Analysis of organic acids by high performance liquid chromatography in urine and plasma and its application
Analysis of organic acids by high performance liquid chromatography in urine and plasma and its application
Ion-moderated partition chromatography using a Bio-Rad Aminex HPX-87H column at 50oC eluted isocratically with sulphuric acid with ultraviolet detection at 210nm, was selected to investigate the potential value of high performance liquid chromatography for screening and management of inherited and acquired organic acid disorders. An anion-exchange extraction procedure was optimised, and used, to improve resolution and reduce interference, especially from urate. Identification of acids was by relative retention time, and confirmed with authentic standards. Analytical recoveries and precision were poor for lactic, pyruvic, propionic, acetoacetic and 3-hydroxybutyric acids, but were acceptable for the remaining eighteen acids studied. Poor sensitivity of the method for dicarboxylic acids and ketones limits its use for the detection of fatty acid oxidation defects. Urine and plasma samples from normal children were analysed to define normal organic acid profiles. For twelve of the fourteen different inherited organic acidurias studied, definite urinary abnormality was found which would have prompted further investigations. However, samples from patients with medium chain acyl-CoA dehydrogenase deficiency and glutaric aciduria type I were not demonstrably abnormal. Plasma profiles were as informative as urine in five different inherited metabolic defects studied. Advantages of the high performance liquid chromatographic procedures are speed, robustness and ease of use, but coelution of compounds, lack of authentic standards and a definite means of identification remain the major drawbacks. Other potentially useful applications of high performance liquid chromatography are: the diagnosis of medium chain acyl-CoA dehydrogenase deficiency using the 3-phenylpropionate loading test; monitoring nitrogen excretion of patients with urea cycle disorders undergoing therapy with sodium benzoate and phenylacetic acid, and monitoring tyrosyluria of babies receiving parenteral nutrition. Coeluting compounds make the technique unacceptable for screening for phaeochromocytoma or neuroblastoma.
University of Southampton
1990
Bulusu, Sudha
(1990)
Analysis of organic acids by high performance liquid chromatography in urine and plasma and its application.
University of Southampton, Doctoral Thesis.
Record type:
Thesis
(Doctoral)
Abstract
Ion-moderated partition chromatography using a Bio-Rad Aminex HPX-87H column at 50oC eluted isocratically with sulphuric acid with ultraviolet detection at 210nm, was selected to investigate the potential value of high performance liquid chromatography for screening and management of inherited and acquired organic acid disorders. An anion-exchange extraction procedure was optimised, and used, to improve resolution and reduce interference, especially from urate. Identification of acids was by relative retention time, and confirmed with authentic standards. Analytical recoveries and precision were poor for lactic, pyruvic, propionic, acetoacetic and 3-hydroxybutyric acids, but were acceptable for the remaining eighteen acids studied. Poor sensitivity of the method for dicarboxylic acids and ketones limits its use for the detection of fatty acid oxidation defects. Urine and plasma samples from normal children were analysed to define normal organic acid profiles. For twelve of the fourteen different inherited organic acidurias studied, definite urinary abnormality was found which would have prompted further investigations. However, samples from patients with medium chain acyl-CoA dehydrogenase deficiency and glutaric aciduria type I were not demonstrably abnormal. Plasma profiles were as informative as urine in five different inherited metabolic defects studied. Advantages of the high performance liquid chromatographic procedures are speed, robustness and ease of use, but coelution of compounds, lack of authentic standards and a definite means of identification remain the major drawbacks. Other potentially useful applications of high performance liquid chromatography are: the diagnosis of medium chain acyl-CoA dehydrogenase deficiency using the 3-phenylpropionate loading test; monitoring nitrogen excretion of patients with urea cycle disorders undergoing therapy with sodium benzoate and phenylacetic acid, and monitoring tyrosyluria of babies receiving parenteral nutrition. Coeluting compounds make the technique unacceptable for screening for phaeochromocytoma or neuroblastoma.
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Published date: 1990
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Local EPrints ID: 458315
URI: http://eprints.soton.ac.uk/id/eprint/458315
PURE UUID: 6d129086-cd24-4f35-b3ea-3c9850554fcc
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Date deposited: 04 Jul 2022 16:46
Last modified: 04 Jul 2022 16:46
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Author:
Sudha Bulusu
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