Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine
Abstract A gas chromatography-mass spectrometry (GC–MS) method was developed and validated in relevant concentration ranges for the simultaneous measurement of l -lysine (Lys, L) and its N ε - and N α -methylated (M), N ε - and N α -acetylated (Ac), N ε -carboxymethylated (CM) and N ε -carboxyethyla...
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Springer Science and Business Media LLC
2021
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Online Access: | http://dx.doi.org/10.1007/s00726-021-03031-6 https://link.springer.com/content/pdf/10.1007/s00726-021-03031-6.pdf https://link.springer.com/article/10.1007/s00726-021-03031-6/fulltext.html |
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crspringernat:10.1007/s00726-021-03031-6 2023-05-15T16:01:49+02:00 Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine Baskal, Svetlana Bollenbach, Alexander Mels, Catharina Kruger, Ruan Tsikas, Dimitrios 2021 http://dx.doi.org/10.1007/s00726-021-03031-6 https://link.springer.com/content/pdf/10.1007/s00726-021-03031-6.pdf https://link.springer.com/article/10.1007/s00726-021-03031-6/fulltext.html en eng Springer Science and Business Media LLC https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 CC-BY Amino Acids ISSN 0939-4451 1438-2199 Organic Chemistry Clinical Biochemistry Biochemistry journal-article 2021 crspringernat https://doi.org/10.1007/s00726-021-03031-6 2022-01-04T16:45:23Z Abstract A gas chromatography-mass spectrometry (GC–MS) method was developed and validated in relevant concentration ranges for the simultaneous measurement of l -lysine (Lys, L) and its N ε - and N α -methylated (M), N ε - and N α -acetylated (Ac), N ε -carboxymethylated (CM) and N ε -carboxyethylated (CE) metabolites in human urine. Analyzed Lys metabolites were the post-translational modification (PTM) products N ε -mono-, di- and trimethyllsine, N ε -MML, N ε -DML, N ε -TML, respectively, N α -ML, N ε -AcL, N α -AcL, and its advanced glycation end-products (AGEs) N ε -CML, N ε -CM-[2,4,4- 2 H 3 ]Lys (d 3 -CML), N ε -CEL and furosine. AGEs of arginine (Arg) and cysteine (Cys) were also analyzed. De novo synthesized trideutero-methyl esters (R-COOCD 3 ) from unlabelled amino acids and derivatives were used as internal standards. Native urine samples (10 µL aliquots) were evaporated to dryness under a stream of nitrogen. Analytes were esterified using 2 M HCl in methanol (60 min, 80 °C) and subsequently amidated by pentafluoropropionic anhydride in ethyl acetate (30 min, 65 °C). The generated methyl ester-pentafluoropropionyl (Me-PFP) derivatives were reconstituted in borate buffer and extracted immediately with toluene. GC–MS analyses were performed by split-less injection of 1-µL aliquots, oven-programmed separation and negative-ion chemical ionization (NICI). Mass spectra were generated in the scan mode (range, m / z 50–1000). Quantification was performed in the selected-ion monitoring (SIM) mode using a dwell time of 50 or 100 ms for each ion. The GC–MS method was suitable for the measurement of Lys and all of its metabolites, except for the quaternary ammonium cation N ε -TML. The Me-PFP derivatives of Lys, Arg and Cys and its metabolites eluted in the retention time window of 9 to 14 min. The derivatization of N ε -CML, d 3 -CML and N ε -CEL was accompanied by partial N ε -decarboxylation and formation of the Me-PFP Lys derivative. The lowest derivatization yield was observed for N ε -DML, indicating a major role of the N ε -DML group in Lys derivatization. The GC–MS method enables precise (relative standard deviation, RSD < 20%) and accurate (bias, < ± 20%) simultaneous measurement of 33 analytes in human urine in relevant concentration ranges. We used the method to measure the urinary excretion rates of Lys and its PTM metabolites and AGEs in healthy black ( n = 39) and white ( n = 41) boys of the Arterial Stiffness in Offspring Study (ASOS). No remarkable differences were found indicating no ethnic-related differences in PTM metabolites and AGEs except for N ε -monomethyllysine and S -(2-carboxymethylcysteine). Article in Journal/Newspaper DML Springer Nature (via Crossref) Amino Acids |
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Organic Chemistry Clinical Biochemistry Biochemistry |
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Organic Chemistry Clinical Biochemistry Biochemistry Baskal, Svetlana Bollenbach, Alexander Mels, Catharina Kruger, Ruan Tsikas, Dimitrios Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
topic_facet |
