Highly Efficient Extraction Procedures Based on Natural Deep Eutectic Solvents or Ionic Liquids for Determination of 20-Hydroxyecdysone in Spinach
Open accessAbstract
A novel, efficient extraction procedure based on natural deep eutectic solvents (NADES) and ionic liquids (ILs) for determination of 20-hydroxyecdysone (20-E) in spinach has been developed. NADES, the first green extraction agent, with different hydrogen bond donors and acceptors are screened in order to determine extraction efficiencies. NADES consisting of lactic acid and levulinic acid at a molar ratio of 1:1 exhibits the highest yields. ILs, the second green extraction agent, with various cations and anions are also investigated, where [TEA] [OAc]·AcOH, χAcOH = 0.75 displays the highest recovery. Moreover, NADES-SLE and IL-SLE (SLE, solid-liquid extraction) parameters are investigated. Using the obtained optimized method, the recoveries of the target compound in spinach are above 93% and 88% for NADES-SLE and IL-SLE procedure, respectively. The methods display good linearity within the range of 0.5–30 μg/g and LODs of 0.17 µg/g. The proposed NADES-SLE-UHPLC-UV and IL-SLE-UHPLC-UV procedures can be applied to the analysis of 20-E in real spinach samples, making it a potentially promising technique for food matrix. The main advantage of this study is the superior efficiency of the new, green extraction solvents, which results in a significant reduction of extraction time and solvents as compared to those in the literature.
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The reported content of 20-hydroxyecdysone in spinach leaves ranges from 17.1 to 885 μg/g.
“The reported content of 20HE in spinach leaves ranges from 17.1 to 885 µg/g [158,159].”
From Ecdysterone and Turkesterone—Compounds with Prominent Potential in Sport and Healthy Nutrition (2024) · cites this paper as [158]
The Conclusions report 20-hydroxyecdysone levels of 17.1–885 µg/g in real spinach samples. Section 2.6 says those samples included leaves, stalks, and seeds, but the available passage does not establish that both endpoints apply to leaves specifically. (LLM, conf=0.86)
Experimental — accuracy not yet measuredDeep eutectic solvents and ionic liquids are a recently introduced method for extracting phytoecdysteroids, with high extraction yields and fast.
“A further, recently introduced method for the extraction of phytoecdysteroids is the use of deep eutectic solvents and ionic liquids [114], which possess high extraction yields and are fast.”
From Ecdysteroids (2021) · cites this paper as [114]
The content of 20E assayed in spinach was of the same order of magnitude as that given in the literature.
“The content of the 20E assayed in spinach was of the same order of magnitude as that given in the literature (Bajkacz et al., 2020;Bakrim et al., 2008;Gorelick et al., 2020).”
From Efficient Extraction and Sensitive HPLC-MS/MS Quantification of Selected Ecdysteroids in Plants (2022) · cites this paper as Bajkacz et al., 2020
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Cites
Natural Deep Eutectic Solvents – Solvents for the 21st Century
Paiva, A., Craveiro, R., Aroso, I., Martins, M., Reis, R. L., & Duarte, A. R. C. (2014) · ACS Sustainable Chemistry & Engineering · doi:10.1021/sc500096j · Cited by 2,458 · see citing works
Recent Applications of Ionic Liquids in Separation Technology
Han, D., & Row, K. H. (2010) · Molecules · doi:10.3390/molecules15042405 · Cited by 560 · see citing works
Green solvents from ionic liquids and deep eutectic solvents to natural deep eutectic solvents
Vanda, H., Dai, Y., Wilson, E. G., Verpoorte, R., & Choi, Y. H. (2018) · Comptes Rendus. Chimie · doi:10.1016/j.crci.2018.04.002 · Cited by 467 · see citing works
Phytoecdysteroids: Biological aspects
