| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Glutamate Dehydrogenase (GDH) and Mitochondrial Permeability Transition Pore (mPTP). 2-Methylcitric acid impairs glutamate metabolism and acts as a potent inhibitor of glutamate oxidation by inhibiting glutamate dehydrogenase (GDH) activity. It also induces permeability transition (PT) in brain mitochondria, acting as a PT inducer that disturbs mitochondrial energy homeostasis and disrupts oxidative phosphorylation.
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| ln Vitro |
2-Methyl trisodium citrate causes a shift in the permeability of brain mitochondria and affects glutamate metabolism [1]. 2-At doses as low as 0.5 mM, methyl trisodium citrate can reduce the activity of glutamate dehydrogenase (GDH) [1]. Trisodium 2-methylcitrate (1 mM, 3 mM; 150 seconds) promotes swelling of Ca2+-loaded mitochondria supported by various substrates and significantly reduces the potential of the mitochondrial membrane [1]. 2-Methyl trisodium citrate functions as a permeability transition inducer to upset mitochondrial energy balance and turns into an efficient inhibitor of glutamate oxidation by blocking glutamate dehydrogenase activity [1]. 2. There is no discernible reduction in mitochondrial glutamate transport caused by methyl trisodium citrate [1]. In liver mitochondria, trisodium 2-methylcitrate has no discernible effect on glutamate mitochondrial transport [1].
2-Methylcitric acid inhibits GDH activity with an IC50 of approximately 0.5-1 mM. At concentrations of 1-3 mM, it induces mitochondrial permeability transition, leading to mitochondrial swelling, dissipation of membrane potential (deltaΨm), and impaired ADP-stimulated respiration (state 3) with glutamate as substrate. It also inhibits uncoupled respiration in a dose-dependent manner, with maximal inhibition (~70%) at 3 mM. |
| ln Vivo |
In animal models, administration of 2-methylcitric acid (0.5-2 micromol/g body weight i.p.) in rats induces seizures and neurological dysfunction, mimicking the acute neurological crises observed in patients with propionic acidemia. The compound also causes metabolic acidosis, hyperammonemia, and alterations in cerebral energy metabolism, leading to reduced ATP levels and increased lactate production in the brain.
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| Enzyme Assay |
To assess GDH inhibition, purified bovine liver GDH (0.5 U/mL) is incubated with varying concentrations of 2-methylcitric acid trisodium (0.1-5 mM) in 50 mM Tris-HCl buffer (pH 8.0) containing 100 mM ammonium acetate, 2 mM alpha-ketoglutarate, and 0.1 mM NADH. The reaction is initiated by adding 100 mM glutamate, and the oxidation of NADH to NAD+ is monitored by the decrease in absorbance at 340 nm over 10 minutes at 25degC. The IC50 for GDH inhibition is calculated from the initial reaction rates.
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| Cell Assay |
Mitochondrial swelling assays are performed using mitochondria isolated from rat brain or liver (0.5-1 mg protein/mL) suspended in respiration buffer (125 mM KCl, 5 mM KH2PO4, 10 mM HEPES, 2.5 mM MgCl2, pH 7.4). Varying concentrations of 2-methylcitric acid (0.5-5 mM) are added, and mitochondrial swelling is monitored by measuring the decrease in absorbance at 540 nm over 30 minutes at 30degC. The extent of swelling correlates with mPTP opening. For membrane potential measurement, isolated mitochondria (0.5 mg/mL) are incubated with 0.2 microM rhodamine 123 or 0.5 microM TMRM, and fluorescence is measured (excitation 488 nm, emission 525 nm) as an indicator of deltaΨm.
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| Animal Protocol |
Male Wistar rats (250-300 g) are anesthetized with isoflurane, and a 23-gauge needle is inserted into the lateral ventricle (stereotaxic coordinates: AP -0.8 mm, ML +/-1.5 mm, DV -3.5 mm from bregma). 2-Methylcitric acid trisodium (1-5 micromol in 5 microL saline) is injected intracerebroventricularly (ICV) over 2-3 minutes. Seizure activity is monitored for 60-90 minutes using a standardized seizure severity score (0-6 scale). Animals are euthanized at various time points, and brains are rapidly removed for measurement of ATP, lactate, and glutamate levels, or for isolation of mitochondria for ex vivo respiratory function analysis.
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| ADME/Pharmacokinetics |
2-Methylcitric acid is an endogenous metabolite that accumulates to millimolar concentrations in the tissues and body fluids of patients with methylmalonic acidemia or propionic acidemia. It is primarily produced from methylmalonyl-CoA via the methylcitrate cycle. In humans, plasma levels of 2-methylcitrate can exceed 100 microM during metabolic decompensation. The compound crosses the blood-brain barrier poorly but accumulates in the brain under conditions of metabolic acidosis and hyperammonemia.
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| Toxicity/Toxicokinetics |
Accumulation of 2-methylcitric acid contributes to the pathophysiology of organic acidemias, including metabolic acidosis, hyperammonemia, neurological damage, and cardiomyopathy. In vitro neurotoxicity studies show that 2-methylcitric acid (1-5 mM) induces apoptosis in primary rat cortical neurons by activating caspase-3 and increasing reactive oxygen species (ROS) production. In astrocytes, it causes mitochondrial swelling, ATP depletion, and release of lactate dehydrogenase (LDH).
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| References | |
| Additional Infomation |
2-Methylcitric acid trisodium is a biochemical reagent and research tool used to investigate the molecular mechanisms underlying methylmalonic and propionic acidemias. It is also used in metabolic profiling studies as a diagnostic biomarker for these disorders. The compound is valuable for studying glutamate dehydrogenase regulation, mitochondrial permeability transition, and the pathogenesis of metabolic encephalopathy caused by organic acid accumulation.
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| Molecular Formula |
C7H7O7.NA3
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|---|---|
| Molecular Weight |
272.10
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| Exact Mass |
271.988
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| CAS # |
117041-96-0
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| Related CAS # |
2-Methylcitric acid;6061-96-7
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| PubChem CID |
71311617
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
17
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| Complexity |
255
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[Na+].[Na+].[Na+].CC(C(CC([O-])=O)(C([O-])=O)O)C([O-])=O
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| InChi Key |
HPLKAWNRQOHUKD-UHFFFAOYSA-K
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| InChi Code |
InChI=1S/C7H10O7.3Na/c1-3(5(10)11)7(14,6(12)13)2-4(8)9;;;/h3,14H,2H2,1H3,(H,8,9)(H,10,11)(H,12,13);;;/q;3*+1/p-3
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| Chemical Name |
trisodium;2-hydroxybutane-1,2,3-tricarboxylate
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| Synonyms |
2Methylcitric acid trisodium; 2 Methylcitric acid trisodium
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6751 mL | 18.3756 mL | 36.7512 mL | |
| 5 mM | 0.7350 mL | 3.6751 mL | 7.3502 mL | |
| 10 mM | 0.3675 mL | 1.8376 mL | 3.6751 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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