Organic Chemistry Clinical Biochemistry Biochemistry |
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Abstract A gas chromatography-mass spectrometry (GC–MS) method was developed and validated in relevant concentration ranges for the simultaneous measurement of l -lysine (Lys, L) and its N ε - and N α -methylated (M), N ε - and N α -acetylated (Ac), N ε -carboxymethylated (CM) and N ε -carboxyethylated (CE) metabolites in human urine. Analyzed Lys metabolites were the post-translational modification (PTM) products N ε -mono-, di- and trimethyllsine, N ε -MML, N ε -DML, N ε -TML, respectively, N α -ML, N ε -AcL, N α -AcL, and its advanced glycation end-products (AGEs) N ε -CML, N ε -CM-[2,4,4- 2 H 3 ]Lys (d 3 -CML), N ε -CEL and furosine. AGEs of arginine (Arg) and cysteine (Cys) were also analyzed. De novo synthesized trideutero-methyl esters (R-COOCD 3 ) from unlabelled amino acids and derivatives were used as internal standards. Native urine samples (10 µL aliquots) were evaporated to dryness under a stream of nitrogen. Analytes were esterified using 2 M HCl in methanol (60 min, 80 °C) and subsequently amidated by pentafluoropropionic anhydride in ethyl acetate (30 min, 65 °C). The generated methyl ester-pentafluoropropionyl (Me-PFP) derivatives were reconstituted in borate buffer and extracted immediately with toluene. GC–MS analyses were performed by split-less injection of 1-µL aliquots, oven-programmed separation and negative-ion chemical ionization (NICI). Mass spectra were generated in the scan mode (range, m / z 50–1000). Quantification was performed in the selected-ion monitoring (SIM) mode using a dwell time of 50 or 100 ms for each ion. The GC–MS method was suitable for the measurement of Lys and all of its metabolites, except for the quaternary ammonium cation N ε -TML. The Me-PFP derivatives of Lys, Arg and Cys and its metabolites eluted in the retention time window of 9 to 14 min. The derivatization of N ε -CML, d 3 -CML and N ε -CEL was accompanied by partial N ε -decarboxylation and formation of the Me-PFP Lys derivative. The lowest derivatization yield was observed for N ε -DML, indicating a major role of the N ε -DML group in Lys derivatization. The GC–MS method enables precise (relative standard deviation, RSD < 20%) and accurate (bias, < ± 20%) simultaneous measurement of 33 analytes in human urine in relevant concentration ranges. We used the method to measure the urinary excretion rates of Lys and its PTM metabolites and AGEs in healthy black ( n = 39) and white ( n = 41) boys of the Arterial Stiffness in Offspring Study (ASOS). No remarkable differences were found indicating no ethnic-related differences in PTM metabolites and AGEs except for N ε -monomethyllysine and S -(2-carboxymethylcysteine). |
format |
Article in Journal/Newspaper |
author |
Baskal, Svetlana Bollenbach, Alexander Mels, Catharina Kruger, Ruan Tsikas, Dimitrios |
author_facet |
Baskal, Svetlana Bollenbach, Alexander Mels, Catharina Kruger, Ruan Tsikas, Dimitrios |
author_sort |
Baskal, Svetlana |
title |
Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
title_short |
Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
title_full |
Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
title_fullStr |
Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
title_full_unstemmed |
Development, validation of a GC–MS method for the simultaneous measurement of amino acids, their PTM metabolites and AGEs in human urine, and application to the bi-ethnic ASOS study with special emphasis to lysine |
title_sort |
development, validation of a gc–ms method for the simultaneous measurement of amino acids, their ptm metabolites and ages in human urine, and application to the bi-ethnic asos study with special emphasis to lysine |
publisher |
Springer Science and Business Media LLC |
publishDate |
2021 |
url |
http://dx.doi.org/10.1007/s00726-021-03031-6 https://link.springer.com/content/pdf/10.1007/s00726-021-03031-6.pdf https://link.springer.com/article/10.1007/s00726-021-03031-6/fulltext.html |
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DML |
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DML |
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Amino Acids ISSN 0939-4451 1438-2199 |
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https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 |
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CC-BY |
op_doi |
https://doi.org/10.1007/s00726-021-03031-6 |
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Amino Acids |
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