Succinic acid d4

Manufactured by Merck Group

Sourced in United States

Succinic acid-d4 is a stable isotope-labeled compound with four deuterium atoms substituted for hydrogen atoms in the succinic acid molecule. It is used as a reference standard or internal standard in analytical techniques such as mass spectrometry and nuclear magnetic resonance spectroscopy.

Automatically generated - may contain errors

Lab products found in correlation

15 protocols using succinic acid d4

Analytical Workflow for Metabolite Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

All water
used was of type 1 (>18 MΩ cm), obtained from a MilliQ ultrapure
water purification system (Merck Millipore, Darmstadt, Germany). Methanol
was obtained from Rathburn Chemicals (Walkerburn, Scotland). Formic
acid (98%) was obtained from Merck.
Tobramycin, acylcarnitines
C2, C12, and C16, D2 glycolic acid, D6 glucose, and acylcarnitines
D3 C2, D3 C12, and D3 C16 were obtained from Larodan (Solna, Sweden).
D4 succinic acid, ¹³C creatine, and uric acid were purchased
from Sigma (Darmstadt, Germany). Vancomycin (1000 mg powder) was obtained
from MIP Pharma GmbH (Blieskastel, Germany). ¹³C₂ guanidinoacetate was obtained from Dr. H Ten Brink (VU University
Medical Center, Amsterdam, The Netherlands). Creatinine was obtained
from Merck. Creatine was obtained from Nutritional Biochemical Corporation
(Cleveland, OH, USA).

Skogvold H.B., Sandås E.M., Østeby A., Løkken C., Rootwelt H., Rønning P.O., Wilson S.R, & Elgstøen K.B. (2021). Bridging the Polar and Hydrophobic Metabolome in Single-Run Untargeted Liquid Chromatography-Mass Spectrometry Dried Blood Spot Metabolomics for Clinical Purposes. Journal of Proteome Research, 20(8), 4010-4021.

+ Open protocol

+ Expand

GC-MS Analysis of Liver Tumor Metabolites

Check if the same lab product or an alternative is used in the 5 most similar protocols

Approximately 50 mg of snap-frozen mouse liver tumors was determined by GC-MS analysis as previously reported (19 (link)). Metabolites were extracted with cold 90% methanol and an internal standard d4-succinic acid (Sigma) was added for every 25 mg of tissue. The samples were homogenized, and supernatant was collected by centrifugation and another internal standard, d27-myristic acid, was added. All samples were then dried en vacuo and resuspended in pyridine containing 40 mg/ml O-methoxylamine hydrochloride (MOX) before GC-MS analysis on an Agilent Zorbax DB-5MS column. Metabolites were identified and their peak area was recorded using MassHunter Quant and analyzed with MassHunter software (Agilent). Statistical analysis was performed using MetaboAnalystR.

Guzzo J, & Dubow M.S. (2000). A Novel Selenite- and Tellurite-Inducible Gene in Escherichia coli. Applied and Environmental Microbiology, 66(11), 4972-4978.

+ Open protocol

+ Expand

Metabolite Extraction from Tissue and Serum

Check if the same lab product or an alternative is used in the 5 most similar protocols

In order to extract metabolites from the tissue, each sample was transferred to 2.0 ml ceramic bead mill tubes (bioExpress). Each sample received 450 ul of 90% cold methanol in diH2O for every 25 mg of tissue. The samples were then homogenized in an OMNI Bead Ruptor 24. Homogenized samples were then incubated at −20 °C for 1 hr. D4-succinic acid (Sigma 293075) was added to each sample as an internal standard. After incubation, all the samples were centrifuged at 20,000 x g for 10 min at 4 °C. 450 ul of supernatant was then transferred from each bead mill tube into a labeled, fresh micro centrifuge tube where another internal standard d27-myristic acid (CDN Isotopes: D-1711). Samples were then dried en vacuo. For metabolite extraction from serum, 90% methanol in diH2O containing D4-succinic acid was added to each sample to give a final methanol concentration of 80%. Samples were vortexed and incubated at −20 °C for 1 hr. After incubation, all samples were centrifuged at 20,000 x g for 10 min at 4 °C. Another internal standard, d27-myristic acid (CDN Isotopes: D-1711), was added to each sample. Process blanks were made using the extraction solvent and went through the same process steps as the real samples. The samples were then dried en vacuo.

Panic V., Pearson S., Banks J., Tippetts T.S., Velasco-Silva J.N., Lee S., Simcox J., Geoghegan G., Bensard C.L., van Ry T., Holland W.L., Summers S.A., Cox J., Ducker G.S., Rutter J, & Villanueva C.J. (2020). Mitochondrial pyruvate carrier is required for optimal brown fat thermogenesis. eLife, 9, e52558.

