Enhanced separation and detection of serum bilirubin species by capillary electrophoresis using a mixed anionic surfactant-protein buffer system with laser-induced fluorescence detection.
The four major bilirubin species in serum are separated by capillary electrophoresis and detected using laser-induced fluorescence detection. The optimum buffer system consists of 40 mM sodium dodecyl sulfate (SDS)-0.012 mM bovine serum albumin (BSA). The use of the SDS-BSA mixture in the mobile phase allows for the separation of four major bilirubin species at physiological pH with untreated capillaries. The results show that the use of BSA as a run buffer modifier in SDS solution improves separation efficiency and increases sample solubility via pH changes of the run buffer. The limits of detection for the bilirubin species using laser-induced fluorescence are between 30 and 150 nM, depending on the bilirubin species; not only is this approximately two orders of magnitude lower than with visible-light absorption methods, it allows the bilirubin species in normal sera to be quantitatively measured without sample pretreatment.
Wu
N
N
Department of Chemistry, University of Illinois at Champaign-Urbana 61801.
Sweedler
J V
JV
Lin
M
M
D000328:Adult; D001663:Bilirubin / Q000097:blood / Q000302:isolation & purification*; D002021:Buffers; D004586:Electrophoresis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D013050:Spectrometry, Fluorescence
D002021:Buffers; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D001663:Bilirubin