HoshMuhammadLashar

i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Cont
ent
s
Cl
ass:
2ndPr
of.Phar
m.D.Cour
seNo:
Phar
m:416Subj
ect
:Phar
macognosy
-IA(
Basi
c)Theor
y2
Ev
aluat
ionofcr
udedr
ugs 2
1.
1Impor
tanceofcr
udedr
ugsf
rom medi
cinal
viewpoi
nt 2
1.
2Consequencesofl
ackofcr
udedr
ugsev
aluat
ion 2
1.
3St
andar
dsf
orev
aluat
ionofcr
udedr
ugs 2
2.
1Conceptofev
aluat
ionofcr
udedr
ugs 3
3.
1Met
hodscommonl
yempl
oyedi
nev
aluat
ingcr
udedr
ugs 3
3.
1.1Or
ganol
ept
icev
aluat
ion 3
3.
1.2Mi
croscopi
cev
aluat
ion 3
3.
1.3Phy
sical
eval
uat
ion 4
3.
1.4Chemi
cal
eval
uat
ion 14
3.
1.5Bi
ologi
cal
eval
uat
ion 17
4Ref
erences 21

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Cl
ass:
2ndPr
of.Phar
m.D.Cour
seNo:
Phar
m:416Subj
ect
:Phar
macognosy
-IA(
Basi
c)Theor
y

Ev
aluat
ionofcr
udedr
ugs
1.
1Impor
tanceofcr
udedr
ugsf
rom medi
cinal
viewpoi
nt
Usingherbsasamedi cineistheoldestform ofheal
thcareknowntohumani t
yandt o
thisdayover80%oft hewor l
dpopul at
iondependsonher balmedi
cinesandpr oduct
s
forhealthylivi
ng.Medici
nalplantconst i
tut
esa sour ce ofr
aw mat eri
alsforboth
tradi
ti
onalsy st
em ofmedi ci
ne and moder n medicine.As such they r
epr
esenta
substant
ialport
ionoft
hedrugmar ket.

1.
2Consequencesofl
ackofcr
udedr
ugsev
aluat
ion
Commer ci
ali
zati
on oft hemanuf actureoft hesemedi cinest o meett heincreasi
ng
demandshasr esul
tedinadecl i
neint hei
rqualit
yandpur i
ty.Oftenherbalrawmat eri
als
areprocur
edandpr ocessedwithoutanysci enti
fi
cevaluati
onandl aunchedont othe
marketwit
houtanymandat orystudi
es.Risei ntheuseofher baldugshasal sogi v
en
ri
setovari
ousformsofadul ter
ati
onsofher balproduct
sresulti
nginfatalconsequences.

1.
3St
andar
dsf
orev
aluat
ionofcr
udedr
ugs
Standardi
zati
onistheprocessofprescri
bingasetofstandar
dsconstantparamet ersof
quali
tat
iveandquant i
tat
iveval
uesthatcarryanassur
anceofqual
ity,
eff
icacyandsaf ety.
Standardi
zati
onofcr udedrugsisactual
lyagreei
ngonspecif
icstandar
dsment ionedi n
standar
dr ef
erencessuchasphar macopeiasandmonographstoensuretheprecisionin
theprocessofev al
uati
ngcrudedrugs.

a)Phar macopeiaancientGr eekword‘’pharmakon-drug’

’‘’
poeia-t
omake’ ’itisa
referencebookforpreparati
onofqualit
ymedicinespublishedbyt heauthor
it
yof
agov er
nmentorconcernedsociet
y.
Or
I
ti sanessent i
alrefer
encef oral
lindivi
dual
sandor ganizat
ionsworkingwithi
n
pharmaceuticalr
esearchanddev el
opment ,manuf act
ureandt esti
ngaroundthe
globe.
 Briti
shpharmacopoeia(BP)

 Eur
opeanphar
macopoei
a(EP)

 Uni
tedSt
atesphar
macopoei
a(USP)

 Br
it
ishher
bal
phar
macopoei
a

 Br
it
ishher
bal
compendi
um

 Uni
tedSt
atesher
bal
phar
macopoei
a

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

 I
ndi
anphar
macopoei
a

b)Monographi tisadet ail

edanddocument edtreat
iseonapar ti
cularsubject.
Monographofcr udedr ugsisaconcer neddocumentt hatdefi
nesabot anical
drugandprov i
desinformationthatal
l
owsf ori
tsproperi
denti
fi
cation.Itcont
ains
basi
c descript
ion incl
uding nomenclature,partused,const i
tuents,range of
appli
cat
ion,contrai
ndicati
ons,sideeff
ects,inter
acti
on,doseandmechani sm of
acti
onofapar ti
cularcrudedrug.

 Monogr
aphsofUni
tedSt
atesHer
bal
Phar
macopoei
a

 Monogr
aphsofEur
opeanSci
ent
if
icCooper
ati
veonPhy
tot
her
apy(
ESCOP)

 Monogr
aphsofGer
manCommi
ssi
onE

 Monogr
aphsofWor
ldHeal
thOr
gani
zat
ion(
WHO)

2.
1Conceptofev
aluat
ionofcr
udedr
ugs
Eval
uati
onofcrudedrugsmeansconf i
rmati
onofitsi denti
tyanddet er
minati
onofits
qual
it
yand pur
ityaccording t
o standardsand detection ofnatureofadulter
ati
on.
I
mprovementsi
nev al
uat
ionmet hodshav edefi
nit
elyl edtol esseni
ngofchancesi n
bi
ochemical
var
iati
onandt headul
terat
ionsandsubsti
tutionsofcrudedrugs.

3.
1Met
hodscommonl
yempl
oyedi
nev
aluat
ingcr
udedr
ugs
3.
1.1Or
ganol
ept
icev
aluat
ion
I
tr eferst oev aluati
onofcr udedr ugsbycol or,odor ,taste,size.shapeandspeci al
features,l
iketouch, t
extureetc.
Or
I
ti sat echnique ofqual it
ative ev aluat
ion based on t he study ofmor phologi
cal
i
mpr essiononor gansofsenses.
Aromat icodorofpepper mintleav esandf ennelf rui
t,Sweett ast
eofliquor
ice,pungent
tasteofcapsi cum andgi nger,spicyt asteofbl ackpepper ,ovoidshapetearsofgum
acacia,wav yshapeofr auwol fi
ar ootsandbr owncol orofci nnamonarei mpor t
ant
diagnosticchar acter
ist
icsofor ganolepticevaluation.

3.
1.2Mi
croscopi
cev
aluat
ion
Thi
smet hodal l
owsmor edet ai
lexami nationofacr udedrugandi tcanbeusedt o
i
denti
fytheorgani
zeddr ugsbyt heirknownhi stol
ogi
cal char
acteri
sti
cs,
Keyaspectsofmicroscopicevaluation
 Mi croscopeisusedbyv i
rtueofi tspr oper
tyt omagni f
ypermitstheminute
struct
ureunderstudyt obemagni fiedt oconf i
rmt hest r
uctur
aldet
ail
soft he
drugs.Forthe ef fect
iver esult
sv ar i
ousstainsorr eagent
scan be used to

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

disti
nguishcellul
arstruct ur
es.
 Mi cr
oscopicst udiesar emadef rom v eryt hin sectionsofcr udedr ugs.The
characteri
sti
cs ofcel lwal ls,cellcont ents,ar rangementofdi fferenttissues
(dermal,groundandv ascul art i
ssues)andt heirinclusionssuchasshapeof
epidermalcell
s, stomat aandt ri
chomesi nder malt i
ssues, ar
rangementofxy lem
andphl oem inv asculartissuesandar rangementofmesophy lli
ngr oundtissues
arespecifi
cdiagnost icchar acteri
sti
csf orqualitativ
emi croscopyofcr udedr ugs.
 Mor eoverotheri mpor tanthi stol
ogicalaspectofmi croscopic ev aluati
on i s
quantit
ativ
emi croscopy .Var i
ousdi agnosti
cpar amet ersst udiedf orquant i
tative
mi cr
oscopy ar e stomat alnumber ,stomat ali ndex,pal i
sade r atio,v ei
ni slet
numberandsi zeofst archgr ainsetc.

