Simultaneous isolation and purification of cucurbitacin D and I from Ecballium elaterium (l.) A. Rich fruit juice

Emir Tosun, Ahmet Baysar

Abstract


The objective of this study was to develop a rapid, economic, and efficient method for simultaneous selective isolation, separation, and purification of cucurbitacin D and I from Ecballium elaterium (L.) A. Rich fruit juice via reversed-phase flash chromatography combined with HPLC. The chloroform extract of the fruit juice was fractionated with flash chromatography using a chloroform, acetone and methanol solvent combination at a 5 ml/min flow rate. Then, a validated HPLC method was utilized for purification of the two targeted cucurbitacins. Cucurbitacin D and I were collected automatically by the fraction collector. The fractions containing the same compounds were pooled and lyophilized. The purified cucurbitacin D and I compounds were identified by NMR, LC-MS, and UV spectra analysis. The results suggest that the applied procedure is simple, quick, and highly efficient. The HPLC method was found to be linear, accurate, precise and rugged for the quantification of the cucurbitacins studied.


Keywords


cucurbitacin D; cucurbitacin I; Ecballium elaterium (L.) A. Rich; isolation; method validation

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References


E. Yesilada, S. Tanaka, E. Sezik, Isolation of an anti-inflammatory principle from the fruit juice of Ecballi-um elaterium, J. Nat. Prod., 51(3), 504–508 (1988).

https://pubs.acs.org/doi/abs/10.1021/np50057a008.

E. G. Attard, A. Scicluna-Spiteri, Ecballium elaterium: an in vitro source of cucurbitacins, Fitoterapia, 72, 46–53 (2001). https://doi.org/10.1016/S0367-326X(00)00256-2.

A. Agıl, M. Miro, J. Jimenez, J. Anerios, M. D. Cara-cuel, A. Garciagranados, M. C. Navarro, Isolation of an anti-hepatotoxic principle from the juice of Ecbal-lium elaterium. Planta. Med., 65, 673–675 (1999).

https://www.thieme-connect.de/DOI/DOI?10.1055/s-2006-960847.

H. G. Gerges, R. A. Khalil, E. A. Mansour, J. Mag-dalou, R. Chahine, N. Ouaini, Cucurbitacins from Ec-ballium elaterium juice increase the binding of biliru-bin and ibuprofen to albumin in human plasma. Chem-Biol Interact., 169, 53–62 (2007).

https://doi.org/10.1016/j.cbi.2007.05.003.

S. Bohlooli, N. Jafari, S. Jahed, Cytotoxic effect of freeze-dried extract of Ecballium elaterium fruit on gastric adenocarcinoma (AGS) and esophageal squa-mous cell carcinoma (KYSE30) cell lines. J Gastroin-test Cancer, 43, 579–583 (2012).

https://link.springer.com/article/10.1007%2Fs12029-012-9383-4.

A. S. Salhab, Human exposure to Ecballium elaterium fruit juice: fatal toxicity and possible remedy. Phar-macol Pharm, 4, 447–450 (2013).

http://file.scirp.org/Html/6-2500327_36044.htm

G. Toker, M. Memişoğlu, M. C. Toker, E. Yesilada, Callus formation and cucurbitacin B accumulation in Ecballium elaterium callus cultures. Fitoterapia, 74, 618–623 (2003).

https://doi.org/10.1016/S0367-326X(03)00165-5.

I. Ielciu, M. Frederich, M. Tits, L. Angenot, R. Paltine-an, E. Cieckiewicz, G. Crişan, L. Vlase, Bryonia alba L. and Ecballium elaterium (L) A Rich -Two related species of the cucurbitaceae family with important pharmaceutical potential. Farmacia, 64(3), 323–332 (2016).

https://www.researchgate.net/publication/304990164.

U. Kaushik, V. Aeri, S. R. Mir, Cucurbitacins - An insight into medicinal leads from nature. Pharmacogn. Rev., 9(17), 12–18 (2015).

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4441156/.

J. Gry, I. Søborg, H. C. Andersson, Cucurbitacins in Plant Food Tema Nord Nordic Council of Ministers; Ekspressen Tryk & Kopicenter; Copenhagen, 2006.

https://www.diva-portal.org/smash/get/diva2:701868/ FULLTEXT01.pdf.

M. Miro, Cucurbitacins and their pharmacological effects Phytother., 9, 159–168 (1995).

https://doi.org/10.1002/ptr.2650090302.

A. A. Alghasham, Cucurbitacins-a promising target for cancer therapy. Int. J. Health Sci., 7(1), 77–89 (2013). https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3612419/.

S. O. Chung, Y. J. Kim, S. U. Park, An updated review of cucurbitacins and their biological and pharmacolog-ical activities (Letter to the editor). EXCLI J, 14, 562–566 (2015). http://www.excli.de/vol14/Park_05052015_proof.pdf.

