Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602, USA.
Appl Spectrosc. 2010 Nov;64(11):1283-8. doi: 10.1366/000370210793335016.
We present a mathematical description of the signal-to-noise ratio (S/N) in a fluorescence-based protein detector for capillary electrophoresis that uses a pulsed ultraviolet (UV) laser at 266 nm as an excitation source. The model accounts for photobleaching, detector volume, laser repetition rate, and analyte flow rate. We have experimentally characterized such a system, and we present a comparison of the experimental data with the predictions of the model. Using the model, the system was optimized for test analytes tryptophan, tyrosine, bovine serum albumin (BSA), and conalbumin, producing detection limits (3σ) of 0.67 nM, 5.7 nM, 0.9 nM, and 1.5 nM, respectively. Based on the photobleaching data, a photobleaching cross-section of 1.4 × 10(-18)cm(2) at 266 nm was calculated for tryptophan.
我们提出了一种基于毛细管电泳的荧光蛋白检测器的信噪比（S/N）的数学描述，该检测器使用 266nm 的脉冲紫外（UV）激光作为激发源。该模型考虑了光漂白、探测器体积、激光重复率和分析物流速。我们已经对这样的系统进行了实验表征，并将实验数据与模型的预测进行了比较。使用该模型，针对色氨酸、酪氨酸、牛血清白蛋白（BSA）和伴白蛋白这几种测试分析物对系统进行了优化，得到的检测限（3σ）分别为 0.67nM、5.7nM、0.9nM 和 1.5nM。基于光漂白数据，在 266nm 处计算得到色氨酸的光漂白横截面积为 1.4×10(-18)cm(2)。
