ArticleJournal of food and drug analysis2026
Advanced hybrid-green ultrasound-infrared-microwave Trifolium repens essential oil isolation with multi-target ethnomedicine bioactivity against neuropathy, inflammation and multidrug-resistant infection.
Article in Journal of food and drug analysis, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Trifolium repens (white clover) is rich in bioactive volatiles with therapeutic potential. This study developed an advanced hybrid ultrasound-infrared-microwave hydro-distillation (UIMHD) method for efficient essential oil (EO) isolation and investigated its active sesquiterpene, pseudoionone, for multi-mechanistic neuroprotection in diabetic neuropathy (DN). The optimized UIMHD system synergistically combined ultrasound cavitation, infrared heating, and microwave hydro-distillation, enhancing yield and reducing time versus conventional extraction. Gas chromatography-mass spectrometry (GC-MS) profiling identified pseudoionone as the dominant constituent. Pharmacological evaluations were performed in alloxan-induced diabetic mice to assess antidiabetic, anti-inflammatory, neuroprotective, and anticonvulsant activities. Cytokine modulation (TNF-α, IL-6, IL-10), oxidative stress markers (CAT, GSH, TBARS), and strychnine-induced seizure assays were conducted to elucidate mechanisms. The UIMHD technique increased T. repens essential oil (TR-EO) yield by 62.5% and shortened isolation time by 50%. Pseudoionone (49.16%) exhibited potent hypoglycemic and insulin-restorative effects, alleviated thermal hyperalgesia and tactile allodynia, and improved antioxidant defenses. Both the TR-EO and pseudoionone significantly down-regulated TNF-α and IL-6 while up-regulating IL-10, indicating anti-inflammatory cytokine balance restoration. Additionally, pseudoionone delayed strychnine-induced tonic seizures, suggesting functional involvement of glycinergic pathways. Strong antibacterial and antibiofilm activities were also observed against multidrug-resistant Shigella dysenteriae. The advanced UIMHD method provides a sustainable platform for isolating bioactive volatiles from T. repens. Pseudoionone emerges as a multi-target natural agent mitigating diabetic neuropathy via TNF-α/IL-6/IL-10 modulation, oxidative- stress restoration, and strychnine-sensitive GlyR potentiation.
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