Dinan, L. (2001) · Phytochemistry · doi:10.1016/s0031-9422(01)00078-4 · Cited by 392 · see citing works
Huang, Y., Feng, F., Jiang, J., Qiao, Y., Wu, T., Voglmeir, J., & Chen, Z.-G. (2017) · Food Chemistry · doi:10.1016/j.foodchem.2016.11.013 · Cited by 366 · see citing works
Fernández, M. D. L. Á., Espino, M., Gomez, F. J. V., & Silva, M. F. (2018) · Food Chemistry · doi:10.1016/j.foodchem.2017.06.150 · Cited by 257 · see citing works
Bajkacz, S., & Adámek, J. (2017) · Food Analytical Methods · doi:10.1007/s12161-017-1118-5 · Cited by 215 · see citing works
Self-citationPhytoecdysteroids and Anabolic Androgenic Steroids--Structure and Effects on Humans
Bathori, M., Toth, N., Hunyadi, A., Marki, A., & Zador, E. (2008) · Current Medicinal Chemistry · doi:10.2174/092986708783330674 · Cited by 201 · see citing works
On the distribution of phytoecdysteroids in plants
Dinan, L., Savchenko, T., & Whiting, P. (2001) · Cellular and Molecular Life Sciences · doi:10.1007/pl00000926 · Cited by 126 · see citing works
Wu, Y., Wu, P., Li, Y. B., Liu, T. C., Zhang, L., & Zhou, Y. (2018) · RSC Advances · doi:10.1039/c7ra13581e · Cited by 93 · see citing works
Gómez, A. V., Tadini, C. C., Biswas, A., Buttrum, M., Kim, S., Boddu, V. M., & Cheng, H. N. (2019) · LWT · doi:10.1016/j.lwt.2019.02.052 · Cited by 87 · see citing works
Absalan, G., Akhond, M., & Sheikhian, L. (2008) · Talanta · doi:10.1016/j.talanta.2008.06.049 · Cited by 77 · see citing works
Ecdysteroids in Spinach (Spinacia oleracea L.): Biosynthesis, Transport and Regulation of Levels
Bakrim, A., Maria, A., Sayah, F., Lafont, R., & Takvorian, N. (2008) · Plant Physiology and Biochemistry · doi:10.1016/j.plaphy.2008.06.002 · Cited by 63 · see citing works
Hunyadi, A., Herke, I., Lengyel, K., Báthori, M., Kele, Z., Simon, A., Tóth, G., & Szendrei, K. (2016) · Scientific Reports · doi:10.1038/srep37322 · Cited by 59 · see citing works
Tarkowská, D., & Strnad, M. (2016) · Planta · doi:10.1007/s00425-016-2561-z · Cited by 56 · see citing works
Occurrence and levels of ecdysteroids in spinach
Grebenok, R. J., Ripa, P. V., & Adler, J. H. (1991) · Lipids · doi:10.1007/bf02536433 · Cited by 52 · see citing works
Wilson, I. D., Morgan, E. D., Lafont, R., Shockcor, J. P., Lindon, J. C., Nicholson, J. K., & Wright, B. (1999) · Chromatographia · doi:10.1007/bf02467609 · Cited by 52 · see citing works
Bogdanov, M. G., Svinyarov, I., Keremedchieva, R., & Sidjimov, A. K. (2012) · Separation and Purification Technology · doi:10.1016/j.seppur.2012.02.001 · Cited by 48 · see citing works
Tan, T., Lai, C., Ouyang, H., He, M., & Feng, Y. (2015) · Journal of Pharmaceutical and Biomedical Analysis · doi:10.1016/j.jpba.2015.12.019 · Cited by 48 · see citing works
Brønsted acids in ionic liquids: how acidity depends on the liquid structure
McCune, J. A., He, P., Petkovic, M., Coleman, F., Estager, J., Holbrey, J. D., Seddon, K. R., & Swadźba‐Kwaśny, M. (2014) · Physical Chemistry Chemical Physics · doi:10.1039/c4cp03217a · Cited by 43 · see citing works
Modlin, R. F., Alred, P. A., & Tjerneld, F. (1994) · Journal of Chromatography A · doi:10.1016/0021-9673(94)80112-6 · Cited by 34 · see citing works
Zhu, H., Fan, Y., Qian, Y.-L., Tang, H.-F., Ruan, Z., Liu, D.-H., & Wang, H. (2013) · Chinese Chemical Letters · doi:10.1016/j.cclet.2013.12.001 · Cited by 24 · see citing works
Svinyarov, I., Keremedchieva, R., & Bogdanov, M. G. (2016) · Separation Science and Technology · doi:10.1080/01496395.2016.1171783 · Cited by 20 · see citing works
HPTLC Method for Determination of 20-Hydroxyecdysone in Sida rhombifolia L. and Dietary Supplements
Jadhav, A. N., Rumalla, C. S., Avula, B., & Khan, I. A. (2007) · Chromatographia · doi:10.1365/s10337-007-0407-3 · Cited by 18 · see citing works