Stomat alnumberi saveragenumberofst omatapersq.mm ofepi dermisoft heleaf.

Stomat alindexi stheper centagewhi cht henumber sofstomat aformt ot het otal
numberofepi dermalcells,eachst omabei ngcount edasonecel l.Stomat alindexcan
becal culatedbyusi ngthef ol
l
owi ngfor mula:
Stomat alI
ndex( S.I)=S/E+S×100
Wher e:S=numberofst omat aperuni tareaandE=numberofepi der malcel l
sin
thesameuni tar
ea.
Vein-isletnumberi sdefinedast henumberofv einisletspersq.mm oft hel eafsurface
midwaybet weent hemi dr i
bandt hemar gin.Itisconst antforagivenspeci esoft he
pl
antandi susedasachar acteri
sti
cf ortheidentif
ication.
Palisade r ati
oi s defi
ned as t he av erage numberofpal i
sade cells beneat h each
epider mal cel
l.Pal
isader atiocanbedet er-minedwi t
ht hepowdereddrug.

3.
1.3Phy
sical
eval
uat
ion
Physi
calev
aluat
ionofcr
udedrugsisperf
ormedwi t
houtr
eact
ingthecr
udedrugswi
th
ot
herchemical
sornosuchchemical
changingsar
emadeincrudedrugmat
eri
al.

3.
1.3.
1Moi
stur
econt
ent
Anexcessofwat eri ncr udedr ugmat erialencouragesmi crobi
algrowt horpresenceof
fungiorinsectswhi chmayl eadt othespoi l
ageordet eri
orati
onofcr udedrugsfollowing
hydrolysi
s.Limitsf ormoi stur econtentshoul dther
eforebesetf oreverycrudedrug.
Met hodsformoi stur econt entdet erminat i
on
 Lossondr yingt het estf orlossondr yi
ngcanbecar ri
edoutei t
herbyheat ingin
anov enat100- 105° Cori nadesi ccatorunderat mosphericorreducedpr essure
atr oom t emper aturef orspeci f
icper iod oftime.Thedesi ccatormet hod is
especiall
y usef ul for mat er
ials t hat contain considerable proport
ion of
substancest hatar ev olat
ileort hoset hatmel ttoast i
ckymassatel evated
temper at
ures( balsamsandr esins).Format eri
alssuchasdi git
ali
sander gotet c.
whichcont ainl itt
leofv ol
atileorst i
ckymat terdirectdryi
ngupt oaconst ant
weightcanbeempl oy ed.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

 Azeotr opicmet hodofmoi sturedet erminati

ontheazeotr
opicmet hodofmoi st
ure
determi nati
ongi vesdir
ectmeasur ementofwat erpresentinthemat er
ialbeing
exami nedwhent hesampl ei sdisti
lledtogetherwit
htheimmi sci
blesolv
entsuch
asxy l
ene,t oluene,orcarbont etrachlori
deet c.Thewaterpresentinthesampl e
i
sabsor bed byt hatimmi scibl
esol vent.Thewat erand solventaredi sti
l
led
togetherandsepar at
edi nar ecei vi
ngf l
askandf i
nal
lytheamountofwat er
received inr eceivi
ng fl
askwi l
lbe t aken into consi
derati
on forcalculat
ing
moi st
ur econt ent
.

Table1.
1crudedr
ugswi
thl
i
mit
sformoi
stur
econtent
Crudedrugs Moi
sturecont
ent(
%)

Al
oe Notmor
ethan10

Di
git
ali
s Notmor
ethan05

Er
got Notmor
ethan08

Acaci
a Notmor
ethan15

St
arch Notmor
ethan15

3.
1.3.
2Opt
icalr
otat
ion
Certaincrudedr ugsampl esarefoundt ocont ai
nthechi ralcar
bonswi thinthechemi cal
str
uct ureofthei
rconst i
tuent’
sduet owhi chtheyexhibitthepropertyofrotat
ingplaneof
polari
z edli
ght(PPL)whent heirsoluti
onsar esubjectedt opolarimeter(devi
ceusedt o
measur ethe opt i
calact i
vi
ty).Such subst ances are descr i
bed as opt i
call
y activ
e
compoundsandt hispropert
yisknownasopt i
calacti
vityoropticalrot
ati
on.
Themol eculesthatrotatesthePPLt orightorclockwi searecal l
edasdext r
orotat
or y
andr epresentedby( d)or(+)andt hemol eculesthatrotatesthePPLt ol
eftoranticl
ock
wisear ecall
edasl evorotat
oryandr epresentedby( l
)or( -)
.

Table1.
2crudedr
ugswi
thl
i
mit
sforopt
icalr
otat
ion
Crudedrugs Anglesofopt
ical
rot
ati
on

Car
awayoi
l +75°t
o+80°

Cast
oroi
l +3.
5°t
o+6.
0°

Cl
oveoi
l 0°t
o–1.
5°

Honey +3°t
o-15°

Chenopodi
um oi
l -
30°t
o-8°

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Eucal
ypt
usoi
l 0°t
o+10°

3.
1.3.
3Sol
ubi
l
ity
Drugs solubil
ity behavi
ort owards solvent
si st aken i
nt o considerati
on fort he
examinati
onofadul t
erants.Fore.
g.castoroili
ssol ubl
eonlyin3partsof90%al cohol
whilet
headul terat
edf or
m mayshowgoodsol ubil
i
tyi nal
cohol
.Simil
arlytherear
emany
otherexampl esofspeci fi
csolubili
tybehavi
orofcr udedrugslikecolophonyisf r
eely
solubl
einlightpetrol
eum andasaf eti
daissol
ublei
ncar bondi
sulphi
deet c.

3.
1.3.
4Mel
ti
ngpoi
nt
Melti
ngpoi ntofsoli
di stemperatureatwhichitchangesstatefrom sol
i
dt oli
qui
d.In
case ofpur e chemicalcompound i solat
ed fr
om naturalsources melt
ing poi
ntis
constantbutincaseofcr udedrugstheycontainthemixtureofmanychemi cal
sthey
aredescribedwithcertai
nr angeofmel ti
ngpoint.Thepurit
yoffoll
owingcrudedrugs
canbeev aluat
edbyusi ngthisparameter
.

Table1.
3crudedr
ugswi
tht
hei
rspeci
fi
crangeofmel
ti
ngpoint
Crudedrugs Mel
ti
ngpoi
nt(°
C)

Col
ophony 75-
85

Cocoabut
ter 39-
42

Beeswax 30-
33

Wool
fat 62-
65

3.
1.3.
5Ref
ract
ivei
ndex
Thespeedofl ightinanymedi um dependsuponpr operti
esoft hatmedi um.Whena
l
ightpassesfrom onemedi um toanot herofdi ffer
entdensity
,itisbentf rom ori
ginal
path.Thusref
racti
veIndexisav aluecalculatedf rom t
her at
ioofthespeedofl i
ghti
na
vacuum tothatinasecondmedi um ofdi fferentdensity.Therefracti
veindexismost
commonl ysymbolizedbytheletternorn' .Ref racti
veindexisconst antf
orl i
quidsand
canbeconsideredasoneoft hecr i
ter
iatocheckt hepurit
yofcrudedr ugs.

Table1.
4crudedr
ugswi
tht
hei
rspeci
fi
crangeofref
ract
ivei
ndex
Crudedrugs Ref
ract
ivei
ndex

Cl
oveoi
l 1.
527t
o1.
535

Cast
oroi
l 1.
4758t
o1.
527

Car
awayoi
l 1.
4838t
o1.
4858

Ar
achi
soi
l 1.
4678t
o1.
470

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

3.
1.3.
6Vi
scosi
ty
Viscosi
tyi
st heresist
ancetoaf l
uidtof l
ow.Thi
sresi
stanceactsagainstt
hemot i
onof
anyobject
.Viscosityofali
quidisconstantatagiv
entemperatur
eandi sanindexofits
compositi
on.Hencei tist
hereforeusedasanev al
uat
ionparameterofcrudedrugs.For
example,l
iquidparaff
inki
nemat i
cviscosi
tyi
s64centi
stokesat37.8°
C.

