6-(Chap 5)-5 Jan Thesis Capsule

8/13/2019 6-(Chap 5)-5 Jan Thesis Capsule

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CHAPTER 5 DEVELOPMENT OF SR CAPSULES OF METOPROLOL TARTRATE

DEVELOPMENT OF SUSTAINED RELEASE CAPSULES CONTAINING

COATED MATRIX GRANULES OF METOPROLOL TARTRATE (A HIGHLY

WATER SOLUBLE DRUG) AND ITS BIOPHARMACEUTICAL EVALUATION.

Development of oral susta ne! release formulat ons for " #"l$ %ater solu&le !ru#s'onstant rate of release "as &e'ome a '"allen#e to t"e p"arma'eut 'al te'"nolo# sts( )#eneral* fast release of !ru# 'auses to+ ' t$ f t s not formulate! as e+ten!e! release !

form( Metoprolol tartrate ,MT-* a. &lo'/er &e n# " #"l$ solu&le* permea&le ,'lasssu&stan'e- s a&sor&e! 'ompletel$ t"rou#" t"e %"ole ntest nal tra'/ % t" n 0.1 "rsu&2e'te! to e+tens ve f rst pass meta&ol sm( )ts lo% & olo# 'al ava la& l t$ ,3 a&sorpt on an! el m nat on ,7.1"r- ne'ess tate t"e a!m n ster n# of 'onvent onal )Rt mes !a l$( To over'ome t" s pro&lem susta ne! release s !evelope! t"at ena&le

fre6uent !os n#( )n t"e present stu!$* matr + #ranules of MT "ave &een forme! &$ su'om& nat on of HPMC an! EC( Eu!ra# t8 RL an! RS %ere '"osen to form 'oat n# on#ranules to e+ten! !urat on of !ru# release( T"e o&2e't ves of t" s %or/ %ere , - to pan! opt m 9e susta ne! release 'apsule 'onta n n# 'oate! matr + #ranules , - to evalu p"$s 'al '"ara'ter st 's of t"e prepare! #ranules , - to elu' !ate t"e effe't of pol$'ompos t on* 'oat n# 'ompos t on an! level on t"e release / net 's , v- to stu!$ in vivo

a&sorpt on of !ru# n ra&& t mo!el(,v- to perform & oe6u valen'e stu!$ n "uman vo

5.1. Ma !"#a$% a&' ! *'%

Ma !"#a$%

Metoprolol tartrate ,:;


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5.+. P"!,a"a #*& *- % a&'a"' a$#/"a #*& 0" ! -*" a&a$2%#% *- ! *,"*$*$ a" "a !

A sto'/ solut on 'onta n n# = m#Bml of metoprolol tartrate %as prepare! &$ ! ssolv!ru# n met"anol( T" s sto'/ solut on %as furt"er ! lute! to =4G#Bml % t" met"anol( A

of t"ese solut ons "a! &een ta/en n a 6uart9 'ell an! s'anne! for ma+ n t"e ran#e of =;4


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mes" s 9e- %ere 'oate! step% se us n# a flu !.&e! 'oater ,Klatt.Po%re+* Po%re+* Kranules %ere f rst spra$ 'oate! % t" an a6ueous et"anol ' solut on of HPMC ?=44form nterme! ate la$er( T" s la$er ma/es 5 %e #"t. #a n to t"e n t al #ranules toan$ n'ompat & l t$ &et%een !ru# an! Eu!ra# t8( MT matr + #ranules ,04 %Bloa! n#- %ere a#a n 'oate! % t" &len!s of Eu!ra# t8 RS an! RL alon# % t" plasttr et"$l ' trate ,TEC- ,5 %B%- n a flu ! 9e! &e! 'oater( All ! spers ons %ere plaovern #"t( T"e follo% n# Eu!ra# t8 RS an! RL &len! rat os %ere nvest #ate! ;5 =:5 5* : (5 0(5 an! =44 4 ,%B%- an! t" s 'auses =4 %e #"t #a n n #ranules( Aftt"ese #ranules %ere mar/e! as &at'" CMK=* CMK0* CMK7* CMK1 an! CMK5( CoCMK1 &at'" "a! susta ne! t"e !ru# release for a lon#er per o! of ; "ours ,Ta&le 5(J&F #(5( (-( )n t"e ne+t step* fres"l$ prepare! CMK1 #ranules %ere 'oate! unt l a %e #

of =5* 05 an! 75 ,%B%- %as a'" eve!( e #"t #a n %as 'al'ulate! on t"e &as s of un'#ranules( T"ese formulat ons %ere mar/e! as CMK=5* CMK05 an! CMK75 respe'tT"e pro'ess parameters %ere as follo%s pre"eat n# temperature 75 C* pre"eat n# m nutes* nlet temperature of 75.14 C* outlet temperature of 70.71 C* pro!u't tempof 70 0 C* spra$ rate . 0.7 #Bm n* atom 9at on pressure.=(0 &ar* volumetr ' flo% =44 m7B"* no99le ! ameter of =(0 mm( After 'oat n# t"e #ranules %ere flu ! 9e! for an! su&se6uentl$ 'ure! for 01B1; " at 14 C an! =5 relat ve "um ! t$ ,RH- an! %s eve! ,=0


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F# . 5.1. P * *% *- %0% a#&!' "!$!a%! *a !' "a&0$!% *- ! *,"*$*$ a" "a !

5.3.+. C a"a !"#6a #*& *- %0% a#&!' "!$!a%! *a !' a "#4 "a&0$!%

F nall$ opt m 9e! #ranules %ere evaluate! for t"e r m 'romer t ' propert es su'" as pas 9e* true !ens t$* tappe! !ens t$* Carr s n!e+* an! flo% propert es( An#le of repos!ens t$ an! tappe! !ens t$ %ere !eterm ne! &$ USP W== 1X met"o!( Tappe! !ens t$ %!eterm ne! &$ us n# tappe! !ens t$ tester ,Ele'trola&* )n! a- an! Carr s n!e+ ,C)- %a'al'ulate!( True !ens t$ %as !eterm ne! us n# a Hel um !ens tometer ,S" ma!9u* ?$otoIapan-( Part 'le s 9e ! str &ut on of t"e #ranules ,CMK05- %as !eterm ne! &$ us n# stan!ar! s eves ,Ele'trola&* )n! a- an! #rap" %as plotte! % t" fre6uen'$ a#a nst s eve Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=41


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,F #(5(:-( T"e !ru# 'ontent n #ranules %as !eterm ne! &$ e+tra't n# an a''uratel$ %e #amount of po%!ere! #ranules ,=44 m#- % t" %ater( T"e solut on %as f ltere! t"rou#" 4Ym mem&rane an! a&sor&an'e %as measure! at 007 nm after su ta&le ! lut on(To furt"er '"ara'ter 9e t"e !ru# release pro'ess* t"e mean ! ssolut on t me ,MDT- an!s m lar t$ fa'tor of ! ssolut on prof les &et%een 'ommer' al pro!u't ,Metolar ZR- an!e+per mental formulat ons %ere 'al'ulate! a''or! n# to t"e follo% n# e6uat on

=

==n

=

n

=m !

[

[@tMDT *%"ere s t"e sample num&er* n s t"e num&er of t me ntervals

'ons !ere!* tm ! s t"e t me at m !po nt &et%een t an! t.=an! \ t"e a!! t onal amount of

!ru# ! ssolve! n t"e per o! of t me t an! t.=(Moore I an! Flanner H H ,Moore =::J- !evelope! t"e follo% n# e6uat on

=44@]T -,R,=Bn-^=54lo#f n

=

4(500 +=

=

%"ere n s t"e sampl n# num&er* R an! T are t"e per'ent of !ru# release! fromreferen'e an! test pro!u'ts at ea'" t me po nt ( f 0 s a measure of s m lar t$ &et%een t%! ssolut on prof les( f 0 value #reater t"an 54 represents e6u valen'e of t"e t%o 'urves an!%"en t"e per'ent error s 9ero , (e( f 0 =44- test an! !ru# referen'e prof les are !ent 'al(Metolar ZR =44. SR %as use! as referen'e su&stan'e an! CMK =5* 05 75 %ere use! as su&stan'es(Fr a& l t$ stu!$ %as also performe! follo% n# stan!ar! pro'e!ure( F ne po%!er %remove! from t"e #ranules samples &$ s"a/ n# t"e samples for 5 m n on a s eve of por

0=0 Ym( A sample of t"e res !ual 'oarse #ranules ,04 #- %as pla'e! n t"e !rum ofEr%e/a fr a&ulator ,Heusenstamm* Kerman$- rotat n# at 04 r(p(m( for =4 m n( T"eforme! !ue to t"e mpa't %as !eterm ne! an! ta/en as n!e+ of fr a& l t$( T"e tes performe! n t"e 'onvent onal* matr + an! 'oate! matr + #ranules* ea'" n tr pl 'ate( %ere s"o%n n ,Ta&le 5(5(-(5.3.3. F#$$#& *- *a !' "a&0$!% #& a a"' !$a #& a,%0$!.

Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=45


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ul/ volume ,ml- per m# of #ranule %as !eterm ne! from ts &ul/ !ens t$ ,5;4 m#Bml-(volume for ;1(7 m# of #ranule t"at 'onta ns =0(5 m# of t"e !ru# %as foun! as 4(=15 mlan mal e+per mentat on* !ose of =0(5 m# !ru# %as re6u re!( S 9e 7 'apsule ,4(7su ta&le for loa! n# ;1(7 m# of #ranule ,4(=15 ml-(Some vo ! spa'e % t" n 'apne'essar$ to ensure eas$ f ll n# an! un form t$ of !ose( To !eterm ne avera#e !ru# 'ont'apsules* J 'apsules %ere '"osen( Ea'" 'apsule %as st rre! for 01 "ours n 4(= N HCl ma#net ' st rrer to ! ssolve out t"e !ru# from #ranules as 'omplete as poss &le( Druanal$9e! &$ a spe'trop"otometer at a %avelen#t" of 007 nm( T"e !ru# 'ontent %as foun!

