The document discusses the methodologies and experiments conducted by the Department of Pharmacy at the University of Rajshahi, focusing on the bioavailability and pharmacokinetics of various substances. It highlights the importance of establishing specific protocols to evaluate drug efficacy and safety. The paper emphasizes the need for rigorous scientific inquiry in pharmacological studies.

1. Written By
Department of Pharmacy
University of Rajshahi
Rajshahi-6205, Bangladesh

2. ~ G ic(>.,S~I L'
';/-' -
0
Q>iocu~v ry-e,rer~ +o tQ rnef:lSWQ.O'Qn- A : ·
I ,, ,_. J - • )~II ri'--
0t
.
~~~ lreccvn.Je. po1-e-ncy ~ ~d Pil'ry of Q.ClTfl~lclS ?t dekrmini~ ·.
pokvitt d} !~ 6.mcJ.H--s1 'Y01_,u.-t'f-ed +o ~ocl~c~-9:~----~~-~-±~.~ .~ffed ·
·.m,fU"'~~I by O"Y °': .su.L-(1',laQ 1e~r o..n~ffio..~ cgq- _fFF'ts&f!r . u.'()de'( sJ-~
Lt.~'-'i"'~ ®di. h(jy)s_. hll experrnm~ ()__f'frms ncu.de _rn_ce .1"f:lh...
VV- NYV.1"1A..,..,. I) b• _ • _J Yahb 1 k , CVh do~<; e}e .
(U..q,NA(V--(1 ~ ~I ~ ~
~~°'n s- Cl ~pi<'.OJ biom9'.y pyoneMe. 1-k 'reS><m~e ~.
vt ~·~(p_ ~ ~ch 1'ndiviclvc:J +est cn1imcd 1s i no·ed und thz ·o-e.SJ..<.ll 1~
,V(m"'1/)' ObtuinQ..c whe.'() Cl ~e.oie5 ~ u.n(110 ..) ro..Q.. ~9'o3ec-ed +o
.Stah.>hct'.A (tY)oJy~1·s +o ealettlcd-Q th.rL roeD..T' ac.f1viby, .S.-CArdrold
eYOY QJC · .
' . .
-/GenQicUj 1
d"6Vi5 g,_ ~nno~
1
bJl Q<;.s°:'f ed _by (::>~y~ico..l
00 Chtn"A c.oJ YYQ.-Yods ()JQ, ,_ 0..'>~ CA"f ed bj b10 L~ I ccJ JrQfhOOS. . '
Dll-1- b oL~ Ic.oJ. 00 s ~ i<. •'.DQYe hr!lP <'.;m5vm.:..~..
I m cne exy~'(~ive. i-~ (',h,tm 1 c.i.c nw-h0cl c,,_~d ess f¥&1se-
I .---
• 1
I'DJ ~wtpo-sg_s o·)· 2., oo..s~o.'I ';.
,.. ~t{~I .CD 1o deJeolflN.. +~ P.--(u-roo.e.o~,ca.l a.ehv1+y <Sf .....
' ' ()e,W ~~1ld l ~l)s--n) ' .
( ~ ® lo d..e.tec~ thR. Q <ff rrew Cfim~t1d (il>,-t>') ~
@ lo t,Q;-eooni..n-e 1-N.· ci2-e.:. Cild d03e 1n}eovoJ..· ···
® 'Toxi' ~-.1Y o(- .env.1Y@ ~OtR p 0)00..n·h:, ~n o.Q
dU-.ecl--ed .-Gy b OLt~~ CL~f- · .
-··-·····----- ..--______ 1·---------··--·--- - ---- -·------ -- ---------- -·-----·-----·: -- -
(~ioa..s.so-y and Micro6o'.011co.' fh~°'y-l(Oj
l'-1D. Shaht:n ~on(Q).
.?_:_r;- 13>o.:-ch
1 / 18
Md.
Imran
Nur
Manik

3. 1 ·
--
~(i. rd -r t>e· ~Y"!. Y'llA__-1cm p ~-e~cy,
(y_;. C~i.)1 po.rlo.hvc b iuClHO-y 'o 0 n -'rii5 biOU~S(}...Y ' .. the.
be d.eJeoYA.Ln~d (n bJ'crLec1'coJ <;yst-em
0 :-re.ftQ"f en Q..Q s.tv"Yd ())-Ld ·
i uch-1iry
. -. e t¥u
(£Y / i1 f,oLtd-c Go o.sso.-:J ~ · ,
g ·11 :--V._> b 0 0-_,S~ 'J > . +n..e.
~~=====-~-=Co.::n bQ ckhmt-ned n ~l,o91>_1 c.CA <;y s--em ~
~ o-he:~ SbndCU.d Su.b~--o:DC:QS
~~- LD~D' ~D~0 ~tc·
......--
··~
C?> o c....~~o..y ~
0 o et.~ s °"Y cY~:n k9 ~ cl~ ~~J . .
/f eqp.ld
if.AA) ~· rL ~ · ....
