Isolation of Piperine from Black Pepper

Introduction
Natural products are compounds/substances that are found freely in nature within a living
organism. These natural products have a wide array of uses within the human world ranging
from foods and dyes to more vital medical and pharmaceutical roles. However, in their nature,
natural products are difficult to use as needed and thus need to be removed. Extraction of natural
products, essentially herbalism, is very important to both the medical and pharmaceutical
industries. Plants, a source of natural products, have a unique ability to synthesize many different
compounds to aid them in preforming biological functions ranging from defense against
predators/pathogens to promoting general health welfare. Because of the significance and
importance of these biological functions, it is important to better study and analyze these natural
products, often done through extraction. With the extraction of these natural products, humans
have been able to effectively fight and treat many different diseases from Alzheimers to
different forms of cancer.
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With more research/analysis, more natural products will come to light
allowing for breakthroughs in the medicinal and pharmaceutical fields.
Extraction of natural products can be done in several different ways. The most common
form of extraction is the use of a compounds solubility to filter out the desired product. Isolation
of piperine from black pepper requires the use of solid-extraction. Piperine along with several
other alkaloids including piperdine, piperetine and piperanine make up 5-9% of ground black
pepper.
Black Pepper, native to south India, is the worlds most traded and used spice. Its
digestive and antioxidant properties have been documented for several decades but it is only
recently that more emphasis has been placed on its natural compounds.
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Piperine, known for
providing most of the spiciness in black peppers, is a compound that has a very distinctive
pungent smell that many associate with peppers. Its use for its antioxidant and anti-asthmatic
properties has been documented from ancient times around the world.
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Piperine uses, however,
dont stop there. New research has found that piperine can speed up energy metabolism, combat
skin depigmentation and has even been known to inhibit enzymes important for drug
metabolism.
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This goal of this experiment is to extract piperine from ground black pepper using solid-
liquid extraction followed by purification by recrystallization. The isolated product will then be
characterized by melting point, IR, MS, 60 MHz
1
H NMR, and 400 MHz
1
H NMR.
Experimental
Ground black pepper (12.5 g) was added to ethanol (50 mL) in a 100 mL round bottom
flask and refluxed for 90 minutes. The resulting solution was cooled to room temperature and
vacuumed filtered. Liquid filtrate was concentrated using a stream of nitrogen and a water bath at
50 C. The residue was dissolved in 10% by weight KOH in ethanol and then cooled in an ice
bath. Water was added to the solution drop-wise and piperine crystals (yellow) were collected
via vacuum filtration. The piperine crystals were dissolved in a hot 3:2 acetone:hexane solution
(5 mL) and left at room temperature for 15 minutes. The solution was then transferred to an ice
bath for 30 minutes and the crystals were collected were collected and washed with cold ether (4
mL) via vacuum filtration. The piperine crystals were air dried for 48 hours (0.315 grams, 2.52
% recovery) mp 132-134C; IR (ATR) V
max
(cm
-1
) 1670-1760, 3000-3100, 3300-3500, 1620-
1680, 1200-1270, 1450-1600, 1410-1440;
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H NMR (60 MHz, CDCI3) (ppm) 6.2-7.2 (m, 7H),
3.4-3.6 (m, 4H), 1.4-1.8 (m, 6H), 6.0-6.4 (d, 2H)

Results/Discussion
The extraction of piperine from black pepper involved the use and knowledge of
refluxing and solid-liquid extraction with the use of several chemicals (ethanol, KOH). The
extracted crude piperine was then purified through recrystallization to yield a final mass of 0.315
(2.52 %). Qualitatively, the product created matched the characteristics of the piperine being
synthesized, with a yield of yellow-colored crystals extracted from a dark black pepper powder.
Qualitative observations alone, such as color, are not sufficient enough to confirm the presence
of piperine. The purified piperine was further characterized thought a series of tests to further
confirm its identity. The accepted melting point value for piperine is 132-134C which is in
agreement with the experimentally determined melting point of 128-132C indicating very little
impurity. Mass Spectrometry of the final product revealed a molecular weight of 285 (Figure 4)
which is also in agreement with the known molecular weight of 285.34. IR analysis provided
greater insight into the products functional groups to provide additional evidence for the
extraction and isolation of piperine. The IR data indicated the presence of the characteristic
functional groups in piperine: amide (1629.9 cm
-1
), aromatic C-H bonds (2938.0 cm
-1
), and C=C
stretch (1612.2). The other more commons signals of piperine were seen and annotated in figure
1.
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H NMR (Figures 2 and 3) was the last test done to confirm the identity of piperine in the
isolated product. The NMR spectra also showed the expected peaks with the 4 hydrogens closest
to the nitrogen in the 3.549-3.5067 ppm range, 2 hydrogens bonded between the oxygen
molecules corresponding to the 6.0440 ppm range, and 7 hydrogens in the 6.9229-7.4228 ppm
range.
The knowledge and use of extraction is beneficial to industries all around the world as
well as providing a fundamental understanding of organic chemistry. The extraction of piperine
from black pepper showed extractions importance with relation to the most globally known and
traded spice. The purpose of this experiment was fulfilled with the extraction and purification of
piperine (0.315g, 2.52% yield) matching up realistically to the literature values listed between
2.5 and 10%. Some possible errors resulting to a low crop yield could be due to insufficient
refluxing time which can be corrected in the future to extract a greater yield of piperine.
Piperines identity was confirmed through the use of melting point, IR, MS, 60 MHz and 400
MHz
1
H NMR. One possible future change for this experiment would be to possibly lengthen to
the time of reflux to improve the percent yield or create another crop of crystals to also improve
the percent yield.
References
1) Cragg, G. M., Newman, D. J., & Snader, K. M. (1997). Natural products in drug
discovery and development. Journal of Natural products, 60(1), 52-60.
2) Srinivasan, K. (2007). Black pepper and its pungent principle-piperine: a review of
diverse physiological effects. Critical reviews in food science and nutrition,47(8), 735-
748.
3) Atal, C. K., Dubey, R. K., & Singh, J. (1985). Biochemical basis of enhanced drug
bioavailability by piperine: evidence that piperine is a potent inhibitor of drug
metabolism. Journal of Pharmacology and Experimental Therapeutics,232(1), 258-262.
4) Rummel, S.A., Lab Guide for Chem 213; Hayden-McNeil Publishing: Plymouth, MI,
2013.