Steroid Analysis in Single Cells by Capillary Electrophoresis with Collinear Laser-Induced Fluorescence Detection

Malek, Amir; Khaledi, Morteza G.

doi:10.1006/abio.1999.3096

Cited by 18 publications

(10 citation statements)

References 29 publications

(14 reference statements)

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“…Cellular analysis by direct MALDI MS is an effective method for determining the peptides and studying the cellular processing in single neurons, and recently, considerable research has been performed utilizing CE for single-cell analyses. − However, enhancements are gained through the coupling of a microseparation technique prior to MS analysis. − In addition to creating an information-rich, two-dimensional analytical technique, a more complete mass profile of the sample is gained by reducing analyte suppression. , There are a variety of examples where CE has been combined with MALDI MS. These methods range from continuous deposition of the CE eluent onto a membrane , to the collection of fractions in small vials or directly onto a MS target. ,,, Here, nanoliter-volume fractions are collected into spots (bins) on a target, each representing a variable (e.g., 15 s) elution window, which allows easier interfacing to a variety of detection modes.…”

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confidence: 99%

Single-Neuron Analysis Using CE Combined with MALDI MS and Radionuclide Detection

2002

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Capillary electrophoresis (CE) has been combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and radionuclide detection to assay mass-limited biological samples. Nanovial sampling techniques enable injections into the CE capillary from 50 to 150-nL volume samples; after the separation, nanoliter fraction collection combines the CE effluent with a MALDI matrix and minimizes sample spreading, thus allowing both MALDI MS and radionuclide detection on the CE fractions. MALDI MS complements the elution time information of CE by providing accurate molecular mass data, and radionuclide detection provides zeptomole limits of detection with quantitative information. While MALDI MS detects all fully processed peptides at sufficient concentration, culturing the neuron in media containing 35S-Met provides selective radionuclide detection of newly synthesized methionine-containing peptides. The analysis and detection of the expected neuropeptides and hormones in a single 40-microm bag cell neuron from Aplysia californica with CE/MALDI MS/radionuclide detection demonstrates the ability of this hyphenated approach to work with chemically complex mass-limited samples.

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confidence: 99%

Single-Neuron Analysis Using CE Combined with MALDI MS and Radionuclide Detection

2002

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“…Steroids were found in single R2C cell while progesterone secretion was monitored from culture cells and subsequently in single cells. The mass detection limit was 10 À18 mol of dansylated steroids [154]. In identified neurons of marine slugs, Arg and citrulline, AA related to the synthesis of nitric oxide [155], and insulin in single pancreatic cells [156], a release of insulin from cell line beta TC3 permeabilized by digititonin [157] were all analyzed by CE-LIFD.…”

Section: Ce and Single Cell Analysismentioning

confidence: 99%

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

Páez

Hernández

2001

Biopharm & Drug Disp

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Capillary electrophoresis (CE) is a high-efficiency analytical technique that has had a great impact as a tool in biomedical research, clinical and forensic practice in the last ten years. Only in one of the applications, the DNA analysis, it has had an explosive exponential growth in the last few years. This impact is expressed in an enormous amount of CE articles and many reviews. The CE advantages with respect to other analytical techniques: the required very small sample volume, rapid analysis, great resolution power and low costs, have made this technique ideal for the analysis of a numerous endogenous and exogenous substances present in biological fluids. The different modes of CE have been coupled to different detection techniques such as UV-absorbance, electrochemical, mass spectrometry and laser-induced fluorescence detection (LIFD) to detect different nature and molecular size separated analytes. This review focuses mostly on the applications of CE-LIFD, to measure drugs and endogenous neuroactive substances such as amino acids and monoamines, especially in microdialysis samples from experimental animals and humans. CE-LIFD trends are discussed: automated faster analysis with capillary array systems, resolution power improvement, higher detection sensitivity, and CE systems miniaturization for extremely small sample volume, in order to make CE easier and affordable to the lab bench or the clinical bed.

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“…Among them, dansylhydrazine (5-dimethylaminonaphthalene-1-sulfohydrazide, DNSH) is a promising choice because of its luminescence features. This label has been widely used for the derivatization of steroids (Appelblad et al 1997;Malek and Khaledi 1999;Visser et al 2000) and carbohydrates (Perez and Colon 1996;Du and Eddington 2002) and to a lesser extent for aldehydes and ketones as a way to carry out LC analysis in air samples (Zhang et al 2000;Pereira, Carrilho, and Tavares 2002;Herrington et al 2005;Herrington and Zhang 2008). On the other hand, DNSH has hardly ever been used for the derivatization of these carbonyl compounds in aqueous samples.…”

Section: Introductionmentioning

confidence: 98%

Determination of Low-Molecular-Mass Aldehydes in Water Samples by Liquid Chromatography and Chemiluminescence Detection Using Continuous in situ Derivatization/Preconcentration with Dansylhydrazine

Baños

Silva

2009

Analytical Letters

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A simple, expeditious, and sensitive method has been developed for the determination of low-molecular-mass aldehydes in water samples by liquid chromatography and peroxyoxalate-chemiluminescence detection. The method is based on continuous solid-phase extraction with in situ derivatization= preconcentration of the aldehydes using dansylhydrazine, which was first adsorbed on an RP-C 18 mini-column. For 10 mL of aqueous sample, the limits of detection (LOD) for C 1 to C 4 aldehydes were 20-30 ng L À1 , except for formaldehyde, which had an LOD of 400 ng L À1 . Application was illustrated by the determination of these aldehydes in water samples; the interday precision was always less than ca. 7%, and relative recoveries were more than 96%.

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Steroid Analysis in Single Cells by Capillary Electrophoresis with Collinear Laser-Induced Fluorescence Detection

Cited by 18 publications

References 29 publications

Single-Neuron Analysis Using CE Combined with MALDI MS and Radionuclide Detection

Single-Neuron Analysis Using CE Combined with MALDI MS and Radionuclide Detection

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

Determination of Low-Molecular-Mass Aldehydes in Water Samples by Liquid Chromatography and Chemiluminescence Detection Using Continuous in situ Derivatization/Preconcentration with Dansylhydrazine

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