=0(5=4( 7 m#( S m larl$* for =44 m# !ose* J 5 m# of #ranules ,&ul/ volume =(=%ere f lle! n 'apsule s 9e 44(C"ara'ter 9at on propert es %ere s"o%n n Ta&le 5(5(Sample 'al'ulat on for opt m 9e! formulat on

544 m# un'oate! #ranules 'onta n metoprolol tartrate.=44 m#* HPMC ? =44M.=05mEt"$l 'ellulose.=05m#* DCP.==0(5m#* HPMC E5.7 (5m#(Su& 'oat n# &$ HPMC ?=44M ,5 - of 544 m# (e( 05 m#(Coat n# &$ Eu!ra# t ,05 - of 544m# (e(=05 m#(,Eu!ra# t RS. =0=(5J Eu!ra# t RLTEC s 5 of 544m# (e( 05m#(Total pol$mer ' 'oat n# = 5m#(T"erefore total %e #"t of 'oate! #ranules 'onta n n# =44m# of !ru# J 5m#

5.7. In Vitro D"0 R!$!a%! S 0'#!% -*" "a&0$!% 8# *'! &0 /!"% MG1 * MG5

T"e in vitro ! ssolut on stu! es %ere performe! us n# USP. 74 t$pe ) ! ssolut on app pa!!le t$pe at 54 rpm for t"e #ranules f lle! n 'apsules 'onta n n# e6u valent amount =of !ru#( T"e ! ssolut on me! um 'ons ste! of 4(=N "$!ro'"lor ' a' ! for f rst 0 "ours an p"osp"ate &uffer % t" pH J(; for t"e ne+t 7 to =0 "ours ,:44 mL- *an! t"e me! um ma nta ne! at 7 C 4(5 C(T" s s mulate! #astro ntest nal pH( An al 6uot ,5% t"!ra%n at spe' f ' t me ntervals an! repla'e! % t" t"e same volume of fres" me!

same temperature( T"e % t"!ra%al sample %as f ltere! t"rou#" 4(15m f lter paper( Ne+t* ts!ru# 'ontent %as !eterm ne! &$ UV.v s &le spe'trop"otometer ,S" ma!9u* ?$oto* Iap0 1 nm( )t %as as'erta ne! t"at none of t"e n#re! ents use! n t"e matr + formul

nterfere! % t" t"e assa$ of !ru# ,MT-( T"e release stu! es %ere 'on!u'te! n tr plPer'ent 'umulat ve !ru# release %as plotte! a#a nst t me of release ,F #(5(J-( Saforement one! pro'e!ure %as use! to perform release stu!$ for 'oate! #ranules 'onta n Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=4J


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Metoprolol tartrate e6u valent to =44 m#( After t"ein vitro !ru# release stu! es ,Ta&le 5(J('F #(5(;(- t"e formulat on CMK05 %as sele'te! for furt"er opt m 9at on &$ e+per

!es #n ,Response surfa'e met"o!olo#$- as t susta ne! for lon#er t me an! ts pol$meran! 'oat n# mater als rat o %ere '"osen as 'entre value(

5.5. O, # #6a #*& *- SR *a !' "a&0$!% /2 "!%,*&%! %0"-a ! ! *'*$* 2 (RSM).

E4,!"# !& a$ '!%# &

T"e opt m 9at on of susta ne! release formulat on of Metoprolol tartrate %as !one &!es #n e+pert soft%are ,!es #n e+pert tr al vers on (4(7 state n'* M nneapol

'entral 'ompos te !es #n ,CCD- % t" = %as emplo$e! as per t"e stan!ar! proto'ol,S n#"* 0445 S n#"* 0440-( ase! on preformulat on stu!$ _t"e rat o of t"e amou

HPMC ? =44MB Et"$l 'ellulose ,A- an! _t"e rat o of t"e amounts of Eu!ra# t8 RSBRL%as sele'te! as t"e n!epen!ent fa'tors* stu! e! at 7 levels ea'"( T"e 'entre po nt ,4* 4- %stu! e! n 6u ntupl 'ate( All ot"er formulat on an! pro'ess n# var a&les %ere /ept t"rou#" out t"e stu!$( Ta&le 5(0 summar 9es an a''ount of t"e =7 e+per mental runs stut"e r fa'tors 'om& nat ons* an! t"e translat on of t"e 'o!e! levels to t"e e+per mentalemplo$e! !ur n# t"e stu!$( Per'ent of !ru# release! n = "our ,rel="r - ,R =-* t me to 54 !ru#release ,t54 - ,R 0-* of !ru# release! n =4 "ours ,rel=4"r - ,R 7- %ere t"e response var a&les(

A''or! n# to !es #n matr + e+per ments %ere run an! t"e response !ata for =7 formula"a! &een ! spla$e! n ,Ta&le 5(0-(T"ese !ata %ere su&2e'te! to Response sumet"o!olo#$ &$ !es #n e+pert soft%are (Soft%are #enerate! ANOVA ,Ta&le 5(;(&.! %asuse! to o&ta n 'orrelat n# mo!el e6uat ons after !elet n# non s #n f 'ant termssoft%are treats t"ese mo!els to pro!u'e response surfa'e(


=4


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Ta/$! 5.+.F*" 0$a #*& "#a$% a""#!' *0 -*" SR *a !' "a&0$!% *- ! *,"*$*$ a" "a !

a% ,!" !4,!"# !& a$ '!%# &.

Tr al no(Co!e! fa'tor levels A

) .= .= )) .= 4 ))) .= = )V 4 .= V 4 4 V) 4 =

V)) = .= V))) = 4 )Z = = Z 4 4 Z) 4 4 Z)) 4 4 Z))) 4 4Translat on of 'o!e! levels n a'tual un tsCo!e! level .= 4 =A HPMCBEC,m#Bm#- = =(5 0

Eu!ra# t8 RSBRL 75 14 15,m#Bm#-

Sample 'al'ulat onFor ever$ =44 m# of un'oate! #ranules total amount of matr + pol$mers s 54 m#(For level .=* rat o of HPMC EC = (e( 05 m# HPMC an! 05 m# EC(For level 4* rat o of HPMC EC =(5 (e( 74 m# HPMC an! 04 m# EC(Total amount of 'oat n# mater alEu!ra# t8 ,RS an! RL- 05 of un'oate! #ranules ,544 m#-( =05m#For level = of Eu!ra# t8 pol$mers *t"e rat o of ,RS RL- 75 (e( =0=(5m# Eu!ra# t87(1 m# RL respe't vel$(For level 4* rat o of Eu!ra# t8 ,RS RL- 14 (e =0=(:5m# an! 7(45m# respe't vel$( Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=4;


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5.9. C* ,0 a #*& *- "!$!a%! :#&! # % *- SR *a !' "a&0$!% *- ! *,"*$*$ a" "a !.

? net 's of !ru# release from t"e formulat ons* CMK=5* CMK05 an! CMK75 %!eterm ne! &$ f tt n# !ata to ! fferent e6uat ons su'" as 9ero or!er ,M /t-* f rste6uat on ,lnM lnM4 /t-* H #u'" mo!el ,M /`t- an! ?orseme$er Peppas e6uat on,M /tn-( A value of n 4(5 n! 'ates 'ase ) ,F '/ an- ! ffus on* 4(5WnW= s for anomanon F '/ an- ! ffus on * n = s for Case )) transport an! nX= n! 'ates super'ase )) trM4 an! M are t"e amount of !ru# , - release! at 9ero t me an! after t me t respe't vel$* t"e release rate 'onstant an! n s t"e e+ponent( T"e me'"an sm of !ru# release %!epen!ent on t"e value of _n ,Ta&le 5(7-(Dru# release follo% n# part 'ular me'"an 2u!#e! &$ t"e l near t$ ,R 0- of plot %"en !ata from F #5(;( s f tte! to a part 'ular mo!e6uat on( Ta&le(5( ( ! spla$s t"e !ata(

Ta/$! 5.3. Va$0! *- & a&' *""!%,*&'#& ! a% *- '"0 "!$!a%!

Va$0! *- & M! a% *- '"0 "!$!a%!n 4(5 Case < ) ,F '/ an- ! ffus on or s6uare root of t me / net 's

4(5 W n W = Anomalous ,non. F '/ an- ! ffus onn = Case < )) transportn > 1 Super Case < )) transport

5.;. S 0'2 *- * ,a #/#$# 2 a&' %0"-a ! *", *$* 2 /2 #&% "0 !& a$ a&a$2%#%

C"an#e n solu& l t$* sta& l t$* ! ssolut on rate an! & oava la& l t$ of !ru#s mto ntera't on amon# t"e a't ve 'onst tuent an! na't ve e+' p ents n sol !Compat & l t$ stu!$ s a len#t"$ pro'ess an! to evaluate t"e 'ompat & l t$ stu!$ t mont"s or $ears( So t s ne'essar$ to '"oose a fast met"o! for t"e evaluat on of sol ! sta& l t$ %" '" prov !es rel a&le nformat on a&out t"e poss &le ntera't on( Amo!ern te'"n 6ues su'" as Four er transform )nfrare! ,FT)R-* D fferent al S'annCalor metr$ ,DSC- an! Z.ra$ D ffra'trometr$ ,ZRD- 'an &e use! to perform 'ompat &stu!$( T"e sele'te! formulat on ,C=- %as su&2e'te! to FT)R* ZRD an! DSC( Surfmorp"olo#$ of !osa#e form ,'oate! #ranules* C=- %as stu! e! &$ S'ann n# Ele'tM 'ros'op$(

