Quantification of phorbol esters content by HPTLC in seeds of Jatropha species genotypes

Authors

Keywords:

Antinutritional Compounds, Genotypes, HPTLC, Jatropha, Phorbol Esters, Toxicity

Abstract

Jatropha belongs to the Euphorbiaceae family and includes species of interest such as Jatropha curcas, J. platyphylla, and J. cinerea, whose seeds have potential for biofuel production due to their valuable oil content (50-60 %) and to produce animal feed due to their high crude protein content (25-30 %). However, Jatropha seeds contain antinutritional and toxic compounds such as phorbol esters, commonly known as cancer promoters, which limits their industrial and food use if they are not properly identified, since their absence or presence determines the need for detoxification processes prior to use. The objective of this study was to develop and validate an analytical method for the quantification of phorbol esters (PE) using high-performance thin-layer chromatography (HPTLC) in Jatropha seeds from different regions of Sinaloa. The results showed that Jatropha curcas-Capule, J. curcas-Imala, and J. cinerea-La Campana presented concentrations of 1.78 mg/g, 0.96 mg/g, and 0.17 mg/g of phorbol esters, respectively. In contrast, in J. curcas-La Campana, J. platyphylla-La Campana, and J. platyphylla-Mocorito, no levels of these compounds were detected. These findings show significant variations in phorbol ester content among genotypes, which is relevant for their potential use and management.

Downloads

Download data is not yet available.

Author Biographies

  • Salazar-Villa E., Facultad de Biología, Universidad autónoma de Sinaloa

    She is a Professor/Researcher at the Faculty of Biology and holds a Ph.D. in Science awarded by the Center for Research in Food and Development A.C.. She is a member of the National System of Researchers and Innovators (SNII) at the Candidate level, an Honorary Researcher of the SSIT, and holds a PRODEP Profile. She has published 10 articles in indexed journals and 5 in peer-reviewed journals, accumulating 120 citations to date and an h-index of 6. She has participated in 23 national and international conferences. Throughout her career, she has supervised 6 undergraduate theses in the area of bioactive compounds. Additionally, she is a member of the Consolidated Academic Body “Research and Innovation in Biological and Health Sciences” and serves as Editor of the RIBIIOS-UAS section. Contact e-mail: edith.salazar@uas.edu.mx

  • Medina-Rodelo D.P., Centro de Investigación en Alimentación y Desarrollo

    She is a Researcher at the Center for Research in Food and Development and holds a Ph.D. in Coordination of Science and Technology of Agricultural Products from Tropical and Subtropical Regions, awarded by the Center for Research in Food and Development. She has published 3 articles in indexed journals, with a total of 22 citations to date and an h-index of 2. She has participated in 10 national and international conferences. Her areas of specialization include solid-state fermentation, animal feed, jatropha, metabolites, HPTLC, and extracts. Contact email: dulce.medina@ciad.mx

  • Rojo-Báez I., Biology Faculty. Autonomous University of Sinaloa

    She is a Professor/Researcher at the Faculty of Biology, Autonomous University of Sinaloa, and holds a Ph.D. in Science (specializing in Phytopathology) awarded by the Center for Research in Food and Development, A.C., Culiacán Coordination. She is a member of the National System of Researchers and Innovators (SNII) at the Candidate level, and of the SSIT as an Honorary Researcher. She has published 11 articles in indexed journals and 11 in peer-reviewed journals, accumulating 140 citations to date and an h-index of 6. She has participated in 16 national and international conferences. Throughout her career, she has supervised 19 undergraduate theses and 1 master’s thesis in the field of Phytopathology. Contact email: indira@uas.edu.mx

  • Báez-Parra K.M., Faculty of Chemical Biological Sciences, Autonomous University of Sinaloa

    She is a Professor/Researcher at the Autonomous University of Sinaloa and holds a Ph.D. in Science awarded by the Center for Research in Food and Development. She is a member of the SSIT as an Honorary Researcher. She has published 12 articles in peer-reviewed journals, accumulating 49 citations to date and an h-index of 4. She has participated in 5 national and international conferences. Contact email: karlabaez.fcqb@uas.edu.mx

