A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle
The identification of endogenous metabolites has great potential for understanding the underlying tissue processes occurring in either a homeostatic or a diseased state. The application of gas chromatography-mass spectrometry (GC-MS)-based metabolomics on musculoskeletal tissue samples has gained tr...
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MDPI AG
2022-05-01
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| Series: | Metabolites |
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| Online Access: | https://www.mdpi.com/2218-1989/12/5/453 |
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| author | Daniela B. Dias Raphaela Fritsche-Guenther Friederike Gutmann Georg N. Duda Jennifer Kirwan Patrina S. P. Poh |
| author_facet | Daniela B. Dias Raphaela Fritsche-Guenther Friederike Gutmann Georg N. Duda Jennifer Kirwan Patrina S. P. Poh |
| author_sort | Daniela B. Dias |
| collection | DOAJ |
| description | The identification of endogenous metabolites has great potential for understanding the underlying tissue processes occurring in either a homeostatic or a diseased state. The application of gas chromatography-mass spectrometry (GC-MS)-based metabolomics on musculoskeletal tissue samples has gained traction. However, limited comparison studies exist evaluating the sensitivity, reproducibility, and robustness of the various existing extraction protocols for musculoskeletal tissues. Here, we evaluated polar metabolite extraction from bone and muscle of mouse origin. The extraction methods compared were (1) modified Bligh–Dyer (mBD), (2) low chloroform (CHCl<sub>3</sub>)-modified Bligh–Dyer (mBD-low), and (3) modified Matyash (mMat). In particular, the central carbon metabolites (CCM) appear to be relevant for musculoskeletal regeneration, given their role in energy metabolism. However, the sensitivity, reproducibility, and robustness of these methods for detecting targeted polar CCM remains unknown. Overall, the extraction of metabolites using the mBD, mBD-low, and mMat methods appears sufficiently robust and reproducible for bone, with the mBD method slightly bettering the mBD-low and mMat methods. Furthermore, mBD, mBD-low, and mMat were sufficiently sensitive in detecting polar metabolites extracted from mouse muscle; however, they lacked repeatability. This study highlights the need for a re-thinking, towards a tissue-specific optimization of methods for metabolite extractions, ensuring sufficient sensitivity, repeatability, and robustness. |
| first_indexed | 2024-03-10T03:26:05Z |
| format | Article |
| id | doaj.art-75f3879b38034311a2629ea7b4a2af29 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-03-10T03:26:05Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-75f3879b38034311a2629ea7b4a2af292023-11-23T12:07:41ZengMDPI AGMetabolites2218-19892022-05-0112545310.3390/metabo12050453A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and MuscleDaniela B. Dias0Raphaela Fritsche-Guenther1Friederike Gutmann2Georg N. Duda3Jennifer Kirwan4Patrina S. P. Poh5Julius Wolff Institute, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyBerlin Institute of Health at Charité—BIH Metabolomics Platform, 10178 Berlin, GermanyBerlin Institute of Health at Charité—BIH Metabolomics Platform, 10178 Berlin, GermanyJulius Wolff Institute, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyBerlin Institute of Health at Charité—BIH Metabolomics Platform, 10178 Berlin, GermanyJulius Wolff Institute, Berlin Institute of Health at Charité—Universitätsmedizin Berlin, 13353 Berlin, GermanyThe identification of endogenous metabolites has great potential for understanding the underlying tissue processes occurring in either a homeostatic or a diseased state. The application of gas chromatography-mass spectrometry (GC-MS)-based metabolomics on musculoskeletal tissue samples has gained traction. However, limited comparison studies exist evaluating the sensitivity, reproducibility, and robustness of the various existing extraction protocols for musculoskeletal tissues. Here, we evaluated polar metabolite extraction from bone and muscle of mouse origin. The extraction methods compared were (1) modified Bligh–Dyer (mBD), (2) low chloroform (CHCl<sub>3</sub>)-modified Bligh–Dyer (mBD-low), and (3) modified Matyash (mMat). In particular, the central carbon metabolites (CCM) appear to be relevant for musculoskeletal regeneration, given their role in energy metabolism. However, the sensitivity, reproducibility, and robustness of these methods for detecting targeted polar CCM remains unknown. Overall, the extraction of metabolites using the mBD, mBD-low, and mMat methods appears sufficiently robust and reproducible for bone, with the mBD method slightly bettering the mBD-low and mMat methods. Furthermore, mBD, mBD-low, and mMat were sufficiently sensitive in detecting polar metabolites extracted from mouse muscle; however, they lacked repeatability. This study highlights the need for a re-thinking, towards a tissue-specific optimization of methods for metabolite extractions, ensuring sufficient sensitivity, repeatability, and robustness.https://www.mdpi.com/2218-1989/12/5/453musclebonemetabolitesGC-MScentral carbon metabolismmetabolomics |
| spellingShingle | Daniela B. Dias Raphaela Fritsche-Guenther Friederike Gutmann Georg N. Duda Jennifer Kirwan Patrina S. P. Poh A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle Metabolites muscle bone metabolites GC-MS central carbon metabolism metabolomics |
| title | A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle |
| title_full | A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle |
| title_fullStr | A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle |
| title_full_unstemmed | A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle |
| title_short | A Comparison of Solvent-Based Extraction Methods to Assess the Central Carbon Metabolites in Mouse Bone and Muscle |
| title_sort | comparison of solvent based extraction methods to assess the central carbon metabolites in mouse bone and muscle |
| topic | muscle bone metabolites GC-MS central carbon metabolism metabolomics |
| url | https://www.mdpi.com/2218-1989/12/5/453 |
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