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| 10mg |
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| 50mg |
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| Targets |
Tempone-H is a spin trap that targets reactive oxygen species (ROS), specifically superoxide radicals and peroxynitrite. Its mechanism of action involves the reaction of the stable nitroxide radical with these highly reactive species. The reaction results in a change in the EPR spectrum of the nitroxide, which can be detected and quantified. This property allows researchers to measure the formation of these radicals in biological systems, providing insights into oxidative stress, aging, and neurodegeneration.
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| ln Vitro |
TEMPONE-H has a sensitivity that is roughly ten times higher than spin traps DMPO or TMIO for the detection of peroxynitrite or superoxide radicals [1].
In vitro, Tempone-H is used as a spin trap to quantify the formation of peroxynitrite and superoxide radicals in chemical and biological systems. Its stable radical properties allow it to provide valuable insights into cellular environments, especially in the context of oxidative stress. It is used in EPR spectroscopy to detect and quantify free radical formation. Ferric and cupric ions are effective oxidants of Tempone-H, which is a consideration in its use. |
| ln Vivo |
In vivo activity of Tempone-H is related to its role as a spin trap for detecting ROS in living systems. Its cell-permeable nature allows it to be used in vivo to detect superoxide radical and peroxynitrite formation. It has been investigated for its potential to provide insights into oxidative stress-related conditions such as aging and neurodegeneration. Its ability to quantify ROS in vivo makes it a valuable tool for studying the role of oxidative stress in disease.
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| Enzyme Assay |
In vitro enzyme/receptor binding (non-cell) assays using Tempone-H are based on its role as a spin trap. A standard protocol for detecting superoxide radicals: a sample containing a superoxide-generating system (e.g., xanthine/xanthine oxidase) is incubated with Tempone-H. The reaction is monitored by EPR spectroscopy, which detects the nitroxide radical signal. The decrease in the EPR signal is proportional to the amount of superoxide produced. This assay is used to measure the activity of superoxide-producing enzymes or to screen for compounds that scavenge superoxide radicals.
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| Cell Assay |
In vitro cell-based assays using Tempone-H involve detecting intracellular ROS production. A standard protocol: cells are incubated with Tempone-H (e.g., 1-10 mM) for a period of time (e.g., 1-2 hours). The cells are then stimulated with an agent that induces ROS production (e.g., phorbol myristate acetate or hydrogen peroxide). The cells are lysed, and the EPR signal of Tempone-H is measured. An increase in the EPR signal indicates the formation of superoxide or peroxynitrite radicals. Alternatively, the cells can be analyzed by EPR spectroscopy directly.
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| Animal Protocol |
In vivo animal experiments using Tempone-H involve administering the compound to animals to measure ROS production in tissues. A standard protocol: Tempone-H is administered intraperitoneally or intravenously to mice. After a period for distribution (e.g., 30-60 minutes), tissues (e.g., brain, heart, liver) are collected and homogenized. The EPR signal of the nitroxide radical is measured in the tissue homogenates. The level of the EPR signal reflects the amount of ROS produced in the tissue. This approach is used to study oxidative stress in various disease models.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of Tempone-H are not well-characterized. The compound is a small molecule (MW 171.24 g/mol) and is cell-permeable. It is water-soluble and stable. Its distribution in vivo would depend on its ability to cross biological membranes. The compound is typically stored as a powder at room temperature. Its reactivity with ROS and its stability are the main considerations for its use as a spin trap.
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| Toxicity/Toxicokinetics |
Toxicity data for Tempone-H is limited. As a spin trap used in research, it is generally considered to have low toxicity at the concentrations used for experiments. However, as with all chemical reagents, appropriate safety precautions should be taken when handling the compound. Inhalation, ingestion, and skin contact should be avoided. The compound is not intended for human use.
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| References |
[1]. Dikalov S, et al. Quantification of peroxynitrite, superoxide, and peroxyl radicals by a new spin trap hydroxylamine 1-hydroxy-2,2,6,6-tetramethyl-4-oxo-piperidine. Biochem Biophys Res Commun. 1997 Jan 3;230(1):54-7.
[2]. Dikalov SI, et al. Amyloid beta peptides do not form peptide-derived free radicals spontaneously, but can enhance metal-catalyzed oxidation of hydroxylamines to nitroxides. J Biol Chem. 1999 Apr 2;274(14):9392-9. |
| Additional Infomation |
Tempone-H is a stable, nitrogen-centered radical compound used as a spin trap in chemical and biological systems. It is used to quantify the formation of peroxynitrite and superoxide radicals. It is also used in MRI and EPR spectroscopy. The compound is cell-permeable. It is not a drug and has no clinical use or approval status. It is commercially available from chemical suppliers for research purposes only.
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| Molecular Formula |
C9H17NO2
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|---|---|
| Molecular Weight |
171.23678
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| Exact Mass |
171.126
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| CAS # |
3637-11-4
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| PubChem CID |
98642
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| Appearance |
Light yellow to brown solid powder
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| Density |
1.02g/cm3
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| Boiling Point |
257.5ºC at 760mmHg
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| Flash Point |
109.5ºC
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| LogP |
1.535
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
12
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| Complexity |
189
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| Defined Atom Stereocenter Count |
0
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| SMILES |
ON1C(C)(C)CC(=O)CC1(C)C
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| InChi Key |
KMEUSKGEUADGET-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C9H17NO2/c1-8(2)5-7(11)6-9(3,4)10(8)12/h12H,5-6H2,1-4H3
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| Chemical Name |
1-hydroxy-2,2,6,6-tetramethylpiperidin-4-one
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 : ~125 mg/mL (~729.97 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 5.8398 mL | 29.1988 mL | 58.3976 mL | |
| 5 mM | 1.1680 mL | 5.8398 mL | 11.6795 mL | |
| 10 mM | 0.5840 mL | 2.9199 mL | 5.8398 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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