0-5-1 l{etht'lsullbnr'ltnctltaneI Dietan' Supplertrents

USP32

TlrbIets I\Icthl'IsuIf'on1'lmethane
' >N { e t h Y l s r - r l l o n v l u r e t h l r t a b l c t s c o l l t a i n t t o t l e s s t h a n Te 9 0 . 0 p e r c e n l a n d n o l t l o r e l h a t t 1 1 0 . 0p c r c e n t o f t h c l a an H b c l c d a r n o i i n i o 1 'n t e t h y " l s r - r l l b n y ' l n t e t h( C :e 6 O ' S ) .
Packaging c o n t a tn d r s . and storage-Preservc in tight. light-resrstlnt

USI' Refcllnce

sl itndards ( l l )-Li.tP,\l

r c t h : l srrl . f t n v l t t t t l tu n t R . \ . c

Idenlil'it'l1ion--1-he retentiotr tirle oi thc' ntrjor peak in the chroo n i a t o g r a l n o 1 ' t h c A : . r a l p r e p u r u t i o t t c o r r L ' s l ) o r l L its t h a l i n t h e c l t r o l n t r t o g r J n r o l L h e S l r . r i t r i . l l tp r c p u n t / i o t t , a s o b t a i r t e d t n t h c ' { r . s r r . r ' . Disintegratitrn altd tlisstrlutiorr i20-i07: tllL-ettlrtr trqtLitcrttetlts t o r D i . t t u t e u , r u l i o l o l t h , : - 1 0r l t n u t c s . Weight varialion (209 1): rreel the requirtments. Assar'ent Di ! trettl. Stundurd p rctttt rul i otr. tnd Clt rrnttttt.tgraphit s:t'st l P r o c e e d a s d i r c - c t e di n t h e . , 1 - r s n u n d e r , l / e l l z r / . r r t l f c t r t ' l n t e t l t a n e . .\ssctt ltrtpttrtiriott \\/eish and finell' pot'dcr not fe\\'er than 20 p Tablets. Dissolve an accuraielr vn'eighed ortion of the finell' pou'd e r e d n a t e r i a l . e q L r i v a l e n tt o a b o u t I T a b l e t " n D i l u e t t t , n t i r . a n d s o n i c a t ef o r 1 5 r n i n u t e sr t - 5 0 ' . A I I o U ' t o c o o l t o r o o n t t e t t t p e r r t u r d , d i l u t e r v i t h D i l u e r t r t o v o l u t . n e .a n d m i x . Q u a n t i t a t i v e l y d i l u t e * i t h D i l u a n r t o o b t a i n a f i n a l c o n c e n t r a t i o t t o f 0 . . 1m g o 1 ' m e t h Y l s u l f o n v l m c t h a n e p e r m L . T r a n s i ' e r a b o u t i n t L o f t h e s u s p e n s i o nt o t a 1 . - 5n ' r l - r . n i c r o c e n l r i l ' u g e u b e . a n d c e n t r i f u s e f o r 2 0 s e c o n d s . U s e the supem.ltant. Prcttecltrre Separatell inject cclualvolunres (about I 1rL) of tbe Startdarcl prepcirutiort and the .4ssut prepctralloi? into the chronratograph. record the chromatogranis. atrd tleasure the responses 1'or the rnajor peaks. Calculate the quantitv. in mg. of lnethvl( - s u l l ' o n r ' l n r e t h a n eC r H r O : S ) i n t h e p o r - t i o n o i - f a b l e t s t a k e n b 1 ' t h c ' formula: lf C(Rr i R.r) in rihich D is the dilution factor: C is the concentralion. ill nt,spL-r nrl.. of USP NIethllsullonvlntethane RS in the Standarclpreparo tion: and Rr and R.\ are the peak response t-atios of nlethl'l) s u l f o r r r ' l n r e t h a n et o d i ( e t h ) i e n e g l , r , ' c o l n t e t h \ ' l e t h e r o b t a i n e d f r o m the Assar' lreparotiotl and lhe Standard preparaticrt, respecti"el)'.