+ Open protocol

+ Expand

Metabolite Extraction and Derivatization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Acetonitrile, isopropanol, and pyridine were of HPLC grade, and methoxyamine hydrochloride (MeOX), 1% TMCS in N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA), adonitol, and d₄-succinic acid were obtained from Sigma-Aldrich, St. Louis, MO, USA.

Misra B.B., Bassey E., Bishop A.C., Kusel D.T., Cox L.A, & Olivier M. (2018). High Resolution GC/MS Metabolomics of Non-Human Primate Serum. Rapid communications in mass spectrometry : RCM.

+ Open protocol

+ Expand

Comprehensive Analytical Standards for Metabolomics

Check if the same lab product or an alternative is used in the 5 most similar protocols

All water used was of type 1 (>18 MΩ cm), obtained from MilliQ ultrapure water purification system (Merck Millipore, Darmstadt, Germany). Methanol was obtained from Rathburn Chemicals (Walkerburn, Scotland). Formic acid (98%) was obtained from Merck.
Tobramycin, acylcarnitines C2, C12, and C16, D2 glycolic acid, D6 glucose and acylcarnitines D3 C2, D3 C12 and D3 C16 were obtained from Larodan (Solna, Sweden). D4 succinic acid, 13 C creatine and uric acid were purchased from Sigma (Darmstadt, Germany).
Vancomycin (1000 mg powder) was obtained from MIP Pharma GmbH (Blieskastel, Germany). 13 C 2 guanidinoacetate was obtained from Dr. H Ten Brink (VU University Medical Center, Amsterdam, The Netherlands). Creatinine was obtained from Merck. Creatine was obtained from Nutritional Biochemical Corporation (Cleveland, OH, USA).

Skogvold H.B., Sandås E.M., Østeby A., Løkken C., Rootwelt H., Rønning P.O., Wilson S.R., & Elgstøen K.B.P. (2021). Bridging the polar and hydrophobic metabolome in single-run untargeted liquid chromatography-mass spectrometry dried blood spot metabolomics for clinical purposes.

+ Open protocol

+ Expand

Extracting Intracellular Metabolites via Rapid Quenching

Check if the same lab product or an alternative is used in the 5 most similar protocols

4 mL cell cultures were mixed with 6 mL cold methanol (−40°C) to arrest metabolism instantaneously. Then, samples were centrifugated at 3000 g for 3 min. Cell pellets were collected and immediately ground to powder in a porcelain mortar, which was pre-cooled to −80°C, under liquid nitrogen for 5 min. Then 100 mg cell powder was mixed thoroughly with 1 ml −40°C 50% methanol (methanol/water, 1:1). The samples were centrifugated at 10000 g for 10 min. The supernatants were collected. Then 10 uL internal standard solution succinic d₄ acid (Sigma, 0.1 mg/ml) was added into the 100 uL extract supernatants before lyophilization. After lyophilization, the derivatization and measurement by GC-MS of these samples were carried out according to a previous method [35 (link)]. Four biological replicates were performed for each sample. The identification and quantification of GC-MS peaks were performed using Agilent software (G1701DA MSD ChemStation ver. D.00.00.38).

Zhang X., Xue C., Zhao F., Li D., Yin J., Zhang C., Caiyin Q, & Lu W. (2014). Suitable extracellular oxidoreduction potential inhibit rex regulation and effect central carbon and energy metabolism in Saccharopolyspora spinosa. Microbial Cell Factories, 13, 98.

+ Open protocol

+ Expand

GC-MS Metabolite Profiling in Drosophila

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The GC-MS assays were performed as previously described (Papadimitropoulos et al., 2018 (link); Tennessen et al., 2014 (link)). Briefly, indicated flies were added with 800 µl prechilled 90% methanol containing 1.25 µg/ml succinic-d4 acid (293075; Sigma) and 6.25 µg/ml U-13C, U-15N amino acid mix (CDNLM-6784; Cambridge Isotope). Samples were then homogenized for 30 s at 6.45 m/s and incubated at −20 °C for 1 h to enhance protein precipitation, followed by centrifugation at 20,000 g for 5 min at 4 °C to remove the resulting precipitate. The supernatant was transferred to a 1.5 ml microfuge tube and the solvent removed with a Speed-Vac (Genevac). The GC-MS detection and analysis software was opened to create the experimental method (6890 Agilent GC with Leco pegasus IV time-of-flight MS instrument). 0.5ul of mixed reagent was drawn with a micro syringe and the sample was injected through the sample inlet of the gas chromatograph. Last, we use the BinBase database software to perform analysis.

