Highly sensitive simultaneous detection of cultured cellular thiols by laser induced fluorescence-capillary electrophoresis

2005 ◽  
Vol 26 (6) ◽  
pp. 1063-1070 ◽  
Author(s):  
Angelo Zinellu ◽  
Salvatore Sotgia ◽  
Anna M. Posadino ◽  
Valeria Pasciu ◽  
Maria G. Perino ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Simultaneous Detection ◽  
Laser Induced Fluorescence ◽  
Highly Sensitive

scholarly journals Amine Analysis Using AlexaFluor 488 Succinimidyl Ester and Capillary Electrophoresis with Laser-Induced Fluorescence

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Christian G. Kendall ◽  
Amanda M. Stockton ◽  
Stephen Leicht ◽  
Heather McCaig ◽  
Shirley Chung ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Fluorescent Probes ◽  
Limit Of Detection ◽  
Sodium Dodecylsulfate ◽  
Laser Induced Fluorescence ◽  
Primary Amines ◽  
Highly Sensitive ◽  
High Concentrations ◽  
Organic Amines ◽  
Long Chain

Fluorescent probes enable detection of otherwise nonfluorescent species via highly sensitive laser-induced fluorescence. Organic amines are predominantly nonfluorescent and are of analytical interest in agricultural and food science, biomedical applications, and biowarfare detection. Alexa Fluor 488 N-hydroxysuccinimidyl ester (AF488 NHS-ester) is an amine-specific fluorescent probe. Here, we demonstrate low limit of detection of long-chain (C9to C18) primary amines and optimize AF488 derivatization of long-chain primary amines. The reaction was found to be equally efficient in all solvents studied (dimethylsulfoxide, ethanol, and N,N-dimethylformamide). While an organic base (N,N-diisopropylethylamine) is required to achieve efficient reaction between AF488 NHS-ester and organic amines with longer hydrophobic chains, high concentrations (>5 mM) result in increased levels of ethylamine and propylamine in the blank. Optimal incubation times were found to be >12 hrs at room temperature. We present an initial capillary electrophoresis separation for analysis using a simple micellar electrokinetic chromatography (MEKC) buffer consisting of 12 mM sodium dodecylsulfate (SDS) and 5 mM carbonate, pH 10. Limits of detection using the optimized labeling conditions and these separation conditions were 5–17 nM. The method presented here represents a novel addition to the arsenal of fluorescent probes available for highly sensitive analysis of small organic molecules.

Simultaneous Detection of Yersinia Enterocolitica and Listeria Monocytogenes in Foodstuffs by Capillary Electrophoresis and Microchip Capillary Electrophoresis Laser-Induced Fluorescence Detector

2018 ◽  
Vol 101 (6) ◽  
pp. 1833-1838 ◽  
Author(s):  
Yongru Li ◽  
Hongwei Su ◽  
Yajia Lan
Keyword(s):  
Capillary Electrophoresis ◽  
Listeria Monocytogenes ◽  
Yersinia Enterocolitica ◽  
Simultaneous Detection ◽  
High Sensitivity ◽  
Laser Induced Fluorescence ◽  
Microchip Capillary Electrophoresis ◽  
Fluorescence Detector ◽  
Sybr Gold ◽  
Pcr Products

Abstract Background: Food safety is one of the most important public health problems in the world, and pathogenic bacterium is a major factor causing serious foodborne diseases. Objective: Two methods of duplex PCR combined with capillary electrophoresis laser-induced fluorescence detector (CE-LIF) and microchip capillary electrophoresis laser-induced fluorescence detector (MCE-LIF) have been developed for the simultaneous detection of Yersinia Enterocolitica and Listeria Monocytogenes in various foods. The specific conservative sequences of these two bacteria were amplified. Methods: After labelled with nucleic acid dye SYBR Gold and SYBR Orange, the PCR products were analyzed by CE-LIF and MCE-LIF, respectively. Under the optimal conditions, the detection of PCR products of the target bacteria was achieved in less than 15 min by CE-LIF and within 6 min by MCE-LIF. Results: The alignment analysis demonstrated that the PCR products had good agreement with the sequences published in GenBank. The CE-LIF method could detect 10 CFU/mL Y. enterocolitica and L. monocytogenes, and the MCE-LIF method could detect 100 CFU/mL Y. enterocolitica and L. monocytogenes. The intraday precisions of migration time and peak area of DNA markers and PCR products were in the range of 1.13 to 1.18% and 1.60 to 6.29%, respectively, for CE-LIF and 1.18 to 1.48% and 2.85 to 4.06%, respectively, for MCE-LIF. Conclusions: The proposed methods could be applied to target bacterial detection infood samples rapidly, sensitively, and specifically. Highlights: Two new methods based on CE and MCE have been developed for the simultaneous detection of Y. enterocolitica and L. monocytogenes in foodstuffs, and they can detect the bacteria directly without any enrichment because of their high sensitivity.

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