Development of New Isolation and Quantification Method of Piperine from White Pepper Seeds (Piper Nigrum L) Using A Validated HPLC

Nindya Kusumorini; Akhmad Kharis  Nugroho; Suwijiyo  Pramono; Ronny  Martien

doi:10.22146/ijp.866

Nindya Kusumorini Faculty of Pharmacy, Universitas Gadjah Mada
Akhmad Kharis Nugroho Department of Pharmaceutics, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta 55281 Indonesia
Suwijiyo Pramono Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta 55281 Indonesia
Ronny Martien Department of Pharmaceutics, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta 55281 Indonesia

Keywords: white pepper, active pharmaceutical ingredient (API), NMR, piperine antioxidant

Abstract

Most of the active pharmaceutical ingredients in the pharmaceutical world are obtained from natural ingredients, one of which is white pepper. White pepper seeds (Piper nigrum L.) contain the main alkaloid compound piperine, which has a broad spectrum pharmacological effect. This research aimed to isolate the piperine compound from white pepper seeds to make piperine more economical, simple, and effective so that it can be applied to large scale production in the pharmaceutical industry. The method of extracting and purifying piperine compounds was carried out using n-hexane and cyclohexane. The results of the isolated piperine were tested for their purity with a set of melting point apparatus, bidimensional TLC, and HPLC. The structure compound was then analyzed using FTIR and NMR. Furthermore, the isolated piperine was tested for its antioxidant activities using DPPH, ABTS, and FRAP. Cyclohexane was successfully used to remove the resinous matter from the n-hexane extract of white pepper to produce isolated piperine with 97.24% of purity. The results of the melting point, FTIR, ¹H-NMR, and ¹²C-NMR of the isolated piperine were similar to that of the piperine reference substance and the literature review. The isolated piperine has weak antioxidant activities in the ABTS and FRAP test with the results of 15.25 ± 0.004 mM and 10.53 mol TE/g of the sample, respectively.

