Gold-Nanoparticle Based Turn-on Fluorometric Sensor for Quantification of Sulfhydryl and Disulfide Forms of Biothiols: Measurement of Thiol/Disulfide Homeostasis
ANALYTICAL LETTERS, cilt.55, sa.4, ss.648-664, 2022 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 55 Sayı: 4
- Basım Tarihi: 2022
- Doi Numarası: 10.1080/00032719.2021.1958830
- Dergi Adı: ANALYTICAL LETTERS
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aerospace Database, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chimica, Communication Abstracts, Food Science & Technology Abstracts, Metadex, Veterinary Science Database, Civil Engineering Abstracts
- Sayfa Sayıları: ss.648-664
- Anahtar Kelimeler: Fluorometric sensor, gold nanoparticles, Rhodamine-6G, thiol, disulfide homeostasis, HIGHLY SELECTIVE DETECTION, FLUORESCENT-PROBE, RHODAMINE 6G, THIOL COMPOUNDS, ENERGY-TRANSFER, GLUTATHIONE, CYSTEINE, ASSAY, HPLC, HOMOCYSTEINE
- İstanbul Üniversitesi-Cerrahpaşa Adresli: Evet
Özet
A simple and sensitive method was developed to determine biothiols in both sulfhydryl (-SH) and disulfide (S-S) forms using a fluorometric sensor composed of citrate-stabilized gold nanoparticles (AuNPs) prepared by electrostatic adsorption of water-soluble fluorophore Rhodamine-6G (Rh-6G) dye onto citrate-stabilized AuNPs resulting in the fluorescence resonance energy transfer in the turn-off mode. Due to the high surface affinities of biothiols to AuNPs, the fluorescence increment at 552 nm (turn-on mode) accompanying the release of Rh-6G indicated the thiol content. This assay also enables the differentiation of oxidized and reduced thiols and the measurement of thiol/disulfide homeostasis as a vital oxidative stress indicator using thiol-alkylating agent N-ethylmaleimide (NEM). After pretreatment with NEM, disulfides were measured in the presence of reduced thiols. The overall biothiol levels and thiol/disulfide ratios were measured using the proposed assay. This method was shown to be selective over various potential interfering compounds (e.g., polyphenols and plasma antioxidants) known to adversely affect redox-based assays. The proposed assay also enabled the naked-eye detection of biothiols by NP agglomeration and may be successfully applied to pharmaceutical samples containing thiols.