Application of totally automated on-line sample clean up system for extraction and high-performance liquid chromatography separation of peptide leukotrienes.
We have developed a fully automated on-line extraction-purification method for peptide leukotrienes in biological fluids using a column-switching technique. Individual leukotrienes in HPLC-collected fractions are determined by immunoassay techniques. Leukotrienes were extracted from nasal lavage samples and eluted from the solid-phase extraction cartridge into the HPLC column with methanol-ammonium acetate buffer (60:40, v/v) pH 5.4, as a mobile phase. This method provides good recoveries, excellent resolution of leukotrienes and is suitable to be combined with off-line radioimmunoassay (RIA) or enzymoimmunoassay (EIA). The most important losses are observed after evaporation-concentration and lyophilization. We also have compared RIA with EIA determination. Significant differences are found between RIA values and EIA values.
Ramis
I
I
Molecular Pathology Unit, Centro de Investigación y Desarrollo, CSIC, Barcelona, Spain.
Hotter
G
G
Roselló-Catafau
J
J
Bulbena
O
O
Picado
C
C
Gelpi
E
E
D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007124:Immunoenzyme Techniques; D017997:Leukotriene C4 / Q000302:isolation & purification*; D017998:Leukotriene D4 / Q000302:isolation & purification*; D017817:Nasal Lavage Fluid / Q000737:chemistry*; D011863:Radioimmunoassay
D017997:Leukotriene C4; D017998:Leukotriene D4