Y ~ ri fVi .Q.. J.> u 'f J - :
'(n 1-oi.)- .
lwo ~f pes f't ~ Yi¥-
.:1 •
(0 10 de}A'.:"fntlytL -the_ olol~ic~ 00- hnn'c {2ft~c~···-·6t
i e11dc:i}tY1 Q].Af) ~v bsJo.n~ es- - !
-1
® GL6Vd JUvel umd Jw5 uol-ivi{y ye/af-icmshi P am
: b Q we.Yl>'i n eci 'oj vi'cJlLSSCLJ .
! @ 3"~ dOJn QQ_ VJ)ed fcrc --Yu deJen"'i"~6Yl a
i ?w~ -h,eyeMe, '*'iv() ckpend5nce . ~
(§) 9.- ,~ N)ed +v <k~ e -h2_ Q,e.ti "'TI ~ .
I • d
; ;_x~hn~ Y9i~.
2 / 18
Md.
Imran
Nur
Manik

4. I! 0..Y
....
---~L°& d ase
Fi X) ~ 6-yo..Qe.d b60--s.su.y
-----------·-·-~ ---···-
1
I
i
I·
• s Y&""}e_)
'I u."'~'T nR.t~H"(Yl~tr.[f.;_
-·I
' tn e.oCZC.SC?
n thQ
-ee.n -hQ..
3 / 18
Md.
Imran
Nur
Manik

5. co, ~~io~''to..-
010 · ~ 'CA~mo.. ITT
0- ~u.~ J;; o- bo1 lG.-J·1l.
I·.•
~ o--~ CAJ;{{d,_ -.;Yit't:/YO-b-i'ol~1·~ N}<Nj,..:_
@) ,. . . '
. ~ °'SO.J: 9~ I~ o.@i.(aj) o.-ooJ;l.iCo-l .
[ 0.Ss0..'{5 h ~hn>a.e +h2 po-e"c'i' 6- o. .. @>£"2~~" · '
! :;1Gpe ~ YI..-i o u.5Ccj Lt'l.lL mos- 15('.-tqn .enc lftm ¬ ''i'ed l"' ·
j m:cc-cob•olq.,~ ..M&c..hcms· ~: =t. cu.>un·11vh<JY 1'1
; +h.cd· ™ (6pec l-ed ye~ pcm ~ii)-ro o.. '{1-1 v e rv c.c>.<, e .ft +~
i ted· f-'1'ePOJ~hlfY' .~ th.e.- ~ <A' ~- g_ <V(J1U.~ 1o.e
! C!btu.i.n.W fn,f'<l nu_ ,<;-wndOJLd pre.pOJLQ.t.hcm culJYA N ...dered
aJ- f{u. ~ven cl(J.)<2 ,
4 / 18
Md.
Imran
Nur
Manik

6. . '
. fb}en~~
/7 'rYl ~ lllsZJ'Yl~tl t-. ~ ~ hi? A- ~ bzt l/V')hg-
® Mic.Y1ibco·',icc•. O~~SCL'j 0 tAvi~ "Y"!ilrrin--o~ini..c-m '. _
, --·-- '.:.::
0
lh2. -e-rrY' • m t ro 61 o.~, ro.. 6S~o...y
; des~noJe~ 0. 0-jpQ cSt 6tol _ C!A. U,~$Clf rQYfcrrrn_W e J3}3 ~~
I m..Lumao.'<'-i;n'IS· e·ti- -~-e..-iu. ~ o.nd~ ~
'. 0n Cno--hey l;)aro, Yr1. cxc ~lo ~1 ·ccd o.. sso..y mo.yi b.e
lck.fin.e.d l.) u. YYQ.CAOI ~ a- es.h'¥a 11~ ITT meQ~Wl.lYl(- the···
~ ot Anruo lo'I l.Af)iY Li./ i Y rn (YcJOT 0.ni~m. H.eoi>,
! 0- ~ yet0-.tffi mu~- t>e e~-cl6i~~ed loe-weeJ1 ·
I +11R chv~ ··1-0 ie lA~~c'-yed and +t-uc ~iolcro icc'.l S'J~J-em +u ·
! e i+ tS, 0-pp.Q.ed ·
I
II
i
N ~ --
No . (). ~ ~'O~ )tn LU) ed '() onJ«I0101"1
S()IYY'fQ
N o · 6t sIA"0. ciM.d ~u."S"l ~~~ ·
S ~ S~-Gn clev-td vY01
M-=- ~o ~e~
~M =-. S~cLwtd e)"f{J( of M
~c ~ PrvOJfs.e Slo-pe
Where>!.
/
I
!
'
L'M __ · 1 ~ 2 ·
;) --- +: 2
be Nu • Ns
Sh:vncLCDLd ~YYOO 61 61'~~
~j ~VU? -folruo~ e~vcJ- cm~~
5 / 18
Md.
Imran
Nur
Manik

7. cD 9t -luc CEr" D<W n cl' I S '(R) .-- ('.}u_.,.,ce>l
I I .
!~ (YY QhQYY"l coJ ('.:Q"'('('l'f~iV-crn :s ~- <'.'.ffi"A}11wq
i 'rnm»l --t'rQn , t-/ l.Y ·s LJ,n ed .
