scholarly journals Highly Sensitive High Performance Liquid Chromatography-Laser Induced Fluorescence for Proteomics Applications

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Ajeetkumar Patil ◽  
K. S. Choudhari ◽  
Vijendra Prabhu ◽  
V. K. Unnikrishnan ◽  
Sujatha Bhat ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Serum Proteins ◽  
Sensitivity Study ◽  
Laser Induced Fluorescence ◽  
Laser Induced Fluorescence Detection ◽  
Highly Sensitive ◽  
Sds Page ◽  
And Performance

This paper describes the sensitivity study and performance evaluation of high-performance liquid chromatography-laser-induced fluorescence detection (HPLC-LIF) system assembled in our laboratory for proteomics applications. The limits of Detection (LOD) of several serum proteins have been estimated with this instrument and are found to be much lower compared to other commonly used proteomics techniques like SELDI, MALDI, 2-D-SDS-PAGE, and so forth. Techniques for improving the LOD still further with similar setup are briefly discussed. Using the system, protein profiles of serum in normal, malignant, and premalignant conditions were recorded for different malignancy situations.

scholarly journals Analysis of neurotransmitter catecholamines and related amines in human urine and serum by chromatography and capillary electrophoresis with 1, 3, 5, 7-tetramethyl-8-(N-hydroxysuccinimidyl propionic ester)-difluoro-boradiaza-s-indacene

2021 ◽  
Author(s):  
Liwei Cao ◽  
Lizhen Wu ◽  
Hailan Zhong ◽  
Hao Wu ◽  
Siyun Zhang ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Capillary Electrophoresis ◽  
Liquid Chromatography ◽  
Fluorescence Detection ◽  
High Performance ◽  
Detection Method ◽  
Laser Induced Fluorescence ◽  
Limits Of Detection ◽  
Laser Induced Fluorescence Detection ◽  
Derivatization Reaction

AbstractTwo sensitive and effective methods were developed for the detection of catecholamines and related biogenic amines (dopamine, epinephrine, norepinephrine, serotonin, levodopa and tyramine) using high performance liquid chromatography with fluorescence detection and capillary electrophoresis with laser-induced fluorescence detection. A BODIPY fluorescent dye, 1, 3, 5, 7-tetramethyl-8-(N-hydroxysuccinimidyl propionic ester)-difluoroboradiaza- s-indacene was used as pre-column derivatization reagent. The separation and derivatization conditions were optimized in detail. In high performance liquid chromatography with fluorescence detection method, the derivatization reaction was completed at 35 °C for 20 min. At the wavelength of λex/λem = 493 nm/513 nm, dopamine, epinephrine, norepinephrine, and levodopa derivatives achieved baseline separation within 15 min. The limits of detection (S/N = 3) were 1.0, 2.0, 5.0, and 0.5 nmol/L, respectively. In capillary electrophoresis with laser-induced fluorescence detection method, the derivatization reaction was completed at 25 °C for 20 min. Serotonin, tyramine and dopamine derivatives reached baseline separation within 10 min at the wavelength of λex = 473 nm. The limits of detection (S/N = 3) for serotonin, tyramine, and dopamine were 0.3, 0.02, and 0.2 nmol/L, respectively. The amino compounds in human serum and urine samples were detected successfully, and the recoveries were 93.3%–106.7% and 91.0%–103.1%, respectively.

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