  • Angulo-Escalante M.A., Centro de Investigación en Alimentación y Desarrollo

    He is a Researcher at the Center for Research in Food and Development and holds a Ph.D. in Pharmacology and Toxicology awarded by the University of Arizona. He is a member of the National System of Researchers and Innovators (SNII) at Level 1 and holds a PRODEP Profile. He has published 48 articles in indexed journals and 48 in peer-reviewed journals, accumulating 1,321 citations to date and an h-index of 20. He has participated in 30 national and international conferences. Throughout his career, he has supervised 13 undergraduate theses, 12 master’s theses, and 7 doctoral dissertations in the area of Bioresources. Contact email: mangulo@ciad.mx

References

Baldini, C., Ferfuia, R., Bortolomeazzi, G., Verardo, V., Pascali, E., Piasentier, L., & Franceschi, L. (2014). Determination of phorbol esters in seeds and leaves of Jatropha curcas and in animal tissue by high-performance liquid chromatography tandem mass spectrometry. Industrial Crops and Products, 59, 268–276. https://doi.org/10.1016/j.indcrop.2014.05.023

Beutler, J., Alvarado, A., McCloud, T., & Cragg, G. (1989). Distribution of phorbol ester bioactivity in the Euphorbiaceae. Phytotherapy Research, 3(5), 188–192. https://doi.org/10.1002/ptr.2650030507

Cavalcante, N. B., da Conceição Santos, A. D., & da Silva Almeida, J. R. G. (2020). The genus Jatropha (Euphorbiaceae): A review on secondary chemical metabolites and biological aspects. Chemico-Biological Interactions, 318, 108976. https://doi.org/10.1016/j.cbi.2020.108976

Chún, D., & Roldan, D. (2013). Cuantificación de ésteres de forbol en partes estructurales de semilla descascarada en ocho accesiones de Jatropha curcas [Tesis de licenciatura]. Escuela Agrícola Panamericana, Zamorano.

Codex Alimentarius Commission. (2017). Guidelines on performance criteria for methods of analysis for the determination of pesticide residues in food and feed (CXG 90-2017).

de Barros, C. R., Ferreira, L. M. M., Fraga, I., Mourão, J. L., & Rodrigues, M. A. M. (2024). Detoxification methods of Jatropha curcas seed cake and its potential utilization as animal feed. Fermentation, 10(5), 256. https://doi.org/10.3390/fermentation10050256

Demissie, A., & Lele, S. (2010). Bioassay-assisted identification of phorbol ester from Jatropha curcas tissue culture. International Journal of Pharma and Bio Sciences, 1(3), 1–7. http://www.ijpbs.net/issue-3/6.pdf

Devappa, R. K., Angulo-Escalante, M. A., Makkar, H. P. S., & Becker, K. (2012a). Potential of using phorbol esters as an insecticide against Spodoptera frugiperda. Industrial Crops and Products, 38, 50–53. https://doi.org/10.1016/j.indcrop.2012.01.007

Devappa, R. K., Sanjay, K. R., Kumar, V., Makkar, H. P. S., & Becker, K. (2012b). Activities of Jatropha curcas phorbol esters in various bioassays. Ecotoxicology and Environmental Safety, 78, 57–62. https://doi.org/10.1016/j.ecoenv.2011.11.002

Dwivedi, J., Gupta, A., & Paliwal, S. (2020). Validated simultaneous HPTLC analysis of scopoletin and gallic acid in the methanolic fraction of Jatropha glandulifera. Journal of Planar Chromatography – Modern TLC, 33, 457–462. https://doi.org/10.1007/s00764-020-00063-8

Duque, L. F. C., & López, F. C. (2022). Propiedades químicas del aceite de cinco genotipos de Jatropha curcas L. en Colombia. Agronomía Mesoamericana, 33(1), 19. http://doi.org/10.15517/am.v33i1.44555