3.
1.3.
7Ashcont
ent
Theashr emai ni ngaf teri gnitionofher balmat eriali sameasur eoft ot alamountof
i
nor ganiccont entofmat er i
al leftaf terbur ning.
Acr udedr ugcont ainsphy siologi calandnon- phy si ol ogi calt ypeofash.Phy siologi cal
ashi sder ivedf rom pl antt i
ssuesandi sbecauseofnat urali norganiccont entofher b.
Whi l
enon- phy si ologi calashconsi st sofr esi dueofext raneousmat t ersand,soi let c.
adheringt ot heher b.Formanycr udedr ugsashv al uegi vesani ndicat ionoft hecar e
takenint hepr epar ationofdr ugs.Adul terat i
onwi thi nor gani cmat terwi l
lgi vehi gherash
valuethanf orunadul terat eddr ugs.
Typesofashv alues
 Tot alashconsi stsmai nlyofcar bonat es,phosphat es,si l
i
cat esandsi li
ca.I t
includesbot h phy siologi calash whi ch i sder iv ed f rom pl anti tsel fand non-
phy siol ogi calashwhi chi sr esi dueofext raneousmat tersuchassand,soi land
li
meet c.t hatmaybepr esent .Tot alashi sdet er mi nedbyi nciner at ing2- 4gofai r
drieddr ugbygr adual lyi ncreasi ngheatupt o450° C,t hisi sbecausev olat i
le
inor gani cmat er i
al may bel ostathi ght emper at ure.
 Sul phat edashi nv olvest het reat mentofcr udedr ugmat er
ialwi thsul phur icaci d
bef or eigni tionduet owhi chal loxi desandcar bonat esar econv er tedt osul phat es
andi gni ti
oncanbecar riedoutat600° C.
 Aci d-insol ubl eashmeanst heashi nsol ublei ndi lutehy drochlor icaci d, whent otal
ashi st reat edwi thdi lut ehy dr ochl or i
caci d,f il
ter edandt hei nsol ubl emat er ials
whi ch ar er etained on ashl ess f ilt
erpaperon i gnition gi v
es ash,whi ch i s
consi der edasaci di nsol ubl eash.Thi smeasur est heamountofsi licapr esent ,
especi allyassandandsi li
ceousear thymat t er .
Inmanyi nst ancest otalashi snotusef uli ndet ect i
ngadul ter at i
onwi thear thy
mat terandi tv ar ieswi thi nawi der angei nsuchcasesaci di nsol ubl eashv alues
isapr efer redpar amet erwhi chwi llindicat et hepr esenceofear thymat teri nt he
root sr hizomesorl eavesofacr udedr ugmat er ial.
 Wat ersol ubl eashwhent otalashi sboi ledwi thwat erandf il
ter edwi t
hashl ess
fil
terpaper ,t hef il
tratet husr ecei v edwi llbecont ainingwat ersol ubl emat er i
al
howev ert her esi duer et ainsov ert hef il
terpaperi sani nsolubler esi duewhi chi s
furtheri gni tedal ongwi thashl essf il
terpaper .Thewei ghtoft hef or medashwi l
l
besubt ract edf rom t otal ashv aluegi vest hewat ersol ubleash.
Wat ersol ubl eashi susedt odet ectmat er i
alexhaust edbywat ere. g.t ea,gi nger
etc.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

3.
1.3.
8Ext
ract
ivev
alues
Theext r
actsobt ai
nedbyt reati
ngcr udedr ugswi t hdiff
er entsolv entsar eappr oximat e
measur eoft hei rchemi calconst i
tuents.Taki ngi ntoconsi der ati
ont hedi versi
tyi n
chemi calnatureandpr oper ti
esofconst ituent spr esentincr udedr ugsv ari
oussol vents
areusedaccor dingt ot hety peofconst i
tuent st obeanal yzed.
Wat er-
soluble ext ract i
v ei s used f orcr ude dr ugs cont ai
ning wat er-soluble( polar
constituents)liket anni ns,muci lages,etc.al cohol -
solubleext ractiveisusedf orcr ude
drugscont ai
ningal coholsol uble( l
esspol ar)const it
uent slikegl ycosides,r esins,et c.
andet her -
solubleext ractivesar eusedf orcr udedr ugscont aininget hersol uble( non-
polar)const i
tuent slikev olat i
leoi l
sandf ats.Ext ractiv
ey ieldofdr ugst osol ventmay be
used t o detectand appr oximat el
ydet ermi ne t he amountofan adul terantwhi ch
dissolvesint hesol v entf orexampl ewhencol ocynthseedsar emi xedwi thi tspulppar t
i
tspet roleum et hersol ubleext racti
v evaluesar ehighert hannor malbecauset heseed
partisusual l
yenr i
chedwi t
hnon- polarconst i
tuentsl i
kefixedoi ls,fatsorv olatil
eoi l
set c.
Butitisnotsof orpul ppar toft hepl ant.

Table1.
5crudedr
ugswit
hthei
rspeci
fi
crangeofextract
iveval
ues
Crudedrug Watersol
ubl
e Alcohol
soluble Ethersol
ubl
e
(W/W%) (W/W%) (W/W%)

Al
oe Notl
esst
han25 Notl
esst
han10 -

Gi
nger Notl
esst
han10 Notl
esst
han4.
5 -

Li
nseed Notl
esst
han15 - Notl
esst
han25

capsi
cum - - Notl
esst
han12

3.
1.3.
9For
eignor
gani
cmat
ter
Foreignmat terisamat erialconsistingofpar tsofcr udedr ugorsomeot hermat er
ial
s
whichar enotment i
onedwi thinthel imi t
sspeci fi
edf orconcer nedcr udedr ugi nthe
respect i
vemonogr aphs.Fort heexami nationofpr esenceoff or
eignmat terwei ghed
quant it
yofcrudedr ugmat eri
aliscar efull
yspr eadinat hinl ay erandt henitisexami ned
byusi ngamagni f
y i
nglens( 6xor10x)t hef oregnmat terdet ect edduringexami nati
onis
separ atedfrom t hecr udedr ugmat eri
ali tist henwei ghedandper centcont entis
calculated.
Her balmaterialshouldbeent i
relyfreef rom visibl
esi gnsofcont aminationbymol dsor
i
nsect sorothercont ami nantsincludingani mal excreta, sincei ti sverydiff
iculttoobt ai
n
plantmat eri
alinanent irel
ypur econdi ti
onphar macopei ast her efor
econt ainst atements
ast ot heper centageofot herpar t
soror ganicmat t
erwhi chmaybeper mit t
ed.Few
exampl esofsuchl i
mitsar e:garli
cshoul dnotcont ainmor et han2%, saffr
onshoul dnot
cont ai
nmor et han2%, satavarishouldnotcont ainmor et han1%, etc.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

3.
1.3.
10Det
ermi
nat
ionofv
olat
il
eoi
lcont
ent
Crudedr ugscont ainingv olatil
eoi l
saremost l
ychar act
eri
zedbyt ypicalodorif
erous
naturebecauset heyar econsi deredasessenceofpl antmat eri
al,whichi swhyt heyare
oft
enr efer
redasessent i
aloils.
Todet erminethev olati
l
eoi lcontentofthecr udedr ugtheplantmat eri
alisdisti
l
ledwith
water,theaqueouspor ti
onal ongwiththev olati
leoilisreceiv
edi nr eceivi
ngf l
ask.For
oil
swi t
hr el
ati
vedensi t
yequalt o orgreatert han watersepar at i
on f r
om wat eris
assist
edbypl acingaknownv olumeofxy leneint her eceiverandr eadingof fthe
combi nedoilandxy l
ene.Int hiswayv ol
ati
leoi lcontentofapar t
icularcrudedr ugcanbe
determined.

