Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection
Indole-containing acids—tryptophan metabolites—found in serum and cerebrospinal fluid (CSF) samples of patients with diseases of the central nervous system (CNS) were determined with the use of microextraction by packed sorbent (MEPS) followed by silylation and gas chromatography–mass spectrometry (...
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MDPI AG
2020-07-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/25/14/3258 |
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| author | Alisa Pautova Zoya Khesina Maria Getsina Pavel Sobolev Alexander Revelsky Natalia Beloborodova |
| author_facet | Alisa Pautova Zoya Khesina Maria Getsina Pavel Sobolev Alexander Revelsky Natalia Beloborodova |
| author_sort | Alisa Pautova |
| collection | DOAJ |
| description | Indole-containing acids—tryptophan metabolites—found in serum and cerebrospinal fluid (CSF) samples of patients with diseases of the central nervous system (CNS) were determined with the use of microextraction by packed sorbent (MEPS) followed by silylation and gas chromatography–mass spectrometry (GC–MS) analysis. MEPS with the following silylation led to the reproducible formation of derivatives with an unsubstituted hydrogen ion in the indole ring, the chromatographic peaks of which are symmetric and can be used for GC–MS analysis without additional derivatization. The recoveries of analytes at the limit of quantitation (LOQ) levels were 40–80% for pooled CSF and 40–60% for serum. The limit of detection (LOD) and LOQ values were 0.2–0.4 and 0.4–0.5 µM, respectively, for both CSF and serum. The precision (the reproducibility, RSD) value of less than 20% and the accuracy (the relative error, RE) value of less than ±20% at the LOQ concentrations meet the Food and Drug Administration (FDA) recommendations. Linear correlations for all analytes were determined over a potentially clinically significant range of concentrations (0.4–10 µM for serum, <i>R</i><sup>2</sup> ≥ 0.9942, and 0.4–7 µM for CSF, <i>R</i><sup>2</sup> ≥ 0.9949). Moreover, MEPS significantly reduced the matrix effect of serum compared to liquid–liquid extraction (LLE), which was revealed in the example of reducing the amount of cholesterol and its relative compounds. |
| first_indexed | 2024-03-10T18:25:58Z |
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| id | doaj.art-970acab472ea4733b8b154c579e05f9b |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T18:25:58Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-970acab472ea4733b8b154c579e05f9b2023-11-20T07:03:23ZengMDPI AGMolecules1420-30492020-07-012514325810.3390/molecules25143258Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS DetectionAlisa Pautova0Zoya Khesina1Maria Getsina2Pavel Sobolev3Alexander Revelsky4Natalia Beloborodova5Laboratory of Human Metabolism in Critical States, Negovsky Research Institute of General Reanimatology, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Petrovka str., 25-2, 107031 Moscow, RussiaA.N. Frumkin Institute of Physical Chemistry and Electrochemistry, Russian Academy of Sciences, GSP-1, Leninsky Prospect, 31, 119071 Moscow, RussiaLaboratory of Human Metabolism in Critical States, Negovsky Research Institute of General Reanimatology, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Petrovka str., 25-2, 107031 Moscow, RussiaLaboratory of Mass Spectrometry, Chemistry Department, Lomonosov Moscow State University, GSP-1, Leninskie gory, 1-3, 119991 Moscow, RussiaLaboratory of Mass Spectrometry, Chemistry Department, Lomonosov Moscow State University, GSP-1, Leninskie gory, 1-3, 119991 Moscow, RussiaLaboratory of Human Metabolism in Critical States, Negovsky Research Institute of General Reanimatology, Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Petrovka str., 25-2, 107031 Moscow, RussiaIndole-containing acids—tryptophan metabolites—found in serum and cerebrospinal fluid (CSF) samples of patients with diseases of the central nervous system (CNS) were determined with the use of microextraction by packed sorbent (MEPS) followed by silylation and gas chromatography–mass spectrometry (GC–MS) analysis. MEPS with the following silylation led to the reproducible formation of derivatives with an unsubstituted hydrogen ion in the indole ring, the chromatographic peaks of which are symmetric and can be used for GC–MS analysis without additional derivatization. The recoveries of analytes at the limit of quantitation (LOQ) levels were 40–80% for pooled CSF and 40–60% for serum. The limit of detection (LOD) and LOQ values were 0.2–0.4 and 0.4–0.5 µM, respectively, for both CSF and serum. The precision (the reproducibility, RSD) value of less than 20% and the accuracy (the relative error, RE) value of less than ±20% at the LOQ concentrations meet the Food and Drug Administration (FDA) recommendations. Linear correlations for all analytes were determined over a potentially clinically significant range of concentrations (0.4–10 µM for serum, <i>R</i><sup>2</sup> ≥ 0.9942, and 0.4–7 µM for CSF, <i>R</i><sup>2</sup> ≥ 0.9949). Moreover, MEPS significantly reduced the matrix effect of serum compared to liquid–liquid extraction (LLE), which was revealed in the example of reducing the amount of cholesterol and its relative compounds.https://www.mdpi.com/1420-3049/25/14/3258tryptophan metabolitesindole-3-acetic acid5-hydroxyindole-3-acetic acidindole-3-propionic acidindole-3-lactic acidmicroextraction by packed sorbent |
| spellingShingle | Alisa Pautova Zoya Khesina Maria Getsina Pavel Sobolev Alexander Revelsky Natalia Beloborodova Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection Molecules tryptophan metabolites indole-3-acetic acid 5-hydroxyindole-3-acetic acid indole-3-propionic acid indole-3-lactic acid microextraction by packed sorbent |
| title | Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection |
| title_full | Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection |
| title_fullStr | Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection |
| title_full_unstemmed | Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection |
| title_short | Determination of Tryptophan Metabolites in Serum and Cerebrospinal Fluid Samples Using Microextraction by Packed Sorbent, Silylation and GC–MS Detection |
| title_sort | determination of tryptophan metabolites in serum and cerebrospinal fluid samples using microextraction by packed sorbent silylation and gc ms detection |
| topic | tryptophan metabolites indole-3-acetic acid 5-hydroxyindole-3-acetic acid indole-3-propionic acid indole-3-lactic acid microextraction by packed sorbent |
| url | https://www.mdpi.com/1420-3049/25/14/3258 |
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