Wang J., Zhu Y., Zhang C., Duan R., Kong F., Zheng X, & Hua Y. (2022). A conserved role of bam in maintaining metabolic homeostasis via regulating intestinal microbiota in Drosophila. PeerJ, 10, e14145.

+ Open protocol

+ Expand

Quantification of NNK and Metabolites in Zebrafish

Check if the same lab product or an alternative is used in the 5 most similar protocols

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) was obtained from LGC-Dr Ehrenstorfer (LGC Standards, Barcelona, Spain) and 4-Hydroxy-4-(3-pyridyl)-butyric acid (HPBA) was purchased from Toronto Research Chemicals (TRC, Ontario, Canada). The metabolites hypoxanthine, cytidine monophosphate (CMP), guanosine monophosphate (GMP), S-adenosylmethionine (SAM), and uridine monophosphate (UMP) were obtained from Sigma-Aldrich (Sigma-Aldrich, St. Louis, MO, USA). Internal standard (IS) solution was prepared with 1 mg mL -1 of succinic-d4 acid and myristic-d27 acid in methanol also from Sigma-Aldrich. All solvents used in the preparation of solutions were LC-MS grade. Fish water was prepared with reverse-osmosis purified water containing 90 µg mL -1 of Instant Ocean ® , 0.58 mM CaSO4•2H2O (Aquarium Systems, Sarrebourg, France).

Merino C., Casado M., Piña B., Vinaixa M., & Ramírez N. (2021). Toxicity of 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NKK) in early development: a wide-scope metabolomics assay in zebrafish embryos.

+ Open protocol

+ Expand

Plasma Metabolomics Analysis Protocol

Cited 3 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Plasma samples for metabolomics were acquired at baseline, 1 h from the start of the clamp and 20 min after the last infusion (50 ng adrenaline kg⁻¹ min⁻¹) using containers containing heparin. Plasma samples (30 μl) were mixed with 400 μl methanol, 10 μl internal standard mixture succinic acid-d4, glutamic acid-d5, valine-d8 and heptadecanoic acid-d33, Sigma Aldrich, USA), and incubated on ice for 30 min. Samples were then centrifuged (10,000 g, 3 min, 4°C), and 180 μl of the filtered extracts were evaporated to dryness and derivatised. A detailed description of the process has been presented previously [13 (link)]. A 7250 GC/Q-TOF instrument (Agilent, USA) equipped with a Gerstel MPS autosampler (Gerstel, Germany) was used to analyse the samples. Features from the quantitative ion peak areas of the data were matched against mass spectral libraries (in-house library, Fiehn library [14 (link)], GOLM DB [15 (link)], GNPS [16 (link)], HMDB [17 (link)] and MassBank Japan [18 (link)]).

She R., Suvitaival T., Andersen H.U., Hommel E., Nørgaard K., Wojtaszewski J.F., Legido-Quigley C, & Pedersen-Bjergaard U. (2024). Metabolic effect of adrenaline infusion in people with type 1 diabetes and healthy individuals. Diabetologia, 67(6), 1095-1106.

+ Open protocol

+ Expand

Metabolite Profiling of Biological Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

All solvents were high purity high-performance liquid chromatography (HPLC) grade purchased from J.T. Baker (Philipsburg, NJ, USA). Distilled water was filtered using a Milli-Q Reagent Water System (Millipore, Billerica, MA, USA). Methyloctanoate, succinic acid-d₄, methoxyamine hydrochloride, pyridine, sodium sulfate, and boron trifluoride-methanol were purchased from Sigma-Aldrich (St. Louis, MO, USA). The certified reference material (CRM), a 37 fatty acid methyl ester mixture, and N-methyl-N-(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane as a silylation reagent were purchased from Sigma-Aldrich (St. Louis, MO, USA). The standards of benzeneethanol, benzaldehyde, 1H-pyrrole-2-carboxaldehyde, γ-butyrolactone, γ-caprolactone, δ-hexalactone, γ-octalactone, ethylene glycol, l-(−)-arabitol, d-mannitol, scyllo-inositol, d-glyceric acid sodium salt, fumaric acid, d-psicofuranose, d-gluconic acid solution, and succinic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). 3-Octen-2-one, pantolactone, malic acid, and azelaic acid were obtained from Supelco (Bellefonte, PA, USA).

Kim H., Jung Y., Jeon S.H., Hwang G.S, & Ahn Y.G. (2019). Rapid Characterization and Discovery of Chemical Markers for Discrimination of Xanthii Fructus by Gas Chromatography Coupled to Mass Spectrometry. Molecules, 24(22), 4079.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!