da Rosa, H. S., Koetz, M., Santos, M. C., Jandrey, E. H. F., Folmer, V., Henriques, A. T., & Mendez, A. S. L. (2018) · Steroids · doi:10.1016/j.steroids.2018.01.014 · Cited by 16 · see citing works
Gallo, M. B. C., Beltrame, F. L., Vieira, P. C., Cass, Q. B., Fernandes, J. B., & da Silva, M. F. D. G. F. (2006) · Journal of Chromatography B · doi:10.1016/j.jchromb.2005.12.006 · Cited by 15 · see citing works
Zhao, C., Lü, Z., Li, C., He, X., Zhao, L., Shi, K., Yang, L., Fu, Y., & Zu, Y. (2014) · The Scientific World JOURNAL · doi:10.1155/2014/283024 · Cited by 15 · see citing works
Muchate, N. S., Kadam, N. S., Rajurkar, N. S., & Nikam, T. D. (2017) · Journal of Planar Chromatography – Modern TLC · doi:10.1556/1006.2017.30.3.7 · Cited by 15 · see citing works
Extraction and Monitoring of Phytoecdysteroids Through HPLC
Ghosh, D., & Laddha, K. S. (2006) · Journal of Chromatographic Science · doi:10.1093/chromsci/44.1.22 · Cited by 11 · see citing works
LIQUID CHROMATOGRAPHIC MONITORING OF PHYTOECDYSTEROID PRODUCTION OF SERRATULA WOLFFII
Báthori, M., Gergely, A., Kalász, H., Nagy, G., Dobos, Á., & Máthé, I. (2000) · Journal of Liquid Chromatography & Related Technologies · doi:10.1081/jlc-100101452 · Cited by 10 · see citing works
Wu, J., Cheng, K., Wang, H., Ye, W., Li, E. T. S., & Wang, M. (2008) · Phytochemical Analysis · doi:10.1002/pca.1097 · Cited by 9 · see citing works
Cao, V. D., Riu, K.-Z., & Boo, K.-H. (2018) · Plant Physiology and Biochemistry · doi:10.1016/j.plaphy.2018.06.027 · Cited by 7 · see citing works
Rybin, V., BOLTENKOV, E. V., & Новожилова, Е. В. (2007) · Natural Product Communications · doi:10.1177/1934578x0700201110 · Cited by 7 · see citing works
Flagged first, then by citations
1 of the 34 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Didion, Y. P., Tjalsma, T. G., Su, Z., Malankowska, M., & Pinelo, M. (2023) · Separation and Purification Technology · doi:10.1016/j.seppur.2023.124147 · Cited by 107 · see citing works
Koh, Q. Q., Kua, Y. L., Gan, S., Tan, K. W., Lee, T. Z. E., Cheng, W. K., & Lau, H. L. N. (2023) · Sustainable Chemistry and Pharmacy · doi:10.1016/j.scp.2023.101218 · Cited by 109 · see citing works
Applications of (natural) deep eutectic solvents in liquid phase microextraction: A review
Mansour, F. R., Bedair, A., Hamed, M., Magdy, G., Ali, I., & Locatelli, M. (2024) · Microchemical Journal · doi:10.1016/j.microc.2024.110178 · Cited by 107 · see citing works
Silica-Based Supported Ionic Liquid-like Phases as Heterogeneous Catalysts
Wolny, A., & Chrobok, A. (2022) · Molecules · doi:10.3390/molecules27185900 · Cited by 65 · see citing works
Self-citationPochivalov, A., Cherkashina, K., Shishov, A. Y., & Bulatov, A. V. (2021) · Journal of Molecular Liquids · doi:10.1016/j.molliq.2021.116827 · Cited by 61 · see citing works
Ionic Liquids for Development of Heterogeneous Catalysts Based on Nanomaterials for Biocatalysis
Wolny, A., & Chrobok, A. (2021) · Nanomaterials · doi:10.3390/nano11082030 · Cited by 53 · see citing works
Self-citationDeep Eutectic Solvents Application in Food Analysis
Ortega-Zamora, C., González‐Sálamo, J., & Hernández‐Borges, J. (2021) · Molecules · doi:10.3390/molecules26226846 · Cited by 52 · see citing works
Lafont, R., Balducci, C., & Dinan, L. (2021) · Encyclopedia · doi:10.3390/encyclopedia1040096 · Cited by 49 · see citing works
Acosta-Vega, L., Cifuentes, A., Ibáñez, E., & Garcia, P. G. (2025) · Molecules · doi:10.3390/molecules30020284 · Cited by 48 · see citing works
Ecdysterone and Turkesterone—Compounds with Prominent Potential in Sport and Healthy Nutrition
Todorova, V., Ivanova, S., Chakarov, D., Kraev, K., & Ivanov, K. (2024) · Nutrients · doi:10.3390/nu16091382 · Cited by 31 · see citing works
“Ecdysteroids improved The reported content of 20HE in spinach leaves ranges from 17.1 to 885 µ g/g [158,159].”