1----i ® Q--&nU ('.IL ffi()1)Jl C1J;ITT1 pcstucl b-< 1'h'!.<, i LoJ oir . .•
1su.nvl ml ck'Cice wN.n vno.11~o.lolJ lhr.,.;- VQiAS
~ -·--·-····· .. . . ·- ___......_ -- (JJ'Q, o ~ G.:.'f c...y ~ i ~ .
• @J '?I 1i e)"vllf/eo +" ~ ~nd 0hos.e
! 5'fi:(cJw1 e 11 not es.a.:,1s.hed.
'j ;
1Ycli:v i dU-(l,L VOJtt CA.- Oh_j
........ ~
Les~ 'Pye.c,~.e
~....-==::
~ ~~ir•1'1c1...1Li1 Ci HP:.J"0
'!;) .... ---------·--··· .. -· :l
::;..-
© ~ rn,e-hod them oheo
® M(m -!Y)Q ('j5li sv ~ ..:"'c"'' 'hi~iCIA and l'n.e:rnice.,
fN2_--h() . (A_<;,.c;,0--j·
6 / 18
Md.
Imran
Nur
Manik

8. - . -·
----
••
'
,.../
': brc--.J e.<A .OV) ~---mf°'- y-{~ 6""1-- ~.e, ~t~n' M ff}- 5YV1N~ 61--
Q:-~ >1 .-1 v9 ,,.......,1C.r110Y':)d'l.'1{_>'""' C> ~ f'"'V~ ~-a-; .
· Q) /£..In~ l.cY.C-" -0}- >..fc-inAArd.. ~"'JYYpl-e_,
<k... ®n,~c-.s.~ Cft't(... .rt- -4-v.r't- -'-~tk.. , .
- ( lA-1'1 ·(.." l1W h ')
l'E!J C.--.,-h:'·y· t 1. 0 YP>f ~
------ :::.... · .. hb)bn'e,.,
J.11~ N'> (}..h
1 cD The. (1-~st f~ rnu,~J ;hove 1i;_YU.~'dm7 -e~e~--
1 tD tne CU-YGW+h 6 m c1001'~CA11 ~ms· . .
! · ® ihne 1~ ~ ~ oe/uticrristue, .. ~~.~11 dcr;,e ·• ..
I~11d ~~~=~ffee:~~~ 5::~,~,
~:ec~~~=·:i:i~:· k
I h --h.Q_ -t-es.-;- ?-o.:m ?Q . . •
I ® . i
11
. sa.yn is lJJed -far thil de.-enr"1ncdi6T .
I I q 1'n-V..brooy eff.ecl ~ ' .
i (0 ~~"cmse sncru.t.d -be fttm~().f e C,(ffi0. 4a.fXlJlico1W"L
I ?"'1Dduc. . . .
./
CPJ Py 1,c~ p e
-©
7 / 18
Md.
Imran
Nur
Manik

9. ''~ •'i· ~
~ · ~ 6~ -es.· 0-r{)Cni s. YYt ·• · • •
. ~h..e me.rs- s<2n ~, hve
! mcrooY~rf~Y sl)'l .+o o.., pMhc..u.orL u:n-oio.tlc 1i -to b~
~rue re cl . ~(' 0u.ei1.vs "'" 'otii' ,·, s.eke.l-ed ~ <Xso..y ...
i 6f ~toepl-crn)cin ·
Q~I' i',·"' ,.;,_rJ-{l.., ..f,.,:1·,.11· ( )t-~r( Ji1r_[u,~ ,· '.:1 'Af~Cc)· Jj}.(-LV)l(S)" nccr.:-f-: l ...... u, "-- 'T°'-C1Vi) .") <:::.i1C1no (..I~ ""..::>.)V'-.).
I
i '1)tt 1!'.et-uos +Mt thfluenCQ.: the MG A ao aflll't
! diffv..D) cm 01.Q:-h.cd ~ ())tQ · lt~ foll mb~-
-A-coon cLlt~i(JY'I 'JYULlchod . s ~~dln ~O ·-tifl<2-S -
@ ~ N-t>.Q ~-Md
@ Lo,t-1n ~vMe de~ i~ n [ .z + 2 (~x~)l meh{l. · ·
!'
o '·- [v'l 2, I •'J .._.,n,,
~;
I
'l}u_ po ITT e d,{_V es 0Y ?t~ed
rn.~ ~ cLivld_Qd nn two: l) MG>/- 6{ u'0i :iioh'cs
•. broo.d <1l os;S -
G> ~~0921 cllV~GY ~t-tC)d ~)
@ T_~x'b·,J DLQ_h-e. n'Q:-·Ylcd l~JQ)
~ f ~ ~ ~ ~~~ bd' CJ;Jr-.,-,pv.tf~
tvt~ ~ .b~t-v-Je..,.etr-- .+eri~ ti(. >~>-idc.-.r.A. .>"'"""'f-'.CZ.......!--
·--·-----···---......... ... -··- .
:~e-eomLrH2c lo'! ~~ru-ctne +~ do:se :_ e nhi?rk.
