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9. A Sample Preparation Method for the Simultaneous Profiling of Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia

Author

He, Yupeng, primary, van Mever, Marlien, additional, Yang, Wei, additional, Huang, Luojiao, additional, Ramautar, Rawi, additional, Rijksen, Yvonne, additional, Vermeij, Wilbert P., additional, Hoeijmakers, Jan H. J., additional, Harms, Amy C., additional, Lindenburg, Peter W., additional, and Hankemeier, Thomas, additional

Published
2022
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10. A Sample Preparation Method for the Simultaneous Profiling of Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia

Author

He, Yupeng, van Mever, Marlien, Yang, Wei, Huang, Luojiao, Ramautar, Rawi, Rijksen, Yvonne, Vermeij, Wilbert P., Hoeijmakers, Jan H. J., Harms, Amy C., Lindenburg, Peter W., Hankemeier, Thomas, He, Yupeng, van Mever, Marlien, Yang, Wei, Huang, Luojiao, Ramautar, Rawi, Rijksen, Yvonne, Vermeij, Wilbert P., Hoeijmakers, Jan H. J., Harms, Amy C., Lindenburg, Peter W., and Hankemeier, Thomas

Abstract

The metabolic profiling of a wide range of chemical classes relevant to understanding sarcopenia under conditions in which sample availability is limited, e.g., from mouse models, small muscles, or muscle biopsies, is desired. Several existing metabolomics platforms that include diverse classes of signaling lipids, energy metabolites, and amino acids and amines would be informative for suspected biochemical pathways involved in sarcopenia. The sample limitation requires an optimized sample preparation method with minimal losses during isolation and handling and maximal accuracy and reproducibility. Here, two developed sample preparation methods, BuOH-MTBE-Water (BMW) and BuOH-MTBE-More-Water (BMMW), were evaluated and compared with previously reported methods, Bligh-Dyer (BD) and BuOH-MTBE-Citrate (BMC), for their suitability for these classes. The most optimal extraction was found to be the BMMW method, with the highest extraction recovery of 63% for the signaling lipids and 81% for polar metabolites, and an acceptable matrix effect (close to 1.0) for all metabolites of interest. The BMMW method was applied on muscle tissues as small as 5 mg (dry weight) from the well-characterized, prematurely aging, DNA repair-deficient Ercc1 Delta/-mouse mutant exhibiting multiple-morbidities, including sarcopenia. We successfully detected 109 lipids and 62 polar targeted metabolites. We further investigated whether fast muscle tissue isolation is necessary for mouse sarcopenia studies. A muscle isolation procedure involving 15 min at room temperature revealed a subset of metabolites to be unstable; hence, fast sample isolation is critical, especially for more oxidative muscles. Therefore, BMMW and fast muscle tissue isolation are recommended for future sarcopenia studies. This research provides a sensitive sample preparation method for the simultaneous extraction of non-polar and polar metabolites from limited amounts of muscle tissue, supplies a stable mouse muscle tissue coll

Published
2022

12. A Single-Step Sample Preparation Method for Probing Signaling Lipids and Polar Metabolites in Small Quantities of Muscle Tissues from a Mouse Model for Sarcopenia

Author

He, Yupeng, primary, van Mever, Marlien, additional, Yang, Wei, additional, Huang, Luojiao, additional, Ramautar, Rawi, additional, Rijksen, Yvonne, additional, Vermeij, Wilbert P., additional, Hoeijmakers, Jan H.J., additional, Harms, Amy, additional, Lindenburg, Peter W., additional, and Hankemeier, Thomas, additional

Published
2022
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13. Profiling acidic metabolites by capillary electrophoresis‐mass spectrometry in low numbers of mammalian cells using a novel chemical derivatization approach

Author

van Mever, Marlien, primary, Willacey, Cornelius C. W., additional, Zhang, Wei, additional, Drouin, Nicolas, additional, Christina, Alphert E., additional, Lindenburg, Peter W., additional, van Veldhoven, Jaco P. D., additional, van der Es, Daan, additional, Harms, Amy C., additional, Hankemeier, Thomas, additional, and Ramautar, Rawi, additional

