piperonal from pepper

piperine from pepper

2002, by 3base


isolation of piperine (black pepper, EtOH, KOH)
isolation of piperine from black pepper (black pepper, EtOH, KOH, soxhlet)
isolation of piperine from black pepper (black pepper, IPA, CaCO3)
isolation of piperine from black pepper (black pepper, CH2Cl2)
isolation of natural piperine (pepper, EtOH, KOH, soxhlet)
extraction and isolation (piper nigrum, EtOH)
piperine and piperiline extraction (black pepper, EtOH, soxhlet, hexane:ether column)
piperine form piper longum (piper longum, CH2Cl2:MeOH, PE:EtOAc column)

oleoresin of pepper
   piperine from resin of black pepper (black pepper resin, EtOH, urea)
   source of piperine (black/long pepper/oleoresin, BuOH, hexane, alumina, charcoal)
   oleoresin of pepper [03] (black pepper, ether)

further recipes and references


isolation of piperine

place 15g (1) of ... ground black pepper in a 250mL round-bottomed flask,
add 150mL of 95% ethanol and 5 boiling chips (2), and heat at reflux for 2h.
filter the mixture by suction filtration and then concentrate the filtrate to a
volume of 10-15mL by simple distillation or by use of a rotary evaporator.
to 10mL of a 10% solution of KOH in 95% ethanol contained ina 125mL
erlenmeyer flask add the concentrated pepper extract. heat the resulting
solution and add water dropwise. a yellow precipitate forms. add water
until no more solid appears to form and then allow the mixture to stand at
least overnight (3). collect the solid by suction filtration and recrystallize it
with 10-20mL of acetone (4).

(1) this extraction may be scaled up to twice the amounts specified without
difficultiy. if a soxhlet extractor is available, this would be an apparatus superior
to a standard reflux setup.

(2) boiling chips are necessary to prevent serious bumping.

(3) it is best to allow piperine to completely precipitate out by allowing the
mixture to stand until the next laboratory period.

(4) in our hands ca. 0.6g of piperine, mp 127-128C, was collected upon

[05, page 527-528]


isolation of piperine from black pepper

10g black pepper is ground to a fine powder and extracted with 150mL 95%
ethanol in a soxhlet extractor for 2h. the solution is filtered and concentrated in
vacuo on a water bath at 60C. 10mL 10% alcoholic KOH solution is added ...
and after a while decanted from the insoluble residue. the alcoholic solution is
left overnight, whereupon 0.3g yellow needles are deposited, mp 125-126C.

TLC of black pepper extract
the crude extract is spotted on a thin-layer plate (silica gel GF254)
and developed with benzene:EtOAc 2:1.
(1) UV365 shows blue fluorescence of piperine
(2) spraying with anisaldehyde-sulfric acid reagent, prepared by mixing 0.5mL
anisaldehyde with 10mL glacial acetic acid, 85mL MeOH, and 5mL concentrated
sulfric acid. this solution is sprayed on the plate, which is then heated at 110C
for 10min. piperine appears as a yellow spot, Rf=0.25.

[06, page 233-236]


isolation of piperine from black pepper

piperine, a very weakly basic substance, can be isolated from a variety of peppers
by extraction with alcohol. ... piperine, along with a small amount of its Z,E isomer,
accounts for about 10% of the weight of black pepper.

place 15g of ground black pepper and 1g of powdered CaCO3 (1) in a 250mL
boiling flask, add 100mL of IPA, and, after fitting the flask with a reflux condenser,
boil the mixture for about 1h on the steam bath. at the end of the heating period,
filter the mixture by gravity into a 125mL erlenmeyer flask, clean the 250mL
boiling flask, and return the filtrate to the boiling flask. fit the boiling flask with a
distillation adapter and condenser, and boil off all but about 10mL of the IPA (2).
transfer the residual solution from the boiling flask to a 25mL erlenmeyer flask,
and set the flask aside to cool for crystallization of piperine (3,4). collect the product
by suction filtration, using small portions of MeOH to rinse the flask and wash
the product. yield: about 0.5g.

