A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols

作者：Zhang, Li-Yun; Tu, Feng-Qin; Guo, Xiao-Feng; Wang, Hong^*; Wang, Peng; Zhang, Hua-Shan

来源：Analytical and Bioanalytical Chemistry, 2014, 406(26): 6723-6733.

DOI：10.1007/s00216-014-8013-3

• 1,7-Dimethyl-3,5-distyryl-8-phenyl-(4 '-iodoacetamido)difluoroboradiaza-s-indacene
• Thiols
• long-wavelength
• Fluorescence detection
• High-performance liquid chromatography

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 C-18 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 %.

• 出版日期2014-10
• 单位武汉大学; 中华人民共和国湖北出入境检验检疫局