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

M. Stumpe, C. Miller, N.S. Morton, G. Bell, D.G. Watson

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.
LanguageEnglish
Pages81-84
Number of pages3
JournalJournal of Chromatography B
Volume831
Issue number1-2
DOIs
Publication statusPublished - 2 Feb 2006

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Plasma Volume
High performance liquid chromatography
Glycoproteins
High Pressure Liquid Chromatography
Newborn Infant
Plasmas
Acids
Quality Control
Calibration
Quality control
Blood Proteins
Assays
Recovery

Keywords

  • anaesthetics
  • alpha1-acid glycoprotein
  • HPLC
  • neonates
  • plasma
  • chromatography

Cite this

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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 {\AA} 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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High-performance liquid chromatography determination of σ1-acid glycoprotein in small volumes of plasma from neonates. / Stumpe, M.; Miller, C.; Morton, N.S.; Bell, G.; Watson, D.G.

In: Journal of Chromatography B , Vol. 831, No. 1-2, 02.02.2006, p. 81-84.

Research output: Contribution to journalArticle

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AU - Bell, G.

AU - Watson, D.G.

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