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Study of metabolic activity of human embryos focused on amino acids by capillary electrophoresis with light‐emitting diode‐induced fluorescence detection
Author(s) -
Celá Andrea,
Mádr Aleš,
Ješeta Michal,
Žáková Jana,
Crha Igor,
Glatz Zdeněk
Publication year - 2018
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201800265
Subject(s) - chromatography , capillary electrophoresis , taurine , amino acid , fluorescence , repeatability , detection limit , chemistry , sodium , fluorescence spectroscopy , histidine , biochemistry , physics , organic chemistry , quantum mechanics
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.