Volume: 2: Issue-2: April-June -2011 ISSN 0976-4550

EXTRACTION AND EVALUATION OF PIPERINE FROM PIPER NIGR! LINN.

Smita R. Kolhe
1
*, Pi!a"#a $oole

,Umi Patel
1
Department of Pharmaceutics, Progressive Education Societys Modern College of
Pharmacy (For adies! Moshi, Pune" #1$1%&'M'S' ((ndia!
A$STRACT% Piperine is the naturally occurring al)aloid that gives the spice, *lac) pepper its
characteristic *iting taste' Piperine is e+tracted from Piper nigrum, inn' (t is medicinally used as
,nti"inflammatory, ,nti"malarial, Cure for Stomach ,che, -eight oss, ,nti"leu)emia, Fever
prevention.reducer, /reatment for Sna)e 0enom Poisoning and ,nti"epileptic' 1ecent medical studies
have sho2n piperine to *e very helpful in increasing the a*sorption of certain vitamins such as
Selenium, 0itamin 3 and 3eta"Carotene' Piperine apparently has the a*ility to increase
thermogenesis'4ence, E+traction and Standardi5ation of piperine is performed' (n Standardi5ation,
Solu*ility, /otal ash content, ,cid insolu*le ash thin layer chromatography, 6ltra"violet
Spectroscopy, 4igh Performance i7uid Chromatography 2ere performed'Piperine 2as e+tracted
successfully, it 2as found to *e solu*le in ethanol and chloroform' /otal ash content and acid
insolu*le content 2as found to *e 182.2 and %'&82.2 respectively' /he 1
f
value 2as found to %'&&
and %'99 for /C 'For 60 the :
ma+
2as found to *e 9;%nm and the a*sor*ance o*served 2as $';&'
4PC results sho2ed that the :
ma+
2as found to *e 9#9nm' /he melting point of piperine 2as found to
*e 1$&
%
C'
Ke!&o'(< Piperine, 3lac) Pepper, E+traction, Standardi5ation
INTRODUCTION
/he common her*al product )no2n as the *lac) pepper is a 2ell )no2n spice around the 2orld '/he
plant itself is a perennial 2oody clim*ing vine 2hich can reach a height of a*out 1& ft or & meters
clim*ing on trees and along man made 2ooden trellises' /he her* *ears characteristic and large oval
shaped leaves, it also has spi)es of many small and 2hite colored flo2ers, the commercial pepper
itself is made from the many clusters of small round fruits *orne on the plant, these small fruits are the
pepper of common use, and these fruits ripen from a green to red coloration as they mature on the
vine' 3lac) pepper has played a ma=or role in shaping historical events in the 2orld' Piperine a ma=or
constituent is a mem*er of the ipids family' ipids are a group defined as consisting of fats or fat
li)e su*stances' ,s an aromatic spice, the *lac) pepper is characteri5ed *y a slight and percepti*le
musty aroma> this smell is *ecause of the presence of many volatile oils mainly in the flesh and s)in
of the fruit' /he very noticea*le and pungent ?*ite@ of the *lac) pepper is due to the presence of t2o
primary al)aloids in the pepper, these are the compounds piperine and the compound piperidine, the
tangy *ite also comes partly *ecause of the many specific plant resins 2hich are found in the seeds of
the her*' ,side from its normal and common use as a spice, these pepper oils are also e+tracted to *e
used in the manufacture of many types of perfumes and commercial food seasoning materials around
the 2orld' Pepper *ased irritants and searing chemicals are utili5ed in a variety of processes' Some of
these chemicals are used in a variety of liniments and mouth gargles' /he pepper *ased compounds
have also seen commercial use as chemical carminatives> they have also *een used for their a*ility to
reduce e+cess gas in the intestines and the stomach' ,side from these, these compounds have also
*een used to stimulate the activity in the human heart and )idneys in medicine' /he specific
compound in pepper )no2n as piperine is commercially utili5ed to prepare different insecticides to *e
used against houseflies and other insect pests'
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MATERIALS AND MET)OD%
