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首页> 外文期刊>Analytical chemistry >Ultrasensitive detection of closely related angiotensin I peptides doing capillary electrophoresis with near-infrared laser-induced fluorescence detection
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Ultrasensitive detection of closely related angiotensin I peptides doing capillary electrophoresis with near-infrared laser-induced fluorescence detection

机译：用近红外激光诱导的荧光检测对紧密相关的血管紧张素I肽进行毛细管电泳的超灵敏检测

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摘要

A novel near-infrared (NIR) fluorescent dye (NN382, LICOR, Inc.) was evaluated as an ultrasensitive peptide-labeling reagent for use with capillary electrophoresis (CE). Six angiotensin I (Ang-I) variants were selected as model peptides for the derivatization and separation studies. The closely related decapeptides were labeled with the NIR dye, separated using CE, and detected by NIR laser-induced fluorescence, Derivatization of the peptides was achieved under aqueous conditions using 2.5-500 pmol of Ang-I in a 50-mu L sample (5 x 10(-8)-1 x 10(-5) M), and between 1.3 and 254 amol of the labeled peptides were injected on column. The fluorescence response was linear over a 200-fold range (correlation r greater than or equal to 0.9986). The limit of detection (SNR = 3, signal/RMS noise) ranged from 100 to 300 zmol,for the six Ang-I variants. Four of six peptides were resolved from each other and excess dye using capillary zone electrophoresis with a simple 50 mM phosphate run buffer, pH 7.2. Two pairs of coeluting peptides were successfully resolved using micellar electrokinetic chromatography with a nonionic surfactant, Triton X-100. The NIR amine-labeling reagent NN382 is a viable alternative to using visible fluorophores for CE methods requiring high sensitivity. [References: 17]
机译：评价一种新型的近红外（NIR）荧光染料（NN382，LICOR，Inc.）作为毛细管电泳（CE）的超灵敏肽标记试剂。选择了六个血管紧张素I（Ang-1）变异体作为模型肽，用于衍生化和分离研究。密切相关的十肽用NIR染料标记，使用CE分离，并通过NIR激光诱导的荧光进行检测。肽的衍生化是在水性条件下使用2.5-500 pmol Ang-I在50μL样品中完成的（ 5 x 10（-8）-1 x 10（-5）M），并在柱上注入1.3至254 amol的标记肽。荧光响应在200倍范围内呈线性关系（相关性r大于或等于0.9986）。对于六个Ang-I变体，检测极限（SNR = 3，信号/ RMS噪声）范围为100至300 zmol。使用毛细管区带电泳和简单的50 mM磷酸盐电泳缓冲液（pH 7.2），将六个肽中的四个彼此分离，并分离出过量的染料。使用胶束电动色谱和非离子表面活性剂Triton X-100，成功分离了两对共洗脱肽。对于要求高灵敏度的CE方法，NIR胺标记试剂NN382是使用可见荧光团的可行替代方法。 [参考：17]

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