BackgroundInfluentialNew phases for analytical scale extraction from plants: Current and future trends
Mastellone, G., Marengo, A., Sgorbini, B., Rubiolo, P., & Cagliero, C. (2021) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2021.116288 · Cited by 31 · see citing works
The Versatility of NADES Across Applications
Freitas, D. S., Ribeiro, A., Cavaco-Paulo, A. M., & Silva, C. (2025) · Molecules · doi:10.3390/molecules30193862 · Cited by 29 · see citing works
Stando, K., Korzeniewska, E., Felis, E., Harnisz, M., & Bajkacz, S. (2022) · Molecules · doi:10.3390/molecules27144378 · Cited by 23 · see citing works
Self-citationSocas‐Rodríguez, B., Mendiola, J. A., Rodríguez‐Delgado, M. Á., Ibáñez, E., & Cifuentes, A. (2022) · Journal of Chromatography A · doi:10.1016/j.chroma.2022.462922 · Cited by 23 · see citing works
Efficient Extraction and Sensitive HPLC-MS/MS Quantification of Selected Ecdysteroids in Plants
Fang, X., Szołtysik, R., Tang, J., & Bajkacz, S. (2022) · Journal of Food Composition and Analysis · doi:10.1016/j.jfca.2022.104580 · Cited by 20 · see citing works
Self-citationImprovement of the Solubilization and Extraction of Curcumin in an Edible Ternary Solvent Mixture
Huber, V., Muller, L., Hioe, J., Degot, P., Touraud, D., & Kunz, W. (2021) · Molecules · doi:10.3390/molecules26247702 · Cited by 17 · see citing works
Urinary Excretion of Ecdysterone and Its Metabolites Following Spinach Consumption
Yuliandra, T., Touvleliou, K., de la Torre, X., Botrè, F., Loke, S., Isenmann, E., Valder, S., Diel, P., & Parr, M. K. (2023) · Molecular Nutrition & Food Research · doi:10.1002/mnfr.202200518 · Cited by 12 · see citing works
Luna, V. M. A., Herrero, M., de la Parra, M. J., Arribas, Á. G., Ibáñez, E., & Montero, L. (2025) · Advances in Sample Preparation · doi:10.1016/j.sampre.2025.100149 · Cited by 8 · see citing works
Todorova, V., Ivanova, S., Yotov, V., Zaytseva, E., Ardasheva, R., Turiyski, V., Prissadova, N., & Ivanov, K. (2024) · Molecules · doi:10.3390/molecules29215145 · Cited by 7 · see citing works
Jędrejko, M., Jędrejko, K., Renaud, V. M. G., Kała, K., & Muszyńska, B. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25073650 · Cited by 7 · see citing works
Cerulli, A., Polcaro, L. M., Masullo, M., & Piacente, S. (2024) · Foods · doi:10.3390/foods13223699 · Cited by 6 · see citing works
Tripathy, S., Ranjan, S., & Srivastav, P. P. (2025) · Journal of Molecular Liquids · doi:10.1016/j.molliq.2025.128980 · Cited by 6 · see citing works
Isenmann, E., Yuliandra, T., Touvleliou, K., Broekmann, M., de la Torre, X., Botrè, F., Diel, P., & Parr, M. K. (2024) · Archiv der Pharmazie · doi:10.1002/ardp.202300689 · Cited by 5 · see citing works
Jędrejko, K., Lazarev, A., Jędrejko, M., & Muszyńska, B. (2024) · Food Reviews International · doi:10.1080/87559129.2023.2238055 · Cited by 4 · see citing works
Carmona-Molero, R., Carbonell‐Rozas, L., Lara, F. J., Ana, M. G., & del Olmo-Iruela, M. (2025) · Microchemical Journal · doi:10.1016/j.microc.2025.115244 · Cited by 3 · see citing works
Isenmann, E., Yuliandra, T., Touvleliou, K., Broekmann, M., Torre, X. D. L., Botrè, F., Diel, P., & Parr, M. K. (2023) · doi:10.1101/2023.10.10.560427 · Cited by 2 · see citing works
Dissemond, J., Havers, T., Held, S., Geisler, S., Kostov, T., Diel, P., Türschmann, S., Parr, M. K., & Isenmann, E. (2025) · Journal of the International Society of Sports Nutrition · doi:10.1080/15502783.2025.2540408 · Cited by 2 · see citing works
“Studies on foods such as spinach or quinoa, which generally contain 20E, show that the concentration is highly dependent on various factors such as origin, genetics and environmental conditions [38,85–90].”
