Determination of formaldehyde in single cell by capillary electrophoresis with LIF detection

Abstract

Determination of formaldehyde in single cell by capillary electrophoresis with LIF detectionAs a small molecule gas, formaldehyde (FA) is the simplest carbonyl active material and plays an important role in the carbon cycle of metabolism. However, due to the volatile nature of the gas, it is difficult to accurately quantify its content, which limits the study of the mechanism of action in life activities. Thus, an efficient approach to quantitative detection of FA in cells especially in single cell is urgent needed. Nevertheless, no method for quantifying FA in single cell has been reported to date. In this work, a fluorescent probe N-propyl-4-hydrazino-naphthalimide (NPHNA), which has highly desirable attributes and has been applied to living biological samples, was chosen as labeling reagent to detect endogenous FA at single cell level. After optimization of separation conditions, fast baseline separation of the FA derivative N-propyl-4-hydrazone-naphthalimide product (NPHNA-FA) and NPHNA was achieved in about 5 min by CE with LIF detection. The detection limit for FA was 5 amol (S/N ratio of 3). The developed method was validated by the measurements of intracellular levels of FA in single cell.

Simultaneous Quantitation of Intra- and Extracellular Nitric Oxide in Single Macrophage RAW 264.7 Cells by Capillary Electrophoresis with Laser-Induced Fluorescence Detection

“…The CE method coupled with LIF provides a powerful and convenient tool for single-cell analysis. ,,− To quantify intra- and extracellular NO in single cells simultaneously, it is of crucial importance to select appropriate fluorescent probes. First, the permeability of the selected two probes to the cell membrane should be disparate in order to ensure that one is retained within the cell and the other is kept outside the cell.…”

“…Among various analytical techniques, capillary electrophoresis (CE) and microfluidics are superior in single-cell analysis because they fit approximately to the size of an individual cell. − Coupled with laser-induced fluorescence (LIF) detection, an ultrasensitive detection method, CE and microfluidics provide potent tools for the determination of trace components in single cells. − Although microfluidics benefit from portability, automation, and high throughput, CE methods bring the advantages of simplicity, convenience, and universality. For nonfluorescent and transient cellular targets, fluorescent probes are usually used as labeling and trapping reagents. − Moreover, the proposal of dual-labeling strategy facilitates the simultaneous detection of two or more components with disparate properties in single cells, such as reactive oxygen species (ROS) and reduced glutathione (GSH), superoxide anion (O 2 · – ) and NO, and hydrogen peroxide (H 2 O 2 ), GSH, and cysteine (Cys) .…”

Challenging Bioanalyses with Capillary Electrophoresis

“…Separations accomplished in under 2 min were used to analyze 10 individual cells . In another report, formaldehyde was detected in single cells using a commercial capillary electrophoresis instrument equipped with laser-induced fluorescence detection . HeLa cells were incubated with propyl-4-hydrazino-naphthalimide, which readily reacted with formic acid.…”

“…175 In another report, formaldehyde was detected in single cells using a commercial capillary electrophoresis instrument equipped with laserinduced fluorescence detection. 176 HeLa cells were incubated with propyl-4-hydrazino-naphthalimide, which readily reacted with formic acid. An individual cell was then manually aspirated into a capillary housed in a commercial cartridge with the aid of an optical microscope.…”

Novel Method of Analysis for the Determination of Residual Formaldehyde by High-Performance Liquid Chromatography

“…Analytical methods have been developed to determine formaldehyde in numerous matrices, such as cosmetics, food, drink, environmental samples, and pharmaceuticals. ese methods are based on various analytical techniques, such as high-performance liquid chromatography ultraviolet detection (HPLC-UV) [8][9][10][11][12][13][14], gas chromatography flame ionisation detection (GC-FID) [15,16], mass spectrometry [17][18][19], fluorescence [20][21][22][23], spectrophotometry [24,25], chromotropic acid spectrophotometry [26], photoluminescence [27], and capillary electrophoresis [28]. Sensors and probes for formaldehyde have also been explored [29][30][31][32][33].…”