Universität zu Köln

He, Yupeng ORCID: 0000-0002-3453-9499, van Mever, Marlien, Yang, Wei ORCID: 0000-0002-3394-7570, Huang, Luojiao ORCID: 0000-0001-5825-5127, Ramautar, Rawi, Rijksen, Yvonne, Vermeij, Wilbert P., Hoeijmakers, Jan H. J., Harms, Amy C., Lindenburg, Peter W. and Hankemeier, Thomas ORCID: 0000-0001-7871-2073 (2022). 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. Metabolites, 12 (8). BASEL: MDPI. ISSN 2218-1989

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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 collection method, and methodologically supports future metabolomic mechanistic studies of sarcopenia.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code He, Yupeng UNSPECIFIED orcid.org/0000-0002-3453-9499 UNSPECIFIED van Mever, Marlien UNSPECIFIED UNSPECIFIED UNSPECIFIED Yang, Wei UNSPECIFIED orcid.org/0000-0002-3394-7570 UNSPECIFIED Huang, Luojiao UNSPECIFIED orcid.org/0000-0001-5825-5127 UNSPECIFIED Ramautar, Rawi UNSPECIFIED UNSPECIFIED UNSPECIFIED Rijksen, Yvonne UNSPECIFIED UNSPECIFIED UNSPECIFIED Vermeij, Wilbert P. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hoeijmakers, Jan H. J. UNSPECIFIED UNSPECIFIED UNSPECIFIED Harms, Amy C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Lindenburg, Peter W. UNSPECIFIED UNSPECIFIED UNSPECIFIED Hankemeier, Thomas UNSPECIFIED orcid.org/0000-0001-7871-2073 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-683527
DOI:	10.3390/metabo12080742
Journal or Publication Title:	Metabolites
Volume:	12
Number:	8
Date:	2022
Publisher:	MDPI
Place of Publication:	BASEL
ISSN:	2218-1989
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language FIBER-TYPE COMPOSITION; SKELETAL-MUSCLE; METABOLOMICS; REPAIR; OPTIMIZATION; STRESS; ROLES; ACID; ATP; AGE Multiple languages Biochemistry & Molecular Biology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/68352