Ali, Z., Yadav, P., Tiwari, A., Dixit, S., Panday, V., & Patel, D. K. (2026) · Journal of Molecular Liquids · doi:10.1016/j.molliq.2026.129768 · Cited by 3 · see citing works
Li, J., Kang, Y., Wang, Y., Liu, J., Wang, Y., Liu, S., Bu, Y., Li, X., Xie, J., & Wang, Z. (2024) · SSRN Electronic Journal · doi:10.2139/ssrn.4946140
Li, J., Kang, Y., Wang, Y., Liu, J., Wang, Y., Liu, S., Bu, Y., Li, X., Xie, J., & Wang, Z. (2025) · Journal of Separation Science · doi:10.1002/jssc.70116
Shirke, H. A., Vasav, A. P., Darshetkar, A. M., Godbole, R. C., Kadam, S. B., Patil, S. S., Naikawadi, V. B., Kishor, P. B. K., Nikam, T. D., & Barvkar, V. T. (2025) · Physiology and Molecular Biology of Plants · doi:10.1007/s12298-025-01656-5
Todorova, V., Ivanova, S., Ardasheva, R., & Ivanov, K. (2026) · Molecules · doi:10.3390/molecules31071090
Ecdysterone: A Component of Dietary Supplements with Ergogenic Potential?
León, S. A., Robayna, B. P., Romero, C. D., & Brito, N. B. (2026) · Nutraceuticals · doi:10.3390/nutraceuticals6020031
Deep Eutectic Solvents for Extraction and Detection of Adulterants in Food Samples
Uddin, M., Muhammad, Z., Saidu, A. K., Zain, N. N. M., & Alahmad, W. (2025) · Springer eBooks · doi:10.1007/978-3-031-99454-8_11
- Authors
- Bajkacz, S., Rusin, K., Wolny, A., Adamek, J., Erfurt, K., & Chrobok, A.
- Affiliations
- Silesian University of Technology (PL)
- Biotechnology Center of Silesian University of Technology, Krzywoustego 8, 44-100 Gliwice, Poland
- Corresponding author
- Bajkacz, Sylwia
- Venue
- Molecules
- Publisher
- MDPI AG
- Published
- 2020-10-15
- Volume
- 25
- Issue
- 20
- Pages
- 4736
- ISSN
- 1420-3049 (ISSN-L)
- PMC
- PMC7587567
- OpenAlex
- W3092796971
- Cited by
- 34
- References
- 34
- Citations by year
- 2026: 3 · 2025: 9 · 2024: 7 · 2023: 5 · 2022: 4 · 2021: 6
- FWCI
- 0.77
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecules (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Physical Sciences › Chemical Engineering › Catalysis › Ionic liquids properties and applications
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical chemistry methods development
- Life Sciences › Agricultural and Biological Sciences › Food Science › Pesticide Residue Analysis and Safety
- Keywords
- 20-hydroxyecdysone, spinach, natural deep eutectic solvents, ionic liquids, solid-liquid extraction, green solvents, lactic acid, levulinic acid, extraction optimization, food analysis, insecticide residues
- MeSH terms
- Biological Products, Ecdysterone, Hydrogen Bonding, Plant Extracts, Solvents, Water, Spinacia oleracea, Ionic Liquids
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