5.;.1. F*0"#!" T"a&%-*" < I&-"a"!' (FTIR)


=4:


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T"e 'ompat & l t$ &et%een !ru# an! pol$mer %as as'erta ne! &$ FT)R stu!$ on t"e pol$mers* !ru#.pol$mer p"$s 'al m +ture an! ne% formulat on ,C=- to nfspe'trop"otometer stu!$ ,Per/ n Elmer* Mo!el.Para#on =444* Sr(No(.7:0J=-( T" s st!ete'ts an$ '"an#e n fun't onal #roups of ea'" su&stan'e( efore 'arr$ n# out FT)R anal$

pure !ru#s* pol$mers* !ru#.pol$mers %ere /ept at room temperature at 55C for 7 %ee/s(F ve m# of t"e !ru# or sample %as ta/en an! t"orou#"l$ tr turate! % t" =44 m# of potas &rom !e( T"e po%!er %as 'ompresse! at 04 ps for =4 m n on ? r press( A pellet %asout of t"e m + an! ntro!u'e! nto t"e nstrument( T"e spe'trum %as re'or!e! n 544.1'm .= an! s"o%n n F #( 5(=0(

5.;.+. X< Ra2 '#--"a #*& (XRD)

ZRD anal$s s %as 'on!u'te! for metoprolol tartrate* pol$mers an! formulate! SR #ranu%" '" %ere f nel$ tr turate! to #et f ne po%!er &efore ta/ n# t"e s'an( A R #a/u M nra$ ! ffra'tometer* Z.ra$ #enerator an! ! ffra'tometer 'ontrol %ere emplo$e! to stu!$ ZRT"e #enerator operat n# volta#e an! 'urrent %ere 74 /V an! =5 mA respe't vel$( Ts'ann n# spee! %as = Bm n an! t"e 0 s'ann n# ran#e %as from 5 to J5 (Result s F #(5(=7(

5.;.3. D#--!"!& #a$ S a&& Ca$*"# ! "2 (DSC)

DSC stu! es %as 'on!u'te! for pure !ru#s* pol$mer* !ru# pol$mer m +ture an! n'ompos t on of SR #ranules of metoprolol tartrate us n# a Per/ n Elmer DSC.74 ! fferent as'ann n# 'alor meter % t" TA 1444 t"ermal anal$s s 'ontroller ,Mettler* S% t9erlansamples %ere run from .=44 to =14 C * at a "eat n# rate of =4 C per m nute* us n# a l 6n tro#en 'ool n# a''essor$ an! a n tro#en DSC 'ell pur#e( T"e result s ! spla$e! n F #

5.;.7. S 0'2 *- %0"-a ! *", *$* 2 /2 S a&& E$! "*& M# "*% *,2 (SEM)

T"e SEM stu!$ s use! to '"e'/ t"e surfa'e te+ture of matr + ta&let* #ranules or an$ sosurfa'e( A smoot" surfa'e # ves un form !ru# release %"ere as uneven or 'ra'/e! surfa'# ves un'ontrolle! an! non.un form !ru# release( Kranules %ere remove! from ! ssolut on apparatus at pre!eterm ne! t me ntervals an! t"e samples for SEM %ere p &$ l #"tl$ spr n/l n# t"e #ranules on a !ou&le.s !e! a!"es ve tape stu'/ to a &rass stu&stu&s %ere t"en 'oate! % t" #ol! to a t" '/ness of 3=;4.044 b un!er a n tro#en atmosp"


==4


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us n# a #ol! sputter mo!ule n a " #" va'uum ,=4.0 to=4


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evaluate t"e ev !en'e of to+ ' t$ 0; !a$s s"ort term su&'"ron ' to+ ' t$ stu!$ %as e+e'a''or! n# to OECD #u !el nes(

M! *'

Ra&'* #6a #*&? &0 /!"#& a&' "*0,#& *- a a$%@

Fort$ e #"t Al& no rats ,a#e! .; %ee/s- (e( 01 male an! 01 female "ealt"$ rats %ere !nto four #roups of J rats per se+ (e( four !ose #roups re'e v n# t"e !ose of 4 m#B

m#B/#* 74 m#B/# an! J4 m#B/#( An mals %ere allo%e! a''l mat on per o! of la&orator$ 'on! t ons pr or to t"e n t at on of !os n#( Ea'" 'a#e 'onta ne! J rats of tse+ % t" a &e!! n# of "us/ an! =0 " l #"tB!ar/ '$'les %ere prov !e!( T"e n! v !ual a%as fur mar/e! % t" p 'r ' a' !( T"e females %ere null parous an! not pre#nant( Pellfee! an! %ater %ere # venad libitum ( An mals %ere /ept n an mal "ouse ma nta ne! atemperature of 00 C 0 C an! a relat ve "um ! t$ of 55 5 ( T"e stu!$ proto'olapprove! &$ t"e )nst tut onal An mal Et" 'al Comm ttee( ,Re# strat onCPCSEABRCPB044;B==J:-( Ta&le 5(=7( ! spla$s allo'at on of an mals nto ! fferent

0% #-# a #*& -*" %!$! #*& *- A$/#&* % "a#& "a % -*" ! % 0'2I. One of t"e ro!ent spe' es re'ommen!e! as test s$stem for t"e use n to+ ' t$ stu! es

II. T" s test s$stem "as &een !emonstrate! to &e sens t ve to to+ ns(III. !el$ use! t"rou#"out n!ustr$ for t"e evaluat on of to+ ' t$ of var ous pro!u'ts(IV. Referen'es prov !e !ata an! ev !en'e(T"e oral route %as sele'te! for !ru# a!m n strat on &e'ause*

I. T"e !osa#e 'an &e a''uratel$ a!m n stere!(II. One of t"e propose! routes for to+ ' t$ test n#(

III. Oral route s 'ons !ere! to &e a potent al route of "uman e+posure(

P"!,a"a #*& *- T!% a !"#a$

T"e test formulat on ,C=- %as 'rus"e! to po%!er &$ mortar an! pestle an! ! spense! suspens on us n# =44ml pur f e! %ater so t"at t"e 'on'entrat on of t"e pure !ru# suspens on %as =m#Bml( T"e HPMC pol$mers n t"e formulat on a'te! as suspen! n#On ea'" !a$ of stu!$ fres"l$ prepare! suspens on %as a!m n stere! to t"e rats &et%een a(m( for 0; !a$s. T"e !ru# %as t"en a!m n stere! to rats at t"e !ose levels of =5m#B/


==0


13/72


74m#B/# an! J4 m#B/# n t"e !ose volume of =mlB =44# &o!$ %e #"t( T"e 'ontrol an mals%ere fe! % t" ve" 'le onl$(

O/%!" a #*&%

All an mals %ere o&serve! !a l$ for 'l n 'al s #ns( T"e t me of onset* ntens t$ anof t"ese s$mptoms* f an$* %ere re'or!e!( All an mals %ere o&serve! t% 'e !amortal t$ !ur n# t"e per o! of t"e stu!$( T"e %e #"t of ea'" rat %as re'or!e! on f rst !a$"r an! at %ee/l$ ntervals t"rou#"out t"e 'ourse of t"e stu!$( _Mean &o!$ %e #"t of #roup %as 'al'ulate!( T"e 6uant t$ of foo! 'onsume! &$ #roups 'ons st n# of s + rats %as re'or!e! %ee/l$ an! t"e foo! 'onsumpt on per rat %as 'al'ulate! for &ot" t"e _'ontro#roup an! _#roups fe! % t" !ose ( Ta&le 5(=1 Ta&le 5(=5 ! spla$ &o!$ %e #"t of

an! t"e r foo! 'onsumpt on(

T!" #&a$ % 0'#!%

La/*"a *"2 #& !% # a #*&%

Follo% n# nvest #at ons %ere 'arr e! out pr or to sa'r f 'e on 'omplet on of !os n#0; !a$s n t"e an mals t"at %ere faste! over.n #"t( loo! samples %ere 'olle'te! from or&s nus on t"e ne+t morn n# &$ us n# "epar n as ant 'oa#ulant(

Ha! a *$* # a$ #& !% # a #*&%T"e "ematolo# 'al parameters "emo#lo& n 'on'entrat on ,H&-* mean 'orpus'ular vol,MCV-* total er$t"ro'$te 'ount ,R C-* ret 'ulo'$te 'on'entrat on ,Rt-* mean 'orpus'ula"emo#lo& n ,MCH-* "emato'r t ,HCT-* an! total an! ! fferent al leu'o'$te 'ount %!eterm ne! us n# "ematolo#$ anal$9er ,Rm! Me! a !s Pvt( Lt!(* Ne% Del" * )n! a-( S%as o&ta ne! &$ 'entr fu# n# at 7444 rpm for =4 m n an! supernatants %ere ta!eterm ne serum #lu'ose* total prote n* al&um n an! &loo! urea n tro#en* so! potass um %ere anal$9e! us n# autoanal$9er ,Rm! Me! a !s Pvt( Lt!(* Ne% Del" * )n! a T"e plasma a't v t es of serum al/al ne p"osp"atase ,SAP-* serum #lutam ' p$transam nase ,SKPT- an! serum #lutam ' o+aloa'et ' transam nase ,SKOT- %ere !eterm ne!(Follo% n# _"ematolo# 'al parameters %ere stu! e! &$ t"e use of S$sma+ < ?05Counter an! results %ere ta&ulate! n Ta&le 5(=Ja(H& Hemo#lo& n ,#m - Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

==7


14/72


R t Ret 'ulo'$te , -Platelets ,Z=45 B'mm-

C " te loo! Corpus'les ,Z=47 Bmm7

B#* ! # a$ #& !% # a #*&%

Follo% n# _& o'"em 'al parameters %ere stu! e! &$ t"e use of Ro&on / ASP.744(Total serum prote n ,#m -@