References

Ahmad, N., Fazal, H., Abbasi, B. H., Farooq, S., Ali, M., & Khan, M. A., 2012. Biological role of Piper nigrum L. (Black pepper): A review. Asian Pacific Journal of Tropical Biomedicine, 2(3), S1945–S1953.
Alves, F. S., Rodrigues Do Rego, J. de A., Da Costa, M. L., Lobato Da Silva, L. F., Da Costa, R. A., Cruz, J. N., & Brasil, D. D. S. B., 2019. Spectroscopic methods and in silico analyses using density functional theory to characterize and identify piperine alkaloid crystals isolated from pepper (Piper Nigrum L.). Journal of Biomolecular Structure and Dynamics, 1–8.
Chaudhri, V. K., 2017. Isolation and Evaluation of Piperine from Black Pepper and White Pepper. World Journal of Pharmacy and Pharmaceutical Sciences, 1424–1430.
Dayan, N., 2008. Skin Aging Handbook: An Integrated Approach to Biochemistry and Product Development. William Andrew.
Dejian Huang, Boxin Ou, § and, & Prior, R. L., 2005. The Chemistry behind Antioxidant Capacity Assays [Review-article]. American Chemical Society.
Floegel, A., Kim, D.-O., Chung, S.-J., Koo, S. I., & Chun, O. K., 2011. Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods. Journal of Food Composition and Analysis, 24(7), 1043–1048.
Ikan, R., 2013. Natural Products: A Laboratory Guide. Elsevier.
Khan, S., Mirza, K. J., Anwar, F., & Abdin, M. Z., 2010. Development of RAPD markers for authentication of Piper nigrum (L.). Environment & We An International Journal of Science & Technology, 5, 47–56.
Kim, H. G., Han, E. H., Jang, W.-S., Choi, J. H., Khanal, T., Park, B. H., Tran, T. P., Chung, Y. C., & Jeong, H. G., 2012. Piperine inhibits PMA-induced cyclooxygenase-2 expression through downregulating NF-κB, C/EBP and AP-1 signaling pathways in murine macrophages. Food and Chemical Toxicology, 50(7), 2342–2348.
Li, L., Liu, H., Shi, W., Liu, H., Yang, J., Xu, D., Huang, H., & Wu, L., 2017. Insights into the Action Mechanisms of Traditional Chinese Medicine in Osteoarthritis. Evidence-Based Complementary and Alternative Medicine.
Mareček, V., Mikyška, A., Hampel, D., Čejka, P., Neuwirthová, J., Malachová, A., & Cerkal, R., 2017. ABTS and DPPH methods as a tool for studying antioxidant capacity of spring barley and malt. Journal of Cereal Science, 73, 40–45.
Nahak, G., & Sahu, R. K., 2011. Phytochemical Evaluation and Antioxidant activity of Piper cubeba and Piper nigrum. Journal of Applied Pharmaceutical Science, 08, 153–157.
Nivas, D., Gaikwad, D. K., & Chavan, P. D., 2010. Antiradical activity of medicinally important Morinda pubescens fruits. International Journal of Pharma and Bio Sciences, 1, 1–4.
Pellegrini, N., Serafini, M., Colombi, B., Del Rio, D., Salvatore, S., Bianchi, M., & Brighenti, F., 2003. Total Antioxidant Capacity of Plant Foods, Beverages and Oils Consumed in Italy Assessed by Three Different In Vitro Assays. The Journal of Nutrition, 133(9), 2812–2819.
Rahman Khan, Z., Moni, F., Sharmin, S., Al-Mansur, M. A., Gafur, A., Rahman, O., & Afroz, F., 2017. Isolation of Bulk Amount of Piperine as Active Pharmaceutical Ingredient (API) from Black Pepper and White Pepper (Piper nigrum L.). Pharmacology & Pharmacy, 08(07), 253–262.
Saha, K., Seal, H., & Noor, M.. 2014. Isolation and characterization of piperine from the fruits of black pepper (Piper nigrum). Journal of the Bangladesh Agricultural University, 11(1), 11–16.
Satyendra, P., Arun, P., Shailendra, P., Neelesh, D., & Neeraj, K., 2018. FORMULATION AND EVALUATION OF TOPICAL CREAM OF PIPERINE FOR VITILIGO. World Journal of Pharmaceutical Research, 7(3), 11.
Shingate, P. N., Dongre, P. P., & Kannur, D. M., 2013. New method development for extraction and isolation of piperine from black pepper. International Journal of Pharmaceutical Sciences and Research, 4(8), 3165.
Shui, G., & Leong, L. P., 2006. Residue from star fruit as valuable source for functional food ingredients and antioxidant nutraceuticals. Food Chemistry, 97(2), 277–284.
Silverstein, R. M., & Bassler, G. C.. 1962. Spectrometric identification of organic compounds. Journal of Chemical Education, 39(11), 546.
Soare, J. R., Dinis, T. C. P., Cunha, A. P., & Almeida, L., 2009. Antioxidant Activities of Some Extracts of Thymus zygis. Free Radical Research, 26, 469–478.
Srivastava, A. K., & Singh, V. K., 2017. Biological Action Of Piper Nigrum—The King Of Spices. European Journal of Biological Research, 7(3), 223–233.
Su, L., Yin, J.-J., Charles, D., Zhou, K., Moore, J., & Yu, L. (Lucy)., 2007. Total phenolic contents, chelating capacities, and radical-scavenging properties of black peppercorn, nutmeg, rosehip, cinnamon and oregano leaf. Food Chemistry, 100(3), 990–997.
Thaipong, K., Boonprakob, U., Crosby, K., Cisneros-Zevallos, L., & Hawkins Byrne, D., 2006. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. Journal of Food Composition and Analysis, 19(6–7), 669–675.
Umar, S., Golam Sarwar, A. H. M., Umar, K., Ahmad, N., Sajad, M., Ahmad, S., Katiyar, C. K., & Khan, H. A., 2013. Piperine ameliorates oxidative stress, inflammation and histological outcome in collagen induced arthritis. Cellular Immunology, 284(1–2), 51–59.
Ying, X., Chen, X., Cheng, S., Shen, Y., Peng, L., & Xu, H., 2013. Piperine inhibits IL-β induced expression of inflammatory mediators in human osteoarthritis chondrocyte. International Immunopharmacology, 17(2), 293–299.
Zarai, Z., Boujelbene, E., Ben Salem, N., Gargouri, Y., & Sayari, A., 2013. Antioxidant and antimicrobial activities of various solvent extracts, piperine and piperic acid from Piper nigrum. LWT - Food Science and Technology, 50(2), 634–641.