GC-MS measurements were performed as described before 11 (link). Ions used for quantification of metabolite levels were as follows: d5–2HG m/z 354; citrate m/z 465; alpha-ketoglutarate m/z 304; succinate m/z 247; fumarate m/z 245; malate m/z 335; aspartate m/z 232; hydroxyproline m/z 332; proline m/z 216; glutamate m/z 246; glutamine m/z 245; lactate m/z 219; pyruvate m/z 174. All peaks were manually inspected and verified relative to known spectra for each metabolite. Peak identification and integration were done with MassHunter software vB.09 (Agilent Technologies). For relative quantification of cell samples, integrated peak areas were normalized to the internal standard d5–2HG and to the packed cell volume of each sample. Absolute quantification of hydroxyproline in tumor acid hydrolysates was performed against a standard curve of commercial trans-4-hydroxy-L-proline (Sigma). In stable isotope tracing experiments, natural isotope abundance correction was performed with IsoCor software v1.2. LC-MS measurements were performed as described before 11 (link). Peak identification and integration were done based on exact mass and retention time match to commercial standards. Data analysis and natural isotope abundance correction were performed with MassHunter Profinder software v10.0 (Agilent Technologies).
Masshunter profinder software v10
Manufactured by Agilent Technologies
MassHunter Profinder software v10.0 is a data analysis tool designed for the processing and interpretation of mass spectrometry data. The software's core function is to enable the identification and quantification of compounds within complex samples.
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Lab products found in correlation
2 protocols using masshunter profinder software v10
1
Quantification of Metabolite Levels by GC-MS and LC-MS
2
Quantification of Metabolite Levels by GC-MS and LC-MS
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GC-MS measurements were performed as described before 11 (link). Ions used for quantification of metabolite levels were as follows: d5–2HG m/z 354; citrate m/z 465; alpha-ketoglutarate m/z 304; succinate m/z 247; fumarate m/z 245; malate m/z 335; aspartate m/z 232; hydroxyproline m/z 332; proline m/z 216; glutamate m/z 246; glutamine m/z 245; lactate m/z 219; pyruvate m/z 174. All peaks were manually inspected and verified relative to known spectra for each metabolite. Peak identification and integration were done with MassHunter software vB.09 (Agilent Technologies). For relative quantification of cell samples, integrated peak areas were normalized to the internal standard d5–2HG and to the packed cell volume of each sample. Absolute quantification of hydroxyproline in tumor acid hydrolysates was performed against a standard curve of commercial trans-4-hydroxy-L-proline (Sigma). In stable isotope tracing experiments, natural isotope abundance correction was performed with IsoCor software v1.2. LC-MS measurements were performed as described before 11 (link). Peak identification and integration were done based on exact mass and retention time match to commercial standards. Data analysis and natural isotope abundance correction were performed with MassHunter Profinder software v10.0 (Agilent Technologies).
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