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| Other Sizes |
| Targets |
The primary target of Arsenazo III is metal ions, particularly calcium (Ca²⁺) and other divalent and trivalent cations. Its mechanism of action is based on its ability to form colored complexes with these metal ions. Upon binding to calcium or other metals, the dye undergoes a conformational change that results in a shift in its absorption spectrum, producing a color change. This property allows it to be used as an indicator for the presence and concentration of these ions in solution. The change in color is directly proportional to the metal ion concentration, enabling quantitative analysis.
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| ln Vitro |
In vitro, Arsenazo III is used as a sensitive reagent for the spectrophotometric determination of various metals. It is employed for the colorimetric determination of thorium and uranium. The dye forms complexes with a wide range of metal ions, including barium, bismuth, calcium, copper, iron, lanthanum, neptunium, palladium, polonium, plutonium, rare earth metals, scandium, thorium, uranium, and zirconium. It is also used as a metallochrome indicator, changing color when complexed to calcium ions under physiological conditions.
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| ln Vivo |
In vivo, Arsenazo III has been used to measure local calcium ion concentrations. Because it changes color upon binding to calcium, it can be injected into tissues or cells, and the color change can be monitored to provide real-time information about calcium dynamics. This application is particularly useful in physiological studies where calcium plays a critical role as a second messenger. However, due to its high molecular weight and the presence of arsenic atoms, its use in vivo is limited to specific research applications.
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| Enzyme Assay |
In vitro enzyme/receptor binding (non-cell) assays using Arsenazo III are typically based on its metal-binding properties. A standard protocol for calcium determination: a sample containing calcium ions is mixed with a solution of Arsenazo III in a buffer at a specific pH (e.g., pH 7.0). The absorbance of the mixture is measured at a wavelength of 650-660 nm using a UV-Vis spectrophotometer. The calcium concentration is determined by comparing the absorbance to a standard curve prepared with known concentrations of calcium. This assay is widely used in clinical chemistry and biological research to measure calcium levels in biological fluids.
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| Cell Assay |
In vitro cell-based assays for Arsenazo III are not common, as it is primarily a reagent for metal detection rather than a cell-active compound. However, it can be used to measure intracellular calcium concentrations. A standard protocol: cells are loaded with a membrane-permeant form of Arsenazo III (e.g., by incubation with the dye in culture medium). After loading, cells are washed and placed in a cuvette or on a coverslip for microscopy. The absorbance or fluorescence of the dye is monitored in response to stimuli that trigger calcium release, such as the addition of a calcium ionophore or a receptor agonist. The change in signal reflects changes in intracellular calcium concentration.
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| Animal Protocol |
In vivo animal experiments using Arsenazo III are limited to specialized physiological studies. A standard protocol for measuring calcium in vivo: Arsenazo III is injected intravenously or directly into a tissue of interest. A fiber optic probe or a microelectrode is used to monitor the absorbance or reflectance of the dye in the tissue. Changes in the signal are correlated with changes in local calcium concentration. This technique has been used to study calcium dynamics in the heart, brain, and other organs. However, due to the presence of arsenic, the use of Arsenazo III in vivo requires careful consideration of toxicity.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of Arsenazo III are not well-characterized. The compound is highly water-soluble and has a high molecular weight (776.37 g/mol). It is not expected to cross biological membranes readily. When injected into the bloodstream, it would likely remain in the vascular space and be cleared by the kidneys. Its distribution would be limited to the extracellular compartment. The presence of arsenic atoms in the molecule raises concerns about potential toxicity and accumulation, which limits its use in vivo.
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| Toxicity/Toxicokinetics |
Toxicity data for Arsenazo III is limited but significant due to its arsenic content. Arsenic compounds are known to be toxic, and Arsenazo III contains two arsenic atoms per molecule. The compound should be handled with extreme caution. Inhalation, ingestion, and skin contact should be avoided. Appropriate personal protective equipment, including gloves and a lab coat, should be worn when handling the compound. Waste should be disposed of according to regulations for hazardous and arsenic-containing waste. The compound is intended for research use only and not for human consumption.
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| References | |
| Additional Infomation |
A chromium indicator that changes color after complexing with calcium ions under physiological conditions. It is used to measure local calcium ion concentration in the body.
Arsenazo III is a metal-binding dye used as a spectrophotometric reagent for the determination of various metals. It is a metallochrome indicator that changes color when complexed to calcium ions under physiological conditions. It is used to measure local calcium ion concentrations in vivo and as a reagent for the colorimetric determination of thorium and uranium. The compound is commercially available from chemical suppliers for research purposes only. It is not a drug and has no clinical approval or therapeutic use. |
| Molecular Formula |
C22H18AS2N4O14S2
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|---|---|
| Molecular Weight |
776.3699
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| Exact Mass |
775.869
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| CAS # |
1668-00-4
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| PubChem CID |
135445682
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| Appearance |
Brown to black solid powder
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| Melting Point |
>320ºC(lit.)
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| LogP |
-5.25
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
18
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
44
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| Complexity |
1300
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
UQHVTNUJRKELCE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H18As2N4O14S2/c29-21-18-11(9-16(43(37,38)39)19(21)27-25-14-7-3-1-5-12(14)23(31,32)33)10-17(44(40,41)42)20(22(18)30)28-26-15-8-4-2-6-13(15)24(34,35)36/h1-10,29-30H,(H2,31,32,33)(H2,34,35,36)(H,37,38,39)(H,40,41,42)
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| Chemical Name |
3,6-bis[(2-arsonophenyl)diazenyl]-4,5-dihydroxynaphthalene-2,7-disulfonic 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 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 : ~10 mg/mL (~12.88 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 | 1.2880 mL | 6.4402 mL | 12.8805 mL | |
| 5 mM | 0.2576 mL | 1.2880 mL | 2.5761 mL | |
| 10 mM | 0.1288 mL | 0.6440 mL | 1.2880 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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