In total, 186 annotated metabolites were quantified using the p180 kit (BIOCRATES Life Sciences AG, Innsbruck, Austria), being 40 acylcanitines (ACs), 21 amino acids (AAs), 19 biogenic amines (BA), sum of hexoses (Hex), 76 phosphatidylcholines (PCs), 14 lyso-phosphatidylcholines (LPCs) and 15 sphingomyelins (SMs). glycerophospholipids were further differentiated with respect to the presence of ester (a) and ether (e) bonds in the glycerol moiety, where two letters denote that two glycerol positions are bound to a fatty acid residue (aa = diacyl, ae = acyl-alkyl), while a single letter indicates the presence of a single fatty acid residue (a = acyl or e = alkyl). In the same company (Biocrates), the European participants had their samples additionally analyzed for the following energy metabolism metabolites: lactate, pyruvate/oxaloacetate, alpha ketoglutarate, fumarate and succinate.
P180 kit
Manufactured by Biocrates
Sourced in Austria
The P180 kits are a metabolite assay product offered by Biocrates. They are designed to provide quantitative analysis of 180 different metabolites in biological samples. The kits utilize mass spectrometry-based techniques to deliver comprehensive metabolite profiling data. Further details on the specific intended use or interpretation of the results are not available.
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Lab products found in correlation
5 protocols using p180 kit
1
Comprehensive Metabolite Quantification Protocol
2
Investigating SM Ratios in Targeted Metabolomics
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For the investigation of SM ratios measured by targeted metabolomics using the Biocrates P180 kit, we used the same cohort data and statistical models used in Toledo et al.20 (link). For the selection of the most informative SM ratio, we first calculated all ratios between short-chain (chain length < C20) and long-chain (≥C20) SMs on metabolite levels not adjusted for medication. For each ratio, we then identified significant medications using backward selection based on the Bayesian Information Criterion. Significant medications were included as additional covariates extending the base models described in Toledo et al.20 (link) for phenotype associations. Using the Pgain criterion, which is defined by the ratio of the minimum association p-value of the constituents of a ratio with the association p-value of the ratio and provides a measure of significance added by the ratio, we obtained the ratio of SM (d34:1) and SM (d43:1) as the one with the largest overall Pgain.
3
Metabolic Profiling of Acylcarnitines
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Serum or plasma samples in study A were analyzed for concentrations of 33 ACs (along with other metabolites, including amino acids) using the Biocrates AbsoluteIDQ p150 or p180 kits and tandem mass spectrometry following the procedure recommended by the vendor [3 (link),[21] (link), [22] (link), [23] (link)]. Participants with missing data on frequently detected ACs such as C3:0 and C18:0 were excluded (n = 28). Only ACs that were present in >80% of subjects were included in the statistical analysis. Details on the data-processing pipeline can be found elsewhere [29 (link)]. Acylcarnitines had a maximum of 35 missing values which were imputed with the median of the corresponding batch. As the metabolomics data originates from controls of several cancer-studies (some of which only included one sex), concentrations were transformed to the residuals of linear mixed-effect models of log-transformed concentrations on study and batch as random effects and sex as a fixed effect. The samples from each study were either all serum or all plasma so normalization by study was also performed to remove differences due to sample type.
4
Metabolomic Extraction of Brain Tissues
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The extraction method applied to the samples was identical to our previously published work [22 (link)] using Biocrates p180 kits and based upon previous optimisation work undertaken by others [34 (link)]. Both human brain and mouse brain samples were collected into individual tubes to avoid cross-contamination, lyophilized, and cryogenically milled to a fine powder. 30 mg (± 0.5 mg) of powdered human PM brain tissue and 15 mg (± 0.5 mg) of powdered mouse brain tissue were weighed and extracted in 100 µL and 50 µL of solvent (85% ethanol and 15% PBS buffer) respectively in a 1.5 mL sterile Eppendorf tube. The samples were shaken (10 min), sonicated (15 min), and centrifuged at (10,000× g; 4 °C; 5 min) and the supernatant retained for analysis.
5
Comprehensive Plasma Lipidomics and Metabolomics
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Saturated fatty acids (SFAs), monounsaturated fatty acids, polyunsaturated fatty acids, industrial trans fatty acids, and natural trans fatty acids were extracted from plasma phospholipid fractions and quantified by gas chromatography.12 (link) For endogenous metabolites, the Biocrates AbsoluteIDQ p150 or p180 Kits were used to measure concentrations of amino acids, biogenic amines, hexose sugars, acylcarnitines, sphingolipids (sphingomyelins), phosphatidylcholines (PC), and lysophosphatidylcholines in serum or plasma, following the recommended procedure.13 (link),14 See the Supplementary Methods section for further details of analytical methodology.
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