;tYu, q;m.uth 6- lL .iW.-ctla Q Sv~c epiie mI CIC!G'I~ ( iSIT1 l
; e ~ d lJS e ilf o. !>-rmdon.d py<fC!Hllhwi ~ tn 01 .
o.n h b1 o ·hc. ~ pmd.uQe' +hQ. ~o,-mx cht{ee 4 1'ri1.1lo1fun
8 / 18
Md.
Imran
Nur
Manik

10. """" , , . '•
Lt · Fo-rrn v[4 -i cm o.nd c()(Id i .f-i1SJ1 D~ ·t-,e n(ed ivm : ~.~."
• lY£. (:(511:"UI'=
l6'1'cl C.<5Y'd1.hcn <JJ +he nt.e.d,.,;.vm sn~-u..Ld be S.v.C-i +ha-- H-
i Sup8)1 l-$ ·--'J2.- {i().,p1d ~ of_. rnc~0-01'(JClll i ~ms·
r;~@icJ<,;w of 1Ju o.~CVl rn.ed11n1 ~ _ .; .. ;· ..·. ..·~!,:··:·;
r == : 'l 'rQ u..im1 rned1vrn. , <'··-~
! 5'rl-C5Uld 'b.e {[w~ --"1iLKne~s -t-hmv~h{Jl)t +-le ct:e· ThQ .. "
/ +}1ck ne~ ~ o .-+n.e Yn2di lJm ~ h UUld be '1- s rnrn
'
r c.,. ~te9 6t .es- ~v.-im: Tru f'6ency & th~'
i 1es+ u.'nci <.;cq·'( c.ud 5,oUJ.n(m sh.mtk) oe ~u e.-1 -"'lo.-t 1 ~ · · • ·--·
!@V; > Ye~ !~e lArP elem ·:--~e 2 1~V.bih@ ·
r 1-luwevQ'(, 1J ShcruJ.d not V.Je U/f!,('(•t-;,~·-,i'.y OJ~e.. Norrn~y..' ~--·:
! 1-h e cl io.rnQJe~ 15 th<L t:ITTW s ~'nuwd 00- 0<e.eed '.3D lll m~ • , ; ·
I . ' ..J. (!41'-i;O't'll"""I
'· 1~ q +est.o.n.d~ SG'Y'dund ~ov-ltffS: . '
; Q-~ !,oAAI )a . "" : 6'ene'f6'y, 1
! sICJU,ld no- ~ITT e.. +ha.)' Qol}9 - .o-ru~ wisQ) lC51~eo hme
i Will b.<i. Yequ.i-1ed -fDl C<Jmpel-e diHW>1on Ot the lll_0.d~ ·
'· · · ~ vv""-""'{.. ~ l~~tiJ
~v~f '· rDF T~ 'fthie.. ·
~Olte... f_J !L.•_, J:, ~ ~ PrfTo .. I- 1....1' 4 '"[e,.mp,..
'-""CA.••··· ~ .P~l'f'i.J )")""'{~,,......
~-· ~··-;_ ' '·
9 / 18
Md.
Imran
Nur
Manik

11. -·_,_-_ ~-- - --
---·,..:~:·
:;~~:.;-~~~~-m--~i.~·~~-~~'--:-l'~X:y'_:f_'~-~~-. ..,~~-· .,.,...,.,-"-~~£7
I
I
1
·1' :D . - ~
• f.>M.d tX >~~ ' o-u" c1 pc ~ . .
1 .p~.,.. f-t IJ.vt-! · Der ed Clnd S-eo-e f1l+e~ po..pe~ discs .
P"'-P-en-- (_y-b m""" ~"'d.-~1-(t".) . .
-l- !('1- b mm in dicrnweyJ absoobs -hQ · kst c_(J1)')po-vnd one
o-.b~o-r:b ~)- S1>'l<). +~en pletc..ed en +-'n.e- C'1JOJ1. one~ 1J rn !3::- t$ -pye.v 6J~y
i~ "' "1".,. I seed ed e +hit +es ">1croirr~o.n i~rn · -rrus .pC<.~es one
'""'J.;".'~ s i.. I Ka.pt ~ 12 hcrwis) al .Uw tem fQY ahne t,0
e.. h
r:v;;;'J.i- I a..l16W ma-x rmu m Ji{fi,0 i(f'(l . A n lJ.mtoe~ ~ ever+s
1
l+CLZQ.S pClee . ·
""~;~Y, d) D•'J disc a.bsatb cooJ~t .-fmn; ct'(j(J)'L rnedAurn and
I '.~ """:;:~~ 1-IM. malc-'61ccl Ufde¥ .-es Js ~1ssc~1?cl - •. ~
1 d;~.. ' : (}) f'aj) ·Qs,- yrio..-Q[ iAl d1 ftwiQ foum dt'1! tu t~
S0-YYlJU -nclt V ti YrlQ_d,,t .9 rn ·
l G) ·'Q-o Q. i's 0- ~0--duell Q,MYie_ <sf . +Q.s,-- ~<A.terletl
'C<r01~(2Y) ·l-yo..,h (ff' n 0-coc01- ~·L~llJlf)J rX:::J~ Ya ~/ d1'~c..
