Simultaneous determination of multiple amino acids in plasma in critical illness by high performance liquid chromatography with ultraviolet and fluorescence detection

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Abstract

There is increasing recognition that the host response to critical illness includes derangement of multiple amino acid pathways, including amino acids (AAs) central to metabolism and immune, endothelial and neurological function. To characterise concentration changes of these plasma amino acid we report the development and validation of a method for the quantification of AAs in small volumes of plasma (50. ?L) using HPLC with simultaneous UV and fluorescence (FL) detection. Protein precipitation and pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is followed by reversed phase HPLC separation. Calibration curves were built with norleucine as an internal standard. Thirty-three (including the 20 proteinogenic) AAs, were selected as standards and their corresponding concentrations in the plasma of healthy human controls and patients with severe falciparum malaria were quantified. This method enables the detection of perturbations in arginine metabolism, aromatic amino acid pathways, methionine transsulfuration and transmethylation pathways and other metabolic pathways. � 2013 Elsevier B.V.
Original languageEnglish
Pages (from-to)53-58
Number of pages6
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume940
DOIs
Publication statusPublished - 2013

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High performance liquid chromatography
Critical Illness
Fluorescence
High Pressure Liquid Chromatography
Plasmas
Amino Acids
Metabolism
Norleucine
Aromatic Amino Acids
Falciparum Malaria
Plasma Volume
Metabolic Networks and Pathways
Phase separation
Methionine
Calibration
Arginine
Proteins

Cite this

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title = "Simultaneous determination of multiple amino acids in plasma in critical illness by high performance liquid chromatography with ultraviolet and fluorescence detection",
abstract = "There is increasing recognition that the host response to critical illness includes derangement of multiple amino acid pathways, including amino acids (AAs) central to metabolism and immune, endothelial and neurological function. To characterise concentration changes of these plasma amino acid we report the development and validation of a method for the quantification of AAs in small volumes of plasma (50. ?L) using HPLC with simultaneous UV and fluorescence (FL) detection. Protein precipitation and pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is followed by reversed phase HPLC separation. Calibration curves were built with norleucine as an internal standard. Thirty-three (including the 20 proteinogenic) AAs, were selected as standards and their corresponding concentrations in the plasma of healthy human controls and patients with severe falciparum malaria were quantified. This method enables the detection of perturbations in arginine metabolism, aromatic amino acid pathways, methionine transsulfuration and transmethylation pathways and other metabolic pathways. � 2013 Elsevier B.V.",
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author = "Hao Wang and Yvette McNeil and Tsin Yeo and Nicholas Anstey",
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TY - JOUR

T1 - Simultaneous determination of multiple amino acids in plasma in critical illness by high performance liquid chromatography with ultraviolet and fluorescence detection

AU - Wang, Hao

AU - McNeil, Yvette

AU - Yeo, Tsin

AU - Anstey, Nicholas

PY - 2013

Y1 - 2013

N2 - There is increasing recognition that the host response to critical illness includes derangement of multiple amino acid pathways, including amino acids (AAs) central to metabolism and immune, endothelial and neurological function. To characterise concentration changes of these plasma amino acid we report the development and validation of a method for the quantification of AAs in small volumes of plasma (50. ?L) using HPLC with simultaneous UV and fluorescence (FL) detection. Protein precipitation and pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is followed by reversed phase HPLC separation. Calibration curves were built with norleucine as an internal standard. Thirty-three (including the 20 proteinogenic) AAs, were selected as standards and their corresponding concentrations in the plasma of healthy human controls and patients with severe falciparum malaria were quantified. This method enables the detection of perturbations in arginine metabolism, aromatic amino acid pathways, methionine transsulfuration and transmethylation pathways and other metabolic pathways. � 2013 Elsevier B.V.

AB - There is increasing recognition that the host response to critical illness includes derangement of multiple amino acid pathways, including amino acids (AAs) central to metabolism and immune, endothelial and neurological function. To characterise concentration changes of these plasma amino acid we report the development and validation of a method for the quantification of AAs in small volumes of plasma (50. ?L) using HPLC with simultaneous UV and fluorescence (FL) detection. Protein precipitation and pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) is followed by reversed phase HPLC separation. Calibration curves were built with norleucine as an internal standard. Thirty-three (including the 20 proteinogenic) AAs, were selected as standards and their corresponding concentrations in the plasma of healthy human controls and patients with severe falciparum malaria were quantified. This method enables the detection of perturbations in arginine metabolism, aromatic amino acid pathways, methionine transsulfuration and transmethylation pathways and other metabolic pathways. � 2013 Elsevier B.V.

KW - 6-aminoquinolyl-N-hydroxysuccinimidyl carbamates

KW - Critical illness

KW - Fluorescence detection

KW - HPLC

KW - Malaria

KW - Precolumn derivatizations

KW - Protein precipitation

KW - Simultaneous determinations

KW - High performance liquid chromatography

KW - Metabolism

KW - Physiology

KW - Plasmas

KW - Precipitation (chemical)

KW - Amino acids

KW - 1 methyl histidine

KW - 4 hydroxy proline

KW - 6 aminoquinolyl n hydroxysuccinimidyl carbamate

KW - amino acid

KW - arginine

KW - asparagine

KW - aspartic acid

KW - citrulline

KW - cystathionine

KW - cystine

KW - diaminobutyric acid

KW - glutamic acid

KW - glutamine

KW - glycine

KW - homoarginine

KW - kynurenine

KW - leucine

KW - methionine

KW - norleucine

KW - ornithine

KW - phenylalanine

KW - serine

KW - taurine

KW - threonine

KW - tryptophan

KW - tyrosine

KW - unclassified drug

KW - amino acid analysis

KW - amino acid blood level

KW - article

KW - controlled study

KW - critical illness

KW - fluorescence

KW - human

KW - human tissue

KW - limit of detection

KW - limit of quantitation

KW - malaria falciparum

KW - precipitation

KW - priority journal

KW - reversed phase high performance liquid chromatography

KW - FL

KW - Adult

KW - Amino Acids

KW - Chromatography, High Pressure Liquid

KW - Critical Illness

KW - Humans

KW - Malaria, Falciparum

KW - Reproducibility of Results

KW - Sensitivity and Specificity

KW - Sepsis

KW - Spectrometry, Fluorescence

KW - Spectrophotometry, Ultraviolet

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U2 - 10.1016/j.jchromb.2013.09.016

DO - 10.1016/j.jchromb.2013.09.016

M3 - Article

VL - 940

SP - 53

EP - 58

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

ER -