consists alnroslcolorol' Niit ro:cciltit The liLrrtu,all epitlerrnis to rt IesspliiisldecellsirrnLnged rilht anglcs the surl'ltce.1'hel have i g r e a l l \ , t l l i c k e r r e c li e rr ' l a l l s . n t o u ' h i l i t t h e l u n l e nc r , l t t i n L rfe \t . ou o tiottt libttveundcrhrgh in :;or.ne disluncc the lbrm of a slit. \/tctt'c-cl n. t, r n a g n i l i c l t r o nh c e c l l ss h o u o n l r a : ; l i t s h l p e dl L r r n u 'T h e l , h a v s ol thickenings the cell u,all ridlcs that appexrirs nodrrlar thickcn..d lavcr The sLrbe'pidr'rnral ol the liuit u'all vnherr vreued 1l'omabove. cells and constithin-ualledparenchvnrel is ntader-rp unliglrilicd o1' o1 cells rnil r:roups cclls ailer-nate lxyer.Coloriess tutesl pign.rcnt this pignrcntcells,thc lattervarvirtgin nutttLrert givesthe fruir r.l'ith rvall Nert appcru'ance. comesthc 1r'r-rit tissue, u'all ris nroltled abont piu'r-nch)'nr:rl elongated ths c.ll: in E cell lrters rhick.uith stippled iat'erof tire iiuit. The cellsof the innerl.tlost arrs longitudinrLl o1'the thc-r'contain larte cigar-shaped lruit uall mav be collapsedr or rnonotljnic crlciLrn oxalateprisnrs.l'he seecicoat epidermrs is formetl from lrigc 1'ellou'pllisrdccellr. Thc cclls have a narro\\' erpandedat each end oi the cell. and the cell llrmen. solneu'hat Thtcellsof the lan-rination. subepiderrlal walls shor.r' conspicuous cel.Ls: their lignifredcell nemslippled ol seedcoatconsist peculiar ("net cells"). rrdges thickenings or close-set haveprominentbrancs oi'celis having toi-rgh. soneivhat Next to therr is a srngleJa1,er {endosperm resrdue). emThe contents ualls and lipophrlic sr.vollen in cellsr'.'hicli. additionto sntali-llands. consists o1'thin-u'alled br1,o and containclunrpso1 c11'stals fat droplets. I dentilicationTesI(20])Idenrifitatirtn A: Tltin-LuterChronatographic prepared directed the as in the Test soluticlt--Use Testsolutiot't. t . C t e : t 1 ^ O r t ' t t l tL l t ' . [ r i l t t , t r t r i r t . Standurdsolution: 1.0mg of USP Siil'dianinRS per nrl. in nretiranol. mixtureof chlorosolvcntsvstent. ireshll'prepared Dctelopinl< a fs a f o r m . a c e t o n e , n d a n h - v d r o uo l n - r i c c i d( 7 , 5 : 1 6 . 5 : 8 . - 5 ) , exceptto dD' the in as Protedure-Proceed directed the chzrpter. rn platc for 30 minutes a currentof cold air. Spral'thc plaic u'ith a in diphenl'lborinate methanol(1 in 100), solutionof 2-aminoethvl of and allorl'to dr1'briefl1'-. tlrenspral'r',itha solutiotr polvcthylene -1000 alcohol(5 in 100t.An hour later.examtne plate the in -ely'co1 of UV underlong-u'avelength light: the chrontatogram thc Testsozoneat an Rr value gr-een-blue fluorescent /ationexhibitsan interlse spot at RFvalue ol'siltbin),and a gray-bJue of about0.5 lprcsen,:e to conespondin-s a spotobsenedin the chrotnatogram of about0.,1. of the Stattdartlsolutiort.The chromatogramof LheTest solution green-blue zore at an may exhibit other coloredzones:an inten-\e and a red-orange o.f Q.25Qtresenr;c stltthrLstint R1 r'alueof abor-rl o/ zone ai an.Rr velueof about0.3 (presence ta,ri.folin'). -sil)'chtisun. timesof the pcaks1br siil'dianin. B: The retention A. silybin A. siiy'binB, isosily,bin and isosill'binB in the chromitogram of tl-relcst solutiort correspondto ihose in the clror.r.lotolr"nl tl'teMilk lhistle stttndortlsolutitttr.as obtairledin the of
lc:1 for Cuttlt rtl o.[ .rilttttrtt'irt.