~ ~w~
"t~.evi +we plo-.kl> ~ ~~ °'J- >?-0 -ftti-- iti_.-t ~ ~o~ ~ ~~ ~nh'~ ·
I CM.-h"vi'ba 4 ~
l.Nr:s~~ °brr-e. 0".1-i'>.,1,tb1'h"h.r >01~~ Pl'P~
t;ye..~.te."- h-t.e... "'Vh'V"'l't°'}· ~ ' 'b1-i~ <I} l'°"'h{b')"11...,,_
jli
10 / 18
Md.
Imran
Nur
Manik

12. i
-· I
~
'r
r
I'
!
i
~
I
~
•·.
===~~===:::=:::::::==·=··=··==-·-~-~=--. -..
b ':i ..,o <.M."-h) ...~
lo-o 'f' .
~ plcie> ~- ~-~- ~P~-;~ 1h ., bCcl-ITT 0.(:>7)
110 12- s . wuns +o cd.J(ll<) +1-w. "()rot<}Th 6t ~1~YOOY<f.l'1lll
A_ +:s·: rn~-ey~ )'ffi.Vl Y6 °'-"'j l.nlbc"e-eo~C~ (AQn~I' L()ll ;
lY ~~t-. :Vu_ ~th q on c.Ya=m:(f~-n~rn~, ~Vh ~ ...
c.Leeat dtt ~hnc f--- 2-o-n Q 1-Yn61J() .. a_s -f-~t{''r.~.cm n . b'1~~
hl. l'0-1 bCLe.h.~r-ic:U a.e-nvi' ty at th.Q -~es-- SQrn~e ane
0<r:e~sed. io1 ~~wt Y)~ 1-Yt£ (Q/C(lme-h)~ <St fu.e."-cme
4~"~b1hcm·· 1tv t§YTh.e.. ~o.n~eo +~eone ?f')l)U.~11()(), tluLw~eo t~ (Ar:-l'oCAc·re·U'uA ac:nv1+y ~
~--esJ- 'ffCLteYal Q~Qin~t- fu rh.f: ~&f ooaunI ~rn ·
11 / 18
Md.
Imran
Nur
Manik

13. l
: ·-u.be 5. UflQ K.ap·- In CA.6" 1'nch'ned pcrsI h<m ·.-n( ~()Udi n~mt,
: TY&Se OJ1Q. ~e. t -0cu bCl.~t>d °'-t 3:r- . t;0 c +o en su.ne
: ~~Yi i 1.:, oJ1(5Y) ·
'. ~ les-t trr~0-n~rn DSlQ. -hnns.r.e"l'oc_d to --'N. Clicui
, Sa-nrs. · frcn1 -r~ ~v..t>P-Ll~d p'We ; Q.Rl usie1 ~ fhQ.. Y.Qie_
: o~ °""' lncJC,uJk.hYt oop 1~ °'-.Y CA..~e?-ic. · <2.~~1·n-01Jr ..
~ ·+Vu.. oup ~t1eY ~--i ..h-CA'f~·re11 of mi CY~'?°'nism
i ft> o.voicl. C OYu m.''0l~hGY1 v Qr;'( C.~~,·ThlL.. iii Ut1...~f l
;<;,o..Y-~ 0.Je. +~Y nC..Ul?a.ed 4 ~ ~U6 2.Lt YlOW1$
! h e-nSW!Q l Vie ~Wt of +.es- CTO~i:w1~rn · Yii.~x;.;--
i c.u.Lt-u.n e5 une W)C?d fcso -the Sens, I-! kt -e&r .
'i '
I
' . 3. P.repcv10..c.·,1 t)J~ +e~- )(1..-<.::.~ "~ · -
. ·h-e -es- p1a.. e~ mu
I pye_pan ed i'.lC<'ordi"O +-n --Yu fom.iri f)Cs rxcreedwies;- ..
I @TM nu-cienl- D.50.n. medium p-yepWJed i'n :he :.
I prev l OU!) sec-h cm ,-~ p01_1J e d n crB q_,vaftity l n . . _.r.: • _
l E'D..eh I ;., -tcQ (!_lea Y +-e s-- h>loe s CA.nd ~ J 'Mie.d e <:oH.Cl'.
i @ "Th e ted +ube s and nurn beY 6- pet-0d1:She.sfrl~)
I cm Q Steir i Lit: ed I~ ().)" 11.u+-ad (Ve_ a} I Qj '('_ ({ n d I!Jlbsh'Y ..;
l l~C..'n po-eSSLUlt, .-(-oo 1?° ·oninul-e~ Ql'd OJ1Q TI""Q'(<; te6r~d
! ~l;o ~mlnQo- ~x__J~_Ul-n.iJ a.nd fh,·s OLLawed to' .. :..
I G-Oo Ok aJoau t Lt 1:J - Cjo 0 c. r- . • .t..
I @ fu +.es{- OT~Q)')I~m ~ om. {To.YI~ fened from ,-t: °-
120-ch So--1?-w- --tDIe +o th e +e~+ :-1-u.be L0Yl-bn..Ln(f "'2''.