3.
1.3.
11Bi
tt
ernessv
alue
Crudedr ugssuchas‘ ’
gentian’’thathaveast r
ongbitt
ert astecalledbittersandar e
employedt herapeuti
call
yasappet i
zi
ngagent s.Theirbi
tternessi ssaidt ost imulate
gastr
icsecretions.Bitt
ernesscanbedet erminedchemi cally,howev ersincet heyar e
composedoft woormor econst it
uentswit
hv ari
ousdegreeofbi tter
nessi tistobe
measur edbyt aste.Thebitt
erpr operti
esofcrudedr ugsaredet erminedbycompar ing
thethresholdbi t
terconcentrati
on( TBC)ofacr udedrugmat eri
alwi ththatofdi lute
solut
ionofquininehy dr
ochloridesoluti
onwhichact sasast andard.Thust hebitter
ness
val
uei sexpressedi nunit
’sequi v
alenttothebi t
ter
nessofasol uti
oncont aini
ng1gof
quini
nehy drochlori
dein2000ml .Themet hodi sident
icalt othatdescr ibedi nt he
EuropeanPhar macopoeia.

Thef
oll
owi
ngpr
ovi
sions,
basedonWHOgui
del
i
nesar
eappl
i
ed:

1.Saf
edrinki
ng-
watershoul
dbeusedf
orext
ract
ingt
heaer
ialpar
tsoft
heher
b,f
or
mouth-
washaftereacht
asti
ng,
andf
ordi
ssol
uti
onoft
hequini
ne.

2.Inordertoobtainaninformedconsentoft hev olunteerstheyar efir

stgivena
seminaronthebitt
ernessv alueandt heWHOmet hodf ordeterminingbit
terness
val
ue.Subsequentl
yadat ef orthetesti
ssel
ect
edsot hatparti
cipantswil
lrefr
ain
fr
om food,dr
inksandmedi cament sanhourbef
oret hetest.

3.Sincesensiti
vit
ytobitt
ernessvar
iesgreat
lybetweenpersonsandatdi
ff
erent
ti
mesi nthesameperson,eachpart
ici
panti
nthetestwil
ltast
ebotht
heherbal
extractandthequi
ninesol
utionwi
thi
nashortspaceoft
ime.

4.Allthepart
ici
pantsaref
ir
strequir
edtotast
ethedr i
nki
ngwatert
obeusedi nthe
testandasol ut
ionof0.
058mgofqui ninehydrochl
ori
dei
n10mLoft hatwater
.
Onlythosewhosensednobi tter
nessi
nthedrinki
ngwater
,butsensedbit
ter
ness
i
nt hequini
nesoluti
onar
eincludedint
hetest.

5.Thet
ast
ingeachser
iesofdi
l
uti
onsoft
heext
ractorqui
ninesol
uti
onmust

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

commencewit
hthel
owestconcent
rat
ioni
nor
dert
oret
ainsuf
fi
cientsensi
ti
vi
ty
oft
hetast
ebuds.

6.Allt
hesol
uti
onsandt
hedr
inki
ngwat
erf
ormout
hwashi
ngshoul
dbekeptat20-
25°C.

Si
ncethecalcul
ati
onofbit
ternessval
uei
sbasedont
hepr
ocedur
eher
edescr
ibed,t
his
mustbefol
lowedwithst
ri
ctlyasfol
l
ows:

Prepar ation ofst andar d quini

nesol uti
on Exact l
y0. 1 g ofqui ninehydrochlori
dei s
dissolvedi nsuf fi
cientdr i
nking-wat ertopr oduce100mL.Subsequent ly
,5mLoft his
solutioni sdi l
uted t o 500 mLwi thsaf edr inki
ng-wat erto givet hestockst andard
solutionofqui ninehy drochloridel abeledSq,andcont ai
ned0. 01mgoft hequinine
standar d/ mL.Ni net est-t
ubesl abeled1t o9ar esetupt ocontain4.2,4.
4,4.6,4.8,5.0,
5.2,5.4, 5.6and5. 8mLof( Sq),respecti
v el
y.Intotubes1t o9isthenadded5. 8,
5.6,5.4,
5.2,5.0,4. 8,4.6,4. 4and4. 2mLofdr i
nking-water,respectiv
ely.Thismeant esttubes1
to9wi llcont ai
n0. 042,0.044,0. 046,0.048,0.050,0. 052,0.054,0.056and0. 058mgof
quininehy drochloride,respectiv
el y.

Preparati
onanddi l
utionsofher balextr
act(t
est)stocksolut
ionExact ly1goft heher
bis
extractedwit
hdr i
nking-watert opr oduce1000mLofaqueousext r
act.Subsequentl
y,5
mLoft heext
racti
sdi lut
edt o500mLwi thdri
nkingwater.Thissol uti
oni slabel
edasthe
stockext r
act(Sh)
.Itcont ains0. 01mgoft heherb/ mL.Tent ubesl abeled1t o10are
thensetupt ocontain1, 2,3, 4,5,6,7,8,9and10mLofSh, respectively
.Intotubes1to
10ist henadded9,8, 7,6,5, 4,3,2,1and0mLofdr i
nki
ng-wat er,r
espect i
v el
y.

Pr
ocedur
e

 Fir
st ,theparti
cipantwil
lri
nsehisorhermout hwi t
hdr i
nki
ng-water,andthenwil
l
taste10mLoft hemostdilut
esolutionbyswirl
i
ngitinthemout hfor30seconds,
not i
cingwhet herornotthesolutiontast
edbitt
er.Heorshehel dt hesoluti
onin
themout hforanot her30s,andt hennotici
ngwhet herornotthereisal ossof
bitterness.Subsequentl
y,thesoluti
onisspitt
edout ,andthemout hri
nsedwith
drinkingwat er
.

 Thepar t
ici
pantwil
lwaitfor10mi nbefor
ethenexthi
gherconcentrat
ionist
ast
ed.
Thepr ocedureaboveisr epeat
eduntilthedi
l
uti
onwi t
hTBC( thati
st hel
owest
concentrat
ionatwhichasol ut
ionconti
nuestotast
ebitt
erafter30seconds)is
att
ainedbyt hepar
ti
cipant.

 Afterthefi
rstseri
esoft
asting(ei
therwit
hthequininesoluti
onortheher
bal
extract
),t
he mout hisr
insed t
horoughl
ywith dr
inking-
waterunt
ilno bi
tt
er
sensati
onremained.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

 Await
ingti
meof10mi
nut
esmustel
apsebef
orecar
ryi
ngoutt
hesecondser
ies
oft
ast
ing.

 Bi
tt
ernessv
alue(
uni
ts/
g)i
scomput
edf
rom Bi
tt
ernessv
alue(
uni
ts/
g)=(
2000x
C)/(AxB)

 whereA=theconcentrati
onoft heherbalst
ocksoluti
on(Sh)in(mg/mL),B=the
vol
umeofSh( i
nmL)i nt hetubewitht hethr
esholdbit
terconcentr
ati
on(TBC)
,
and C =thequant i
tyofqui ninehydrochl
ori
de( i
nmg)i nt hetubewiththe
thr
eshol
dbit
terconcentr
ation.

3.
1.3.
12Swel
l
ingi
ndex
Crudedr ugscont
aininganappr eci
ableamountofhet er opol
ysacchar i
dessuchasgums,
mucilage’ sandpectinareofspeci fi
ct her apeuti
corphar maceuti
calut i
lit
yandt heyare
charact eri
zedbyt heirswel l
ingpr oper t
ies.Theswel lingindexi st hev olume( inml)
occupi edby1gofadr ugaf terithasswol leninanaqueoussol utionf or4hour s.The
drugi st r
eatedwi
th1mLet hanol(96%)and25mLwat erinagraduat edcy li
ndershaken
every10mi nfor1hourandal lowedt ost andasspeci fied.
Insomecasesaswi thli
nseedandpsy li
um seedswher ethemuci l
agei si nal ayernear
thesur faceofcrudedr ugt henwhol edr ugi ssubj ectedt otheprocedur e.Howev erin
othercasessuchasmar shmal low r
oot swher et hemuci l
ageisdi stri
butedt hroughout
thetissuest hedr
ugi spowder edandt heni tissubjectedt otheprocedur e.

