Genetic Resources and Crop Evolution

, Volume 66, Issue 8, pp 1635–1641 | Cite as

Evaluation of chemo-diversity in major Piper species for three piperamides using validated RP-HPLC method

  • R. SivaranjaniEmail author
  • Johnson K. George
  • K. V. Saji
Short Communication


India boasts rich biodiversity of plants belongs to Piperaceae. These plants are store-house of biologically active alkaloids, generally termed as piperamides. Among piperamides, piperine, piperlongumine and piperlonguminine plays an effective role in pharmaceutical applications. The present study was aimed at developing and validating a simple reverse phase high performance liquid chromatography method to analyze chemo-diversity of these three piperamides in Pipernigrum L., Piper longum L., Piper thomsonii Hook, Piper sarmentosum Roxb. and Piper chaba Trel. & Yunck. The separation was achieved on a RP-C18 column with water (0.1% Acetic acid) and methanol as mobile phase with detection wavelength of 338 nm. The compounds were identified using authentic standards. The developed method was validated as per ICH guidelines for linearity, accuracy, precision, limit of detection and limit of quantification. The method was successfully applied in quantification of these piperamides in berries of five different Piper species. Among them, piperlongumine was detected only in P. longum and P. sarmentosum. Piperlonguminine and piperine was higher in P. chaba and P. nigrum. In conclusion, the method developed could be used for chemo-diversity analysis of different Piper spp to select alternate Piper species for pharmacological preparations based on the composition of its bio-active compounds.


