Rapid analysis of trace levels of flavins by pressurized capillary electrochromatography-laser induced fluorescence detection with sulfonated N-octadecyl methacrylate monolith.
In this paper, pressurized capillary electrochromatography (pCEC) with laser induced fluorescence detection (LIF) was demonstrated as a viable approach for the separation and determination of trace flavins in human plasma, where flavins tend to be degraded ex vivo. Using a sulfonated N-octadecyl methacrylate monolithic column in isocratic pCEC separation, symmetrical peak shapes and rapid separation could be obtained in a weakly acidic mobile phase. Baseline separation of riboflavin, flavin mononucleotide and flavin adenine dinucleotide could be achieved within 4.5 min in a mobile phase containing 60% (v/v) acetonitrile and 40% (v/v) of 20 mmol L(-1) phosphate buffer (pH 4.0), with -22 kV of applied voltage and 290 psi of supplementary pressure and 0.02 mL min(-1) of flow rate. Based on a 473 nm laser diode double pumped solid state source, flavins could be determined by LIF with the detection limit (LOD) as low as 0.5 nmol L(-1) (S/N=3). The concentration ranges were 0.005-2 micromol L(-1) for RF and FMN, and 0.02-40 micromol L(-1) for FAD. Owing to the weakly acidic condition selected in this experiment, the high fluorescence quantum yields and good stability of flavins contributed to a preferable analysis. Combined with a simple clean-up procedure, this method has been proved to be effective for the rapid and selective analysis of trace levels of flavins in human plasma without sample preconcentration.
Wu
Yimin
Y
Department of Chemistry, Fuzhou University, Fuzhou 350108, China.
Lin
Jian
J
Wu
Qingzheng
Q
Wu
Xiaoping
X
Lin
Xucong
X
Xie
Zenghong
Z
D053801:Capillary Electrochromatography / Q000379:methods*; D005415:Flavins / Q000032:analysis* / Q000097:blood; D005456:Fluorescent Dyes; D006801:Humans; D007834:Lasers; D008689:Methacrylates / Q000032:analysis* / Q000737:chemistry; D011108:Polymers / Q000032:analysis / Q000737:chemistry; D013997:Time Factors
D005415:Flavins; D005456:Fluorescent Dyes; D008689:Methacrylates; D011108:Polymers