title: A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols.
abstract: A new long-wavelength fluorescent probe, 1,7-dimethyl-3,5-distyryl-8-phenyl-(4'-iodoacetamido)difluoroboradiaza-s-indacene (DMDSPAB-I), was designed and synthesized for thiol labeling in high-performance liquid chromatography (HPLC). The excitation and emission wavelengths of DMDSPAB-I are 620 and 630 nm, respectively, with a high fluorescence quantum yield of 0.557, which is advantageous in preventing interference of intrinsic fluorescence from complex biological matrices and enabling high sensitivity HPLC. Based on DMDSPAB-I, a reversed-phase HPLC method was developed for measuring low-molecular-weight thiols including glutathione, cysteine, homocysteine, N-acetylcysteine, cysteinylglycine, and penicillamine. After the specific reaction of DMDSPAB-I with thiols, baseline separation of all six stable derivatives was achieved through isocratic elution on a C18 column within 25 min, with the limits of detection (signal-to-noise ratio = 3) from 0.24 nmol L(-1) for glutathione to 0.72 nmol L(-1) for penicillamine. The proposed method was validated in part by measuring thiols in blood samples from mice, with recoveries of 95.3-104.3%.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China.
authors.forename: Li-Yun
authors.initials: LY
authors.lastname: Zhang
authors.forename: Feng-Qin
authors.initials: FQ
authors.lastname: Tu
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Guo
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Hua-Shan
authors.initials: HS
authors.lastname: Zhang
mesh_terms: D000111:Acetylcysteine / Q000097:blood / Q000302:isolation & purification; D000818:Animals; D001896:Boron Compounds / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D003545:Cysteine / Q000097:blood / Q000302:isolation & purification; D004151:Dipeptides / Q000097:blood / Q000302:isolation & purification; D005456:Fluorescent Dyes / Q000737:chemistry*; D005978:Glutathione / Q000097:blood / Q000302:isolation & purification; D006710:Homocysteine / Q000097:blood / Q000302:isolation & purification; D007460:Iodoacetamide / Q000031:analogs & derivatives* / Q000737:chemistry; D057230:Limit of Detection; D008297:Male; D051379:Mice; D010396:Penicillamine / Q000097:blood / Q000302:isolation & purification; D013438:Sulfhydryl Compounds / Q000097:blood* / Q000302:isolation & purification
chemical_list: C000598431:1,7-dimethyl-3,5-distyryl-8-phenyl-(4'-iodoacetamido)difluoroboradiaza-s-indacene; C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D004151:Dipeptides; D005456:Fluorescent Dyes; D013438:Sulfhydryl Compounds; D006710:Homocysteine; C028505:cysteinylglycine; D005978:Glutathione; D010396:Penicillamine; D003545:Cysteine; D000111:Acetylcysteine; D007460:Iodoacetamide