Bowser M T, Kennedy R T
University of Florida, Department of Chemistry, Gainesville, USA.
Electrophoresis. 2001 Oct;22(17):3668-76. doi: 10.1002/1522-2683(200109)22:17<3668::AID-ELPS3668>3.0.CO;2-M.
Microdialysis sampling was coupled via a flow-gated interface on-line to capillary electrophoresis with laser-induced fluorescence (LIF) detection for in vivo monitoring of neuroactive amino acids and amines. In the instrument, analytes are derivatized precolumn with o-phthaldehyde and beta-mercaptoethanol to form fluorescent isoindole products. The instrument was improved over previous designs by incorporating a sheath-flow cuvette for reduced background in LIF detection which improved sensitivity by 15-fold. The methodology was improved by utilizing a voltage ramped injection which allowed generation of 500000 theoretical plates with 20 s separations. Resolution of the isoindole derivatives was further improved by addition of hydroxypropyl-modified beta-cyclodextrin to the electrophoresis buffer. The new instrumentation and methods allow resolution and detection of glutamate, gamma-aminobutyric acid, glycine, aspartate, serine, taurine, glutamine and dopamine (if levels are elevated) collected from in vivo sampling probes every 20 s. The technique is suited to continuous monitoring for dynamic measurements of these compounds in vivo.
微透析采样通过流动门控接口与带有激光诱导荧光(LIF)检测的毛细管电泳在线联用,用于体内神经活性氨基酸和胺类的监测。在该仪器中,分析物在柱前用邻苯二甲醛和β-巯基乙醇衍生化,形成荧光异吲哚产物。该仪器在先前设计的基础上进行了改进,采用了鞘流比色皿以降低LIF检测中的背景,灵敏度提高了15倍。通过采用电压梯度进样改进了方法,在20秒的分离时间内可产生500000理论塔板数。通过向电泳缓冲液中添加羟丙基修饰的β-环糊精,进一步提高了异吲哚衍生物的分离度。新的仪器和方法能够每20秒从体内采样探针收集的样品中分离并检测谷氨酸、γ-氨基丁酸、甘氨酸、天冬氨酸、丝氨酸、牛磺酸、谷氨酰胺和多巴胺(如果含量升高)。该技术适用于对这些化合物在体内的动态测量进行连续监测。