Table1.
6crudedr
ugswi
tht
hei
rspeci
fi
crangeofswel
l
ingi
ndex
Crudedrugs Swel
li
ngi
ndex(mL)

Agarwhol
edr
ug 10

Fenugr
eekwhol
edr
ug 06

I
sapghul
ahuskpowder
eddr
ug 40

I
sapghul
aseedwhol
edr
ug 09

Li
nseedwhol
edr
ug 04

Li
nseedpowdr
eddr
ug 4.
5

Mar
shmal
l
owr
ootpowder
eddr
ug 10

3.
1.3.
13Foami
ngi
ndex
Saponinri
chplantdrugscancauseper si
stentf
oam whenanaqueousdecocti
onis
shaken.The foami
ng abil
i
tyofan aqueous decocti
on ofcr
ude drug mater
iali
s
measuredint
ermsoff oami
ngindex
.
 1gofcrudedrugmateri
alist
ransf
erredtoa500mLconi
calf
laskcont
aini
ng100

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

mLofboi
l
ingwat
er.

 After30minofmoderat
eboi
l
ing,coolandfi
l
t eri
ntoa100mLf
laskandmake
thevolumeoff
il
tr
ateupt
o100mLwi thsuf
fi
cientwat
er.

 Transfersuccessi
veport
ionsof1mL,2mL,3mLup t o 10 mLofdecocti
on
respecti
vel
yinto10mLstopper
edt estt
ubesandmakev olumeupto10mLwith
water,st
opperandshaket
hem for15secondsandsetasidefor15mi
nut
es.

 Measuretheheightofthefoam i
fitisl essthan1cm inalltubesthefoaming
i
ndexistakenasl esst
han100,ifi
ti smor ethan1cm inalltubesthefoaming
i
ndexisover1000andt esti
stober epeatedbydil
uti
onsuchthatfoam hei
ghtof
about1cm isgotinatl
eastoneofthet ubes.

 Thevol
umeinmLofthedecoct
ionusedf
orpr
epar
ingt
hedi
l
uti
onint
het
esttube
wher
ef oami
ngof1cm heightisobser
vedi
susedtodeter
minet
hefoaming
i
ndexusingt
hef
ormul
a1000/a.

3.
1.3.
14Ut
il
izat
ionofchr
omat
ogr
aphi
candspect
roscopi
ctechni
quesi
nphy
sicalev
aluat
ion
Theut i
li
zati
onofchr omat
ographi
candspect roscopictechni
quesismor econvinci
ng
wellunderst
oodandsy st
ematical
l
ypracti
cedmet hodi nquali
tycontr
olofcrudedrugs.
Dif
ferentchromatogr
aphicandspect
r oscopi
cmet hodsofanalysisarealsophy si
cal
methodsofev al
uati
on.

3.
1.3.
14.
1Chr
omat
ogr
aphi
ctechni
ques
Chromat ographyisessent i
al l
yagroupoft echniquesforthesepar ati
on,identif
icati
on
andpur if
icati
onoft hecompoundsofmi xturebyt hei
rcontinuousdistri
butionbet ween
twophasesoneofwhi chismov i
ng(mobil
ephase)andot herisfixed(stat
ionaryphase) .
Classif
icati
onofchr omatogr aphi
ctechni
que’s
Ont hebasisoft ypeofst ati
onaryphasechr omatographictechniquesar ebr oadl
y
classif
iedintot womechani sms:
 Sol id stati
onary phase and a li
quid orgaseous mobi l
e phase ( adsorpti
on
chromat ography)
.

 Li
quid stat
ionar
y phase and a l
i
qui
d orgaseous mobi
l
e phase (
par
ti
ti
on
chr
omat ogr
aphy).

Onthebasi sofgeomet r
yofchr omatogr
aphysystem chromat ogr
aphytechniquesare
br
oadl
ycl assif
iedas:
 Pl anarchromatographyi
sachr omatogr
aphictechniquei nwhi
cht hesupport
ing
medi um isaflatbedorpl anesur
facerat
herthanacol umn.Substancestobe
separ at
ed are appli
ed as a spots on tot he pl ane sur
face.E.g.paper

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

chromatography(
PC),t
hinlayerchromat
ogr
aphy(
TLC)andhi
ghper
for
mance
thi
nlayerchromat
ogr
aphy(TLC)etc.

 Columnchr omatographyi satechni

quei nwhicht hesubst ancestobeseparated
areintroduced ontot het op ofacol umn packed wi t
h an stati
onar
yphase,
substancesar esupposedt opassedt hroughthecol umnatdi ff
erentr
atesthat
dependont heaffi
nit
yofeachsubst ancewi t
ht hestationaryphaseandwi t
ht he
mobile phase e.g.high-perf
ormance li
quid chromat ography( HPLC)and gas
chromat ogr
aphy(GC)et c.

3.
1.3.
14.
2Ter
msusedi
nchr
omat
ogr
aphy
 Retent i
onI tisnot hingbutameasur eofspeedatwhi chacompoundmov esi na
chromat ographicsy stem.I ncolumnchr omat ographysy st
emsl ikeHPLCandGC
ther etenti
oni susual l
ymeasur edasr etenti
ont i
meRtwhi chist het i
mebet ween
i
nject i
onanddet ect ionorr etenti
ontimeRti sameasur eofthet i
met akenf ora
salut etopasst hr oughachr omatographycol umn.I nplanarchr omat ography
syst emsr etentioni smeasur edast heRet enti
onf actorRfwhi chi sther atioof
distancet ravell
edbysol utedividedbydistancetravell
edbysol v
ent .
 Chr omat ogram and chr omatograph a chr omat ogram i st he out put/resul
t
(productoft hatt echni que).Thechromat ographi stheinstr
umentt hatisusedt o
gener atethechr omat ogram.
 Nor mal Phase and r ever
se phase chr omat ography in a nor mal-phase
chromat ography ,apol arstat
ionaryphase( suchassi l
ica)andanon- polarmobi l
e
phase ( e.g.chl or ofor m and hexane)ar e empl oyed whereas r eversed-phase
chromat ography ,incont rast
,hasanon- polarstati
onar yphaseandmoder atel
y
polarmobi lephase.

3.
1.3.
14.
3Chr
omat
ogr
aphi
cfi
nger
pri
nti
ngofcr
udedr
ugs
Cr
ude drugscont ain a myr
iad ofchemi calconsti
tuent
s,which are consi
dered as
marker
sofcr udedr ugusedforstandardi
zati
onorevaluati
onpurpose.
Marker
sar ebasicall
yoffoll
owingtypes:
 Act iveconst i
tuent
’soneoraf ew const
it
uentsspecif
ictoparticul
arcr
udedr ug
responsiblefortheclaimedact i
vi
tye.g.cur
cumi nfortur
mer i
candment holfor
peppermi ntetc.