Edrisi, S., Dubey, R., Tripathi, V., Bakshi, M., Srivastava, P., Jamil, S., Singh, H., & Abhilash, P. (2015). Jatropha curcas L.: A crucified plant waiting for resurgence. Renewable and Sustainable Energy Reviews, 41, 855–862. https://doi.org/10.1016/j.rser.2014.08.082

Faria-Machado, A. F., Licurgo, F. M., Pires, J. M., da Silveira-Campos, R., Wilhelm, A. E., de Lourdes, M., & Antoniassi, R. (2019). Method validation for analysis of phorbol esters from Jatropha curcas. Industrial Crops and Products, 140, 111627. https://doi.org/10.1016/j.indcrop.2019.111627

Francis, G., Makkar, H. P., Carle, R., Mittelbach, M., Wink, M., Martinez Herrera, J., & Becker, K. (2021). Critique on conclusions regarding toxic compounds in Jatropha curcas kernel cake. Communications Biology, 4(1), 1348. https://doi.org/10.1038/s42003-020-0919-z

Fujiki, H., Suttajit, M., Rawangkan, A., Iida, K., Limtrakul, P., Umsumarng, S., & Suganuma, M. (2017). Phorbol esters in seed oil of Jatropha curcas L. and their association with cancer prevention. Journal of Cancer Research and Clinical Oncology, 143(8), 1359–1369. https://doi.org/10.1007/s00432-017-2341-6

Govaerts, R., Frodin, D., Radcliffe-Smith, A., & Carter, S. (2012). World checklist and bibliography of Euphorbiaceae (with Pandaceae) (Vol. 4). Royal Botanic Gardens.

Halim, A. H. A., Zamberi, M. M., Husin, M. H. M., Haminudin, N. F., Idral, F., & Ghani, S. A. (2022). Physicochemical properties of Jatropha curcas oil as a potential feedstock for biodiesel production. Proceedings of Mechanical Engineering Research Day, 283–284.

International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use. (2005). ICH Q2(R1): Validation of analytical procedures: Text and methodology. https://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1__Guideline.pdf

Jaspal, N., Sharma, M., Prashar, D. B., Sharma, R., & Sharma, M. (2023). Jatropha curcas L.: A sustainable resource for biofuel feedstock with medicinal and commercial attributes. Journal of Innovative Agriculture, 10(3), 1–13. http://doi.org/10.37446/jinagri/ra/10.3.2023.1-13

Jonas, M., Ketlogetswe, C., & Gandure, J. (2022). Quantification of phorbol-12-myristate-13-acetate in Jatropha seed oil and cake at different stages of fruit maturity. International Journal of Environmental Studies, 79(1), 88–97. https://doi.org/10.1080/00207233.2021.1903672

Kongmany, S., Truong, H., Le, H., Imamura, K., Maeda, Y., & Van Boi, L. (2016). Semi-preparative HPLC separation followed by HPLC/UV and tandem mass spectrometric analysis of phorbol esters in Jatropha seed. Journal of Chromatography B, 1038, 1–27. https://doi.org/10.1016/j.jchromb.2016.10.021

Leyva-Acuña, M. A., Márquez-Zequera, I., García-Estrada, R. S., Soto-Landeros, F., Montes-Avila, J., & Angulo-Escalante, M. A. (2023). High performance thin layer chromatography fingerprint profile and antifungal activity of methanolic extracts of Jatropha platyphylla. International Journal of Agriculture and Biology, 29, 164. https://doi.org/10.17957/IJAB/15.2015

Leyva-Padrón, G., Vanegas-Espinoza, P. E., Evangelista-Lozano, S., Del Villar-Martínez, A. A., & Bazaldúa, C. (2020). Chemical analysis of callus extracts from toxic and non-toxic varieties of Jatropha curcas. PeerJ, 8, e10172. https://doi.org/10.7717/peerj.10172