Mateial(
Pepper (*lac)! 2as procured from local mar)et 2hile remaining all chemicals 2ere o*tained
from college la*oratory'
Metho'
(' EXTRACTION OF PIPERINE FROM $LACK PEPPER
E+tract po2ered drug 2ith ethanol (A&8! in a So+hlet apparatus for 9 hours' Filter the
solution and concentrate under vacuum on 2ater *ath at ;%BC' ,dd 1% 8 alcoholic
potassium hydro+ide 2ith constant stirring and filter' ,llo2 the alcoholic solution to
stand overnight 2hereupon needles of piperine separate out'
*. EVALUATION OF PIPERINE%
I. Chemi+al Chaa+tei(ti+(%
1' Crganoleptic properties of dried 3lac) pepper and Piperine li)e colour, odour, taste 2ere
determined'
$' /he e+tract content of the piperine 2as determined *y ta)ing 82.2
9' /he Solu*ility of piperine in various solvents li)e petroleum ether, chloroform, ethanol,
methanol and 2ater 2as carried out'
#' Moreover, the determination of melting point of piperine 2as also carried out'
II. Ph!(i+o+hemi+al A"al!(i(%
,' /otal ash content
3' ,cid insolu*le ash contents are analy5ed and compare 2ith standards
III. Thi" La!e Chomato,a-h! .TLC/ A"al!(i(<
Sample Details< Piperine
,dsor*ent < Precoated Silica gel
Solvent System< /oluene< Ethyl acetate
D% < 9%
Sample Preparation< Piperine (1 mg! 2as dissolved in methanol (1 ml! and this solution
2as applied on the /C plate 2ith the aid of capillary tu*e'
Detection< Saturated (odine Cham*er
IV. )i,h Pe0oma"+e Li12i' Chomato,a-h! .)PLC/ A"al!(i(
Mo*ile phase< Methanol< -ater
;A < 91
Detection < 9#9 nm
Flo2 rate < 1'& ml.min
0' Ulta Violet3 S-e+to-hotomete A"al!(i(
/he %'%18 2.v solution of piperine in ethanol 2as prepared and :
ma+
2as determined'
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RESULTS
I. Chemi+al Chaa+tei(ti+(
1' Craganoleptic Characteristics<
Ta4le 1% O,a"ole-ti+ +haa+tei(ti+(%
*. E5ta+t Co"te"t% /he e+tract content of oleoresin is found to *e 1'%;82.2
6. Sol24ilit!< /he solu*ility of oleoresin in various solvents 2as found to *e as follo2s<
Ta4le *% Sol24ilit! o0 -i-ei"e i" 7aio2( (ol7e"t(.
8. Melti", Poi"t< Melting point 2as found to *e 1$A
%
C
II. Ph!(i+o+hemi+al A"al!(i(<
/he total ash value and the acid insolu*le ash value of the oleoresin 2ere found to *e as
in ta*le 9'
Ta4le 6% Ph!(i+o+hemi+al a"al!(e(%
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Colo 3lac)
O'o2 ,romatic
Ta(te ,romatic and Pungent
Petole2m ethe Solu*le
Chloo0om Solu*le
Etha"ol Solu*le
Metha"ol Solu*le
9ate (nsolu*le
Properties Standard value Practical value
/otal ash Content E ;'%82.2 18 2.2
,cid (nsolu*le ash Content E 1'%82.2 %'&8 2.2
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III. Thi" La!e Chomato,a-h! .TLC/ A"al!(i(<
/he 1
f
value 2as found to *e %'&&, %'99, as t2o spots 2ere produced in /oluene< Ethyl
acetate system'
Fi,2e 1% TLC A"al!(i( o0 e5ta+t
III. )i,h Pe0oma"+e Li12i' Chomato,a-h! .)PLC/ A"al!(i(%
Fi,2e *% )PLC A"al!(i( o0 e5ta+t
File "ame% ) -i-ei"e :; --m
MP< 2ater< MeC4 F9%<D%G
Column< 4iH Sil C1I-
Flo2 rate< 1 ml.min
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Ta4le 8% Re(2lt( o0 )PLC
max 1/ ,reaFJ0'SecG Plates 1esolution 4eightFJ0G ,symmetry 8 ,rea
9#9nm 1&'%#9 #$$ID1D'%## 9#99'A;A %'%% 1%$$%; 1'D9 1%%'%%
IV. Ulta Violet3 S-e+to-hotomete A"al!(i(

Ta4le :% Re(2lt( o0 UV
,*sor*ance :
ma+
$';&9&I 9;%nm
Fi,2e 6% UV S-e+to(+o-i+ a"al!(i(
DISCUSSION
/here are numerous types of (ndian *lac) pepper ,that is 2hy pepper as 2ell as piperine is
characteri5ed *y variations in colour,odour and taste 2hich sho2ed characteristic 3lac) and yello2
colour,aromatic odour,aromatic and pungent taste for pepper and piperine respectively'
Piperine 2as found to *e solu*le in various nonpolar solvents *ut insolu*le in polar solvent li)e
2ater'
Piperine 2as also characteri5ed for its melting point 2hich sho2ed that it undergoes melting at 1$ABC
2hich is 2ithin the standard range (1$A"19%BC!