I. T. Silva, F. C. Geller, L. Persich, S. E. Dudek, K. L. Lang, M. S. B. Caro, F. J. Duran, E. P. Schenkel, S. Ludwig, C. M. O. Simoes, Cytotoxic effects of natural and semisynthetic cucurbitacins on lung cancer cell line A549. Invest New Drug, 34(2), 139–148 (2016). https://link.springer.com/article/10.1007%2Fs10637-015-0317-4.

D. H. Lee, G. B. Iwanski, N. H. Thoennissen, Cucur-bitacin: ancient compound shedding new light on can-cer treatment. Scientific World Journal, 10, 413–418 (2010).

https://www.hindawi.com/journals/tswj/2010/565972/abs/.

L. H. Mary, Lakshmi, F. Tilton, J. Joseph, S. S. So-losan, D. Sudarsanam, Phytochemical antioxidant and cytotoxic properties of the fruit extract from Cucur-bita digitate. Int J. Pharm. Pharm. Sci., 6(4), 353–356 (2014).

https://innovareacademics.in/journal/ijpps/Vol6Issue4/9124.pdf.

K. Dhiman, A. Gupta, D. K. Sharma, N. S. Gill, A. Goyal, A review on the medicinally important plants of the family cucurbitaceae. Asian Journal of Clinical Nutrition, 4(1), 16–26 (2012).

https://scialert.net/abstract/?doi=ajcn.2012.16.26.

N. S. Gill, M. Bali, Isolation of anti ulcer cucurbitane type triterpenoid from the seeds of Cucurbita pepo. Res J Phytochemistry, 5(2), 70–79, (2011).

https://scialert.net/abstract/?doi=rjphyto.2011.70.79.

L. Dinan, J. Harmatha, R. Lafont, Chromatographic procedure for the isolation of plant steroids. J. Chro-matogr. A 935, 105–123 (2001).

https://doi.org/10.1016/S0021-9673(01)00992-X.

J. Bartalis, F. T. Halaweish, Relationship between Cu-curbitacins reversed-phase high-performance liquid chromatography hydrophobicity index and basal cyto-toxicity on HepG2 cells. J Chromatogr B, 818, 159–166 (2005). https://doi.org/10.1016/j.jchromb.2004.12.020.

W. A. Oleszek, Chromatographic determination of plant saponins. J. Chromatogr A, 967, 147–162 (2002).

https://doi.org/10.1016/S0021-9673(01)01556-4.

C. A. Rice, K. A Raymal, O. L. Chambliss, F. A. John-son, Chromatographic and mass spectral analysis of Cucurbitacins of three Cucumis sativus cultivars, J. Agr Food Chem.,29, 194–196 (1981).

https://pubs.acs.org/doi/abs/10.1021/jf00103a051.

N. Hassan, J. Ahamad, S. Amin, M. Mujeeb, S. R. Mir, Rapid preparative isolation of erythrocentaurin from Enicostemma littorale by medium-pressure liquid chromatography its estimation by high-pressure thin-layer chromatography and its α-amylase inhibitory ac-tivity, J. Sep. Sci., 38, 592–598 (2015).

https://doi.org/10.1002/jssc.201401030.

J. Tao, R. Yan, L. Zhao, D. Wang, X. Xu, Separation and purification of two taxanes and one xylosyl-containing taxane from Taxus wallichiana Zucc: A comparison between high-speed countercurrent chro-matography and reversed-phase FC. J. Sep. Sci., 40(6), 1273–1282 (2017).

https://doi.org/10.1002/jssc.201601066.

S. Sturm, H. Stuppner, Analysis of cucurbitacins in medicinal plants by high pressure liquid chromatog-raphy-mass spectrometry, Phytochem Analysis, 11(2), 121–127 (2000).

https://doi.org/10.1002/(SICI)1099-1565(200003/04)11:2<121:AID-PCA493>3.0.CO;2-9.

G .I. Kaya and M. F. Melzig, Quantitative determina-tion of cucurbitacin E and cucurbitacin I in homoeo-pathic mother tincture of Gratiola officinalis L by HPLC. Pharmazie, 63, 851–853 (2008).

https://doi.org/10.1691/ph.2008.8197.

F. T. Halaweish, D. W. Tallamy, Quantitative determi-nation of cucurbitacins by high performance liquid chromatography and high performance thin layer chromatography, J. Liq. Chromatogr. 16, 497–511 (1993).

https://doi.org/10.1080/10826079308020927.

X. S. Feng, D. C. Wang, H. Cai, X. M. Deng, Y. R. Liu, Determination of the cucurbitacins from Cucubita pepo cv dayangua by HPLC, Zhong Yao Cai 30(4), 418–420 (2007). https://www.researchgate.net/publication/6164198_Determina-tion_of_the_cucurbitacins_from_Cucubita_pepo_ cv_dayangua_by_HPLC.