 Generalchemicalconsti
tuent’
scompoundswi del
ypresentinmanypl antssuch
asgall
icacid,l
upeolandflavonoidsetc.
Thesemarkersar eempl oyedt oassesst heov erallqual
it
yorpur i
tyofcr udedrug
thr
oughchromatographicfi
ngerpri
nting.Chromat
ogr aphi
cfingerpr
intofaplantextr
act
i
si ts chr
omat ographi
c pattern dev el
oped under a st andardr eput
able set of
exper
iment
al condi ti
ons. Dev el
oping such r eproduci
ble and hi ghly specifi
c

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

chromat ographi cpat t

ernsf orcr udedr ugshasbeenasi gnif
icantmi lestonei ncr ude
drugev aluation.l ikewiseTLCandHPTLCf i
nger pri
nti
ngi sofkeyi mpor tancei ncr ude
drugsst andar dizat i
onwher et hecr udedr ugext r
actsareanal yzedwi t
hr espectt ot heir
patternunderspeci f
icexper i
ment alcondi ti
onanddat acanber ecordedi nt ermsof
specificRfv aluesofmar kercomponent spr esentinapar ti
cularcr udedr ugext ract.
Thust hei nformat i
ongener atedf rom t heTLCandHPTLCchr omat ogram haspot ential
applicati
oni nt hei dentif
icati
onofanaut henti
ccr udedrugbyexcl udingadul terant sand
alsoi nmai ntainingt hequal i
tyandconsi stencyofcr udedr ugsampl es.Si milarlyHPLC
andGCf i
nger printingincludesr ecordi ngofchr omat ogramsi nt ermsofr etentiont i
me
ofindi v
idualmar kercomponent spr esenti nacr udedr ugsampl es.Inanut shel lthe
resultsf r
om t hese sophi sti
cated t echniques pr ovi
de f i
ngerprints ofchemi calor
i
mpur i
ti
espr esenti nacr udedr ugsampl e.

3.
1.3.
15Spect
roscopi
ctechni
ques

3.
1.3.
15.
1Basi
csofspect
roscopy
 Waveawav ecanbedescri
bedasadi st
urbancethattr
avel
sthroughamedium,
t
ransport
ingenergyf
rom onelocat
iontoanot herl
ocati
onwithouttr
anspor
ti
ng
matter
.

 El ect
romagnet i
cradiati
ons( EMR)arewaveswhi
chareassociatedwit
hmagneti
c
fi
eldandel ectr
icf
ield.Orelect
romagnet
icwavesarewavesthatarecreat
edasa
resul
tofvibrati
onsbet weenanelectr
icf
iel
dandamagneticfi
eld.
Characteri
sti
csofEMR
Crestandt r
ought hehighestpointofawav eisknownas' crest
'whereast
he
l
owestpoi ntisknownas' tr
ough'.

Wavelengt
hiti
sthedist
ancebet
weent woadjacentcr
ests(C-C)ort
roughs(T-
T)
i
napar ti
cul
arwave.I
tisdenotedbyl et
terλ( l
ambda).Itcanbeexpressedin
uni
tssuchasangst
rom,nanometer
,centi
meterandpicometer.

Frequencyiti
sdefinedasnumberofwaveswhichcanpassthr
oughar ef
erence
pointinonesecond.Iti
sdenot
edbylet
terV(nu)i
tisexpr
essedinuni
tssuchas
cyclespersecondorHertz(
Hz)
.

ForanEMRFrequencyi
sdi
rectl
ypropor
ti
onalt
oener
gy,
great
ert
hef
requencyof
EMRgreat
erwil
lbeener
gyofthatEMR.

Fr
equencyand ener
gyarei nver
sel
ypropor
ti
onalt
o wav
elengt
h gr
eat
ert
he
wavel
engt
hsmall
erthef
requencyandenr
gyofEMR.

Wavenumberi
tisdef
inedasnumberofwaveswhichcanpasst hr
oughaspace
ofonecm.iti
sdenot
edbylet
terύ(nubar
)iti
sthereci
procalofwavel
engt
hand

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

i
tsuni
tispercent
imet
er.

Wavenumberi
sinver
sel
yproport
ionalt
owav
elengt
hgr
eat
ert
hewav
enumberof
EMRshort
erwil
lbeit
swavelengt
h.

3.
1.3.
15.
2Spect
roscopyt
hest
udyofst
ruct
ureandpr
oper
ti
esofat
om bymeansoft
he
spect
r alinformationobtainedfr
om interacti
onofEMRwi thatom orinsi mpleter
msi tis
thestudyofi nteractionbetweenmatterandEMR.
Inspect roscopict echni
quesEMRi nt hef orm ofmonochromaticli
ghti spassedf r
om
anal
yteorsampl esol ut
ion.Theanalytesoluti
onabsorbsoremi t
st hoser adi
ati
onst he
magnit udeoft hoseabsor bedoremitt
edr adiat
ionsbyanaly
tesolut
ionismeasur edwith
thehelpofi nstrumentspect r
ometerli
kespect rophot
ometerorspectrofurometeretc.

3.
1.3.
15.
3Rel
evancyofabsor
pti
onandemi
ssi
onspect
roscopywi
thcr
udedr
ugev
aluat
ion
Theener gylev el
sofamol ecul
eateachst atearequanti
zed.Toexciteamol eculefrom
groundst at
et oexcitedstatetheexactamountofener gyequalt
ot hedistancebet ween
twostateshast obeabsor bed.Thev alueofenergylevel
svari
eswithsubst ances.
Molecularabsor pti
onspect raaremeasur eofamountofEMRataspeci ficwav elength
absorbedbyasubst ancewhileincaseofemi ssionspectr
oscopyt hemagni tudeof
radi
ati
oni ntensit
yemi t
tedbyasubst ance.Theisolat
edandpur i
fi
edpl antconstituents
canbei dentifi
edbyt hei
rspectralcharacter
isti
cs.Spectr
oscopyisusedt oanal yzet he
spectr
aofcr udedrugsaf t
erabsorptionoremi ssi
onofEMR.

Tabl
e1.
7ty
pesofspect
roscopi
ctechni
que’
sont
hebasi
sofEMR

Ty
peofEMR Typeof Ef
fectonmol
ecul
e I
nformati
on
spectr
oscopi
c obtai
ned
techni
que

Radi
owav
es Nuclearmagneti
c Changeinthe Str
uct
ure
resonance magneti
c det
erminat
ion
spectroscopy pr
opert
iesof
at
omicnulei

I
nfr
a-r
edwav
es Inf
rar
ed Changei nthe Det
ectionofal
most
spect
roscopy vibrational
and al
lfuncti
onal
rotational gr
oups
mov ementsof
mol ecules

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Vi
si
ble(
380-
780 Uv/Vi
s Changei n Numberofdoubl e
nm) spect
roscopy el
ectroni
cenergy bondsor
l
evelwithint
he unsaturati
onand
Ul
tr
avi
olet(
180-
380 molecule aromatic
nm) conjugati
onwi t
hin
themol ecule.
Detecti
onof
functi
onal groups.

3.
1.3.
15.
4Fl
uor
escenceanal
ysi
s
Theor ganicmol eculesabsorbli
ghtoveraspecifi
cr angeofwav elengthandmanyof
them r eemitsuchr adi
ati
onswhent hereemissi
onofabsor bedl i
ghtlastonl ywhilst
substancei sr
ecei vi
ngtheexcit
edray
st hephenomenoni sdefinedasf l
uorescence.
Fluorescenceist hephenomenonexhi bi
tedbothinv i
sibl
eandUV-l i
ghtbyv ari
ous
chemi calconstituents pr
esentinthe plantmaterial
.Some cr ude drugs are of
ten
assessedqual i
tativelyi
nthiswayanditisanimportantparamet erofpharmacognost ic
evaluation.