(1) the addition of CaCO3 should prevent the extraction of acidic
components of pepper

(2) IPA boils at 80C, only a little below the maximum temperature attainable on
the steam bath. to make the distillation proceed quickly, clamp the boiling flask so
that it is well down in the rings of the steam bath, and drape a towel over the flask
and the steam bath to make a tent that will hold steam around the top of the flask.

(3) crystallization occurs slowly, and the flask must be allowed to stand for
at least 24h.

(4) alternatively, add 25mL of water to the IPA solution of piperine, allow the mixture
to stand for at least 24h so that precipitation will be complete, collect the solid by
sucction filtration, and recrystallize it from either IPA or acetone.

[04, page 340-341]


isolation of piperine from black pepper

add 5g of pure ground pepper and 10mL of CH2Cl2 to 50mL round-bottomed flask.
use the round-rottomed flask as the basis for a reflux apparatus having a water-cooled
condenser and heating mantle. heat the sample to reflux, and then maintain a gentle
reflux for 20min. after the required reflux period, lower the heating mantle and allow the
reflux apparatus to cool for 5min. suction filter the slurry with the aid of a 4.5cm buchner
funnel, washing the pepper grounds once with 5mL of CH2Cl2. remove 2 or 3 drops of
the extract and place it in a capped vial for use in the TLC analysis.

transfer the extract obtained above to a clean 25mL round-bottomed flask and
concentrate in vacuo. the resulting olive-brown, viscous oil should be cooled in an
ice-bath and then 3mL of cold ether added to the oil while gently stirring for 3-4min.
some piperine may percipitate at this point, but remove the solvent in vacuo anyway.
once again coole the resulting oil in an ice-bath and then add 3mL of cold ether to the
oil while gently stirring to promote the percipitation of piperine. allow the flask to cool
for an additional 10min with occasional stirring. isolate the straw-yellow crystals of
crude piperine by suction filtration with the aid of a 1.5cm hirsch funnel. wash the
crystals twice with 2mL portions of cold ether. place a small portion of the filtrate in a
capped vial for use in the TLC analysis.

place the crude piperine isolated above into a 13x100mm test tube and dissolve it in a
minimum amount of hot 3:2 acetone:hexane solution. once all the solid has dissolved,
allow the test tube to sit undisturbed for 15min at room temperature. rod-like, yellow
crystals of piperine should be present. cool the solution for an additional 30min in an
ice-bath before isolating the purified piperine by suction filtartion with the help of a 1.5cm
hirsch funnel. wash the crystals once with a 2mL portion of cold ether, allow them to air
dry for several minutes. typically, yields of approximately 2% or 100mg are obtained.
the melting point of the purified piperine now can be determined and the identity of the
product confirmed by mixed melting point, TLC analysis, or spectral analysis.

TLC analysis
transfer a small portion of the purified crystals to a small vial and dissolve them in a
drop or two of acetone. prepare a silica-gel TLC plate for the spotting of 4 samples
(kieselgel 60 F254 plates having a 0.2mm coating thickness are recomended).
with the aid of a capillary, place a sample of the crude oil remaining from the extraction
procedure, a sample of the filtrate from the trituration/isolation procedure, the sample
prepared from the purified piperine, and a sample of a piperine standard on separate
points of the thin-layer plate. develop the plate using 3:2 acetone:hexane.
visualize under UV illumination and stain in an iodine chamber.



isolation of natural piperine

grind 25g fresh peppercorns to a fine powder, place in a soxhlet thimble, and extract
with 100mL ethanol for 90 minutes. cool the resulting solution, filter if necessary, and
concentrate on the rotary evaporator. keep the water bath below 60C during the
concentration. dissolve the residue in 25mL 10% alcoholic KOH. decant the solution
if any residue remains. cool the solution in an ice bath, and add water dropwise
(about 30mL will be required) to precipitate the piperine. collect the piperine on a
sintered glass funnel, and dry it on the vacuum pump.
... recrystallization from acetone:hexane 3:2 will produce cleaner material.