UN loo! urea N tro#en ,m# -SKPT Serum Klutam ' P$ruv ' Transam nase ,)UBL-SKOT Serum Klutam ' O+aloa'et ' Transam nase ,)UBL-Results %ere ta&ulate! n Ta&le 5(= a(N! "*,%2

All an mals %ere sa'r f 'e! on !a$ 0:* us n# C40 asp"$+ at on te'"n 6ue ,&o!$ %e #"tsment one! n t"e ta&le se't on are fast n# &o!$ %e #"ts-( Ne'rops$ of all an mals %aout an! t"e %e #"ts of t"e follo% n# or#ans %ere re'or!e! L ver* / !ne$s an! "eartor#an %e #"ts %ere re'or!e! as a&solute values an! t"e r relat ve values , (e( per 'ent &o!$ %e #"t- %ere 'al'ulate!(Results %ere ta&ulate! n Ta&le 5(=;a(A0 *,%2 a&' #% *,a *$* 2

o!$ of anaest"et 9e! an mal %as ! sse'te! an! all or#ans %ere o&serve! ma'ros'op 'a

an! sele'te! or#ans %ere e+' se! an! %e #"ts of or#ans %ere re'or!e! an! '"an#e n &o%e #"t %as 'al'ulate!( T"en t"ese or#ans %ere f +e! n =4 &uffere! neutral formsolut on( Paraff n se't ons %ere prepare! an! sta ne! % t" "emato+$l n an! eos" stopat"olo# 'al e+am nat on( All sl !es %ere e+am ne! &$ a pat"olo# st(Follo% n# t ssue samples of or#ans from 'ontrol an! an mals treate! % t" t"e " #"eslevel of J4 m#B/# %ere preserve! n =4 formal n for " stopat"olo# 'al e+am nat onA!renals* "eart* / !ne$s* l ver* lun#s an! stoma'"(A!renals* "eart* / !ne$s* l ver* lun#s an! stoma'" of _lo% an! nterme! ate !ose #an mals %ere preserve! for poss &le " stopat"olo# 'al e+am nat on* n '" stopat"olo# 'al e+am nat on of " #" !ose #roup an mals s n! 'at ve of a&noasso' ate! % t" t"e treatment(F #(5(=J( ! spla$s " stopat"olo# 'al se't ons of !or#ans(

S a #% # a$ ! a$0a #*&


==1


15/72


1 #roups of =0 rats ,J male J female- ea'" %ere ass #ne! four treatments (e( 'ontrol*!ose* m !!le !ose an! " #" !ose( After 0; !a$s all pat"olo# 'al an! & o'"em str$ parame"a! &een evaluate! stat st 'all$( Anal$s s of var at on ,ANOVA- ena&les to 'ompameans of t"ree or more var a&les( T"e test 'ompares t"e var at on ,var an'e- n t"e &et%een treatments % t" t"ose % t" n treatments( T"e rat o of var at ons ,var anmean &et%een treatments % t" t"ose % t" n treatments !eterm nes t"e F value( Ta&l5(= & an! 5(=;& ! spla$ ANOVA t"at "elp to stu!$ stat st 'al s #n f 'an'e(

5.1 . BIOPHARMACEUTICAL EVALUATION ON RABBITS

T"e in vivo stu!$ %as performe! a''or! n# to t"e #u !el nes approve! &$ t"e Comm ttee_Purpose of Control an! Superv s on of E+per ments on An mals ,CPCSEA- * M

So' al Iust 'e an! Empo%erment* Kovernment of )n! a( T"ein vivo stu! es %ere 'on!u'te!n "ealt"$ male al& no ra&& ts %e #" n# 0(0 to 0(5 /#( Ra&& ts %ere /ept for =

an mal "ouse to a''l mat 9e t"em an! fe! a f +e! stan!ar! ! et( T%elve ra&& ts %ere nto 0 #roups of J ea'" an! %ere faste! for 01 "ours( T"e f rst #roup %as fe! % t" =0((e( e+a'tl$ "alf of Metolar 05 m# ta&let ,C pla l m te!* )n! a- an! SR 'apsules e6u v

=0(5 m# of MT %as fe! to t"e se'on! #roup of an mals( ater %as allo%e!ad libitum !ur n#fast n# an! t"rou#"out t"e e+per ment( T"e formulat ons %ere a!m n stere! to t"e r

% t" an oral 'annula( T"e ra&& ts s%allo%e! t"e formulat on % t"out an$ ! ff 'u'apsules %ere put &e" n! t"e ton#ue to prevent 'rus" n# !ue to & t n#( loo! samples %ere % t"!ra%n 'arefull$ from t"e mar# nal ear ve n an! transferre! nto "epar'entr fu#e tu&es after a!m n strat on of !ose at 4(5* =* =(5* 0* 7* 1* 5* J* ;* =4"ours !ur n# t"e stu!$( T"e total &loo! 'olle't on from a ra&& t ! ! not e+'ee! mu'" mt"an ma+ mum safe &lee! (e( J(5. (5 mlB/# &o!$ %e #"t ,M '"ael*0440% t"!ra%n &loo! samples %ere transferre! to a ser es of #ra!uate! 'entr fu#e t'onta n n# 4(1 ml of 0(5 %Bv so! um ' trate solut on( T"e samples %ere 'entr fu#e!rpm for 5 m nutes( T"e plasma %as transferre! nto anot"er set of sample tu&es an! funt l assa$e!( One plasma sample ,% t"out !ose of !ru#- %as /ept as &lan/( T"e samplef ltere! t"rou#" 4(05.Ym mem&rane f lter ,M ll pore-( T"e MT 'on'entrat on n &loosamples %as anal$9e! us n# HPLC a''or! n# to t"e follo% n# met"o!( A total 044 Y plasma sample %as m +e! % t" = mL of a'eton tr le* an! 'entr fu#e!( T"e supernataevaporate! un!er n tro#en stream* an! t"e res !ue %as ! ssolve! % t" 744 YL of t"e Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

==5


16/72


mo& le p"ase an! t"en t %as n2e'te! nto t"e HPLC 'olumn> a reverse.p"ase 'olumn ,AM* 1(Jc=54 mm* dMC* Iapan- %as use!( UV !ete'tor %as use! to anal$9e( A m +tu p"osp"ate &uffer ,pH 7* 'onta n n# 4(5 tr et"$lam ne- met"anol a'eton tr le ,:4run as mo& le p"ase at a flo% rate of =(1 mLBm n ,Re$nol!s* =::;-( Ta&le 5(=:(! p"arma'o/ net ' parameters n ra&& t mo!el( F #(5(=; ! spla$s )V)VC 'orrelat on(

P a" a *:#&! # a&' % a #% # a$ a&a$2%#%

Plasma 'on'entrat on %as plotte! a#a nst t me n F # 5(= (T"e ma+ mum 'on'entrat on ,Cma+- an! t"e t me to rea'" t"e ma+ mum 'on'entrat on ,Tma+- %ere ! re'tl$o&ta ne! from t"e o&serve! values T"e area un!er t"e 'urve upto 01 " after a!m n str,AUC- %as 'al'ulate! &$ t"e trape9o !al rule from t"e o&serve! values( Stat st 'al ana

%as performe! &$ t"e Stu!ent s t test an! PW4(45 %as use! to n! 'ate stat st 'al s #n

5.11 . BIOE UIVALENCE STUDY OF SR COATED GRANULES CONTAINING

1 MG METOPROLOL TARTRATE IN COMPARISON OF REFERENCE

PRODUCT ON HEALTHY HUMAN VOLUNTEERS

T"e a m an! o&2e't ve of t"e present stu!$ %as to evaluate t"e p"arma'o/ net ' paraman! to 'ompare t"e oral & oava la& l t$ of s n#le !ose _SR 'oate! #ranules e6u valent to =44m# metoprolol tartrate ,TEST su&stan'e. C=- % t" t"e follo% n# REFERENCE su&stMetolar ZR =44. SR ta&lets of C pla lt!(* )n! a

( No( < 4:=0 4Mf#( Date Iune 044:E+p( Date Ma$ 044==#) E # % "! #!8 ,"* !'0"!

T"e in-vivo stu!$ %as 'arr e! out as per #u !el nes of DCK) ,Dru#s Control Keneral of )n Ne% Del" -( T"ese #u !el nes met t"e re6u rements of t"e U(S( Co!e of Fe!eral Re#u

,T tle 0=* Part 5J-* t"e De'larat ons of Hels n/ an! t"e Cana! an MRC Ku !el n proto'ol an! t"e nforme! 'onsent form from t"e volunteers %ere su&m tte! to _)nst tut onal Et" 'al Comm ttee of Ia!avpur Un vers t$* )n! a pr or to t"e n tstu!$( T"e stu!$ %as not starte! unt l t"e approval of t"e Et" 'al Comm ttee "a! &re'e ve!(


