Polarity-Directed Extraction and Metabolomic Profiling of Saffron Petals for Reproducible Anti-Inflammatory Activity

Authors

  • Eric Neubauer Vickers Harvard University, United States Author
  • Devin Bowen Wan University of Hong Kong, Hong Kong SAR, China Author
  • Thembisa Monki University of Cape Town, South Africa Author

Keywords:

Crocus sativus, extraction, metabolomics, anti-inflammatory, macrophages, standardisation

Abstract

Background: Variation in solvent composition can substantially alter the biological performance of botanical extracts and complicate reproducibility.

Objective: To optimise saffron-petal extraction for enrichment of anti-inflammatory constituents and link chemical profiles to cell-based activity.

Methods: Eighteen solvent systems varying in aqueous, ethanolic and non-polar proportions were compared using a response-surface design. Untargeted LC-MS profiling generated metabolomic fingerprints, and anti-inflammatory activity was assessed in lipopolysaccharide-stimulated macrophages by nitric oxide, TNF-α and IL-6 production. Cytotoxicity was measured in parallel.

Results: Extraction yield ranged from 8.4% to 31.7% by dry weight, but high yield did not predict biological activity. A 20:60:20 non-polar/ethanol/water system generated the strongest multivariate separation and reduced nitric-oxide production by 48% at a non-cytotoxic concentration. This fraction was enriched in seven discriminant features provisionally annotated as flavonoid glycosides and apocarotenoid-related metabolites. TNF-α and IL-6 were reduced by 35% and 29%, respectively, compared with stimulated controls. Re-extraction on three separate days produced coefficients of variation below 8% for the five most abundant marker peaks, supporting procedural reproducibility.

Conclusion: Polarity-directed extraction produced a chemically reproducible saffron-petal fraction with stronger anti-inflammatory activity than yield-maximising methods. The study supports metabolomic marker panels as a practical bridge between botanical processing and biological standardisation.

Publication format: research abstract. No full-text manuscript accompanies this record.

References

Alam SR, Nasrin N, Chowdhury NS, Akter R, Rahman MA, Chowdhury RN, Tahsin MR, Arnob TA, Morshed N, Mia MSH, Opo FADM, Aktar F, Miah MM, Büyüker SM, Reza R, Ayubee MS, Reza MS, Chowdhury JA, Chowdhury AA, Kabir S, Amran MS. Exploring the abundance of de-oiled non-resin analgesic and anti-inflammatory compounds in Crocus sativus through polarity-directed quaternary solvent extraction in both diabetic and non-diabetic conditions: Integrating in silico and in vivo analysis. Acta Pharm Sci. 2026;64(2):181-232. doi:10.23893/1307-2080.APS64012.

Forman HJ, Maiorino M, Ursini F. Signaling functions of reactive oxygen species. Biochemistry. 2010;49(5):835-842.

Nathan C, Ding A. Nonresolving inflammation. Cell. 2010;140(6):871-882.

Medzhitov R. Origin and physiological roles of inflammation. Nature. 2008;454(7203):428-435.

Hotamisligil GS. Inflammation and metabolic disorders. Nature. 2006;444(7121):860-867.

Donath MY, Shoelson SE. Type 2 diabetes as an inflammatory disease. Nat Rev Immunol. 2011;11(2):98-107.

Srinivasan K, Ramarao P. Animal models in type 2 diabetes research: an overview. Indian J Med Res. 2007;125(3):451-472.

OECD. Test No. 425: Acute Oral Toxicity: Up-and-Down Procedure. OECD Guidelines for the Testing of Chemicals, Section 4. Paris: OECD Publishing; 2008.

Daina A, Michielin O, Zoete V. SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci Rep. 2017;7:42717.

Banerjee P, Kemmler E, Dunkel M, Preissner R. ProTox 3.0: a webserver for the prediction of toxicity of chemicals. Nucleic Acids Res. 2024;52(W1):W513-W520.

Hollingsworth SA, Dror RO. Molecular dynamics simulation for all. Neuron. 2018;99(6):1129-1143.

