Micellar Electrokinetic Capillary Chromatography Determination of Guanidino Compounds in Serum of Uremic Patients by using Pyridoin as Derivatizing Reagent.

摘要

An analytical procedure has been developed for the determination of 5 guanidino compounds; guanidine (G), guanidinoacetic acid (GAA), guanidinopropionic acid (GPA), guanidinobutyric acid (GBA), and guanidinosuccinic acid (GSA) by micellar electrokinetic capillary chromatography (MEKC) within 5 min. using pyridoin as derivatizing reagent. Sodium dodecyl sulfate (SDS) (0.03 M) was used as a micellar medium in sodium tetraborate (0.1 M) buffer at pH 9. Uncoated fused silica capillary was used with effective length 39 cm and 75 mu m id. Applied voltage was 25.2 kV and photo diode array detection was at 228 nm. Linear calibrations were obtained from 0.70 to 122 mu mol/L and limits of detection (LODs) were within 0.023-0.032 mu mol/L. The derivatization and separation were repeatable with relative standard deviation (RSDs) within 1.2-3.1%. Serum of healthy volunteers and uremic patients was analyzed and amounts found in uremic patients were G 1.71-2.64, GAA 4.56-6.61, GPA 1.15-1.94, GBA 1.25-1.98, and GSA 6.81-8.56 mol/L with RSD (n=4) within 1.2-3.3%. The amount of guanidino compounds in uremic patients was found higher than healthy volunteers.