High-performance liquid chromatography and sensitive detection of amino acids derivatized with 7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole

✍ Scribed by Yoshihiko Watanabe; Kazuhiro Imai

• Publisher: Elsevier Science
• Year: 1981
• Tongue: English
• Weight: 139 KB
• Volume: 116
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(81)90390-0

✦ Synopsis

7-Fluoro+nitrobenzo-2-oxa-I .3-diazole is used as a precolumn fluorescent labeling reagent for high-performance liquid chromatography of amino acids. including proline and hydroxyproline.

The reaction is run at pH 8.0 at 60°C for 5 min. The fluorophors (Asp, GIL Hyp. Ser, Gly. Thr, Ala, Pro) are separated on a reversed-phase column (FBondapak C,,) with 0.1 M phosphate buffer (pH 6.0) containing 6.75% methanol and 1.8% tetrahydrofuran. and are detected at the level of IO fmol with excitation at 470 nm and emission at 530 nm.