Premium
The analysis of human amniotic fluid using capillary electrophoresis
Author(s) -
Stewart Claire J.,
Iles Ray K.,
Perrett David
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683()22:6<1136::aid-elps1136>3.0.co;2-a
Subject(s) - capillary electrophoresis , amniotic fluid , chromatography , electrophoresis , chemistry , capillary action , materials science , biology , fetus , pregnancy , genetics , composite material
Abstract This study has investigated the composition of amniotic fluid (AF) using capillary electrophoresis (CE). A detailed optimisation investigation was undertaken to obtain the best resolution of the major peaks in amniotic fluid. In the final method, capillary zone electrophoresis (CZE) of AF was performed on a Hewlett Packard 3D CE instrument using a fused‐silica capillary of 44 cm total length (36 cm to the detector) with in internal diameter of 50 μm. The background electrolyte was 20 m M sodium tetraborate containing 0.8 m M EDTA adjusted to pH 9.0. AF was diluted 1 plus 1 with deionised water prior to hydrodynamic injection for 3 s at 50 mbar. The separation was performed at + 22.5 kV and resulted in a current of 65 μA. The capillary temperature was 28°C. Using this CZE method, some eight peaks were consistently resolved in AF samples and several other more transient peaks have been separated from AF in less than 10 min. A scheme for the identification of peaks once they had been separated was also developed. Four peaks have been identified as proteins, i.e., gamma‐globulin, α1‐antitrypsin, transferrin and albumin. Surprisingly, one major peak was shown to be the purine catabolite, xanthine.