Characterization and HPLC Quantification of Piperine in Various Parts of Piper Guineense

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Medicinal plants have been used since medieval times for treating ailments. The extended use have led to high market value and adulteration of their herbal products. Piper guineense, commonly known as West African Black Pepper, is a climbing plant up to 12m high by its adventitious rootlets. It has a corky lower stem and simple leaves which have five principal nerves. The fruits which are aromatic are red- brown when ripe and black when dry. Various parts of the plant have medicinal uses such as antibacterial, anticancer, etc. This project seeks to develop an HPLC method to standardize the various parts of Piper guineense using isolated piperine as a biomarker and a secondary reference. All samples of P. guineense parts were obtained and authenticated from the Physique garden of Pharmacognosy Department, KNUST. Piperine was isolated from the dried fruits of P. guineense with ethanolic KOH and recrystallized from acetone: hexane 3:2. The melting point was determined to be 128ºC - 130ºC and the crude yield was 2.07%w/w. TLC gave Rf values of 0.285 ± 0.01291 for hexane: ethylactate: glacial acetic acid 3: 1: 0.3 and 0.70 ± 0.009574 for chloroform: ethylacetate 1: 1. It was characterized by melting point as well as Ultraviolet, Nuclear Magnetic Resonance and Mass Spectroscopy. A validated reverse phase HPLC with methanol: water 80: 20 at a flow rate of 1.40ml/min on a Phenomenex Kromosil 5 C8 (250mm x 4.6mm 5 micron id) column with detection at 343nm gave a retention time of 3.78 ± 0.05656 min. Piperine was found to be 5.44 x10-3 ± 9.24 x 10-5%w/w in the dried leaves (DL), 0.0437 ± 0.000816%w/w for fresh leaves (FL), 0.115 ± 0.00228%w/w for dried stem (DS) and 3.345 ± 0.0339%w/w for dried fruits (DF).
A thesis submitted to the School of Graduate Studies, Kwame Nkrumah University of Science and Technology in partial fulfilment of the requirement for the award of Master of Philosophy in Pharmaceutical Chemistry, October-2012