High-performance liquid chromatography determination of alpha1-acid glycoprotein in small volumes of plasma from neonates.
In order to investigate how the alpha1-acid glycoprotein (AGP) concentrations of neonates change in response to surgical stress, a simple high-performance liquid chromatography (HPLC)-assay for the measurement of alpha1-acid glycoprotein levels was developed. A fraction containing alpha1-acid glycoprotein was isolated from the bulk of plasma protein by addition of 0.6M perchloric acid and was then analysed directly on a short PLRP-S 4000A reversed phase column column. The method was validated by analysis of pooled plasma from healthy adults both in comparison with a calibration curve and by standard additions. The procedure was able to isolate alpha1-acid glycoprotein rapidly (<30 min) and required only 50 microl of plasma. The mean extraction recovery was 79.1% (CV 6.4%). The within-run precision for the analysis of three replicates of quality control sample ranged from +/-1.2 to +/-3.8% and the between-run precision was +/-6.1%. The method was linear (r(2)=0.988) over a concentration range from 6 to 100.0 mg/100 ml. The AGP levels in neonatal samples ranged from 25 to 93 mg/100 ml.
Stumpe
M
M
Department of Pharmaceutical Sciences, Strathclyde Institute of Biomedical Sciences, 27 Taylor St., Glasgow G4 0NR, UK.
Miller
C
C
Morton
N S
NS
Bell
G
G
Watson
D G
DG
D000328:Adult; D000767:Anesthesia, Epidural; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D007231:Infant, Newborn; D009961:Orosomucoid / Q000032:analysis*; D015203:Reproducibility of Results
D009961:Orosomucoid