extraction and isolation

10kg dried fruits of p nigrum l ... were crushed and extracted 5x with EtOH at room
temp. after removal of solvent in vacuo, the syrupy residue was left overnight at room
temp. when a white crystallizate sept out, which was filtered. on recrystallization from
MeOH it formed 50.54g fine needles, mp 128-129, and was identified as piperine.



piperine and piperiline extraction

750g dry fruits of black pepper were ground and extracted with 2.5L ethanol, in a
soxhlet apparatus for 72h. the extract was concentrated under reduced pressure
in a rotary evaporator to leave 8g of a dark brown oil. this material was fractionated
on a silica gel column, eluting with hexane + diethyl ether (1 + 2 by volume).
the fractions obtained were combined according to their similarities as analysed by
thin layer chromatography (TLC) and this led to the isolation of 2g piperine and 0.3g
piperiline. these compounds were recrystallised with a mixture of dichloromethane
and hexane and the physical and spectroscopic data obtained were in accordance
with those reported in the literature.



piperine form piper longum

p longum ... 500g dried fruits were powdered and extracted 120h with CH2Cl2:MeOH
1:1. the concentrated extract (1.45g) was chromatographed over silica gel, the column
being eluted with pet. ether and pet. ether-EtOAc mixture. the following compounds were
eluted according to the increasing order of polarity: 120mg tridecyldihydro-p-coumarate,
126mg eicosanyl-(E)-p-coumarate, 132mg pellitorine, 67mg piperlongumine and 148mg



oleoresin of pepper

... black pepper ... ground ... berries must be extracted repeatedly(percolated) with
volatile solvents: alcohol, acetone or ether. concentration of the solutions and removal
of the solvent in vacuo yields the socalled oleoresin of pepper.
quantitative composition of the oleoresin depends upon the solvent used. oleoresin
of pepper generally contains these compounds: 1. piperine, ... 2. chavicine, ... an
isomer of piperine ... 3. other piperidides, ... 4. the volatile oil, ... 5. a volatile alkaloid,
... present in samll quantities only. 6. resins, not identified.

[08, page 144]


process to recover piperine from resin of black pepper

a reaction vessel was charged with 0.9kg urea, 3.7kg ethanol (or 2.7kg methanol)
and 0.3kg of the resin of black pepper to form a mixture. the mixture was refluxed
for approximately 1h. the refluxed mixture was cooled overnight to 0-5C. the cooled
mixture was then filtered to remove solids (e.g wax, urea, fatty acids). the filtrate was
then concentrated to obtain 180g of piperine and some unreacted urea. the 180g of
piperine was recrystallized by adding ethanol. the piperine crystals were collected
by filtration and dried to yield approximately 75g of 98% pure piperine.



source of piperine

black pepper oleoresin or long pepper oleoresin is used as the source of piperine.
ground up black pepper or long pepper can also be used

to 35L buthanol/hexane mixture, 35kg black pepper oleoresin is added and
heated to 40C. the mixture is then cooled and filtered. the percipitate is washed
with buthanol/hexane mixture to obtain crude piperine. the crude piperine is
dissolved in ethanol at 60C and treated with alumina and charcoal by stirring.
it is then filtered and concentrated under vacuum to obtain a pale yellow crystalline
powder, melting range: 128-131C, min.98% pure piperine (by HPLC).



further recipes and references

with EtOH [26, (1)]

with MeOH [27]

with EtOAc: recipe 1, recipe 2

with CHCl2: recipe 1 [10], recipe 2

with acetone: [hive post 282850 on 2002-03-15 by Antoncho]

microwave-assisted extraction of piperine from piper nigrum [15]

extraction of piperine from piper nigrum (black pepper)
by hydrotropic solubilization [31]

process for extraction of piperine from piper species [07]

extraction and description of the piperines in pepper [16]

isolation of piperine from black pepper

... extracted ground black pepper with acetone ...
agricult food chem, 1981 29, 115; su, horvat
(mentioned in [21])

... piper nigrum fruit extract and its main alkaloid, piperine [17]

... pepper raw spice, oleoresins, and soluble seasonings.
piperine is extracted into ethylene dichloride ... [11]

