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Simultaneous determination of acylcarnitines, fatty acids, and amino acids by 3-nitrophenylhydrazine derivatization and ultra-high performance liquid chromatography/tandem mass spectrometry

Ardalani, Hamidreza LU ; Dannarm, Srinivas Reddy LU and Spégel, Peter LU (2026) In Journal of Chromatography A 1782.
Abstract

Acylcarnitines (ACs) play a pivotal role in metabolism, most notably by facilitating the transport of fatty acids (FAs) into mitochondria for β-oxidation, a key step in cellular energy production. Dysregulation of AC, FA, and amino acid (AA) levels has been linked to various metabolic disorders, including cardiovascular diseases, neurodegenerative conditions, and cancer. Consequently, monitoring these metabolites in blood samples provides valuable insights into metabolic health and disease progression.In this study, we developed a method for the quantification of carnitine, seven ACs, fifteen FAs, and thirteen AAs in human serum using reversed-phase ultra-high performance liquid chromatography coupled with tandem mass spectrometry... (More)

Acylcarnitines (ACs) play a pivotal role in metabolism, most notably by facilitating the transport of fatty acids (FAs) into mitochondria for β-oxidation, a key step in cellular energy production. Dysregulation of AC, FA, and amino acid (AA) levels has been linked to various metabolic disorders, including cardiovascular diseases, neurodegenerative conditions, and cancer. Consequently, monitoring these metabolites in blood samples provides valuable insights into metabolic health and disease progression.In this study, we developed a method for the quantification of carnitine, seven ACs, fifteen FAs, and thirteen AAs in human serum using reversed-phase ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). By employing 3-nitrophenylhydrazine (3-NPH) derivatization, we achieved high detectability for ACs, with limits of detection (LODs) ranging from 0.01–0.27 ng/mL for ACs, 0.22–1.76 ng/mL for FAs and 0.17–18.25 ng/mL for AAs. Recovery rates ranged from 92–126% for ACs, 56–116% for FAs and 86–115% for AAs. Inter- and intra-day precision were below 20% for all metabolites except two FAs.This method provides a reliable and sensitive tool for the simultaneous analysis of ACs, FAs, and AAs in serum, with potential applications in clinical diagnostics and metabolic research.

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author
; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Acylcarnitine, Amino acids, Fatty acids, LC-MS/MS, Nitrophenylhydrazine
in
Journal of Chromatography A
volume
1782
article number
467105
publisher
Elsevier
external identifiers
  • pmid:42166804
  • scopus:105039333685
ISSN
0021-9673
DOI
10.1016/j.chroma.2026.467105
language
English
LU publication?
yes
id
73db2f1b-088c-4716-ad04-6f37d161502e
date added to LUP
2026-08-24 14:42:41
date last changed
2026-09-07 15:33:18
@article{73db2f1b-088c-4716-ad04-6f37d161502e,
  abstract     = {{<p>Acylcarnitines (ACs) play a pivotal role in metabolism, most notably by facilitating the transport of fatty acids (FAs) into mitochondria for β-oxidation, a key step in cellular energy production. Dysregulation of AC, FA, and amino acid (AA) levels has been linked to various metabolic disorders, including cardiovascular diseases, neurodegenerative conditions, and cancer. Consequently, monitoring these metabolites in blood samples provides valuable insights into metabolic health and disease progression.In this study, we developed a method for the quantification of carnitine, seven ACs, fifteen FAs, and thirteen AAs in human serum using reversed-phase ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). By employing 3-nitrophenylhydrazine (3-NPH) derivatization, we achieved high detectability for ACs, with limits of detection (LODs) ranging from 0.01–0.27 ng/mL for ACs, 0.22–1.76 ng/mL for FAs and 0.17–18.25 ng/mL for AAs. Recovery rates ranged from 92–126% for ACs, 56–116% for FAs and 86–115% for AAs. Inter- and intra-day precision were below 20% for all metabolites except two FAs.This method provides a reliable and sensitive tool for the simultaneous analysis of ACs, FAs, and AAs in serum, with potential applications in clinical diagnostics and metabolic research.</p>}},
  author       = {{Ardalani, Hamidreza and Dannarm, Srinivas Reddy and Spégel, Peter}},
  issn         = {{0021-9673}},
  keywords     = {{Acylcarnitine; Amino acids; Fatty acids; LC-MS/MS; Nitrophenylhydrazine}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Chromatography A}},
  title        = {{Simultaneous determination of acylcarnitines, fatty acids, and amino acids by 3-nitrophenylhydrazine derivatization and ultra-high performance liquid chromatography/tandem mass spectrometry}},
  url          = {{http://dx.doi.org/10.1016/j.chroma.2026.467105}},
  doi          = {{10.1016/j.chroma.2026.467105}},
  volume       = {{1782}},
  year         = {{2026}},
}