==J


17/72


##) D!%# & *- !4,!"# !&

T"e stu!$ proto'ol %as approve! &$ t"e )nst tut onal Et" 'al Comm ttee of Ia!avUn vers t$* ?ol/ata* )n! a pr or to t"e 'ommen'ement of t"e stu!$( T"e & oe6u valen'of TEST preparat on an! REFERENCE preparat on %as assesse! ut l 9 n# a t$p per o!* ran!om 9e!* t%o.%a$ 'omplete 'rossover !es #n n J "ealt"$* male* "volunteers( T"ere %as 0 !os n# sess ons % t" a %as"out per o! of !a$s &et%een sess ons( All t"e volunteers part ' pate! n t%o !os n# sess ons( )n ea'" !os n# svolunteers re'e ve! e t"er t"e test preparat on or referen'e preparat ons onl$ on t"e s!a$* as per t"e ran!om 9at on 'o!e( Des #n of e+per ment %as s"o%n n Ta&le 5(04(t"e volunteers nor _t"e p"$s ' an an! nurs n# staff n '"ar#e of t"e 'l n 'al aspe'ts*

nforme! re#ar! n# t"e se6uen'e of a!m n strat on( T"e )nvest #ator prepar n# t"e !r

a!m n strat on s t"e onl$ person %"o %as a%are of t"e 'o!e( Total s + male non smo/ n#volunteers %ere enrolle! for t"e stu!$ &ase! on t"e r la&orator$ tests ,serum '"em"ematolo#$* an! ur ne anal$s s-* me! 'al " stor$* p"$s 'al e+am nat on an! H)V s' No al'o"ol or 'on'om tant me! 'at on %as allo%e! 0 "rs pr or to t"e n t al a!m nof !ose an! for t"e ent re 'ourse of t"e stu!$( Su&2e'ts faste! for =0 "rs pr or a!m n strat on of !ru#s( Formulat ons %ere # ven to t"e volunteers a''or! n# to t"e pr

loo! samples %ere o&ta ne! at seventeen t me po nts from pre !ose ,4 "r- unt l 01 "r!ose ,4* 4(5* =* =(5*0* 0(5* 7* 7(5*1* ;* =4*=0* =;* 01-( T"e plasma samples %e.04 C unt l assa$e!( T"e p"arma'o/ net ' parameters for metoprolol tartrate %!eterm ne! &$ _9ero.moment non 'ompartmental met"o! ( Results %ere s"o%n n Ta&

5(00( T"e ma+ mum plasma 'on'entrat on ,Cma+- an! t me to rea'" ma+ mum plasma'on'entrat on ,Tma+- %ere ! re'tl$ o&ta ne! from t"e plasma 'on'entrat on vs( t me !ata( Aun!er t"e plasma 'on'entrat on


18/72


sem .lo#ar t"m #rap" paper an! Ct s last 6uant f a&le 'on'entrat on( T"e el m nat on "a,t=B0- %as 'al'ulate! as 4(J:7B? e D"0 a' #% "a #*& * ! 0 a& *$0& !!"% T"e volunteers %ere ran!om 9e! on t"e prev ous !a$ of P"ase )( )n P"ase )* ea'" volunteer re'e ve! e t"er t"eTEST preparat on ort"e REFERENCE preparat ons as a s n#le !ose at a f +e! t me( )n P"ase ))* t" s or!erreverse! as per t"e ran!om 9at on( For t"e purpose of a''urate sampl n# t me for all samstu!$ me! 'at ons %ere a!m n stere! at ntervals of 0 m nute to #roups of 0 su&2e'tsme! 'at on %as # ven % t" 014 ml %ater at room temperature(B$**' *$$! #*& -"* 0 a& *$0& !!"%@All t"e volunteers %ere assem&le! at J(44 a(m(on t"e stu!$ !a$ = of ea'" sess on* after overn #"t fast n# of at least =4 "rs( T"e r TP%ere re'or!e! an! an n!%ell n# ntravenous 'annula %as ntro!u'e! % t" str 't ase

pre'aut ons n t"e ant 'u& tal ve n for &loo! 'olle't on( T"e volunteers re'e ve! e t"estu!$ preparat ons ,REFERENCE B TEST- a''or! n# to t"e r 'o!e nos( % t" 014.ml( of %ater(T"e e+a't 'lo'/ t me %as 'al'ulate! a''or! n# to t"e !ru# a!m n strat on s'"e!ule( T"e f rs &loo! sample ,t 4- %as 'olle'te! mme! atel$ pr or to !ru# a!m n strat on( T"e e+a't of 'olle't on of all &loo! samples %as re'or!e! an! reporte! for ea'" su&2e't( An$ !ev atfrom t"e sampl n# s'"e!ule %as re'or!e! n t"e su&2e't s sampl n# t me s"eet( A total &loo! samples %ere 'olle'te! from ant 'u& tal ve n at 4 "r( ,&efore !ru# a!m n strat=(4* =(5* 0(4* 0(5* 7(4* J(4* :(4* =0(4* =5(4* =;(4 an! 01(4 "r n 'o!e!* 'entr f'onta n n#EDTA( loo! samples %ere 'entr fu#e! mme! atel$* t"e plasma separate! !upl 'ate pol$prop$lene tu&es 'onta n n#EDTA an! store! n a !eep free9er ma nta ne! at . O, # 0 "!$!a%! ,"*-#$! *- ! *,"*$*$ a" "a !

R!$!a%! ,"*,!" 2 Ra& !T me for 54 release 5(5.J Release n =4 "ours ;5.:4

5.17.5. Va$#'a #*& *- RSM "!%0$ % *- SR *a !' a "#4 "a&0$!% *- ! *,"*$*$ a" "a !.

Response surfa'e met"o!olo#$ #ave us ; solut ons( T"e results of t"e p"$s 'al evaluatan! 'apsule assa$ %ere foun! % t" n l m ts( Ta&le 5(=4 l sts t"e 'ompos t on* t"e r

an! e+per mental values of all response var a&les an! t"e per'enta#e error(


=14


41/72


Ta/$! 5.1 . C ! :,*#& -*" 0$a #*&% *- ! *, # #6!' SR *a !' a "#4 "a&0$!% *-

! *,"*$*$ a" "a ! (CMG+5) a&' a$#'a #*& *- !4,!"# !& a$ '!%# &.

T"e formulat on C= an! CJ as s"o%n n Ta&le 5(=4( pro!u'e! least error( T"e formulaC= %as '"osen as t"e &est opt m 9e! formula for SR 'oate! matr + #ranules of metoprololtartrate as t"e error %as m n mum an! rat o of pol$mers s least( T"e 'ompos t onformula s # ven &elo%(Ta/$! 5.11. C* ,*%# #*& *- -#&a$ *, # #6!' SR a "#4 "a&0$!% *- ! *,"*$*$ a" "a !

1 (C1)

Ma "#4 #& "!'#!& % A *0& ? K C*a #& #& "!'#!& % A *0& ? (K)a

MT =44 04 HPMC ? =44M 05 5HPMC ? =44M =05 05 Eu!ra# t8 ,RS RL- ,=0=(5J 7(11- 05Et"$l 'ellulose =05 05 TEC 05 5DCP ==0(5 00(5 Total = 5

HPMC E5 7 (5 (5 Sum of ma n n#re! ents 'oat n# n#re!J 5 m#(Total 544 =44

a* per'ent on t"e &as s of 'ore #ranule* (e(,05B544-@=44 5(Rat o of Eu!ra# t8 RS RL =0=(5J 7(11 75(775.17.9. S a/#$# 2 % 0'2


S.N*. HPMC

1 M EC

E0'"a #

J RS RL

5 K R!$!a%! #& 1 "%

P"!'# !' E4,!"# !& a$

K

E""*" P"!'# !' E4,!"# !& a$

K

E""*"C= =(45 75(1 5(:: 5(J; .5(=; :4(44 ;:(;1 .4(=;C0 =(=4 75(7 5(:: 5(5J . (=; :4(44 ;;(=1 .0(4C7 =(=4 75(0 5(: 5(55 . (41:4(44 :4(=5 4(=C1 =(05 75(5 J(44 5(51 . (J ;:(;: ;:(JJ .4(0JC5 =( 7 7J(J J(44 5(J7 .J(= ; (71 ; (5; 4(0

CJ =( 7 7J(5 J(44 5(:; .4(77 ; (7 ; (J0 4(0:C =( 7 7J(5 J(44 5(J0 .J(77 ; (7: ; (=; .4(01C; =( 1 7 (1 J(44 5(J0 .J(77 ; (4 ; (04 4(=5

=1=


42/72


43/72


%i".5.#&. %!' spectra of metoprolol tartrate (a)* +,- /#00- ( ) et$1l cellulose (c)+,- 25 (d) E0'"a # J S(e) E0'"a # J 3(f) po4der mi ture(")S "ranules #

($).

X


44/72


T"e !ru#.pol$mer an! !ru#.e+' p ent 'ompat & l t$ %as as'erta ne! &$ 'arr$ n# out t"stu!$ of t"e follo% n# metoprolol tartrate(a), HPMC K100M (b) ethyl cellul !e (c) HPMC"# (d) Eu!ra# t8 $%(e) Eu!ra# t8 $L(&) p 'de mi tu e(g) ptimi*ed %$ g anule! (h).T"e spe'trums o&ta ne! "ave &een llustrate! n F #(5(=7a to 5(=7"( No !ru#.po

ntera't on %as foun!(

%i".5.# 7 D %,! "0 % of metoprolol tartrate (a)* +,- /#00- ( ) et$1l cellulose

(c) +,- 25 (d) E0'"a # J S(e) E0'"a # J 3(f) po4der mi ture(")S "ranules

# ($).

D#--!"!& #a$ S a&& Ca$*"# ! "2 (DSC) % 0'2 *- *a !' "a&0$!% *& a#& 1

! *,"*$*$ a" "a !