S. Sasidharan, Y. Chen, D. Saravanan, K. M. Sun-dram, L. Y. Latha, Extraction isolation and characteri-zation of bioactive compounds from plants' extracts. Afr. J. Tradit. Complem., 8(1), 1–10 (2011).

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218439/.

P. B. Krepsky, M. O. Cervelin, D. Porath, R. R. Peters, R. Ribeiro-do-Valle, M. R. Farias, High performance liquid chromatography determination of cucurbitacins in the roots of Wilbrandia ebracteata Cogn. Braz. J. Pharmacog, 19(3), 715–719 (2009).

http://dx.doi.org/10.1590/S0102-695X2009000500011.

E. Sezik, Research on the Turkish medicinal plant, Chem. Nat. Compound, 33(5), 541–542 (1995).

https://link.springer.com/article/10.1007/BF02254799.

E. Attard, Rapid detection of Cucurbitacins in tissues and in vitro cultures of Ecballium elaterium (L.) A. Rich Cucurbit Genet. Coop Report, 25, 71–75 (2002). http://cuke.hort.ncsu.edu/cgc/cgc25/cgc25-25.pdf.

C. C. Wang, L. G. Chen, T. L. Chang, C.T. Hsieh, Extracts of aquilaria hulls and use thereof in the treat-ment of cancer, US Patent 20110160152 A1, June 30, 2011.

https://patents.google.com/patent/US20110160152A1/en.

M. M. Saker, M. M. Farid, A. A. Fahmi, S. A. El-Mekkawy, H. S. Taha, A. I. Amin, Large scale produc-tion of antitumor cucurbitacins from Ecballium elate-rium using bioreactor, Afr. J. Biotechnol., 11(66), 12974–12982 (2012). http://dx.doi.org/10.5897/AJB12.878.

ICH Q2 (R1). Proceedings of International Confer-ence of Harmonization. Geneva, 2005.

http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/09/WC500002662.pdf.

S. Hu, Y. Wang, B. Avula, M. Wang, I. A. Khan, Sep-aration of cucurbitane triterpenoids from bitter melon drinks and determination of partition coefficients us-ing vortex-assisted dispersive liquid-phase microex-traction followed by UHPLC analysis, J. Sep. Sci, 40, 2238–2245 (2017). https://doi.org/10.1002/jssc.201700023.

Cucurbitacin D.

https://pubchem.ncbi.nlm.nih.gov/compound/Cucurbitacin-D, accessed September 2019.

Cucurbitacin I.

https://pubchem.ncbi.nlm.nih.gov/compound/Cucurbitacin-I, accessed September 2019.

V. Subbiah, Method of isolating cucurbitacin. US Pa-tent 5925356, July 20, 1999.

https://patents.google.com/patent/US5925356A/en.

C. Che, X. Fang, C. H. Phobe, A. D. Kinghorn, N. R. Farnsworth, High-field 1H-NMR analysis of some cu-curbitacins, J. Nat. Prod., 48(3), 429–434 (1985). https://pubs.acs.org/doi/abs/10.1021/np50039a010.

C. Seger, S. Sturm, M. E. Mair, E. P. Ellmerer, H. Stuppner, 1H and 13C NMR signal assignment of cu-curbitacin derivates from Citrillus colocynthis (L) Schrader and Ecballium elaterium L (Cucurbitaceae). Magn. Reson. Chem., 43, 489–491 (2005).

https://doi.org/10.1002/mrc.1570.

C. Seger, S. Sturm, E. Haslinger, H. Stuppner, NMR signal assignment of 22-deoxocucurbitacin D and cu-curbitacin D from Ecballium elaterium L (Cucurbita-ceae). Monatshefte für Chemie, 136, 1645–1649 (2005).

https://link.springer.com/article/10.1007/s00706-005-0347-2.

M. Wen-Li, L. Feng, Z. Wen-Jian, W. Hui, D. Hao-Fu, Cucurbitacins from fruits of Aquilaria sinensis. Chin. J. Nat. Med., 10(3), 234–237 (2012).

https://doi.org/10.3724/SP.J.1009.2012.00234.

A. Alsayari, L. Kopel, M. S. Ahmed, H. S. M. Soliman, S. Annadurai, F.T. Halaweish, Isolation of anticancer constituents from Cucumis propheratum var. propheratum through bioassay-guided fractionation. BMC Complem. Altern M, 18, 1–12 (2018).

https://doi.org/10.1186/s12906-018-2295-5.

R. Chawech, R. Jarraya, C. Girardi, M. Vansteelandt, G. Marti, I. Nasri, C. Racaud-Sultan, N. Fabre, Cucurbitacins from the leaves of Citrullus colocynthis (L.) Schrad, Molecules, 20, 18001–18015 (2015).

DOI:10.3390/molecules20101800




DOI: http://dx.doi.org/10.20450/mjcce.2019.1648

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