Tabl
e1.
8cr
udedr
ugswi
tht
hei
rspeci
fi
cfl
orescencechar
act
eri
sti
c

Cr
udedr
ug Col
oroff
lor
escence

Ci
nchona Pur
plebl
ue

Rhubar
b Vi
olet

Quassi
a Whi
ti
shbl
ue

Gent
ianr
oot Whi
ti
shbl
ue

I
pomoea Deeppur
plev
iol
et

3.
1.4Chemi
cal
eval
uat
ion
Deter
minat
ionofchemicalconsti
tuent
sinacr
udedr
ugbyspeci
fi
cchemi
calt
est
sis
ref
err
edtoaschemical
evaluat
ion.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

3.
1.4.
1Stepsinvol
vedi
nsyst
emat
icapproacht
owardschemi
calev
aluat
ion
3.
1.4.
1.1Procur
ementofr
awmateri
alandqual
it
ycontr
ol
Iti
sthefirststeptowardschemicaleval
uat
ionofcrudedrugitinv
olvesthearr
angement
ofplantoritspartther
eofwhichwillser
veasar awmat er
ialwhosechemicalevaluat
ion
i
ssupposedt obeexecut ed;fur
thermoreauthenti
cati
onoft hatplantmaterial(r
aw
material
)shoul dbedonebef oreperfor
mingext r
acti
on.Anyf or
eignmat t
ershouldbe
completelyeli
minated.
3.
1.4.
1.2Pr
epar
ati
onofext
ract
Oncetheaut hent icat i
onofr awmat erialisest abl i
shed, themat erialisfurthersubj ected
t
oextract i
on.
 Ext ractioni nv olvest hesepar ationofmedi ci nallyact i
vepor t
ionofpl antorani mal
tissuesf r
om t hei nact i
v eori nertcomponent sbyusi ngsel ectivesol vent sin
st andar dext ractionpr ocedur esorExt ract i
oni sapr ocessofsepar atingsol uble
mat eri
al from ani nsol ubl eresi duebyt reatmentwi thasui t
abl esolv ent.
 Pur poseofext r
act i
ont hepur posesofst andar dizedext ractionpr ocedur esf or
cr udedr ugsar et oat tai
nt het herapeut icallydesi redpor t
ionandt oel i
mi nat ethe
iner tmat erialbyt reat mentwi thasel ectivesol vent.Theext r
actt husobt ained
mayber eadyf oruseasamedi cinalagent ,ori tmaybef urtherpr ocessedf orthe
ev aluationofi ndiv idual chemi cal entit
iespr esentwi thin
 Gener almechani sm ofext ract iont hepr i
nci pleofsol i
d- l
iquidext ractioni st hat
whenasol idmat erialcomesi ncont actwi thasol v
entofsui t
abl epol ari
ty,the
sol ublecomponent si nt hesol idmat er i
almov et ot hesol ventr esultingi nt he
masst ransf erofsol ubleact ivepr i
nciple( chemi calconst ituents)t ot hesol vent
.
Thi stakespl acei naconcent rat i
ongr adi ent.Ther ateofmasst r
ansf erdecr eases
ast heconcent rationofact ivepr i
nciplei nt hesol ventincreases, unt ilequi l
ibri
um
isr eached.
 Sel ectionofsol vent si fthet her apeut i
cv aluel iesi nnon- polarconst i
tuent s,anon-
pol arsol ventmaybeused.Forexampl e,l upeoli st heact iv
econst i
tuentof
Cr ataev anur valaand,f ori tsext racti
on,hexanei sgener allyused.Li kewi se,for
plant sl ikeBacopamonni er iand Cent ell
aasi aticat heact iveconst i
t uentsar e
gly cosidesandhenceapol arsol ventlikemet hanoloraqueousmet hanolmaybe
used.

Tabl
e1.9Li
stofcommonl
yempl
oyedsol
vent
salongwi
tht
hei
rpol
ari
ty
Sol
vent
s Pol
ari
ty

Hexane 0.
1

Pet
rol
eum et
her 0.
1

Cy
clohexane 0.
2

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Benzene 2.
7

Di
chl
oromet
hane 3.
1

I
sopr
opy
lal
cohol 3.
9

Chl
orof
orm 4.
1

Et
hyl
acet
ate 4.
4

Acet
one 5.
1

Met
hanol 5.
1

Et
hanol 5.
2

Acet
oni
tr
il
e 5.
8

Di
met
hyl
sul
foxi
de 7.
2

Wat
er 10.
2

St
epwi
sepr epar at
ionofext ract
a)Sizer educt i
on t hedr ied plantmat eri
alisdi sint
egrat
ed byf eeding itintoa
hammermi lloragr i
nderthepl antmat eri
alisreducedtoasi zebet ween30and
40mesh, butthi scanbechangedi ft heneedar ises.Theobjecti
vef orpowder ing
thepl antmat erialist orupt urei t
sor gan,ti
ssueandcel lstr
ucturessot hati ts
medi cinalingredi entsar eexposedt ot heextracti
onsolvent.Fur t
hermore,si ze
reductionmaxi mi zest hesur facear ea, whichint ur
nenhancest hemasst r
ansf er
ofact ivepr i
nci plef rom plantmat er ialtothesol vent
.The30- 40meshsi zei s
optimal ,whilesmal lerparticl
esmaybecomesl i
mydur i
ngext r
act i
onandcr eate
diff
icultyduringf il
tration.
b)Extractionoft hepl antmat erialiscar ri
edoutbyasui tableextracti
onmet hod,
someoft hecommonl yadopt edextr actionmethodsar eli
stedbelow.
 Macer at i
on

 Per
col
ati
on

 Sox
hletext
ract
ion

 Super
cri
ti
cal
flui
dext
ract
ion(
SCFE)

 Mi cr
owav eassi
stedextr
acti
on( MAE)
I
fthe consti
tuentsarethermolabi
le,extr
act
ion methodslike cold macerat
ion and
per
col
ati
onarepr ef
err
edforthermostabl
econsti
tuent
s,Soxhl
etextract
ionareuseful
.

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

c)Fil
tr
ati
ont heext r
actsoobt ainedi ssepar at
edoutf rom t heexhaust edpl ant
materialbyal l
owingi tt opasst hroughaf i
lt
eringmedi um ( f
il
terclothorf il
ter
paper).Theexhaust edmat erialisretainedov ert hefilteri
ngmedi um andt he
extr
actisreceivedint heholdi ngtank.
d)Concentration and drying theenr i
ched ext r
actt husobt ained isf ed i nt
o an
evaporator(usuall
y,rotaryev aporator)
,wher eiti
sconcent ratedunderv acuum t o
produceat hickconcent rat
edext r
act.Theconcent ratedext ractisfurtherdr i
edt o
produce a sol i
d mass f r
ee f r
om sol vent.The sol ventr ecovered f rom t he
evaporatorisrecycl
edbackf ort heextracti
onofnextbat chofpl antmat erial
.

3.
1.4.
1.3Qual
i
tat
iveanal
ysi
sofpr
epar
edcr
udedr
ugext
ract
Theextract
st husobtai
nedaresubject
edtochemicalt
est
sfort
heconf
ir
mat
ionof
var
iouschemicalconst
it
uent
spresenti
nacr
udedrug.

Tabl
e1.10speci
fi
cqual
it
ati
vechemi
calt
estandt
hei
robservat
ion
Chemical Chemi
calTest Observat
ion
group

Legal
’
stest Pi
nkcol
or

Gl
ycosi
des Kel
l
er-
Kil
l
ani
’
stest Reddi
shbr
ownl
ayeri
sfor
med,

whi
cht
urnsbl
uishgreenaf
terst
andi
ngduet
o
pr
esence

ofdi
git
oxose.

Bor
ntr
ager
stest Pi
nkt
oredcol
or

Mol
i
scht
est Pur
ple/
viol
etcol
orr
ing

Bar
foed’
stest Reddi
shbr
ownppt
s

Car
bohy
drat
es Benedi
ct’
stest Reddi
shbr
own

Fehl
i
ng’
stest Redppt
s

May
er’
stest Cr
eamywhi
teppt
s

Al
kal
oids Hager
’st
est Yel
l
owi
shppt
s

Wagner
’st
est Reddi
shbr
ownppt
s

Dr
agendr
off
’st
est Or
angebr
ownppt
s

Fer
ri
cchl
ori
det
est Bl
uecol
or

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Tanni
ns Gel
ati
ntest Whi
teppt
s

Fl
avonoi
ds Al
kal
i
ner eagent I
ntensey
ell
owcol
or
test

Phy
tost
erol
s Li
ber
mann- Gr
eenbl
uecol
or
Bur
chardt
est

Ter
penes Sal
kowski
test Resi
dhbr
owncol
or

3.
1.4.
1.4Quant
it
ati
veanal
ysi
sofpr
epar
edcr
udedr
ugext
ract
Acrudedrugext racti
sassayedquanti
tat
ivel
yf orapart
icul
argroupofconsti
tuent
,for
exampletotalalkaloi
ds,t
otalflav
onoid’
s,totalphenol
ics,tot
alcarbohydr
ates,t
otal
sennosi
desetc.foll
owingmethodsareadaptedtocarr
youtquanti
tat
iveanal
ysi
s.
 Gravi
met r
icassay

 Aci
dbaset
it
rat
ionbasedassay

 Spect
rophot
omet
ri
cmet
hods

 Chr
omat
ogr
aphi
cmet
hods

3.
1.5Bi
ologi
cal
eval
uat
ion
Iti
sanev al
uat
ionofcr udedrugorcrudedr ugextr
actbyvar
iousbiologi
calmet
hodsto
determineit
stoxici
tyandpharmacologicalact
ivi
ty.
Int he l
ightofdef i
nit
ion st
ated abovet here ar
etwo maj oraspects ofbi
ologi
cal
evaluat
iontoxi
cit
ytesti
nganddeterminationofpharmacol
ogi
cal act
ivi
ty.

