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PreprintCC BYChemRxiv

Simultaneous quantification and natural abundance δ15N analysis of protein-bound amino acids in human lung tissue by GC-C-IRMS

  • Fatmeh Al Rahal1
  • Anthony Anchisi1
  • Youssef Bakkour2,3
  • Illa Tea1
  • Illa TEA
  1. 1Université Lyon 1, CNRS, ISA, UMR
  2. 2College of Applied Medical Sciences
  3. 3King Khalid University
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Abstract

Compound specific isotope analysis in natural abundance provides a tracer free approach to investigate metabolism, but its application to amino acids in complex human tissues remains limited by derivatization requirements and the need for high precision isotope ratio measurements. Here, we developed and validated a Gas Chromatography–Combustion–Isotope Ratio Mass Spectrometry (GC-C-IRMS) method using methyl ester–pentafluoropropionyl (Me-PFP) derivatization for simultaneous quantification and δ15N analysis of protein-bound amino acids in human lung tissues. The method provided 12 measurable chromatographic signals corresponding to 13 amino acids, with satisfactory isotopic precision from standards to biological matrices, and derivative stability for 22 days in standards and for approximately 28 days in lung cancer tissue sample at −20 °C. Application to 12 paired tumor and adjacent non-tumor samples showed negative median tumor–adjacent δ15N shifts for glycine and hydroxyproline. Exploratory histology stratified analysis further suggested opposite δ15N shifts for alanine and tyrosine between adenocarcinoma and squamous cell carcinoma. These results demonstrate the suitability of a Me-PFP GC-C-IRMS workflow for combined quantitative and natural abundance nitrogen isotopic profiling of amino acids in human lung tissues.

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Cite

BibTeXRIS
BibTeX
@misc{rahal2026simultaneous,
  title = {{Simultaneous quantification and natural abundance δ15N analysis of protein-bound amino acids in human lung tissue by GC-C-IRMS}},
  author = {Fatmeh Al Rahal and Anthony Anchisi and Youssef Bakkour and Illa Tea and Illa TEA},
  howpublished = {ChemRxiv},
  year = {2026},
  month = sep,
  doi = {10.26434/chemrxiv.15009234/v1},
  url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009234/v1},
  note = {CC BY},
}
RIS
TY  - UNPB
TI  - Simultaneous quantification and natural abundance δ15N analysis of protein-bound amino acids in human lung tissue by GC-C-IRMS
AU  - Fatmeh Al Rahal
AU  - Anthony Anchisi
AU  - Youssef Bakkour
AU  - Illa Tea
AU  - Illa TEA
PY  - 2026
DA  - 2026/09/22
PB  - ChemRxiv
DO  - 10.26434/chemrxiv.15009234/v1
UR  - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009234/v1
AB  - Compound specific isotope analysis in natural abundance provides a tracer free approach to investigate metabolism, but its application to amino acids in complex human tissues remains limited by derivatization requirements and the need for high precision isotope ratio measurements. Here, we developed and validated a Gas Chromatography–Combustion–Isotope Ratio Mass Spectrometry (GC-C-IRMS) method using methyl ester–pentafluoropropionyl (Me-PFP) derivatization for simultaneous quantification and δ15N analysis of protein-bound amino acids in human lung tissues. The method provided 12 measurable chromatographic signals corresponding to 13 amino acids, with satisfactory isotopic precision from standards to biological matrices, and derivative stability for 22 days in standards and for approximately 28 days in lung cancer tissue sample at −20 °C. Application to 12 paired tumor and adjacent non-tumor samples showed negative median tumor–adjacent δ15N shifts for glycine and hydroxyproline. Exploratory histology stratified analysis further suggested opposite δ15N shifts for alanine and tyrosine between adenocarcinoma and squamous cell carcinoma. These results demonstrate the suitability of a Me-PFP GC-C-IRMS workflow for combined quantitative and natural abundance nitrogen isotopic profiling of amino acids in human lung tissues.
ER  -