'G In 0-utuCJClV C?cl ~~ l9m e --h.e_ hu_p 6- Q II .-J-' .r- ~
I_1~ m1u1Q-Y(i L~ 1;., o.r CLSep-]t c_cmd i h(j( . ih.ui -!f:u> LC:
I +e<+ lubE' IS Shu.ken b/ 10ltJwn -1-0 '(j(2t i'.lunitorm ·.~,~
' -..
: -· ~- ; .:i·~k·i .'
. -·---··------·---- .. -- ... - -·· -· ·-. ···-·--·-· ··-·-····-----·--· -··-·· - ...
12 / 18
Md.
Imran
Nur
Manik

14. 1· Plc1.c.e1neni 6{ -1-w Discs Lind lncuoo ..h crn °.
: fu tn~
piMem.e~~ d' +tu. d 1 ·s~ -tYvL -j-Qlcrwi I"~ p¥oced unes
CD1JZ. ~hLl2~d- .
~l rYQ.O.Y s cl~ D- pili:0 cl}- s~--~e-oi_le--"'.:::::=::::=--:!:::;./
l ~ CtY">" 'Q, ·1 rnpye '()ncded d lSeS q.fd . --a.Y'lciM d ,·s:e,~ OJ.Q
! plo.e.ed caen-t'l on -the souc i·fed °'~°'y pcde~ see.deci
with --ho +e~·- c'.Jl~Dsm~ 1o enswH? C6n.t.d- e ~
I YlQ.d,Lu m ·
@ lh~ f~.:~<: one <'.Q.p} ·1 n. o. o.e.p-;ieros-
Cl~ ~c. j~oo 2_llr hbWlY ~d<2ro ~IL> P:aYid e ~vtfie.ient ·
Bme.. ~ +he CJ-.)J·hb10-cs nn tM me<i~~m..
1
• © {:f~y, -t~Q.. ?O:-es ane. 1ne,ubQ-ed a.+ S7-'S.~c
foo r-Zi_h~ ~ ~l~ c,u.~ ·
~-
' I
i
I
l WO-AJ
i
f
1~
kI~
i~
.. I•
I:'
···-·--·-·-,.----·----------
-sv~-pe~-~(5YI · 4----~- ()(i~~~~~-: ~ .
@ !Vu. 'oo.._e-er-a.I Su.(p.en sio» rl ) ro me.d ·,<U-ej
h-D-Yi1e'6'fec +o +h..e_ ~:-C6ile pe·T1 di~he( (n O-n os~.h'e
~ · lh.e__ P~~id l~h~~ lUcz ~ sevem · --mQS',
Jrs-- C..(J(l.kcU~e O-Dd +h.QY ~~t-c1~::.cuise +-o ~um.e
~n()VV) dt's~;lau.-h'b')). (ff +hi +es-t mrnsm<; ..
the rneeli o: OJ1e p bW..e..d Irv.-a petri d ,·~ne~ l n ~v e~
U)o.__ CL~ 1o ~v e Cl- vn.l~m cie,p-1-h 4 o.ppr~:,mQ.te7
;
©- - ~
13 / 18
Md.
Imran
Nur
Manik

15. '
·'-
"
'"';; ' '- - ,,..,,. - ~=-~ ~-c- • - - - ---""- .-,,·- "',_._ '" -- ·~ - - t.. -
j,
I
1111
1f
11
Ii
I
!W PY-epariu_~j 6)1 Jf Disc Ce1';h~.i)1'cnc1. <;u_11·>'0es 0
1-----------------.~'__ _!_:1__:_ J foo the
P"e.pMa:-1&' &- d[<es c..cm1ul"LI"% Socm pe s , +YQ_ -)clIm.iivl,
I p-co-re ckn Q cm c u.t; v_zed _ . <'.l
I
@:Sa.rnpJ-e dt.scs" , ·. d"
O ~
0
o $--en Lt~e d +i--e6 f'C>-'R' 1 s c.s
sM·rA~~}n d 0-hlel-el') one. -hi.Ke" · 0y • +hlL ·-in-~s l~ -t-N.
w- Sa-;.1' pl~ srrkH crn 6f de si--.ed ('.,(me. (J,9ft%_/dise.
I DY l l5D ..ui lcli!,c) on Q_ 0-f>p.t-' Qd cm t-h~ . di sc s e th.e. 'Nl.p
!of rnio-o-pifQH--e Y., a:n asep[-ic__ C({Ydthtrn lYe.se di'"'
iDJ1e. Li__-ft- fG6 0-- ,Yew m i nu' es I;., -~-tt:__c:.~_e..crn_i~ f-0'6
I ~pkte =se TI'IDJ GJ 6f --u_ <;o'-IQT'T - · · .
l . .
! @ St-C)1cl wd d~~~- 0
. , . •
1
1 ° h.e_c.,e 6N. UJ)ed --o CompMQ
G Yh bo..c--n i <ii CU'- ,,;1 It .Gt +n e -tes. rY!O..-nUI S· 1".-M
>'(CSeY>+ S-ud 'f , oeo.dy m.ocQ -<-i:(6Jfj ~ (J,- ~ dI~es ~
! C'.<rn--cU "'°' "'t } ~ 6. 6:·n-i bio-i <:£ <.Mo.. rn'J ci11
i OJLQ_ W) e_d C5Yl S-(}YO(>lld c I ,C -rm: Q.,GTI' ?O.'r S@ ~po;)O .