)ntera't ons n t"e sample are !er ve! or !e!u'e! from DSC &$ '"an#es n t"e t"ermevents* su'" as el m nat on of an en!ot"erm or e+ot"erm pea/* or appearan'e of a ne% Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=15


45/72


Ho%ever* some &roa!en n# of pea/s lea! n# to '"an#es n pea/ temperature o''urs s!ue to m + n# of t"e 'omponents % t"out n! 'at n# an$ s #n f 'ant ntera't on( )f afeatures more or less rema n t"e same* 'ompat & l t$ 'an &e e+pe'te! as %ell as t"er'omple+ format on( T"e ! fferent al s'ann n# 'alor metr$ ,DSC- pattern of pure metoprolol tartrate(a), HPMC K100M (b) ethyl cellul !e (c) HPMC "# (d) Eu!ra# t8$%(e) Eu!ra# t8$L(&) p 'de mi tu e(g) ptimi*ed %$ g anule! (h) are # ven n F #(5(=1ato 5(=1"( )t %as o&serve! t"at t"e DSC pattern of t"e pure !ru# ! ! not alter n SR 'oa#ranules(

%i".5.#8 DSC T !" * "a % of metoprolol tartrate (a)* +,- /#00- ( ) et$1l

cellulose (c) +,- 25 (d) E0'"a # J S(e) E0'"a # J 3(f) po4der mi ture(")S

"ranules # ($).5.17.=. S0"-a ! *,* "a, 2 *- SR *a !' "a&0$!% *- ! *,"*$*$ a" "a ! /2 % a&&

!$! "*& # "*% *,! (SEM)

S'ann n# ele'tron m 'ros'op$ ,SEM- %as performe! to !eterm ne t"e surfa'e topo#rap"$ SR #ranules( F #(5(=5 s"o%s p"oto#rap" of #ranule ,C=- &$ s'ann n# ele'tron m 'ro%" '" &u the c n&i med b th di&&u!i n and e !i n mechani!m! t be pe ati+e du ing d ug


=1J


46/72


elea!e & m the ptimi*ed batch & %$ g anule! (C=). %"M ph t mic g aph & the g anule!ta en at di&&e ent time inte +al! a&te the di!! luti n e pe iment !h 'ed that p e! had been

& med th ugh ut the mat i . %"M ph t mic g aph! & g anule !u &ace at di&&e ent time

inte +al! al! !h 'ed that the e !i n & p lyme inc ea!ed 'ith time. %"M ph t mic g aph

& the !u &ace & & e!h g anule! (F #(5(=5a) did n t !h ' much p e!. Ph t mic g aph!e+ealed p e! 'ith inc ea!ing diamete at -, #, and 10 h u !. he!e ph t mic g aph! al!

e+ealed & mati n & gelling !t uctu e indicating the p !!ibility & !'elling & mat i

g anule! ( F #(5(=5c d). Hence, the & mati n & b th gelling !t uctu e and p e! in g anule!indicate the in+ l+ement & b th e !i n and di&&u!i n mechani!m! t be e!p n!ible &

!u!taining the elea!e & M & m the & mulati n.

%i".5.#5. S2- p$otomicro"rap$s of optimi9ed S "ranules ( C1 ) s$o4in" surface

morp$olo"1 after 0 $ours (a)* & $ours ( )* 5 $ours (c)* and #0 $ours (d) of dissolution

stud1.

5.15. SUBCHRONIC ORAL TOXICITY STUDY (+= DAYS) ON RATSSu&'"ron ' oral to+ ' t$ stu!$ %" '" s man!ator$ for reformulate! !ru# &eforein vivo stu!$%as a''ompl s"e!( T"e allo'at on of 1; rats %" '" %e #" n# =44 #m to =54 #m %as tr

n 1 ! fferent !oses n'lu! n# 'ontrol #roup s s"o%n n Ta&le 5(=7(


=1


47/72


Ta/$! 5.13. A$$* a #*& *- a a$% * a"#*0% "*0,%

C$# a$ %# &% *- "a %@ A$$ a a$% (7=)%ere free of nto+ 'at n# s #ns t"rou#"out t"e !os per o! of 0; !a$s( M*" a$# 2 *- a a$%@Male an! Female an mals from 'ontrol an! ! fferent !ose #roupssurv ve! t"rou#" t"e !os n# per o! = of 0; !a$s n! 'at n# no mortal t$ an! !ru# safet$(Va"#a #*& *- /*'2 8!# *- !4,!"# !& a$ "a % @

ot" male an! female an mals from 'ontrol an! t"e ! fferent !ose #roups e+" & te! no &o!$ %e #"t #a n t"rou#"out t"e !os n# per o! of 0; !a$s as llustrate! n Ta&le5(=1(

Ta/$! 5.17.G"*0, !a& /*'2 8!# ( ) *- a a$%

G".

N*.

D*%!

:

D a 2

; 17 +1 +=

) Control ==4(1JJ(5

==0(J71(;

==;( ; (07

==:(71J(14 =04(0;5(;0

)) =5==0(0:J(1

5==7(=15(J

;== (5J (=

0=04(501(5

=00(=05(17


=1;


48/72


))) 74 =4;(;JJ(71

=4:(J (15

==7(10J(5J

==;(7=5(=0 ==:(=;1(5=

)V J4 =4 (J:J(J0

=4;(0 5(71

==:(7=5(;=

==5(4;1(5 =07(705(0=

,p W4(45-

F**' C*&%0 , #*& *- E4,!"# !& a$ a a$% (Ma$! a&' F! a$!)@ Dur n# t"e !os n# per o! an! at term nat on t"e 6uant t$ of foo! 'onsume! &$ &ot" mfemale an mals from ! fferent !ose #roups %as foun! to &e 'ompara&le % t" t"at of 'oan mals %" '" s ta&ulate! n Ta&le 5(=5(

Ta/$! 5.15. G"*0, !a& -**' *&%0 , #*& ( a a$)

G". N*.D*%!

:

D a 2

; 17 +1 +=

) Control Mean =5(7 =(: =J(= =(; =J(1 =( = (4 =(; = (; =(J)) =5 Mean =1(0 =(1 =5(= =(5 =J(4 =(5 =J(7 0(4 =J(: =(5))) 74 Mean =1( 0(= =5(4 =(1 =5(7 =(7 =J(4 0(0 =J(5 =(7)V J4 Mean =7( =(: =1(4 =(7 =1(5 =(0 =5(: =(: =J(0 =(=

,p W4(45-

Ha! a *$* # a$ #& !% # a #*&% *- /$**' ,a"a ! !"% Ta/$! 5.1=a 1=/ T"e "aematlo# 'al parameters of male an! female . at term nat on of !os n# on !a$ 0:t"* nos #n f 'ant '"an#es %ere o&serve! n t"e values of ! fferent parameters stu! e! ,Ta&le%"en 'ompare! % t" 'ontrols an! values o&ta ne! %ere % t" n normal & olo#la&orator$ l m ts(T"e ANOVA %as appl e! to evaluate t"e stat st 'al ! fferen'e of t"e mean "aematolo# parameters amon# four n!epen!ent !ru# treatments( S n'e t"e value for F o&ta ne!

present e+per ment s less t"an t"e re'or!e! value* t %as 'on'lu!e! t"at t"e ! fferen &et%een t"e treatment means s not s #n f 'ant ,pW4(45-(Ta/$! 5.19a. G"*0, !a& a! a *$* 2 *- a a$%

G". N*. D*%! : H/ ( K)P$a !$! %

(1 5 3)R ( K )

T* a$ WBC

( 4 1 3 3)


=1:


49/72


) Control =J(= 4(;; (74(7; =(J4(=J (4J4(1;)) =5 =J(404(1: (154(70 =(5=4(= (4;4(04))) 74 =J(7:4( = (554(7 =(5;4(=; J(:; 4(11

)V J4 =5(:14(7J (114(17 =(=;4(=5 (044(J;

Ta/$! 5.19/.ANOVA *- H! a *$* # a$ ,a"a ! !"% *- 7 '#--!"!& "*0,% (N*. *-

%0/Q! %? 7=) 0&'!" %0/ "* *4# # 2 % 0'#!%

Na ! *- !

,a"a ! !"

M!a& % 0a"!

(/! 8!!&

T"!a !& )

M!a& % 0a"!

(8# #&

"!a !& )

F< Va$0!

S a #% # a$

%# -# a& !

Hemo#lo& n 4(1J; 4(1=0 =(=7; NS

Platelets

, + =45Bmm7-

4(==; 4(=14 4(;7; NS

Rt , - 4(40J 4(407 =(=70 NSTotal C

, + =47Bmm7-4(=4 4(07 4(1= NS

B#* ! # a$ I& !% # a #*&% *- a$! a&' -! a$! a a$% 0&'!" %0/ "* *4# # 2 % 0'2 Ma$! a&' F! a$! At term nat on on !a$ 0:* all & o'"em 'al parameters stu! e! (e( Total serum prote n

SKOT* UN %ere foun! to &e 'ompara&le % t" 'ontrols an! %ere % t" n t"e n & olo# 'al an! la&orator$ l m ts( T"e ANOVA %as appl e! to evaluate t"e sta! fferen'e of t"e mean & o'"em 'al parameters amon# four n!epen!ent !ru# treatmeS n'e t"e value for F o&ta ne! n t"e present e+per ment for ! fferent & o'"em 'al pa

s lesser t"an t"e re'or!e! value* ) 'on'lu!e t"at t"e ! fferen'e &et%een t"e treatment meas not s #n f 'ant ,pW4(45-(

Ta/$! 5.1;a. G"*0, !a& /$**' ! #% "2

G". N*. D*%! ( : )

T* a$ S!"0

P"* !#&

( K)

BUN

( K)

SGPT

( IU L )

SGOT

( IU L )

) Control (=14(1; =;(5;0(J= =;(4;=(=J 01(=:=(4)) =5 J(:; 4(74 = (J=(J= = (77=(=5 01( 04(;5

))) 74 (44(11 = (1==(4; = (J=(07 05(4J=(4J


=54


50/72


)V J4 (04(J; = (5=(4 = (;7=(5: 00( J=(4

Ta/$! 5.1;/. ANOVA *- /#* ! # a$ ,a"a ! !"% *- 7 '#--!"!& "*0,% (N*. *- a a$%?

7=) 0&'!" %0/ "* *4# # 2 % 0'#!%.

Na ! *- !,a"a ! !"

M!a&

% 0a"!(/! 8!!&

"!a !& )

M!a&

% 0a"!(8# #&

T"!a !& )

F< Va$0! S a #% # a$%# -# a& !

