Determination of biogenic amines in HeLa cell lysate by 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein and micellar electrokinetic capillary chromatography with laser-induced fluorescence detection.
An MEKC-LIF method using 6-oxy-(N-succinimidyl acetate)-9-(2'-methoxy-carbonyl) fluorescein (SAMF) newly synthesized in our lab as a labeling reagent for the separation and determination of eight typical biogenic amines was proposed. After careful study of the derivatization condition such as pH value, reagent concentration, temperature, and reaction time, derivatization reaction was accomplished as quickly as 10 min with stable yield. Optimal separation of SAMF-labeled amines was achieved with a running buffer (pH 9.3) containing 30 mM boric acid, 25 mM SDS, and 20% v/v ACN. The proposed method allowed biogenic amines to be determined with LODs as low as 0.25-2.5 nmol/L and RSD values from 0.4 to 4.5%. The present method has been successfully used to monitor biogenic amines in HeLa cells and fish samples. This study exploits the potential of MEKC-LIF with SAMF labeling as a tool for monitoring biogenic amines involved in complex physiological and behavioral processes in various matrices.
Cao
Liwei
L
Department of Chemistry, Wuhan University, Wuhan, PR China.
Wang
Hong
H
Ma
Ming
M
Zhang
Huashan
H
D001679:Biogenic Amines / Q000032:analysis* / Q000737:chemistry; D002457:Cell Extracts / Q000737:chemistry; D020374:Chromatography, Micellar Electrokinetic Capillary; D019075:Electrophoresis, Capillary; D005396:Fish Products / Q000032:analysis*; D005453:Fluorescence; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013388:Succinimides / Q000032:analysis / Q000737:chemistry*
C500508:6-oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl)fluorescein; D001679:Biogenic Amines; D002457:Cell Extracts; D005456:Fluorescent Dyes; D013388:Succinimides; D012967:Sodium Dodecyl Sulfate