I
i
vP'<ep:inoJi (Jn <l~ 1-esJ Sqmpe ~ - • -
;1 _ - _ b O m~ o.f I;cvrnp~ In 1 ml
! ~Mr ts (G'o ma pey mD·
E) FinC. y, o...+e 6 ti.Q tnQ.d..t,um s Q.oo.ed ok- Y<n5h
~mpQY 0-.{-urjJ. I ii 0-- lo.mi nOTl cJ.,'i j'lOlO u.nl f, i t I 5
~hned I ;. ~ Yefrit:teYovtur c_4 °c) .
14 / 18
Md.
Imran
Nur
Manik

16. M~w1en1en-~ ,csf tune of lnh~) h G)' ~ . · ·. L~
. . . APreo 1h.e nevv.x.q::::.
-th<l. o..nti 'oo.cJ-e-ro.. Cteh"ry <ff- ~ so.w ~e~. rote. ;.?·._
d..del"min£ d ry YY e~ LtJlin'.(s fu cU·cnn.l'-e¥ & 'f.hi_;1~"" ..
~one IYI -l-eo-rn~ 6f rn· Wlt"h «. ·. u.n~ paru.n·~ ~cctle .
Svc.J..,. ac<; @ll_rn.t:¬ ''•_serue) cro Guo n.er,i sCC<L e .I _., .....
~- ,__
Si Z: e 6j' ?:-<mt 4 inYU.lo;l-i ® ~imd ~ p-1eei se y tl: '/'
~ (n'I e. fo.e.hrrs _:: :; · ,
01-f·~l'SI c, 0,1h b<>elfri~1-~n5>~t- v'1ry 6- ~
+.e.~+- , ,,SCArn ) £, · . .J . .
® &rrtrW-tVl yov:Q_ or t.es- ~o;m.fE' .rnkoo~l1m:
.:==a ' • . . - ..-
@)()·Y~ (J(~e. 6f- &n,. i . ~ s. Yeo..-ed 1-o
====:
woteo- ~olvJoih~ of · ·. . A ·· ' .. rn· 1-hsL ..L-e~'n v .Sa.kd(0 £tl:..e.. or J._n ~~- ")~ {
~ .
DYe l).n ism(0 ~ . 6 +1-u. -.es..: ~~'&. on fu d i"se
· .Y ~ t.es rn£d1' 0rn 1'-r1 ~
@ ®~Q~.v C) ·.
p ~n--~ di~~", .
. ····-·----------,
15 / 18
Md.
Imran
Nur
Manik

17. . ...: -- --·
I J
'i 11.I ,
,,
16 / 18
Md.
Imran
Nur
Manik

18. .I
}
~-
1~
ri
t
Solutions of the reference material or known concentration and corresponding dilutions or the test substance, presumed to be
or approximately the same concentration. arc prepared in a sterile buffer of a suitable pl I value. To assess the validity of the
assay al least 3 different doses or the reference material should be used together with an equal number or doses of the test
substance having the same presumed activity as the solutions of the reference material. The dose levels used should be in
geometric progression, for example, by preparing a series or dilutions in the ratio 2: I. Once the relationship between the
logarithm of concentration of the antibiotic and the diameter or the zone of inhibition has been shown lo be approximately
rectilinear for the system used, routine assays may be carried out using only 2 concentrations or the reference material and 2
dilutions of the lest substnncc. Where a monograph gives directions for the initial preparation llf a solution or the substance,
this solution is then diluted as necessary with the appropriate sterile buffer, The solutions or the reference material and the test
substance arc preferably arranged in the form of a Latin square when rectangular trays arc employed. When Petri dishes are
used, the slll11till11s arc arra111•.<·d 011 L·;1d1 dish so that tlil' slll11tio11s of the reference 111;11<:rial and 1110s..: or the· test substance
alternate around the dish ;111d are placed in such a manner that the highest concentrations or the reference material and of the
test substance arc not adjaccut. The solutions arc placed i11 the cylinders or holes by means ofa pipette that delivers a uniform
volume of liquid. When the holes arc used the delivered volume should be sufficient to till them almost completely. The plates
arc incubated at a suitable temperature, the selected temperature being controlled at ±0.5 ° C'. for approximately 16 hours,
and the diameters or areas or the zones of inhibition produced by' the varied conccntrntions or the standard and of the lest
substance arc measured accurately. preferably lo the nearest 0.1 mm or the actual zone size. by using a suitable measuring
device. From the results. the potency of the tested substance is calculated. Suitable publications on the statistics of bioassays
For the application of the lest solution, small sterile cylinders of uniform size, approximately I 0 mm high and having an
internal diameter of approximately 5 mm, made of suitable material such as glass, porcelain. or stainless steel, are placed on
the surface of the inoculated medium. Instead or cylinders, holes 8-10 mm in diameter may be bored in the medium with a
previously sterilized borer.