NIilk Thistle
'l-lristle of consisls the driedripe fruit of Silvbwn " Mrlk (l-.) the tnoriortLuTt Gaertn.(Fam. Aster-accac). pappus ltot of It havingbeenremoved. contains iessthan2.)cic ) a s i l y r n a r i n c a i c u l a t e d s s i l y b i n ( C : s H : : O r r , o.n t h e . dried basis.
Packaging and storage-Preset've in u'ell-closed c()ntriller\. protected flonr light and tncrislute. larbe states the Latin brnomial and. follou'ing the I I-abeling-The o{ficial narle. the pan of t}re planl conlained in the artrclc. USP Referencc standnrds (l l)-L1SP Povdcrcd trlill; Tltistle E.t' t r u c t R S . U S P S i l v h i n R . t . U . l P S i / r z / l o r r u rR 5 . R o l a n i e c h a r a t ' l t ' t ' i s il t s M u c r o . s t o l t i c T h c f r u i t s ( a c i t c t t c s )a r e e l o n g a t e d o v o i d . s l i g h t l l c r o o k c d . s o m e r . l ' h a ti ' l a t t c n e d . r o u - l h l 1 ' 6 t o 7 n r l t r i n l e n g t h . L r p t c t s 3 r n n t i n u , i d t h . a n d l . - 5 n r n r i n t h i c k n e s s .a n c l l . i a v ea p r o . i L - e t i r tc r - r l c t i l a g i n o u s g l o s s 1 ' r ' e l l o r . v i s h d g e o n t h c L t p p c rs u r f a c c a n d a g r o o v e d hilunr at thc basc. Thc lrLrjt coal is glossv brou n black or lttat grayb r o r v n u ' i t h c l a r l . o r p a l e g l u y s t r e a k s t i t c l t c l o s c r st h e s t r a i g h t e n l bryo har,ing t$'o thick. flattcned cotllcdons thrl contain {'attv oil a n d a l c L r r t r n eq r a n L r l c s .

not countdoes Nlicrobial cnunreration(202l)-The totalbacterial coun! nloldsand yeasts cfu exceed10.000 per g. the total conrbined oI thc doesnot excecdI 00 cfu per g. and it nleets reqr:irernents me coli ancl ol testsfor the absence Salminellospecies Erc'harichia and at!reu.\. of for absence Slnplttlococ{Lt.t Foreign orsanic matter (561): not nrorethan 2.092. o T o t a l a s h ( 5 6 I ) : n o t r r o r e t h a n 8 . 0 9 i .d c t c r n r i n e d n 1 0 g o f lvlilk Thistlc. finely por',dered the Pcsticideresidues(561): rlreets requtretttents. l\liik Thrsde Loss on drf ing (73l)-Dry 1.0-go1'fincll' pou'ciereci of at 105" lbr 2 hours:it losesnot trore tharlE.O?o its r'reight' Ileavr nretals(23l): not more than 0.(X)l%. Content of sill'nrarinnetltanrixture u'ater" of anclclcgassei-l A ,Soltrtiott I.lsea fllterecl acid (80 : 20 : 0.5). nol. and phoslrlroric tllixture of metbanol' ancidcgassecl B-tlsc a fllrerecl Solt.rtiorr acid (80 : 20 : 0.5). phosphoric water.ar.rd A nrirturer>fSolu/irtn nd Solutiorl Mohilc phu.sc Use a vari:rble tI ad.iusttnc-ltts .tt.t/crir. N'lakc IJ rs direcicdin the Chrrtmtttogrrtphit (seeSr'.r1cirr Ch,r,rrtrtlrt.qrtrtth.v \6ll)) Suitubiiitt nt'tclcl ncce ssar\'