... pepper or piper longum ... is extracted ... [34]

process for preparing piperine [02]

immersing extraction of pepper [32]

CO2 extraction, up to 9.2% piperine from black pepper(madagascar) [23]

liquid CO2 extraction of piperine ... of pepper [33]





journal of chemical education 1993 70(7): 598-599
"isolation of piperine from black pepper"
william w epstein, david f netz, jimy l seidel

patent CN1294127, 2001-05-09 (chinese)
"process for preparing piperine"
xiaoming lin, zaifeng shi, zhenfan sun
(Lutianyuan Hi-Tech Development Co., Ltd., Hainan, Peop. Rep. China)
Faming Zhuanli Shenqing Gongkai Shuomingshu
abstract (from esp@cenet): a process for preparing piperine includes
immersing extraction of pepper is mixed organic solvent, distilling to
separate out organic solvent to obtain liquid mixture of pepper oil resin
and piperine, cooling to separate out the pepper oil resin, and further
purifying. Its advantages are simple process, high extraction rate and
separation effect, and high purity.
abstract (from CAS): the process comprises extg. pepper with mixed
solvent at 25-40 for 1-6 h, filtering, concg., crystg., and recrystg. with
dichloroethylene or Et acetate. the mixed solvent is ethanol, methanol,
acetone, Et ether, Et acetate, dichloromethane, benzene, and/or chloroform.
keywords (from CAS): piperine isolation pepper

king's american dispensatory 1898
m d harvey wickes felter, phr m ph d john uri lloyd

"techniques and experiments for organic chemistry" 6.edition 1998
addison ault (cornell college); universityscience books, susalito, california
ISBN 0-935702-76-8

"laboratory experiments in organic chemistry" 3.edition 1979
jerry r. mohrig, douglas c. neckers
d. van nostrand company, new york
ISBN 0-442-25471-7

"natural products - a laboratory guide" 2.edition 1991
raphael ikan; academic press, inc; ISBN 0-12-370551-7

patent US6365601, 2002-04-02
"process for extraction of piperine from piper species"
abstract: the present invention relates to a process for extraction
of piperine of formula I from the fruits of piper species, comprising
the steps of:contacting the fruit of piper species with aqueous
hydrotrope solution at a temperature in the range of 0-100 C. and
separating the solution from the solid residue by known methods, and
recovering piperine from the solution by known methods.

"the essential oils" volume five 1952, ernest guenther
d van nostrand company, toronto - new york - london

pest management science 2000 56(2): 168-174
"synthesis and insecticidal activity of new amide derivatives of piperine"
vanderlcia fde paula, luiz c de a barbosa, antnio j demuner,
dorila pil-veloso, marcelo c picano
abstract: the natural lipophilic amides piperine and piperiline were isolated
from piper nigrum l (piperaceae). piperine was hydrolysed into piperic acid
(85% yield) which was converted into 16 amides (28-89% yield). the contact
toxicity of all synthetic amides, and also that of piperine and piperiline, at the
dose 10 g per insect, was evaluated for the brazilian economically important
insects ascia monuste orseis latr, acanthoscelides obtectus say, brevicoryne
brassicae l, protopolybia exigua de saus and cornitermes cumulans kollar.
the results demontrated that the insects have different sensivities to the various
amides, with mortality ranging from 0 to 97.5% according to the compound
and insect species.

phbcan99.rtf from http://www.uni-bayreuth.de/departments/oc1/teaching/
"isolierung von piperin aus schwarzem pfeffer und abbau zur piperinsaeure"
ANALYTISCHES PRAKTIKUM fuer Biochemiker (Stand: September 1999), page 16-18

j assoc off anal chem 1987 70(1): 112-3
"UV spectrophotometric determination of piperine in pepper preparations:
collaborative study."