Li, C. Y., Devappa, R. K., Liu, J. X., Makkar, H. P. S., & Becker, K. (2010). Toxicity of Jatropha curcas phorbol esters in mice. Food and Chemical Toxicology, 48, 620–625. https://doi.org/10.1016/j.fct.2009.11.042

Liu, S., Sporer, M., Wink, J., Jourdane, R., & Ruppel, A. (1997). Anthraquinones in Rheum palmatum and phorbol esters in Jatropha curcas with molluscicidal activity. Tropical Medicine & International Health, 2(2), 179–188. https://doi.org/10.1046/j.1365-3156.1997.d01-240.x

Makkar, H. P. S., Aderibigbe, A. O., & Becker, K. (1998). Comparative evaluation of non-toxic and toxic varieties of Jatropha curcas. Food Chemistry, 62(2), 207–215. https://doi.org/10.1016/S0308-8146(97)00183-0

Makkar, H. P. S., & Becker, K. (1997). Potential of Jatropha curcas seed meal as a protein supplement to livestock feed, constraints to its utilization and possible strategies to overcome constraints. Biofuels and Industrial Products from Jatropha curcas, 23-27.

Makkar, H. P. S., & Becker, K. (2009). Jatropha curcas, a promising crop for biodiesel. European Journal of Lipid Science and Technology, 111(8), 773–787. https://doi.org/10.1002/ejlt.200800244

Makkar, H. P. S., Kumar, V., Oyeleye, O., Akinleye, A., Angulo-Escalante, M. A., & Becker, K. (2011). Jatropha platyphylla, a new non-toxic Jatropha species. Food Chemistry, 125(1), 63–71. https://doi.org/10.1016/j.foodchem.2010.08.037

Neu, P., Schober, S., & Mittelbach, M. (2018). Quantification of phorbol esters in Jatropha curcas by HPLC-UV and HPLC-ToF-MS. European Journal of Lipid Science and Technology, 120(4). https://doi.org/10.1002/ejlt.201700293

Nishshanka, U. H., Jayasuriya, C., Chattopadhaya, P. J., Kijak, P. S., Chu, R., Reimschuessel, R., Tkachenko, A., Ceric, O., & De Alwis, H. G. (2016). Screening for toxic phorbol esters in jerky pet treat products using LC–MS. Journal of Chromatography B, 1020, 90–95. https://doi.org/10.1016/j.jchromb.2016.03.018

Olloqui, E. J., Castañeda-Ovando, A., Evangelista-Lozano, S., Alanís-García, E., Ramírez-Moreno, E., Valadez-Vega, C., & Añorve-Morga, J. (2022). Measurement of nutrients and minor components of a non-toxic variety of Jatropha curcas. Journal of Food Measurement and Characterization, 16(2), 1029–1037. https://doi.org/10.1007/s11694-021-01229-6

Oskoueian, E., Abdullah, W. Z., Saad, A., Omar, S., Ahmed, W. B., Kuan, N. A., Zolkifli, R., Hendra, R., & Ho, Y. W. (2011). Bioactive compounds and biological activities of Jatropha curcas kernel meal extract. International Journal of Molecular Sciences, 12(9), 5955–5970. https://doi.org/10.3390/ijms12095955

Patel, R. B., Patel, M. R., Bhatt, K. K., & Patel, B. G. (2010). HPTLC method development and validation. Analytical Methods, 2(5), 525–531. https://doi.org/10.1039/B9AY00259D

Pelletier, G., Padhi, B. K., Hawari, J., Sunahara, G. I., & Poon, R. (2015). Development of a sensitive in vitro assay to quantify the biological activity of pro-inflammatory phorbol esters in Jatropha oil. In Vitro Cellular & Developmental Biology – Animal, 51(6), 644–650. https://doi.org/10.1007/s11626-014-9851-x

Ramadan-Hassanien, M. F. (2023). Bioactive phytochemicals from Jatropha oil processing by-products. In Bioactive phytochemicals from vegetable oil and oilseed processing by-products (pp. 485–503). Springer.