Genheden S, Ryde U. The MM/PBSA and MM/GBSA methods to estimate ligand-binding affinities. Expert Opin Drug Discov. 2015;10(5):449-461.

Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem. 2010;31(2):455-461.

Hopkins AL. Network pharmacology: the next paradigm in drug discovery. Nat Chem Biol. 2008;4(11):682-690.

Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498-2504.

Szklarczyk D, Gable AL, Lyon D, Junge A, Wyder S, Huerta-Cepas J, et al. STRING v11: protein-protein association networks with increased coverage. Nucleic Acids Res. 2019;47(D1):D607-D613.

Srivastava R, Ahmed H, Dixit RK, Dharamveer, Saraf SA. Crocus sativus L.: a comprehensive review. Pharmacogn Rev. 2010;4(8):200-208.

Melnyk JP, Wang S, Marcone MF. Chemical and biological properties of the world's most expensive spice: saffron. Food Res Int. 2010;43(8):1981-1989.

Hosseinzadeh H, Nassiri-Asl M. Avicenna's (Ibn Sina) the Canon of Medicine and saffron (Crocus sativus): a review. Phytother Res. 2013;27(4):475-483.

Abdullaev FI. Cancer chemopreventive and tumoricidal properties of saffron (Crocus sativus L.). Exp Biol Med. 2002;227(1):20-25.

Bukhari SI, Manzoor M, Dhar MK. A comprehensive review of the pharmacological potential of Crocus sativus and its bioactive apocarotenoids. Biomed Pharmacother. 2018;98:733-745.

Pop-Busui R, Boulton AJM, Feldman EL, Bril V, Freeman R, Malik RA, et al. Diabetic neuropathy: a position statement by the American Diabetes Association. Diabetes Care. 2017;40(1):136-154.

Feldman EL, Callaghan BC, Pop-Busui R, Zochodne DW, Wright DE, Bennett DL, et al. Diabetic neuropathy. Nat Rev Dis Primers. 2019;5(1):41.

Callaghan BC, Cheng HT, Stables CL, Smith AL, Feldman EL. Diabetic neuropathy: clinical manifestations and current treatments. Lancet Neurol. 2012;11(6):521-534.

Colloca L, Ludman T, Bouhassira D, Baron R, Dickenson AH, Yarnitsky D, et al. Neuropathic pain. Nat Rev Dis Primers. 2017;3:17002.

Finnerup NB, Haroutounian S, Kamerman P, Baron R, Bennett DLH, Bouhassira D, et al. Neuropathic pain: an updated grading system for research and clinical practice. Pain. 2016;157(8):1599-1606.

Treede RD, Jensen TS, Campbell JN, Cruccu G, Dostrovsky JO, Griffin JW, et al. Neuropathic pain: redefinition and a grading system for clinical and research purposes. Neurology. 2008;70(18):1630-1635.

Tesfaye S, Boulton AJM, Dyck PJ, Freeman R, Horowitz M, Kempler P, et al. Diabetic neuropathies: update on definitions, diagnostic criteria, estimation of severity, and treatments. Diabetes Care. 2010;33(10):2285-2293.

Chaplan SR, Bach FW, Pogrel JW, Chung JM, Yaksh TL. Quantitative assessment of tactile allodynia in the rat paw. J Neurosci Methods. 1994;53(1):55-63.

Hargreaves K, Dubner R, Brown F, Flores C, Joris J. A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia. Pain. 1988;32(1):77-88.

Published

2026-10-01

Issue

Vol. 1 No. 1 (2026): October 2026 — Inaugural Issue

Section

Research Papers

License

Copyright (c) 2026 Nur Aisyah Rahman, Nattaporn Srisuk, Ama Mensah

Creative Commons License

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

Authors retain copyright. This work is licensed under a Creative Commons Attribution 4.0 International Licence (CC BY 4.0). Sharing and adaptation are permitted with appropriate attribution, a link to the licence and an indication of changes. Third-party material may be subject to separate terms stated in its credit line.