High-performance liquid chromatography determination of σ1-acid glycoprotein in small volumes of plasma from neonates

Stumpe, M. and Miller, C. and Morton, N.S. and Bell, G. and Watson, D.G. (2006) High-performance liquid chromatography determination of σ1-acid glycoprotein in small volumes of plasma from neonates. Journal of Chromatography B, 831 (1-2). pp. 81-84. ISSN 1570-0232 (http://dx.doi.org/10.1016/j.jchromb.2005.11.039)

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Abstract

In order to investigate how the α1-acid glycoprotein (AGP) concentrations of neonates change in response to surgical stress, a simple high-performance liquid chromatography (HPLC)-assay for the measurement of α1-acid glycoprotein levels was developed. A fraction containing α1-acid glycoprotein was isolated from the bulk of plasma protein by addition of 0.6 M perchloric acid and was then analysed directly on a short PLRP-S 4000 Å 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 α1-acid glycoprotein rapidly (<30 min) and required only 50 μl 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 (r2 = 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.

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