High temporal resolution coupling of low-flow push-pull perfusion to capillary electrophoresis for ascorbate analysis at the rat vitreoretinal interface
High temporal resolution coupling of low-flow push-pull perfusion to capillary electrophoresis for ascorbate analysis at the rat vitreoretinal interface
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DOI：
10.1039/b813887g
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发表时间：
2009-01-01
期刊：
影响因子：
4.2
通讯作者：
Shippy, Scott A.
中科院分区：
文献类型：
--
作者：
Patterson, Eric E., II;Pritchett, Jeanita S.;Shippy, Scott A.
一个系统，展示了高时间分辨率耦合低流量推挽式灌注采样（LFPS）毛细管电泳抗坏血酸在大鼠玻璃体视网膜界面的吸光度测量。该系统将所有分离组件保持在低压下，作为LFPS期间抽取样品的手段。该系统使用流量门控接口将LFPS探针的提取毛细管直接连接到分离毛细管，无需任何离线样品处理。该系统的时间分辨率受到注射时间的限制，小于16 s。这种高的时间分辨率被应用于在大鼠玻璃体视网膜界面的体内抗坏血酸水平的监测。在玻璃体视网膜界面处，抗坏血酸盐的基线浓度为86 μ M +/- 18 μ M。基线浓度与尸检大块玻璃体分析获得的浓度匹配良好。在用145 mM K+刺激后，观察到基线值的最大增加在32-107%之间（n = 3）。该系统展示了抗坏血酸在大鼠玻璃体视网膜界面的第一个体内时间研究。
A system is presented demonstrating the high-temporal resolution coupling of low-flow push-pull perfusion sampling (LFPS) to capillary electrophoresis for the absorbance measurement of ascorbate at the rat vitreoretinal interface. This system holds all separation components at a low pressure as the means for withdrawing sample during LFPS. The system uses a flow-gated interface to directly couple the withdrawal capillary from the LFPS probe to a separation capillary and eliminates the need for any offline sample handling. The temporal resolution of the system was limited by injection time and is less than 16 s. This high temporal resolution was applied to the monitoring of in vivo ascorbate levels at the rat vitreoretinal interface. Baseline concentrations of ascorbate were found to be 86 mu M +/- 18 mu M at the vitreoretinal interface. Baseline concentrations matched well with those obtained for the postmortem bulk vitreous analysis. Upon stimulation with 145 mM K+, a maximum increase in baseline values between 32-107% for n = 3 was observed. This system demonstrates the first in vivo temporal study of ascorbate at the rat vitreoretinal interface.