{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T12:44:48Z","timestamp":1792586688247,"version":"4.5.0"},"reference-count":0,"publisher":"The Journal of Food and Drug Analysis (JFDA), Food and Drug Administration, Taiwan (TFDA)","issue":"2","license":[{"start":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T00:00:00Z","timestamp":1783296000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Food and Drug Analysis"],"abstract":"<jats:p>Trifolium repens (white clover) is rich in bioactive volatiles with therapeutic potential. This study developed an advanced hybrid ultrasound\u2013infrared\u2013microwave 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\u2013mass spectrometry (GC\u2013MS) 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-\u03b1, 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-\u03b1 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-\u03b1\/IL-6\/IL-10 modulation, oxidative-stress restoration, and strychnine-sensitive GlyR potentiation.<\/jats:p>","DOI":"10.38212\/2224-6614.3595","type":"journal-article","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T08:16:11Z","timestamp":1783584971000},"source":"Crossref","is-referenced-by-count":0,"title":["Advanced Hybrid-Green Ultrasound\u2013Infrared\u2013Microwave Trifolium repens Essential oil Isolation with Multi-Target Ethnomedicine Bioactivity against Neuropathy, Inflammation and Multidrug-Resistant Infection"],"prefix":"10.38212","volume":"34","author":[{"given":"Sara Al","family":"Ayoubi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, Faculty of Pharmacy, Beirut Arab University, Beirut, Lebanon."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ibrahim","family":"Abdelwahab","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Faculty of Pharmacy, Pharos University in Alexandria, Alexandria, Egypt."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maha","family":"Aboul-Ela","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Sciences, Faculty of Pharmacy, Beirut Arab University, Beirut, Lebanon"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdalla","family":"El- Lakany","sequence":"additional","affiliation":[{"name":"Department of Pharmacognosy, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karim","family":"Raafat","sequence":"additional","affiliation":[{"name":"Department of Pharmacognosy and Natural Products, Faculty of Pharmacy, Pharos University in Alexandria, Canal El Mahmoudia Street, Beside Green Plaza Complex 21648 Alexandria, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"25385","published-online":{"date-parts":[[2026,7,6]]},"container-title":["Journal of Food and Drug Analysis"],"original-title":[],"language":"en","deposited":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T12:14:26Z","timestamp":1792584866000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.jfda-online.com\/journal\/vol34\/iss2\/7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,6]]},"references-count":0,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,7,6]]}},"URL":"https:\/\/doi.org\/10.38212\/2224-6614.3595","relation":{},"ISSN":["2224-6614"],"issn-type":[{"value":"2224-6614","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,6]]}}}