Total serum prote n

4(=4=;7(=:= 4(445

NS

UN ,m# - 7(1 0(;: =(=:; NSSKPT =(=; =(J : 4( 4 NSSKOT =0(700 =7( 4(;:1 NS

Va"#a #*& *- *" a& 8!# % *- a$! a&' -! a$! a a$% 0&'!" %0/ "* *4# # 2 % 0'2

(5.+ A 5.+ B)

Ma$! a&' F! a$! T"e an mals from 'ontrol an! t"e ! fferent !ose #roups e+" & te! normal or#an %e #"t"e sa'r f 'e at 0:t" !a$( T"e ANOVA %as appl e! to evaluate t"e stat st 'al ! fferen'e of tmean %e #"t of v tal or#ans amon# four n!epen!ent !ru# treatments( S n'e t"e valueo&ta ne! n t"e present e+per ment for ! fferent or#an %e #"ts s less t"an t"e r

value* %e 'on'lu!e t"at t"e ! fferen'e &et%een t"e treatment means s not s #n f,pW4(45-Ta/$! 5.1=a G"*0, !a& "!$a # ! a$0!% (K) *- '#--!"!& *" a& 8!# % #& '#--!"!&

"*0,%.


G"*0, N*.D*%! : L# !"

M!a& SD

( )

#'&!2%

M!a& SD

( )

H!a"

M!a& SD

( )) Control 1(J: 4(70 4(;5 4(41 4(5; 4(47

)) =5 1( ; 4(05 4(;1 4(4J 4(J4 4(41

))) 74 1(;J 4(01 4(; 4(47 4(J= 4(41

)V J4 1(:5 4(01 4(; 4(47 4(5: 4(47

=5=


51/72


Ta/$! 5.1=/ ANOVA *- #% *$* # a$ ,a"a ! !"% *- 7 '#--!"!& "*0,%.

Na ! *-

!

,a"a ! !"

N*.*-

%0/Q! %

M!a&

% 0a"!(/! 8!!&

T"!a !& )

M!a&

% 0a"!(8# #

&

T"!a !& )

F


52/72


)t %as o&serve! t"at t"e an mals fe! % t" t"e formulat on ,C=- suspens on %ere "ealtunusual '"an#es n &e"av or or n lo'omotor a't v t$* no ata+ a* an! no s #ns of n%ere o&serve! !ur n# t"e 0;.!a$ per o!( No ! fferen'es %ere foun! n #ro%t" &et%e

'ontrol #roup an! t"e test an mals( T"e foo! 'onsumpt on of male an! female rats of 'ontan! e+per mental #roups %as s m lar* n! 'at n# t"at t"e fee! nta/e an! ut l 9at oaffe'te!(

O&servat ons of #ross pat"olo#$ mme! atel$ after ! sse't on* on rats of all #roupfoun! to &e un forml$ "ealt"$* s"o% n# no apparent pat"olo# 'al a&normaH stopat"olo# 'al e+am nat on of t"e l ver an! / !ne$s n t"e 'ontrol an! t"e test #s"o%e! no ! fferen'es* n! 'at n# t"at formulat on ! ! not result n an$ a!verse to+ 'oeffe't on t"ese or#ans ,F #(5(=J a.f-( Haematolo# 'al parameters* "emo#lo& n 'on'entratotal an! ! fferent al er$t"ro'$te 'ount* total an! ! fferent al leu'o'$te 'ount* "emato'r t*mean 'ell "emo#lo& n 'on'entrat on* n &ot" 'ontrol an! e+per mental rats* n! 'as #n f 'ant ! fferen'e ,P W 4(45- &et%een t"e t%o #roups( All values %ere foun! to &t"e normal ran#e for rats , r 2* =: - an! t"ere %ere no ! fferen'es &et%een t"e #roups( T"elevels of plasma anal$tes* su'" as total prote n* &loo! urea n tro#en , UN-* #l & l ru& n* al&um n* 'reat ne* '"olesterol* '"lor !e* 'al' um* p"osp"orus* so

potass um ons %ere not s #n f 'antl$ ! fferent &et%een t"e 'ontrol an! t"e e+per#roups of rats ,PW4(45-( No s #n f 'ant ! fferen'es %ere o&serve! n en9$me a' &et%een t"e 'ontrol an! test an mals( No a&normal '"an#es %ere o&serve! n or#an% t" respe't to &o!$ mass of t"e test rats n 'ompar son % t" 'ontrols(

5.19. BIOPHARMACEUTICAL EVALUATION OF SR CAPSULES OF

METOPROLOL TARTRATE ( 1+.5 ) ON RABBITS

T"e opt m 9e! 'oate! #ranules f lle! n 'apsules %ere nten!e! for tsin vivo test n ra&& t

mo!el( Pla!ma c ncent ati n and pha mac inetic pa amete ! a&te al admini!t ati n && mulated %$ c ated mat i g anule! and c n+enti nal tablet 'e e !umma i*ed in ig.#.1

and able #.12. 3 !u!tained bl d le+el & M 'a! e+ident a&te al admini!t ati n & the

c n+enti nal & mulati n. he & mulated mat i g anule! !h 'ed !igni&icantly l 'e C ma

than c n+enti nal tablet (p40.0#) and e5ui ed !igni&icantly m e time t each C ma (tma

0(1 4(5 h u !) a! c mpa ed 'ith c n+enti nal tablet! (t ma 4(; 4(7 h u !). T"e area un!er t"e Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=57


53/72


'urve ,AUC- n'rease! from =05: ==0(J n#("rBmL to 0=0 (; 07=(J n#("rB mL ,nearlt mes more- for SR 'apsules( Values of Cma+ an! tma+ 'learl$ n! 'ate! t"at t"e !ru# release%as susta ne! to a&out =0 "ours after oral a!m n strat on n ra&& ts ,n J-( T"e formulat on maintained c n!tant pla!ma c ncent ati n up t ab ut 10 h u !. hemaintenance & pla!ma c ncent ati n & l nge du ati n a&te admini!t ati n & ne'

& mulati n indicated it6! !m th and e tended in vivo ab! pti n. In vitro-in vivo

c elati n 'a! dete mined by pl tting a g aph ( ig.#.17) !h 'ing the & acti n & d ug

ab! bed in vivo +e !u! the & acti n & d ug elea!edin vitro . 8 high +alue & c elati n

c e&&icient (- 9 0.2:;:) !ugge!ted g d c elati n bet'een in vitro-in vivo data. he

!light +a iati n in IVIVC may be att ibuted t the di&&e ence in .0, >.7, .# at +a i u! !tage!, cecum >.>, c l n .- and that

& human i!= !t mach 1 -, du denum #.#,!mall inte!tine > , di!tal ileum 7,c l n .#

,E!%ar!*=::=-( hu! n the ba!i! & the c mpa ati+e !tudy &


54/72


0 1 2 3 4 5 6 7 8 9 10 11 120

50

100

150

200

T#(! #& )*0"%

M ! a & , $ a % a . * & . ! &

" a # * & * - ! * , " *

$ * $ a "

" a !

( & 5 D

L )

F# .5.1;. P"*-#$! % *8 !a& ,$a% a *& !& "a #*& *- ! *,"*$*$ a" "a ! a a#&% # !?

-*$$*8#& *"a$ a' #% "a #*& *- MT * "a//# %. Data are represented as mean SD (n =

6). ( ) mmediate "!$!a%! a/$! *- MT ( ) Ca,%0$! *& a#& %0% a#&!' "!$!a%!coated

matri "ranules of -! .

y = 0.9175x

R2 = 0.9434

0

0.2

0.4

0.6

0.8

1

1.2

0 0.2 0.4 0.6 0.8 1 1.2

Fraction of r!g release

F r a c

t i o n

o f

r !

g a

b s o r

b e

F# .5.1=. F"a #*& *- '"0 a/%*"/!' !"%0% -"a #*& *- '"0 "!$!a%! ( invitro- invivo

*""!$a #*& -*" ! *, # #6!' -*" 0$a #*&? C1).

5.1;. PILOT BIOE UIVALENCE STUDY OF SR COATED GRANULES IN

HEALTHY HUMAN VOLUNTEERS

S!$! #*& *- *$0& !!"%


=55


55/72


Ta/$! 5.+ . D!%# & *- /#*a a#$a/#$# 2 % 0'2 *- SR *a !' "a&0$!% *& a#& 1

! *,"*$*$ a" "a ! /* -*" REFERENCE (A) a&' TEST %a ,$!

(CMG+5 C1)(B).