Published
2021
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14. Capillary Electrophoresis-Mass Spectrometry at Trial by Metabo-Ring: Effective Electrophoretic Mobility for Reproducible and Robust Compound Annotation

Author

Drouin, Nicolas, primary, van Mever, Marlien, additional, Zhang, Wei, additional, Tobolkina, Elena, additional, Ferre, Sabrina, additional, Servais, Anne-Catherine, additional, Gou, Marie-Jia, additional, Nyssen, Laurent, additional, Fillet, Marianne, additional, Lageveen-Kammeijer, Guinevere S.M., additional, Nouta, Jan, additional, Chetwynd, Andrew J., additional, Lynch, Iseult, additional, Thorn, James A., additional, Meixner, Jens, additional, Lößner, Christopher, additional, Taverna, Myriam, additional, Liu, Sylvie, additional, Tran, N. Thuy, additional, Francois, Yannis, additional, Lechner, Antony, additional, Nehmé, Reine, additional, Al Hamoui Dit Banni, Ghassan, additional, Nasreddine, Rouba, additional, Colas, Cyril, additional, Lindner, Herbert H., additional, Faserl, Klaus, additional, Neusüß, Christian, additional, Nelke, Manuel, additional, Lämmerer, Stefan, additional, Perrin, Catherine, additional, Bich-Muracciole, Claudia, additional, Barbas, Coral, additional, Gonzálvez, Ángeles López, additional, Guttman, Andras, additional, Szigeti, Marton, additional, Britz-McKibbin, Philip, additional, Kroezen, Zachary, additional, Shanmuganathan, Meera, additional, Nemes, Peter, additional, Portero, Erika P., additional, Hankemeier, Thomas, additional, Codesido, Santiago, additional, González-Ruiz, Víctor, additional, Rudaz, Serge, additional, and Ramautar, Rawi, additional

Published
2020
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15. Profiling acidic metabolites by capillary electrophoresis‐mass spectrometry in low numbers of mammalian cells using a novel chemical derivatization approach.

Author

van Mever, Marlien, Willacey, Cornelius C. W., Zhang, Wei, Drouin, Nicolas, Christina, Alphert E., Lindenburg, Peter W., van Veldhoven, Jaco P. D., van der Es, Daan, Harms, Amy C., Hankemeier, Thomas, and Ramautar, Rawi

Subjects
CAPILLARY electrophoresis, CARBOXYLIC acids, MASS spectrometry, METABOLITE analysis, CHEMICAL derivatives
Abstract

The simultaneous analysis of a broad range of polar ionogenic metabolites using capillary electrophoresis‐mass spectrometry (CE‐MS) can be challenging, as two different analytical methods are often required, that is, one for cations and one for anions. Even though CE‐MS has shown to be an effective method for cationic metabolite profiling, the analysis of small anionic metabolites often results in relatively low sensitivity and poor repeatability. In this work, a novel derivatization strategy based on trimethylmethaneaminophenacetyl bromide was developed to enable CE‐MS analysis of carboxylic acid metabolites using normal CE polarity (i.e., cathode in the outlet) and detection by mass spectrometry in positive ionization mode. Optimization of derivatization conditions was performed using a response surface methodology after which the optimized method (incubation time 50 min, temperature 90°C, and pH 10) was used for the analysis of carboxylic acid metabolites in extracts from HepG2 cells. For selected metabolites, detection limits were down to 8.2 nM, and intraday relative standard deviation values for replicates (n = 3) for peak areas were below 21.5%. Metabolites related to glycolysis, tricarboxylic acid cycle, and anaerobic respiration pathways were quantified in 250,000 cell lysates, and could still be detected in extracts from only 25,000 HepG2 cell lysates (∼70 cell lysates injected). [ABSTRACT FROM AUTHOR]

Published
2022
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