Study of metabolic activity of human embryos focused on amino acids by capillary electrophoresis with light-emitting diode-induced fluorescence detection.
Andrea CeláAleš MádrMichal JešetaJana ŽákováIgor CrhaZdenĕk GlatzPublished in: Electrophoresis (2018)
Assisted reproduction is a quickly developing field of reproductive medicine whose importance is growing every year due to the increasing number of patients suffering from infertility. As a result, there is a need for the continuous development and/or improvement of assisted reproductive technologies. This paper presents a new method for the in vitro measurement of the amino acid turnover of developing embryos based on capillary electrophoresis with light-emitting diode-induced fluorescence detection. Amino acids were derivatized with naphthalene-2,3-dicarboxaldehyde/NaCN, and the resulting fluorescent derivatives were baseline resolved within 25 min in a background electrolyte comprised of 50 mM sodium tetraborate, 73 mM sodium dodecyl sulphate, 5 mM sodium deoxycholate and 2.5 mM (2-hydroxypropyl)-β-cyclodextrin (pH ≈ 9.3). The migration time and the peak area repeatability (n = 10) were below 0.5 and 4.3%, respectively. The limits of detection ranged from 12.6 nM (histidine) to 39.3 nM (taurine). The developed method, which only requires 2 μL of raw sample, was successfully applied for determining the metabolic activity of human embryos exposed to different environmental stress conditions.
Keyphrases
- capillary electrophoresis
- light emitting
- amino acid
- mass spectrometry
- endothelial cells
- high glucose
- label free
- loop mediated isothermal amplification
- real time pcr
- end stage renal disease
- diabetic rats
- photodynamic therapy
- single molecule
- ejection fraction
- induced pluripotent stem cells
- newly diagnosed
- ionic liquid
- drug induced
- chronic kidney disease
- living cells
- pluripotent stem cells
- energy transfer
- type diabetes
- quantum dots
- prognostic factors
- climate change
- sensitive detection
- life cycle