Roach, J. S., Devappa, R. K., Makkar, H. P. S., & Becker, K. (2012). Isolation, stability, and bioactivity of Jatropha curcas phorbol esters. Fitoterapia, 83(3), 586–592. https://doi.org/10.1016/j.fitote.2012.01.006

Roque, J., Dias, L., & Teófilo, R. (2017). Multivariate calibration to determine phorbol esters in seeds of Jatropha curcas using NIR and UV spectroscopies. Journal of the Brazilian Chemical Society, 28(8), 1506–1516. https://doi.org/10.21577/0103-5053.20160333

Salazar-Villa, E., Alcaraz-Meléndez, L., León-Félix, J., Heredia, J. B., Soto-Landeros, F., & Angulo-Escalante, M. A. (2020). Morphological variability and oil content of Jatropha platyphylla germplasm. Scientia Horticulturae, 261, 108968. https://doi.org/10.1016/j.scienta.2019.108968

Shrivastava, A., & Gupta, V. B. (2011). Methods for determination of limit of detection and limit of quantitation. Chronicles of Young Scientists, 2(1), 21–25. https://doi.org/10.4103/2229-5186.79345

Sosa-Segura, M., Dave-Oomah, B., Drover, J., Heredia, J., Osuna-Enciso, T., Valdez-Torres, J., Salazar-Villa, E., Soto-Landeros, F., & Angulo-Escalante, M. (2014). Physical and chemical characterization of three non-toxic oilseeds from the Jatropha genus. Journal of Food and Nutrition Research, 2(1), 56–61. https://doi.org/10.12691/jfnr-2-1-10

Soto-León, S., López-Camacho, E., Milán-Carrillo, J., Sánchez-Castillo, M. A., Cuevas-Rodríguez, E., Picos-Corrales, L. A., & Contreras-Andrade, I. (2014). Jatropha cinerea seed oil as a potential feedstock for biodiesel. Revista Mexicana de Ingeniería Química, 13(3), 739–747.

Vega-Ruiz, Y. C., Hayano-Kanashiro, C., Gámez-Meza, N., & Medina-Juárez, L. A. (2021). Determination of chemical constituents and antioxidant activities of leaves and stems from Jatropha cinerea and Jatropha cordata. Plants, 10(2), 212. https://doi.org/10.3390/plants10020212

Vischi, M., Raranciuc, S., & Baldini, M. (2013). Evaluation of genetic diversity between toxic and nontoxic Jatropha curcas accessions using SSR markers. African Journal of Biotechnology, 12(3), 265–274. https://doi.org/10.5897/AJB12.2155

Wakandigara, A., Nhamo, L. R. M., Kugara, J., & Mushonga, P. (2020). Mechanisms of phorbol ester toxicity, determined by molecular modelling. International Research Journal of Pure & Applied Chemistry, 21(18), 10-24. https://doy.org/10.9734/irjpac/2020/v21i1830267

Yamunadevi, M., Wesely, E., & Johnson, M. (2011). Phytochemical studies on terpenoids of Aerva lanata using HPTLC. Asian Pacific Journal of Tropical Biomedicine, 1(2), S220–S225. https://doi.org/10.1016/S2221-1691(11)60161-0

Salazar

Downloads

Published

2026-05-12

Data Availability Statement

The data supporting the findings of this study are available within the article and its supplementary material.

Issue

No. 5 (2026): QUIBIOUAS Revista de Ciencias Químico Biológicas

Section

Scientific articles

Categories

License

Copyright (c) 2026 QUIBIOUAS, Journal of Biological Chemical Sciences

Creative Commons License

This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License.

How to Cite

Salazar-Villa, E., Medina-Rodelo, D. P., Rojo-Báez, I., Báez-Parra, K. M., Soto- Landeros, F., & Angulo-Escalante, M. A. (2026). Quantification of phorbol esters content by HPTLC in seeds of Jatropha species genotypes. QUIBIOUAS, Journal of Biological Chemical Sciences, 5, 8-19. https://revistas.uas.edu.mx/index.php/QBU/article/view/1362