Suzuki S, Shimotsu N, Honda S, Arai A, Nakanishi H
Faculty of Pharmaceutical Sciences, Kini, University, Higashi-osaka, Japan
Electrophoresis. 2001 Oct;22(18):4023-31. doi: 10.1002/1522-2683(200110)22:18<4023::AID-ELPS4023>3.0.CO;2-P.
Microchip electrophoresis for the short-time analysis of amino sugars is described. D-Glucosamine, D-galactosamine and their reduced forms were labeled with 4-nitro-2,1,3-benzoxadiazole 7-fluoride (NBD-F) at pH 6.0 and the fluorescent derivatives were purified on an octadecyl silica (ODS) gel plate. The derivatives were analyzed by electrophoresis on a microfabricated chip with a 33 mm long separation channel with argon laser-induced fluorescence detection. Under the established conditions, these amino sugarderivatives were well separated from each other within 60 s. Amino sugars of as small an amount as 0.5 fmol could be detected with a signal-to-noise (S/N) ratio of 3, and peak response showed good linearity between at least 0.8 and 8 fmol of samples with a relative standard deviation (RSD) of ca. 4%. This method was also applied to the analysis of amino sugar composition of O-linked glycans released from bovine submaxillary mucin with alkali in the presence of borohydride. The result of amino sugar composition analysis for individual O-glycans fractionated by high-performance liquid chromatography was quite useful for their identification.
本文描述了用于氨基糖短时间分析的微芯片电泳法。D-葡萄糖胺、D-半乳糖胺及其还原形式在pH 6.0条件下用4-硝基-2,1,3-苯并恶二唑-7-氟化物（NBD-F）进行标记，荧光衍生物在十八烷基硅胶（ODS）凝胶板上进行纯化。通过在具有33 mm长分离通道的微制造芯片上进行电泳，并采用氩激光诱导荧光检测对衍生物进行分析。在既定条件下，这些氨基糖衍生物在60 s内能够很好地相互分离。最低可检测到0.5 fmol的氨基糖，信噪比（S/N）为3，在至少0.8至8 fmol的样品之间，峰响应呈现良好的线性关系，相对标准偏差（RSD）约为4%。该方法还应用于在硼氢化钠存在下用碱从牛下颌粘蛋白中释放的O-连接聚糖的氨基糖组成分析。通过高效液相色谱法分离的各个O-聚糖的氨基糖组成分析结果对其鉴定非常有用。
