Haj-Yehia A I, Benet L Z
Department of Pharmacy, University of California, San Francisco 94143-0446, USA.
J Chromatogr B Biomed Appl. 1995 Apr 7;666(1):45-53. doi: 10.1016/0378-4347(94)00548-j.
The use of 2-(4-N-maleimidophenyl)-6-methoxybenzofuran as a fluorogenic pre-column derivatizing agent for sensitive high-performance liquid chromatographic (HPLC) determination of aliphatic thiols is described. The maleamic acid, its methyl ester and the N-maleimide derivatives of the common amine precursor were easily synthesized and characterized according to conventional methods. These derivatives lack fluorescent properties in their native form but display strong fluorescence upon reaction with aliphatic thiols. The reaction is rapid and highly selective for thiols over a wide pH range (7.1-8.8). Following derivatization, the thiol adducts were separated on a reversed-phase column (Ultrasphere-ODS) using 0.1% hexanesulfonic acid in 10 mM potassium hydrogen phosphate-acetonitrile (65:35, pH 4.5) and were detected fluorimetrically (excitation at 310 nm; emission at 390 nm). The method is highly sensitive (femtomole range) and is easily applied to the determination of SH-containing drugs and endogenous thiols in biological samples.
描述了使用2-(4-N-马来酰亚胺基苯基)-6-甲氧基苯并呋喃作为荧光柱前衍生剂，用于高效液相色谱(HPLC)灵敏测定脂肪族硫醇。根据常规方法，可轻松合成并表征马来酰胺酸、其甲酯以及常见胺前体的N-马来酰亚胺衍生物。这些衍生物本身不具有荧光特性，但与脂肪族硫醇反应后会显示出强烈的荧光。该反应快速，在较宽的pH范围(7.1 - 8.8)内对硫醇具有高度选择性。衍生化后，硫醇加合物在反相柱(Ultrasphere-ODS)上进行分离，流动相为含0.1%己烷磺酸的10 mM磷酸二氢钾-乙腈(65:35, pH 4.5)，并采用荧光检测(激发波长310 nm；发射波长390 nm)。该方法具有高灵敏度(飞摩尔范围)，且易于应用于生物样品中含硫药物和内源性硫醇的测定。