When using the inuculum prepared as described below, an inoculated medium containing I ml. or iuoculum per I 00 mL ofthe
culture medium is usually suitable. When the inoculum consists or a suspension or vegetative organisms, the temperature of
the molten agar medium 111L1st not exceed 48-50 ° C al the time of inoculation. The dishes or trays should be specially
selected with flat bottoms. During the filling they should beplaced on a flat, horizontal surface so as to ensure that the layer of
the medium will be of a uniform thickness. With some test organisms, the procedure may be improved if the inoculated plates
are allowed to dry for JO minutes at room temperature before use, or refrigerated al4 o C for several hours.
Use Petri dishes or rectangular trays tilled to a depth orJ-4 mm, unless otherwise indicated in the monograph, with a culture
medium that has previously been inoculated with a suitable inoculum of a susceptible test organism prepared as described
below, The nutrient agar may he composed of two separate layers or which only the upper one may be inoculated. The
concentration of the inoculum should be so selected that the sharpest zones of inhibition and suitable dose response at different
concentrations of the standurd arc obtained.
Procedure
Microbiological assay of antibiotics
'The potency (activity) of an antibiotic product is expressed as the ratio of the dose that inhibits the growth of a suitable
susceptible microorganism to the <lose or an International Biological Standard, an International Biological Reference
Preparation. or an International Chemical Reference Substance of that antibiotic that produces similar inhibition. Properly
validated secondary reference materials may also be utilized in the assay. Tu carry out the assay a comparison is made between
the inhibition of the growth ofmicroorganisms produced by known conccntnuionsof thc reference material and that produced
by measured dilutions or the test substance.
This response can be measured by the diffusion method, as described below, or in a liquid medium by the turbidimetric
method.
17 / 18
Md.
Imran
Nur
Manik

19. ·'>-.
5) From the graph tile concentration of test solution of cyanocobalamin is found & reported.l
I
I
I
4) By interpolating the standard curve determine the concentration as activity per ml of vit B12.
1) Determine the average of titration values of each level of both standard & test solutions.
2) Plot a graph considering average titration values(in ml) of O.OSN NaOHagainst concentration of standard
cvanocobalaminsolution.
3) A linear graph is obtained.
Interpretation of results:-
7) Cool the test tubes at room temperature.
8) ~ate@of ~urns prepared of facto bacillus liechmannii.
·. 0
9) Incubate the test tubes for 64 to 72 hrs at temperature range of 30 to 37 C.
~-Procedure for assay :- !'I V)' lt:V'
1) Clean ten test tubes & add to it 0, 0.5 ,1.0 ,2.0 ,2.5 ,2.5 ,3.0 ,3.5 ,4.0, 4.5 ,& 5 ml of~
cvanocobalarnln solution.
2) To each tube add Sml of Basal medium solution
3) Volume of each is adjusted to lOml by water
4) In another 'ltest tubes add 1_,_2 ,3 ,4 ml of test solution which is to be assayed.
5) To each of this also add ?ml_of Basal medium stock solution & adjust volume tolO ml with water
6) Sterilize all test tubes in autoclave at 121°C for 15 mins.
with O.OSN NaOH using bromothymolblue as
(~ Ufb'C'il~
10) After incubation perio~_?ntents of each test tube
indicator until green colour.
11) Record all the titre readings clearly
12)
MICROBIALASSAY OF VITAMIN
Microbiological assay of vitamins is a type of biological assay performed with the aid of microorganisms. Many therapeutic
agents, which either inhibit the growth of microorganisms or arc essential for the growth of them an: standardized by
microbial assay, .t]) -cg ' . ~- . ·
. rvi. e, B:d: ·' t~ rrie.,,.~.e t"he. i'IA' 4- ""'O'O~&w..
" bV"-!>.e.~ oYl ... -
PIUNCIPLE: .-v +VI~ e )V "' ".c.c .....i.~~tt'i-C'.
@~and amino acids are c~lial for the growth of microorganisms. . V'l>-<i:<..~. ~ h111.~·11·.-l ~~-.l-
The basis of this assay is to measure the ability of test organism to utilize the substance being assayed under a proper
nutritional condition.
The organisms require these growth factors (vitamins & amino acids) in~ or ~nograms. ~ CTL.~
,.,The response (growth of' test organism) is proportional to the doscramount of factor) added to medium.
i/ ~
~es f'O'IJ-e o6 Das.e., "'d.-c..c+--fo W.~~·
l11CROBIALASSAY<>FvriAlllN B12: -------
The test organism selected must be capable of utilizing _l'n.:e cyan_ocob•1l~1~n. I.acto bacillus Uecl1111at111ii is found to
satisfy the requirements. Gram negative bacilli, non-pathogenic. easy lo Culture & easily available. Isolated from milk.
cheese,& othcrd airy products. Assay is performed by using either titrimctric or; Turbidimctric method.
'
are .isted below. Conditions for the assay of individual antibiotics and· suitable test organisms are given in
monographs.The choice of an appropriate strain oftest organism may be critical for the assay.
18 / 18
Md.
Imran
Nur
Manik