t lupina, h cripps
abstract: eight collaborating laboratories performed replicate analyses for piperine
on 5 samples representing pepper raw spice, oleoresins, and soluble seasonings.
piperine is extracted into ethylene dichloride and measured at maximal
absorbance 342-345 nm with a UV light source. piperine content is calculated
using an absorbance factor derived from piperine. Intralaboratory coefficients
of variation (CVo) ranged from 0.5 to 3.1%; interlaboratory coefficients of
variation (CVx) ranged from 3.0 to 5.8%. the method has been adopted as an
official method of the american spice trade association and as an official first
action method by AOAC.
[pubmed, PMID: 3558260]

patent US6054585, 2000-04-25
"process for making high purity piperine for nutritional use"
muhammed majeed, vladimir badmaev; sabinsa corp
abstract: the present invention relates to a process of making high purity
piperine for nutritional and nutraceutical application. this process recovers
piperine from piperine-containing oleoresin by using isourea, urea or a
urea derivative to remove organic matter other than piperine from the
oleoresin. preferrably, the process recovers piperine from oleoresin of fruit
of piper nigrum or piper longum. more preferably, the process recovers
piperine from oleoresin of fruit of piper nigrum, i.e. black pepper.

patents US5744161, US6054585, US5972382, US5536506

http://www.chemistry.uvic.ca/chem465-66/465e03piperine.pdf from

ind eng chem res 2002 41(10): 2521-2528
"microwave-assisted extraction of piperine from piper nigrum"
girija raman, vilas g gaikar
abstract: a novel microwave-assisted technique (MAE) for the extraction of
piperine from coarsely powdered black pepper (Piper nigrum) was studied.
the parameters such as nature of the solvent, microwave energy input, and
solid loading were optimized. the mechanism of the enhancement of extraction
rates was investigated by microscopic studies of the irradiated material.
studies have revealed that dielectric heating of the polar cellular matrix resulted
in remarkable swelling and coalescence of the oil cells and other constituents.
the resulting pressure, built-up within the cell, breaks open the cell, releasing
the constituents and providing easy access for solvent penetration and
subsequent solubilization of piperine and other substances. because the cell wall
is mainly composed of slightly ionic cellulose, there is a marked increase in the
dielectric heating rates. rapid degradation of the cellulosic cell wall occurs,
thereby further increasing the permeability of the wall toward solvent penetration.
selective extraction of piperine in nonpolar solvents to the extent of 94% with a
purity of 85% has been achieved. the MAE procedure is simple, rapid, and reliable.

bull soc chim 1877 27(2): 290
"the extraction and description of the piperines in pepper"
p caseneuve, o caillol

planta medica 1999 65: 600-3
"stimulation of mouse melanocyte proliferation by
piper nigrum fruit extract and its main alkaloid, piperine"

zhixiu lin, j r s hoult, dorothy c bennett, amala raman
abstract: during a herbal screening programme to find potential repigmenting agents
for the treatment of vitiligo, piper nigrum l. fruit (black pepper) extract was found to
possess growth-stimulatory activity towards cultured melanocytes. its aqueous extract
at 0.1mg/ml was observed to cause nearly 300 % stimulation of the growth of a cultured
mouse melanocyte line, melan-a, in 8 days (p < 0.01). piperine (1-piperoylpiperidine),
the main alkaloid from piper nigrum fruit, also significantly stimulated melan-a cell growth.
both piper nigrum extract and piperine induced morphological alterations in melan-a cells,
with more and longer dendrites observed. the augmentation of growth by piperine was
effectively inhibited by RO-31-8220, a selective protein kinase C (PKC) inhibitor,
suggesting that PKC signalling is involved in its activity. this is the first full report on such
an activity of black pepper and piperine.

phytochemistry 1997 45(8): 1617-1619
"an amide from fruits of piper nigrum"
bina s siddiqui, sabira begum, tahsin gulzar, farhat noor, fatima noor
abstract: a new amide, N-isobutyl amide of octadeca-trans-2-cis-4-dienoic acid
has been isolated from the dried and crushed fruits of piper nigrum, along with the
known alkaloid, piperine. complete assignments of the protons and carbons of the
new amide and piperine have been made based on 2D NMR studies.

