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| 5mg |
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| 10mg |
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In the NAD+ biosynthesis pathway, nicotinic acid mononucleotide triethylamine is the product of the first committed step of the Preiss-Handler pathway. It is formed from nicotinic acid (NA) by the enzyme Nicotinic Acid Phosphoribosyltransferase (NAPRT). Following its formation, NAMN is then adenylylated by Nicotinamide Mononucleotide Adenylyltransferase (NMNAT) to form Nicotinic Acid Adenine Dinucleotide (NAAD). NAAD is subsequently amidated by NAD+ Synthase to produce the final product, NAD+. By providing this intermediate, the compound allows researchers to bypass upstream enzymatic steps in the pathway and specifically study the activity of downstream enzymes like NMNAT. As a substrate for these enzymes, it is a critical tool for metabolic pathway dissection.
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| ln Vitro |
The in vitro activity of nicotinic acid mononucleotide triethylamine is defined by its role as a substrate for NMNAT. In an in vitro enzyme assay, it is used to measure the kinetic parameters of NMNAT enzymes. The conversion of NAMN to NAAD by NMNAT is coupled with a colorimetric or fluorometric readout. As a building block for NAD+ synthesis, it can also be used in cell culture to boost NAD+ levels in NAPRT-expressing cells. In a cell-based context, supplementing the media with nicotinic acid (NA) leads to the production of this intermediate; however, the direct addition of this charged mononucleotide may not penetrate the cell membrane efficiently. The triethylamine salt is provided primarily as a cell-free or biochemical assay standard to quantify NAMN levels in biological samples.
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| ln Vivo |
The in vivo effects of nicotinic acid mononucleotide (NAMN) are extrapolated from its role as an NAD+ precursor in the Preiss-Handler pathway. While it is not typically administered directly in animals due to its charged nature, increasing the flux through this pathway by supplementing with niacin (Vitamin B3) leads to elevated NAD+ levels. Elevated NAD+ has been shown to activate sirtuins, improve mitochondrial function, and protect against metabolic diseases in animal models. The triethylamine salt is primarily for use as an analytical standard for quantifying NAMN levels in tissue extracts, plasma, and urine by LC-MS/MS. This is crucial for understanding the metabolic flux through the NAD+ salvage pathways in various disease states.
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| Enzyme Assay |
Cell-free enzyme assays are used to characterize nicotinic acid mononucleotide triethylamine as a substrate. For a NMNAT activity assay: A 96-well plate is prepared with a reaction mixture containing 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, and the substrate (Nicotinic acid mononucleotide) at concentrations ranging from 0 to 500 uM. The reaction is initiated by adding recombinant NMNAT1-3 enzyme (e.g., 10 ng/well) and ATP (1-5 mM). The plate is incubated at 37degC for 30-60 minutes. The enzymatic activity can be monitored in real-time using a coupled enzyme assay or by terminating the reaction with EDTA and using a NAD/NAAD detection kit. Alternatively, the product (NAAD) is quantified using HPLC or LC-MS/MS by comparing the peak area to standard curves generated with known concentrations of the NAMN TEA salt. This allows for the calculation of Km and Vmax values for NMNAT enzymes.
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| Cell Assay |
NAMN levels in cell or tissue samples can be quantified using LC-MS/MS. For a typical experiment, cells (e.g., HEK293 or primary hepatocytes) are seeded in 6-well plates and treated with nicotinic acid (NA, 10-1000 uM) for 24-48 hours. The media is aspirated, and the cells are washed with cold PBS. Metabolites are extracted by adding 500 microL of ice-cold 80% methanol/20% water (v/v) containing an internal standard (e.g., ¹3C-NAMN). Cells are scraped, transferred to a microcentrifuge tube, and incubated at -80degC for 10 minutes. The lysates are centrifuged at 15,000 rpm for 10 minutes at 4degC, and the supernatant is transferred to a new tube and dried down using a SpeedVac. The dried extract is reconstituted in a suitable LC-MS mobile phase. The reconstituted sample is injected onto a C18 reverse-phase column coupled to a triple quadrupole mass spectrometer operating in positive or negative ion mode. The concentration of NAMN is determined by comparing the peak area ratio (analyte/internal standard) to a calibration curve generated from known amounts of the nicotinic acid mononucleotide triethylamine standard.
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| Animal Protocol |
A typical in vivo protocol involves the administration of nicotinic acid (NA) as a precursor, followed by the detection of NAMN (this compound) as a biomarker. Adult C57BL/6 mice are administered nicotinic acid (50-200 mg/kg) by oral gavage or intraperitoneal injection. Blood is collected via retro-orbital bleeding or tail vein at various time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24 hours). Plasma is separated by centrifugation and stored at -80degC. Liver tissue is also collected and flash-frozen in liquid nitrogen. Metabolites are extracted from plasma (100 uL) or tissue (50 mg) using a methanol-based extraction method. The extracted samples are analyzed by LC-MS/MS specifically for NAMN levels. The TEA salt of NAMN is used as the primary standard to generate calibration curves and validate the specificity of the mass spectrometry method. The data is used to calculate the PK parameters of NA in terms of its conversion to downstream metabolites.
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| ADME/Pharmacokinetics |
The nicotinic acid mononucleotide triethylamine salt (molecular formula: C17H29N2O9P, MW: 436.4 g/mol) is a stable, research-grade compound. It should be stored as a powder at -20degC, protected from moisture, where it is stable for up to 3 years. The triethylamine counterion is used to improve the aqueous solubility of the highly polar mononucleotide. It is soluble in water and DMSO. It is not intended for in vivo administration as a therapeutic. As a natural metabolite, it is expected to have a very short half-life in circulation due to rapid cellular uptake and further metabolism by NMNAT enzymes. It is primarily used as a high-quality analytical standard and a substrate for in vitro biochemistry.
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| Toxicity/Toxicokinetics |
This product is a research tool and is not for human consumption. As it is an intermediate in the NAD+ biosynthesis pathway, which is essential for life, it has no known direct toxicity at research doses. However, the solvents or vehicles used for reconstitution (DMSO, methanol) may be toxic. Standard laboratory safety practices should be followed. The TFA salt form is not applicable; this is a triethylamine (TEA) salt, which is generally considered less toxic than TFA. The primary potential hazard is the TEA component, which can be an irritant in high concentrations, but it is present in stoichiometric amounts. The compound is for research use only, and its hazards are generally limited to the physical hazards of handling fine powders (inhalation, eye contact).
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| References | |
| Additional Infomation |
This compound is an important tool for metabolomics and biochemistry research. The Preiss-Handler pathway is one of three pathways for NAD+ synthesis in mammals (the others are the De Novo pathway from tryptophan and the salvage pathway from nicotinamide). It is the primary pathway for converting dietary niacin (vitamin B3) into NAD+. Nicotinic acid mononucleotide (NAMN) is a specific and quantifiable biomarker of the activity of the Preiss-Handler pathway. Researchers use this compound as a standard to understand how metabolic diseases, aging, or drug treatments affect NAD+ homeostasis. The triethylamine salt form is a common synthetic counterion used to improve the solubility and handling of nucleotide intermediates for research applications.
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| Molecular Formula |
C11H14NO9P.1.7C6H15N
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| Related CAS # |
Nicotinic acid mononucleotide;321-02-8
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| Appearance |
Off-white to light yellow solid powder
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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) |
H2O :~125 mg/mL (~372.91 mM)
DMSO :~100 mg/mL (~298.33 mM ) |
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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.) |
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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