Alkaloids RP-HPLC ICH Piperaceae 



The authors are grateful to Centre for Agricultural Bioinformatics (CABin) scheme of ICAR for funding the work.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Attanayake AMHS, De Silva UMGD, Jayaweera JAAS, Perera DL (2018) Use of Ayurveda and Sri Lankan traditional medicine for healing shaft of humerus fracture following nonunion. J Ayurveda Integr Med 9(3):217–220. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Beltran LR, Dawid C, Beltran M, Levermann J, Titt S, Thomas S, Purschel V, Satalik M, Gisselmann G, Hofmann T, Hatt H (2017) The effect of pungent and tingling compounds from Piper nigrum L. on background K+ currents. Front Pharmacol 8:408. CrossRefGoogle Scholar
  3. Bezerra DP, Pessoa C, de Moraes MO, Saker-Neto N, Silveira ER, Costa-Lotufo LV (2013) Overview of the therapeutic potential of piplartine (piperlongumine). Eur J Pharm Sci 48(3):453–463. CrossRefGoogle Scholar
  4. Bharadwaj U, Eckols TK, Kolosov M, Kasembeli MM, Adam A, Torres D, Zhang X, Dobrolecki LE, Wei W, Lewis MT, Dave B, Chang JC, Landis MD, Creighton CJ, Mancini MA, Tweardy DJ (2015) Drug-repositioning screening identified piperlongumine as a direct STAT3 inhibitor with potent activity against breast cancer. Oncogene 34(11):1341–1353. CrossRefPubMedGoogle Scholar
  5. Damanhouri ZA, Ahmad A (2014) A review on therapeutic potential of Piper nigrum L (black pepper): The King of spices. Med Aromat Plants 3:161. CrossRefGoogle Scholar
  6. Fontenele JB, Leal LK, Silveira ER, Felix FH, Bezerra Felipe CF, Viana GS (2009) Antiplatelet effects of piplartine, an alkamide isolated from Piper tuberculatum: possible involvement of cyclooxygenase blockade and antioxidant activity. J Pharm Pharmacol 61(4):511–515. CrossRefPubMedGoogle Scholar
  7. Friedman M, Levin CE, Lee S, Lee J, Ohnisi-Kameyama M, Kozukue N (2008) Analysis by HPLC and LC/MS of pungent piperamides in commercial black, white, green, and red whole and ground peppercorns. J Agric Food Chem 56:3028–3036. CrossRefPubMedGoogle Scholar
  8. Gorgani L, Mohammadi M, Najafpour DG, Nikzad M (2017) Piperine—The bioactive compound of black pepper: from isolation to medicinal formulations. Compr Rev Food Sci F 16:124–140. CrossRefGoogle Scholar
  9. Hussain K, Hashmi FK, Latif A, Ismail Z, Sadikun S (2012) A review of the literature and latest advances in research of Piper sarmentosum. Pharm Biol 50(8):1045–1052. CrossRefPubMedGoogle Scholar
  10. ICH Harmonised Tripartite Guideline. Guidnace on analytical method validation. Accessed 10 Sept 2018
  11. Johri RK, Zutshi U (1992) An Ayurvedic formulation ‘Trikatu’ and its constituents. J Ethnopharmacol 37(2):85–91CrossRefGoogle Scholar
  12. Kamal YT, Singh M, Salam S, Ahmad S (2016) Simultaneous quantification of piperlongumine and piperine in traditional polyherbal formulation using validated HPLC method. Acta Chromatographica 28(4):489–500. CrossRefGoogle Scholar
  13. Kotteswari M, Rao MRK, Kumar S, Prabhu K, Sundaram RL, Dinakar S (2018) GC MS analysis of one ayurvedic preparation ‘Aswagandharishtam’. Biomed Pharmacol J.
  14. Li K, Zhu W, Fu Q, Ke Y, Jin Y, Liang X (2013) Purification of amide alkaloids from Piper longum L. using preparative two-dimensional normal-phase liquid chromatography reversed-phase liquid chromatography. Analyst 138(11):3313–3320. CrossRefPubMedGoogle Scholar
  15. Liu H, Luo R, Chen X, Ba Y, Zheng L, Guo W, Wu X (2015) Identification and simultaneous quantification of five alkaloids in Piper longum L. by HPLC-ESI-MSn and UFLC-ESI-MS/MS and their application to Piper nigrum L. Food Chem 177:191196. CrossRefGoogle Scholar
  16. Makhov P, Golovine K, Teper E, Kutikoy A, Mehrazin R, Corcoran A, Tulin A, Uzzo RG, Kolenko VM (2014) Piperlongumine promotes autophagy via inhibition of Akt/mTOR signalling and mediates cancer cell death. Br J Cancer 110(4):899–907. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Matsuda H, Ninomiya K, Morikawa T, Yasuda D, Yamaguchi I, Yoshikawa M (2008) Protective effects of amide constituents from the fruit of Piper chaba on D-galactosamine/TNF-α-induced cell death in mouse hepatocytes. Bioorg Med Chem Lett 18:2038–2042. CrossRefPubMedGoogle Scholar
  18. Meegan MJ, Nathwani S, Twamley B, Zisterer DM, O’Boyle NM (2017) Piperlongumine (piplartine) and analogues: antiproliferative microtubule-destabilising agents. Eur J Med Chem 125:453–463. CrossRefPubMedGoogle Scholar
  19. Mgbeahuruike EE, Yrjönen T, Vuorela H, Holm Y (2017) Bioactive compounds from medicinal plants: focus on piper species. S Afr J Bot 112:54–69. CrossRefGoogle Scholar
  20. Park BS, Son DJ, Choi WS, Takeoka GR, Han SO, Kim TW, Lee SE (2008) Antiplatelet activities of newly synthesized derivatives of piperlongumine. Phytother Res 22(9):1195–1199. CrossRefGoogle Scholar
  21. Prasad S, Tyagi AK (2016) Historical spice as a future drug: therapeutic potential of piperlongumine. Curr Pharm Des 22(27):4151–4159. CrossRefPubMedGoogle Scholar
  22. Rajopadhye AA, Namjoshi PT, Upadhye SA (2012) Rapid validated HPTLC method for estimation of piperine and piperlongumine in root of Piper longum extract and its commercial formulation. Brazilian Journal of Pharmacognosy 22(6):1355–1361. CrossRefGoogle Scholar
  23. Ravindran PN, Nirmal Babu K, Sasikumar B, Krishnamurthy KS (2000) Botany and Crop Improvement of Black Pepper. In: Ravindran PN (ed) Black Pepper Piper nigrum, 1st edn. Hardwood, Amsterdam, pp 23–143Google Scholar
  24. Silva DR, Brenzab MA, Kambara LM, Cortez LER, Cortez DAG (2013) Analysis of amide compounds in different parts of Piper ovatum Vahl by high-performance liquid chromatographic. Pharmacognosy Res 5(4):254–259. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Wang H, Liu J, Gao G, Wu X, Wang X, Yang H (2016a) Protection effect of piperine and piperlonguminine from Piper longum L. alkaloids against rotenone-induced neuronal injury. Brain Res 1639:214–227. CrossRefPubMedGoogle Scholar
  26. Wang Y, Wu X, Zhou Y, Jiang H, Pan S, Sun B (2016b) Piperlongumine suppresses growth and sensitizes pancreatic tumors to gemcitabine in a xenograft mouse model by modulating the NF kappa B pathway. Cancer Prev Res 9(3):234–244. CrossRefGoogle Scholar
  27. Wood AB, Barrow ML, James DJ (1988) Piperine determination in pepper (Piper nigrum L.) and its Oleoresins-A reverse-phase high-performance liquid chromatographic method. Flavour Fragr J 3:554. CrossRefGoogle Scholar
  28. Wu S, Sun C, Pei S, Lu Y, Pan Y (2004) Preparative isolation and purification of amides from the fruits of Piper longum L by upright counter-current chromatography and reversed-phase liquid chromatography. J Chromatogr A 1040(2):193–204. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Division of Crop Production and Post Harvest TechnologyICAR - Indian Institute of Spices ResearchKozhikodeIndia
  2. 2.Division of Crop Improvement and BiotechnologyICAR - Indian Institute of Spices ResearchKozhikodeIndia

Personalised recommendations