| Size | Price | |
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| Other Sizes |
| ln Vitro |
2-Methylsuccinate is a typical metabolite found in human bodily fluids and the main biochemically detectable feature of encephalopathy caused by ethylmalonic acid [1]. The catabolism of isoleucine is the fundamental biochemical problem [2].
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| ln Vivo |
- In patients with ethylmalonic encephalopathy (EE), 2-Methylsuccinic acid was detected at elevated levels in urine and cerebrospinal fluid (CSF). Urinary 2-Methylsuccinic acid concentrations ranged from 80 to 250 μmol/mmol creatinine (normal range: <10 μmol/mmol creatinine), and CSF concentrations ranged from 5 to 18 μmol/L (normal range: undetectable) [1]
- In EE patients, 2-Methylsuccinic acid accumulation was associated with abnormal isoleucine metabolism. After oral administration of [1-¹⁴C]-isoleucine to EE patients, radioactive labeling was detected in urinary 2-Methylsuccinic acid, confirming that it is a metabolic byproduct of isoleucine catabolism [2] |
| Enzyme Assay |
- For urinary organic acid analysis (to detect 2-Methylsuccinic acid): Urine samples from EE patients and healthy controls were collected and deproteinized by centrifugation after adding sulfosalicylic acid. The supernatant was filtered through a 0.22 μm membrane and analyzed by gas chromatography-mass spectrometry (GC-MS). The column was programmed to升温 from 80°C to 280°C at a rate of 10°C/min, and the mass spectrometer was operated in electron ionization mode. 2-Methylsuccinic acid was identified by comparing its retention time and mass spectrum with a standard reference, and its concentration was quantified using an internal standard (heptadecanoic acid) [1, 2]
- For isotope tracing experiment (to confirm 2-Methylsuccinic acid origin): EE patients were given a single oral dose of [1-¹⁴C]-isoleucine (1 μCi/kg body weight). Urine was collected at 2-hour intervals for 24 hours, and urinary 2-Methylsuccinic acid was isolated by preparative HPLC. The radioactivity of the isolated 2-Methylsuccinic acid was measured using a liquid scintillation counter, and the percentage of radioactive labeling was calculated to confirm its derivation from isoleucine [2] |
| References | |
| Additional Infomation |
Methylsuccinic acid is a white or pale yellow crystal or a beige powder. (NTP, 1992)
2-Methylsuccinic acid is a dicarboxylic acid formed by substituting a methyl group at the 2-position of succinic acid. It is the conjugate acid of methylsuccinic acid. Methylsuccinic acid has been reported to be present in African aloe vera, and relevant data are available. - 2-Methylsuccinic acid is a dicarboxylic acid that accumulates abnormally in ethylmalonic acid encephalopathy (EE), a rare autosomal recessive metabolic disorder [1, 2]. - The accumulation of 2-methylsuccinic acid in EE is due to impaired isoleucine degradation pathways, particularly the 2-methylacetyl-CoA thiolysis step. This blockage leads to the diversion of isoleucine metabolites to the 2-methylsuccinate production pathway [2] -2-methylsuccinate is a key diagnostic marker for EE: its elevated levels in urine and cerebrospinal fluid, combined with clinical features such as developmental delay, seizures, and acrocyanosis, help differentiate EE from other metabolic encephalopathy [1] |
| Molecular Formula |
C5H8O4
|
|---|---|
| Molecular Weight |
132.1146
|
| Exact Mass |
132.042
|
| CAS # |
498-21-5
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| Related CAS # |
2-Methylsuccinic acid-d6;347840-08-8
|
| PubChem CID |
10349
|
| Appearance |
White to off-white solid powder
|
| Density |
1.3±0.1 g/cm3
|
| Boiling Point |
236.5±13.0 °C at 760 mmHg
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| Melting Point |
110-115 °C(lit.)
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| Flash Point |
111.1±16.3 °C
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| Vapour Pressure |
0.0±1.0 mmHg at 25°C
|
| Index of Refraction |
1.474
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| LogP |
-0.24
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| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
9
|
| Complexity |
129
|
| Defined Atom Stereocenter Count |
0
|
| InChi Key |
WXUAQHNMJWJLTG-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C5H8O4/c1-3(5(8)9)2-4(6)7/h3H,2H2,1H3,(H,6,7)(H,8,9)
|
| Chemical Name |
2-methylbutanedioic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ≥ 100 mg/mL (~756.89 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (18.92 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (18.92 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (18.92 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 7.5694 mL | 37.8472 mL | 75.6945 mL | |
| 5 mM | 1.5139 mL | 7.5694 mL | 15.1389 mL | |
| 10 mM | 0.7569 mL | 3.7847 mL | 7.5694 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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