patent US4820517, 1989-04-11
"process for obtaining a pepper extract with insecticidal activity"
abstract: PCT No. PCT/EP85/00333 Sec. 371 Date Jul. 24, 1986 Sec. 102(e) Date
Jul. 24, 1986 PCT Filed Jul. 6, 1985 PCT Pub. No. WO86/01981 PCT Pub. Date Apr.
10, 1986.An insecticidally active fraction is obtained from black pepper by a process
comprising the following steps: (a) extraction of black pepper in ground form with CO2
at 30 to 70C and 150 to 500 bar; (b) removal of sharp tasting fractions therein in a first
expansion step at 25 to 35C and 70 to 150 bar; (c) removal of an oily fraction containing
the insecticidally active components as well as most of the essential oils in a second
expansion step at 15 to 30C and 40 to 70 bar; (d) removal of essential oils by steam
distillation, and if desired; (e) hydrogenation of the insecticidally active components.

patent EP0023680, 1981-02-11
"process for the production of spice extracts"
abstract: 1. aprocess for the production of spice extracts by extraction with a
physiologically acceptable gas as a solvent in two stages, characterised in that the
ethereal oils acting as odour component are removed from the spices in the first stage
using a liquid gas of which the temperature is in the subcritical range and the pressure
in the supercritical range whilst the flavour components are removed in the second
stage using the same gas of which the temperature and pressure are both in the
supercritical range, the extracts are separated off from the solutions obtained by
changing the pressure and/or temperature and optionally mixed with one another.

(1) piperine from black pepper (2) hydrolysis of piperine to piperic acid

"piperine to piperonal procedure", the cook

ind eng chem res 2002 41(12): 2966-2976
"extraction of piperine from piper nigrum (black pepper)
by hydrotropic solubilization"

girija raman, vilas g gaikar
abstract: hydrotropes, such as sodium alkyl benzene sulfonates and sodium butyl
monoglycol sulfate, were used for the selective extraction of piperine by cell
permeabilization of piper nigrum fruits. penetration of the hydrotrope molecules into
the cellular structures and subsequent cell permeabilization were hypothesized to
explain the enhanced extraction rates of aqueous hydrotrope solutions. hydrotrope
molecules, after adsorption on a cell wall, cause disorder in its structure and in the
bilayered cell membrane to facilitate the rapid extraction of piperine. the hydrotrope
solution showed selective and rapid extraction of piperine from black pepper.
the recovered piperine was ~90% pure and substantially free from oleoresins. the
type and nature of the hydrotrope, the hydrotrope concentration, the temperature,
and the particle size all had significant effects on the extraction process.

patent CN1294127, 2001-05-09
"process for preparing piperine"
abstract: A process for preparing piperine includes immersing extraction of pepper is
mixed organic solvent, distilling to separate out organic solvent to obtain liquid mixture
of pepper oil resin and piperine, cooling to separate out the pepper oil resin, and further
purifying. Its advantages are simple process, high extraction rate and separation effect,
and high purity.

patent US4985265, 1991-01-15
"process for extraction of spices"
abstract: a process for extracting spices in liquid carbon dioxide in the presence
of an alimentarily acceptable cosolvent which does not degrade the flavors of the
spices. the process is particularly applicable to the extraction of piperine and
aromatic compounds of pepper.

patent CN1298936, 2001-06-13
"process for preparing health-care peperic acid wine"; li yueting
abstract: the present invention relates to a preparation technology of piperic-acid
health-care wine which uses pepper or piper longum as raw material, and it is
extracted and mixed with wine to obtain the product which not only possesses wine
flavour, but also possesses the medicinal health-care active of pepper or piper longum.
it can prevent the rise of blood serum cholesterol and formation of choleithiasis.

boll chim farmaceutico 1998 137(8): 319-320
"long chain esters and alkamides from piper longum"
b das, a kashinatham, p madhusudhan
abstract: from the fruits of piper longum two long chain esters,
tridecyldihydro-p-coumarate and eicosanyl-(E)-p-coumarate and three alkamides,
piperine, piperlongumine and pellitorine have been isolated. the first ester is a
new compound and the second ester is reported for the first time from this plant.