/otal ash method is designed to measure the total amount of material remaining after ignition 2hile
the acid insolu*le ash is the residue o*tained after *oiling the total ash 2hich measures the amount of
silica present ',mount of total ash 2as found to *e 18 2.2 (E;82.2! and acid insolu*le ash 2as
found to *e %'&82.2 (182.2!'
/hin ayer Chromatography a particularly valua*le 7ualitative method for determination of small
amount of impurities and fre7uently used for evaluating medicinal plant materials' Different samples
have different 1f value 2hich 2as determined for piperine and 2as found to *e %'&&9I and %'99I#
(%'$& and %';! 2hich sho2ed good separation of piperine'
Piperine in Piper nigrum 2as separated from other compounds *y 4PC' /he separated compounds
2ere compared 2ith the standard Piperine pea)'
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CONCLUSION
/he proposed method for e+traction and characteri5ation lead to rapid, accurate and sensitive method
re7uiring less amount of solvent' arge num*er of samples can *e suita*ly employed for routine
7uality control analysis'
ACKNO9LED<EMENT
/he authors are than)ful to the Principal, P'E'Ss Modern college of Pharmacy, Moshi, for providing
necessary facilities for the pro=ect 2or)' ,lso 2e than)ful to 3otonical survey of (ndia for
authentifying the given sample and to 3om*ay College of Pharmacy for 4PC analysis'
REFERENCES
1' ,'K' Mu)har=ee ($%%$!' Huality Control of 4er*al drugs, 3usiness hori5on
Pharmaceutical Pu*lication< 1
st
Edition, page no' 9I$";%&'
$' C'K' Ko)ate, ,'P' Purohit , S'3' Lo)hale ($%%I!' Pharmacognosy, #$nd Edition, Mirali
Pra)ashan, Page no' 11'&;"11'&I
9' 4' -agner, S' 3ladt ($%%D!' Plant Drug ,nalysis, , /C ,tlas, ((
nd
Edition, page no'
$AI"9%%
#' http<..222'her*s$%%%'com.her*s.her*sN*lac)Npepper'htm
Dated on< %1"$%"$%1%
&' http<..iher*al'*logspot'com.$%%;.%$.piperine"uses"effects'html'
;' (ndian 4er*al Pharmacopoeia, 1evised Me2 Edition ($%%$!' Page no'91D"9$#
D' K'1' Khandel2al ($%%9!'Practical Pharmaconosy, /echni7ues and E+periments,
Mirali Pra)ashan, page no' 1&D
I' /he -ealth of (ndia, 1a2 Materials ($%%&!'Ph"1e, , Dictionary of (ndian 1a2 Materials
and (ndustrial Products, CS(1, Me2 Delhi, volume 0(((, page no' I9"11I'
A' 0'D'1angari ($%%9!' Pharmacognosy and Phytochemistry, Part ((, First Edition, Career
Pu*lication, page no' $&D" 991'
1%' -'C'Evans ($%%%!'/rease and Evans> Pharmacognosy,1&
th
Edition, Page no'
9&9"9&#
In"ern#"ion#l Journ#l o$ Applie% &iolo'( #n% P)#rm#*eu"i*#l +e*)nolo'( P#'e:149
A,#il#-le online #" .../i0#-p"/*om