Mo!e of treatment

S0/Q! N*. P!"#*' I P!"#*' II

= A0 A7 A1 A5 AJ A

P"*'0 I&-*" a #*&%

1) REFERENCE sample ,A-Metolar ZR =44. SR ta&lets of C pla lt!(* )n! a

( No( < 4:=0 4Manufa'tur n# !ate Iune 044:E+p r$ !ate Ma$ 044==+) TEST sample , -

SR 'oate! #ranules 'onta n n# metoprolol tartrate =44 m#(

Manufa'tur n# !ate Ma$ 044:

Manufa'ture! at Department of P"arma'eut 'al Te'"nolo#$* Ia!avpur Un vers t$* ?ol/at44 470* )n! a(

at'" No( C=S 0'2 '!%# &

T" s %as a s n#le !ose* ran!om 9e!* t%o treatment an! t%o.%a$ 'ross over stu!$* %as"out per o! of !a$s &et%een t"e t%o !os n# sess ons( )n ea'" !os n# sess on* vore'e ve! e t"er of t"e TEST or t"e REFERENCE preparat on onl$ on t"e stu!$ !a$ at a ft me(

Dates of t"e loo! Colle't on for p"arma'o/ net ' prof les ,01 "r-SESS)ON ) 7=(45(04=4SESS)ON )) 4;(4J(04=4

Va$#'a #*& *- HPLC ! *' -*" !% # a #*& *- ! *,"*$*$ #& 0 a& ,$a% a@ Development of susta ne! release !ru# !el ver$ s$stems an! ts & op"arma'eut 'al evaluat on

=5J


56/72


A " #".performan'e reverse!.p"ase l 6u ! '"romato#rap" ' met"o! for 6uant f 'at ometoprolol tartrate ,MT- n "uman plasma s use! a''or! n# to met"o! !es'r &e! n l ter

,A6 l* 044 -( A C=; 'olumn %as use! % t" a'eton tr le


57/72


un!er plasma 'on'entrat on t me 'urve up to nf n t$ ,AUC4. - == (;55 = (47J ,Ta&le5(01(-(A!m n strat on of t"e REFERENCE preparat on pro!u'e! t"e plasma el m natl fe* ,t=B0- J(10= 4(==J "r %"ereas a!m n strat on of t"e TEST preparat on pro!u'e plasma el m nat on "alf.l fe ,t=B0- 5(=15 =(=41 "r( A!m n strat on of t"e REFEREN preparat on pro!u'e! t"e plasma el m nat on 'onstant ,? el- 4(=4; 4(440"r .= %"ereasa!m n strat on of t"e TEST preparat on pro!u'e! t"e plasma el m nat on 'onstant ,/ el- 4(=14

4(474 ="r .=( On t"e &as s of 'ompar son of t"e AUC4. * t"e relat ve & oava la& l t$metoprolol tartrate n t"e TEST preparat on %as ::(0J to t"at of t"e REFEREN preparat on(T"e a!vanta#e of ne% formulat on s t"at t s n 'apsule form %" '" fle+ &le t"an ta&let !osa#e form(S a #% # a$ #&-!"!& ! -*" ,#$* /#*! 0# a$!& % 0'2

ANOVA ,Su&2e't* per o!* treatment- %as appl e! to t"e Cma+* lnCma+* AUC4.t an! lnAUC4.

values( No s #n f 'ant ! fferen'e %as foun! stat st 'all$ for t"e treatments* su&2e't anvalues of Cma+* ln Cma+* AUC4.t* an! ln AUC4.t* AUC4. * an! ln AUC4. ( Ta&le5(0; s"o%s'onf !en'e l m t an! ANOVA ta&le(A' !"%! "!a #*&%

None of t"e volunteers 'ompla ne! of an$ a!verse rea't on on t"e p"arma'o/ net ' prof!a$s(


=5;


58/72


Ta/$! 5.+1. B#*a a#$a/#$# 2 % 0'2 *- SR *a !' "a&0$!% *& a#& 1 ! *,"*$*$ a" "a !@

P$a% a *& !& "a #*& *- REFERENCE ,"!,a"a #*& -*" 9 *$0& !!"%.

V*$0& !!"

N*.

T# ! ( ")

.5 1. 1.5 + +.5 3 3.5 7 =. 1 . 1+ 1= +7Plasma 'on'entrat on ,n#Bml-

1 4 J7(07 (=4 :;(0= ::(=7 == (44 =04(=4=0=(04 == (:4 J5(07 5=( ; 14(=0 =0(;4 7(=4

+4 J7(07 (=4

=40(=0 =47(04 =0=(=4 =00(=4

=07(=4 ==J(=4 JJ(04 50(=4 1=(=4 =7(04 0(:4

34 JJ(14 5(0=

=40(04 =41(44 == (44 =0=(=0

=07(04 =47(04 J (=0 54(=0 7:(=0 =0(;4 0(;0

74 J5(;4 1(=4 :;(=4 =44(;4 ==J(74 =0=(44

=00(74 ==;( 4 J;(0= 5=( ; 14(=0 =0(;4 7(=4

54 J=(04 5(14 ::(=4 ::(=7 == (44 =04(=4

=0=(04 == (:4 J (07 54(=0 7:(=0 =0(04 0(

94 J5(=0 1(04 : (=4 :;(0= ==;(=4 =0=(=4

==:(74 ==;(04 J1(74 1:(70 7 (0= ==(:; 0(:4

M!a&

4 J1(=J 5(50 ::(1 =44( 5 == ( 5 =04(:0

=0=(

0 ==5(77 JJ(7; 54(; 7:(1 =0(J7 0(:0S.D. 4 =(:J =(77 0(=; 0(7; =( 1 4( 5 =(1 J(4= =(11 =(=J =(77 4(15C.V. 4 7(45 =( J 0(=: 0(7J =(1; 4(J0 =(0= 5(0= 0(=J 0(0 7(7 7(5

Development of susta ne! release !ru# !el ver$ s$stems an! t s & op"arma'eut 'al evaluat on

=5:


59/72


60/72


Ta/$! 5.+3. B#*a a#$a/#$# 2 % 0'2 *- SR *a !' "a&0$!% *& a#& 1 ! *,"*$*$ a" "a !@ P a" a *:#&! #

,a"a ! !"% *- ! *,"*$*$ a" "a ! #& REFERENCE (A) a&' TEST (B) ,"!,a"a #*&%.

Development of susta ne! release !ru# !el ver$ s$stems an! t s & op"arma'eut 'al evaluat on

V*$0& !!"

N*.

!$

( "


61/72


Ta/$! 5.+7. B#*a a#$a/#$# 2 % 0'2 *- SR *a !' "a&0$!% *& a#& 1 ! *,"*$*$

a" "a !@ P a" a *:#&! # ,a"a ! !"% *- ! *,"*$*$ a" "a ! 8# !

REFERENCE (A) a&' TEST (B) ,"!,a"a #*&.

P a" a *:#&! # ,a"a ! !"%

REFERENCE

P"!,a"a #*& (A)

M!a& S.D

TEST P"!,a"a #*&

(B)

M!a& S.D

C a4 (& $) =00(4= 4(:;7 =07(:54 =(007a4 ( ") 7(1= 4(041 7(777 4(05;

AUC < (& . ". $) == (45J =5(:41 == 0(:J0 =1(01JAUC < (& . " $) ==;J(J77 =J(45 == (;55 = (47J

: !$ ( "


62/72


4 4 4 44(5 J5(;44 ,,J5(; 4-B0-@,4(5.4- =J(15 =J(15= 1(=44 ,, 1(= J5(;-B0-@,=.4(5- 71(: 5 ,=J(15 71(: 5- 5=(

=(5 :;(=44 17(45 :1(1 50 =44(;44 1:( 05 =11(0

0(5 ==J(744 51(0 5 =:;(1 57(4 =0=(444 5:(705 05 (;7(5 =00(744 J4(;05 7=;(J051(4 ==;( 44 J4(05 7 ;(; 51(5 =45(444 55(:05 171(;5(4 :;(=04 54( ; 1;5(5;J(4 5(0=4 ;J(JJ5 5 0(015

(4 J(0=4 5( = J1 (:55;(4 J;(0=4 0(0= 04(=J5=4(4 5=( ;4 ==:(:: ;14(=55=0(4 14(=04 :=(: :70(455=; =0(;44 =5;( J =4:4(;=501 7(=44 1 ( ==7;(5=5

1;(4 =(= 54(1 ==;;(:=5

F# . 5.+ . P$a% a '"0 *& !& "a #*& (& $) *- *$0& !!" N*.1 %. # ! ( )

No% for 'al'ulat on of el m nat on rate 'onstant last s + plasma 'on'entrat ons 'ons !ere! as s"o%n n Ta&le 5(0J( Ne+t ln 'on'entrat on !ru# vs( t me %as plotte! 5(0=(- an! t"e slope s 'ons !ere! el m nat on rate 'onstant e 4(=4;(

Ta/$! 5.+9. La% %#4 ,$a% a '"0 *& !& "a #*&% *- *$0& !!" N*.1


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F# . 5.+1.D! !" #&a #*& *- !$ *- a/$! -*" 0$a #*&

AUC4. %as 'al'ulate! &$ t"e formula

AUC4. AUC4.t CtB/ el ==;;(:=5 =(=B4(=4; ==::(4:F nall$* t=B0 %as 'al'ulate! a''or! n# to t"e follo% n# formula t=B0 4(J:7B4(=4; J(7JJ

Ta/$! 5.+;. > K *&-#'!& ! #& !" a$ -*" ! *,"*$*$ a" "a ! 8# ! TEST a&'

REFERENCE ,"!,a"a #*&.

C a4 Untransforme! Data =(44J .=(401:

ln transforme! Data =(44=1.=(445=

AUC < Untransforme! Data 4(:;77.=(44:J


T# !( ) P$a a '"0 *& . (& $) $& P$a% a '"0 *& .J(0= 1(7771:0J;;

; J;(0= 1(0005:==;0=4 5=( ; 7(:1 447: 1=0 14(=0 7(J:=; 1:J7=; =0(; 0(51:115= =01 7(= =(=7=140===1; =(= 4(4:57=4=;

=J1


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ln transforme! Data 4(:: J.=(44=7

AUC < Untransforme! Data 4(: J:.=(44;0

ln transforme! Data 4(::J .=(44=0

Ta/$! 5.+=. ANOVA %0 a"2 -*" , a" a *:#&! # ,a"a ! !"% *- ! *,"*$*$ a" "a !

Pa"a ! !" S*0" ! '.-. S.S M.S.S. F P

Cma+ Su&2e'tsTreatmentPer o!Error Total

5==1==

J(: J==(0=774(;5771(1 7707(5=J

=(7:57==(0=774(;577=(==;7

=(01=4(40J;4( J74

N(SS

N(S....


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lnCma+ Su&2e'tsTreatmentPer o!Error Total

5==1==

4(44454(4444(444=4(44474(44=J

4(444=4(4444(444=4(444=

=(05;1=4(=4J=4( 14

N(SS

N(S

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6-(Chap 5)-5 Jan Thesis Capsule