Chan K C, Muschik G M, Issaq H J
Analytical Chemistry Laboratory, SAIC Frederick, NCI-Frederick Cancer Research and Development Center, MD 21702, USA.
Electrophoresis. 2000 Jun;21(10):2062-6. doi: 10.1002/1522-2683(20000601)21:10<2062::AID-ELPS2062>3.0.CO;2-Z.
Two solid-state UV lasers were applied to the laser-induced fluorescence (LIF) detection of various groups of compounds after separation by capillary electrophoresis. These lasers are thermoelectric-cooled, highly compact, and inexpensive. Such lasers provide few mW of quasi-continuous wave (CW) power which are sufficient and stable for LIF detection. Native fluorescence detection of tryptophan-containing proteins and peptides and related indoles was achieved at the nM level with the laser operating at 266 nm. Detection of fluorescamine-labeled amino acids and peptides was also possible at the nM level with the laser operating at 355 nm. Amino acids at a concentration as low as 10 ng/mL could be labeled with fluorescamine. Solid-state UV-LIF detection of the tryptic digest of cytochrome c after fluorescamine derivatization was demonstrated.
两台固态紫外激光器被应用于毛细管电泳分离后各类化合物的激光诱导荧光（LIF）检测。这些激光器采用热电冷却，高度紧凑且价格低廉。此类激光器可提供几毫瓦的准连续波（CW）功率，足以满足LIF检测且功率稳定。在激光器工作波长为266 nm时，实现了对含色氨酸的蛋白质、肽及相关吲哚的天然荧光检测，检测限可达纳摩尔水平。在激光器工作波长为355 nm时，也能够在纳摩尔水平检测荧光胺标记的氨基酸和肽。浓度低至10 ng/mL的氨基酸可用荧光胺进行标记。展示了荧光胺衍生化后细胞色素c胰蛋白酶消化产物的固态紫外-LIF检测。
