Hapuarachchi Suminda, Janaway Gordon A, Aspinwall Craig A
Department of Chemistry, University of Arizona, Tucson, AZ 85721, USA.
Electrophoresis. 2006 Oct;27(20):4052-9. doi: 10.1002/elps.200600232.
We report the utilization of a high power UV light-emitting diode for fluorescence detection (UV-LED-IF) in CE separations. CE-UV-LED-IF allows analysis of a range of environmentally and biologically important compounds, including PAHs and biogenic amines, including neurotransmitters, amino acids, proteins, and peptides, that have been derivatized with UV-excited fluorogenic labels, e.g., o-phthalic dicarboxaldehyde/beta-mercaptoethanol (OPA/beta-ME). The 365 nm UV-LED was used as a stable, low cost source for detection of UV-excited fluorescent compounds. UV-LED-IF was used with both zonal CE separations and MEKC. Native fluorescence detection of PAHs was accomplished with detection limits ranging from 10 nM to 1.3 microM. Detection limits for OPA/beta-ME-labeled glutamic acid and aspartic acid were 11 and 10 nM, respectively, for off-line labeling, and 47 and 47 nM, respectively, for on-line labeling, comparable to UV-laser-based systems. Analysis of OPA/beta-ME-labeled proteins and peptides was performed with 28 and 47 nM detection limits for BSA and myoglobin, respectively.
我们报道了在毛细管电泳分离中利用高功率紫外发光二极管进行荧光检测(紫外发光二极管 - 间接荧光检测法)。毛细管电泳 - 紫外发光二极管 - 间接荧光检测法能够分析一系列对环境和生物具有重要意义的化合物,包括多环芳烃和生物胺,其中生物胺包括神经递质、氨基酸、蛋白质和肽,这些化合物已用紫外激发的荧光标记物进行了衍生化处理,例如邻苯二甲酸二醛/β - 巯基乙醇(OPA/β - ME)。365nm的紫外发光二极管被用作检测紫外激发荧光化合物的稳定、低成本光源。紫外发光二极管 - 间接荧光检测法可用于区带毛细管电泳分离和胶束电动毛细管色谱分析。多环芳烃的天然荧光检测限范围为10 nM至1.3 μM。对于OPA/β - ME标记的谷氨酸和天冬氨酸,离线标记的检测限分别为11 nM和10 nM,在线标记的检测限分别为47 nM和47 nM,与基于紫外激光的系统相当。对OPA/β - ME标记的蛋白质和肽进行分析时,牛血清白蛋白和肌红蛋白的检测限分别为28 nM和47 nM。