UsP32

Dierury Monographs/ Ntilk Thistle

1055

culatethe contentof silym:rrin. percentage, the portionof Milk in in IIilk rlistle stantkrnl solutiort Dissolve an rccurately weighed 'I'histle takenby addingthe individualpercenta_qes. quantityof USP PowdcredNlilk Thistie Extr:rctRS in methanol. to and dilute lvith methanol obtain a solt-tsonicltetirr l0 minutes, of concentration about0.7 rng of extrilctper don hlving a kno',vn and ml-. Transt-cr1 mL of this solution to a 5-mL volumetric f-lask, dillrte lvith methanolto volume. Passthrougha membranctllter u h u ri n l u u . - 1 5 m o r l ' i n c rp o r u . i t \ . Silybin standttrtl solutions-Dissolve an trccuratelyweighcd and diltrtewith methanol of ctuantit) L'SPSillbin RS in methanol, > Powdered Milk Thistle is Milk Thistle reducedto a of havingknorvnconcentrations about0.20 mg to obtrin solutions not fine or very fine powder.It contains less than2.0Va Pass mg per mL, 0.01 mg per mL, and 0.00'1 per mL, respectively. of silymarin,calculatedas silybin (C:.II::Oro),on the or l membrlne iilt.'r having a 0.:15-ptm finer porosity. ihrough dried basis. Tesr solutiott-Transferabout l0 g of tinely powderedNIilk Thisweighed.to an ertractionthimble.and cover with a tle, accLlretelv proin well-closed contrtiners, the cotton ball. Transf'er thimble to a continuoLls-e,\traction Packaging and storage-Presen'e sma.ll tected fiom light and moisture. iitted $ith a 250-mL round-bottomflask containing aDDaratus Labeling-The label statesthe Latin binomial and. tbllowing the on and heat the f-lask a heatingmantle nL 1-50 ol solventhexane. ofTicial name, the pa-rtof the plant source tiom which the article ibr rl hours. Following the extraction, separatethe round-bottom rvas derived. apparaextractlrom the extraction solventhexane flaskcontaining Remole the adherent tus.:rnddiscardthe solventheranesoltttion. 'lIilk Tltistle ErUSP Reference standards (11)-USP Povr.tlered thimble by drying,and transl-er trctct RS. USP SiL1bin solrenthexanefrom the extraction RS. LTP Siltdiurtin RS. suitable hot ertractionand tbr apparatus thethimbleto an ertraction is Botanic characteristics-The por.r,'der characterizedby fragfitted l'ith a 250-mL round-bottomflask containing100mL of 75 mentsof colorless cells,up to approximately pm in lengthand 8 if acetate.[NoTE-Adjust the volume ol ethy'lacetate. neceseth.yl um in width, from the palisadelayer of the epiderrnisof the tn-rit Heat the flaskon a heating extraction.] a to sustain continllous sary. pigmentlayer (they assume rcd color in wall. with their adherent a mantleto allow the solventto rellux gently. After 8 hours, transi-er chloral hydrate preparations),and by gray pieces. viewed iiom into a 100-mL volumetrictlask, dilute rhe extract quantitatively above. r.r'iththe siitlike lumen produced by the pronounced wall to with methanol volume.and mix. Transfer1.0mL of this solution thickenin-g the nodeson the cell *'all formed by the thickened or 25-mL volumetric ilask. and dilute wirh methanolto volume. to a ridges; fragmentsof the pigment layer viervediiorn above.with red t s ChronttLog rap h i c s-.,rem (see Ch r omatographv (621))-The liqcolorationditfusing out of them in chloral hvdrate prl'prrttions. and a:1.6rvith a 28S-nmdetector is uid chromatograph equipped parenchymal pigmentcellsalternating with colorless cells:consprc5-gm packingL1 ard is mainmm x 15-cmcolumn that.'ontains prgment lls are visice uouslystippled colorless cells throughr.vhich oi tainedat a temperature 40'. The i-lorvrate is about 1.0 ml. per or ble when viewed from abovelciear-shaped monocliniccalciurn is minute.The chromatograph programniedas follorvs. tiagments of'cells:nLlrncrous oxalateprisms.lying tiee or in groups cells of the seedcoat. up to rou-ehly of the lemon yellow palisade 150 pm in length.having a very narrowlumen and conspicuously A SolutionB Solutiort Time stippledwhen viewed fiom above:pale ycllowish tiagmentsfiom (7c) (minrires) (cic\ Elution of ihc net cell laler together with portions the embrlo consisting of 1socratlc 15 thin-wailedcells with small glandsand lipophilic substances. 