3.
1.5.
1Toxi
cit
ytest
ing si
mpl
yst
ated,al
lchemi
cal
sar
etoxi
cwhatdi
sti
ngui
shesone
chemi calf r
om t henexti sthedosewhi chpr oducesbi ologicalresponseandadv erse
eff
ects.Anynew chemi calwhi chappear sint hemar ketorf i
ndsi tswayi nt ot he
dri
nkingwat erorfoodsuppl ymustbecl assifi
edt oitspot entiall
eveloft oxici
ty.
Toxici
t ytestsut i
l
izedi nthet oxicological assessmentpr ocess
 Acut et oxi
citytestinacut et oxici
tyt estingsingl eexposur eofchemi cali
sgivent o
testanimalsandobser vationper iodi s14day s.
 Sub- acutet oxi
cityinsub- acutet oxi ci
tyt est
ingt hedosageusedi sextendedf or
theper i
odof28day s.
 Sub- chronictoxici
tyt estsi nsub- chr oni
ct oxi
ci tytesti
ngt hedosageusedar el ess
thanacut et oxici
tydosagei nor der st heani mal stosur v i
vef orent i
reper i
od.
Dosageoft hetestchemi calsisext endedf ort heper i
odof90day s.
 Chr onict oxici
tyt esti n chr onict oxi
ci tytest i
ng r epeated exposur e oft est
chemi calisgiv enandobser v at
ionper i
odi supt ot woy ears.Chr onictoxicit
y
studi
esar eusual l
yempl oy edi fther eisaconcer nt hatpeopl ewi l
lbeexposedt o

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

thechemi cal forasi gni fi

cantpor t
ionofl i
f et i
me.
 Repr oduct i
v e and dev elopment alt oxici t
yt estt hist ype oft oxici
tyt est ing
eval uat es whet her chemi cal wi ll af fect r eproductive success or i nduce
terat ogeni cef f
ect s.
 Ocul arandski ni rrit
ati
ont estusesr abbi tst odet erminewhet herchemi cal swi l
l
damaget heset issues.Theey e/ ocul art esti nvolvesadmi nisteringaf ixeddoseof
achemi calt ooneey eofar abbit,theot herey eservesasacont rol.Ther eact i
on
i
nt heey ei sev aluat edat24,48and72hour s.Forski nt esti
ngt hebackoft he
rabbi ti sshav edandt het estchemi cali sappl i
eddi rectlyt ot heski nandt he
effectont heski ni seval uat edat24, 48and72hour s.
 Hy per sensi tiv
ityt esti sanext ensi onofski nt esti
ngt oev al uatechemi calswhi ch
maynotdi rectlydamaget heski nbutmayel ictani mmunol ogical response.
 Phot o-toxi cityt esti susedt odet er mi newhet hersunl ightwi l
lact ivat
et het est
chemi cal andpr oducet heski nirritationorhy persensitivi
ty.
 Toxi co- kinet ict esti susual lyundert akenonl ywheni nconsi st entr esul tsar ise.
Manyt i
messpeci esdi fferencesi nt oxi cityar eobser vedf orexampl ebet ween
ratsandmi ce,int hesecasest hedi fferencescanusual lybeaccount edont he
basi s ofdi ff
er ences i n ki net i
c par amet er s such as absor pt i
on,di stribution,
met abol ism andexcr etionoft het estchemi cal.
 Behav iort oxicityt estisempl oyedt oev aluatepot entialneur ologi calef fect sof
chemi cal s ei ther di rect l
y or t hrough neur ological damage dur ing f et al
dev elopmenti nt hel at
t ercasepr egnantr atsar eexposedt ot estchemi cal sand
behav iorpat ternsofneonat esar ef oll
owedatear lystagesofdev elopment .

3.
1.5.
2Det
ermi
nat
ionofphar
macol
ogi
calact
ivi
ty
Thepharmacologicalactiv
ityofadr ugisdeter
mi nedi ntermsofi tseffi
cacyand
pot
ency.
a)Pot encyi
tisameasur eofamountofdr ugnecessar ytopr oduceanef fectofa
giv
enmagni tude.Theconcentrati
onofthedrugt hatproducesar esponseequal
to50%oft hemaxi malresponseisknownasEC50.Thepot encyofdrugcanbe
comparedusi ngtheEC50thesmal l
ert
heEC50t hemorepot entthedrug.

b)Effi
cacyabi l
ityoft hedrug to el
ictther esponsewheni tint
eractswiththe
receptor.Ef
fi
cacyi sdependentonnumberofdr ugr
eceptorcomplexesfor
med
andt heeffi
ciencyofcoupli
ngofr eceptoracti
vati
ont
ocellul
arresponse.Adrug
withgr eat
eref f
icacyismoret herapeuti
call
ybenefi
cialt
hanonet hati
smor e
potent.

3.
1.5.
3Bi
oassay
s
Bi
oassayisthemeasur eofsampl
ebeingt
estedcapabl
eofpr
oducingbiol
ogi
calef
fect
asthatofst
andardprepar
ati
on.
 I nvit
robiassayswhensomethi
ngisperf
ormedinvi
tr
o,ithappensout
sideofa

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

l
i
vingorganism wi
thmolecul
esand/orcel
lsinst
eadofwhol
eorganisms.
 I
nv i
vobioassaysi
tref
erstoworkthat
’sperf
ormedinawhole,
li
vingorgani
sm.

Bat
ch2k23
HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

Bat
ch2k23
 HoshMuhammadLashar
i,Depar
tmentofPhar
macognosy
,Facul
tyofPhar
macy
,U.
O.SJamshor
o

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erences
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http:/ /downl oadsphar macy .blogspot .
com/ 2011/ 07/phar
macognosy -ckkokat e_21.ht ml .
2.Evans,W. C.,Tr easeandEv ansPhar macognosy .6ed.2009:Edi nbur ghLondon
New
Yor kPhiladelphiaSt Loui sSy dney Tor ont o.ht
tp: /
/downl oadsphar macy .blogspot .co
m/ 2011/ 01/ pharmacognosy -by-treese- and-evans_ 4513. html.
3.Shah, B.andA. K.Set h, Tex tbookofPhar macognosyandPhy tochemi stry.Firsted.
2010, ReedEl sev i
er India Private Limi t
ed: El sev i
er . 587.
https://www. pdfdr i
v e.com/ textbook- of -
phar macognosy -
and-phy tochemi stry-
d184620437. html.
4.AbdulHakeem Memon,MadeehaLaghar i
,andN.Memon,Fol kMedi cinal&
Ethnomedi cinalusesofsi xtyPl ant sofSi ndh Paki stan 2012:LAP LAMBERT
Academi cPubl i
shi ng( December9, 2012) .
5.Qual i
tyCont rolMet hodsf orMedi cinal PlantMat eri
als.WHO, Genev a;1998.128p.
http://aps.who. i
nt /medi ci
nedocs/ collect/medi cin edocs/ pdf/h1791e. pdf .
Accessed2009Oct2.
6.Har borneJB.Phy tochemi calMet hods.2ndEdi ti
on.ChapmanandHal l
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