85 0-5 Heavy metals (23 l): 20 pg per g. lincrrsrrdicnt l5 -+5 5-20 85- 55 in 45 1sOcratIC Other requirements-lt meets the requirements the tests tbr 20-40 00 IderLtificatiort. Foreign organic inatter. Totul ctsh,Pesticitle resiI5 1incar gradicnt 45- 40-41 55-85 and Content of' dues, Loss on drying. Mic'robittLenurnerutiott. 15 __egL1.1ibratiorl ~ 41-55 85 sil"'tnrin under Milk Thistle. and record the Chromatosraph :Wilk thistle stuntlat'dsoLtttittnrhe peak rcsponses directcd for Proceclure.the chromatogramobas tained similarto the Rcference providedwith the Chromatogram is USPPowdered retention timcs ivlilk ThrstleExtractRS: the relative areabout0.68 fbr siiychristin.0.T3 sil-vdianin. 1.00lbr sillbin for A, 1.0-5 silybinB. 1.09tirr dehl'drosilybin, 1.15fbr isosilybinA, for and 1.19for isosilybrn the resolutron factor,R. betw'een silybin B: > Porvdered Miik Thistle Extractis preparedfiorn Nlilk A andsilr.'bin is not lessthan 1.0:the tailingfactoris not lessthan B Thistle fruits or seedsby fat reinoval and subseqlleilt 0.8 anclnottnor. than 1.0: rnrl thc relaLivr*t,rn.irr.l deviationfbr It extractionwith suitablesolvents. containsnot less lhe sum o1'peakresponses to silybin A and silvbin B is not due rlore than J.07c. than 90.0 percentand not more thtut I 10.0 percentof (aboLit $Ll of the as Procedure-Separatel,v equal';olumcs l0 the labeledamountof silymarincalcLrlated silybin inject illilk rhi.stle.stundard solLtsolution. each of the .lilr,bit stiutul(rft| (C::H::O'o), the dried basis, of consistitlg not less on ;loiri. and the fa.il .sr:lution and into the chrornatograph, record the than 20.0 percentand not more than ,15.0pc'rccntfor chromatosrrms. srlydienin. Identifl, the perks clueto silvchristin. not and silychristin. lessthan40.0 ' t l y h i n, \ . s i l l b i n B . i s o s i l y b i n . a n t i i s o s i l ; - b iR b _ 'c c m p a t i s o r t the sumof silydianin n " A 'rrth rhc --hrtrrrrrr[t)sftrnl the 1,/l/Atlti.stle percentand not more than 65.0 percentfbr the sum of and stundord solutiott. oi lneasurc peak arels of the rclcvnnipeaks.Plot tite areasof the the not lesstlian 10.0percent silybinA and silybinB. ancl \ u m o t ' s i l r b i n A a n d s i i v b i nB p e e k s e r s u s h e c o n c e n t r a t i oo f v t n and not n.Iore than 20.0 percent for titc sunl of U S PS i l y h i nR S i n t h e 5 i h L r i trrt u , ' J r t r ,t!r t ! t d i t t t ta.n d o b t a i nr r e t isosilybin and isosilybin B. A qression Lri calculate pcrcr'ntJge the St-par:rteiv line for calibration. tach relevantconrponent sillmliin es silybin (C:.il::Oro)in thc of eontxlnPackagingand storlge-Prcsene in tight. light-resisrirrtt pt-rttion Ililk Thistlernkenbv the forrnula: oi ers. in a cool place. Lahcling-The iabel statesthe Latin binonrirl end. fbilorving the l50o'l,1' otTicialname.the prrrtoi thc plrrntfrom whrch thc articlc rvas prein v;hichC is the conccntratrcrn.mg crer r!-levant commL. of thc in pared. it meets the requircmcntsfor Lubeling under Botitttit'nl E,r' ponint in the lest srtltriitn inte4rtillit.-d the celibration gt.tr.,i'i: in as tracts (-565). andltr/ ihe ueight. in g. of the ponronoi lfilk l-histletrrkcn. Celrs

PoryderedMilk Thistle

Powdered Milk Thistle Extract

:^ ~

100c of t a s d e f i n e d t l r e r e i n C a l c L r l a t e h e c o n t e n t 1c in which thc terlr. C, rs rclding the porrion of capsules talierr Lrv l;'*!.'in ,;r;;;t;: anoullts individLral