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| Other Sizes |
| Targets |
TPTZ has no biological target; it is a chemical chelator. Its "target" is ferrous ions (Fe2+). The three pyridine nitrogen atoms serve as electron donors, coordinating to the Fe2+ ion to form a 2:1 ligand-to-metal complex, [Fe(TPTZ)2]2+. This complex is highly stable and absorbs light in the visible region (595-600 nm). The intensity of the purple color is directly proportional to the Fe2+ concentration, according to the Beer-Lambert law. This is the basis for its use as a chromogenic agent. It is also used to prepare electrode sensors.
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| ln Vitro |
TPTZ is not a drug and has no pharmacological activity. Its in vitro activity is analytical: the formation of a purple chromophore with Fe2+. In the standard ferric reducing antioxidant power (FRAP) assay, TPTZ is used to measure the total antioxidant capacity of a sample. In acidic conditions (pH 3.6), the ferric-iron (Fe3+)-TPTZ complex is reduced to the ferrous (Fe2+)-TPTZ complex by antioxidants, leading to a blue color (λmax ~593 nm). The change in absorbance is measured and correlates with the reducing power of the sample.
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| ln Vivo |
TPTZ is not administered in vivo for therapeutic effect. It is used as a staining agent to detect iron in histological sections. For example, tissue sections are incubated with a solution of TPTZ and a reducing agent (e.g., hydroxylamine) to convert all iron to Fe2+. The sections are then stained, and sites of iron accumulation (e.g., in the liver or spleen in iron overload disorders) appear purple. This is a diagnostic staining technique. TPTZ is also used in clinical chemistry analyzers for measuring serum iron and total iron-binding capacity (TIBC).
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| Enzyme Assay |
In a non-cellular FRAP assay, TPTZ (10 mM) is dissolved in 40 mM HCl. The FRAP reagent is prepared by mixing 25 mL of 0.3 M acetate buffer (pH 3.6), 2.5 mL of 10 mM TPTZ solution, and 2.5 mL of 20 mM FeCl3 solution. A 10 uL sample is mixed with 300 uL of freshly prepared FRAP reagent. After a 4-30 min incubation at 37degC, the absorbance at 593 nm is measured. A standard curve is prepared using FeSO4 solutions (100-1000 uM). This method measures the ability of a sample to reduce Fe3+ to Fe2+, reflecting its antioxidant capacity.
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| Cell Assay |
TPTZ is not used in live cell assays as a treatment. It can be used in cell lysates or biological fluids to measure total iron or antioxidant capacity. A typical protocol: a cell pellet (1×10⁶ cells) is lysed in RIPA buffer. The lysate is cleared by centrifugation. To measure ferric reducing antioxidant power (FRAP), 10 uL of the lysate is added to 300 uL of FRAP reagent (containing TPTZ). After 10-20 minutes at 37degC, the absorbance is read at 593 nm. A parallel set of samples is used to create a protein standard curve for normalization. This assay provides a measure of the total reducing capacity of the cell lysate.
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| Animal Protocol |
For iron staining in animal tissue, a rat model of iron overload is used. Male Sprague-Dawley rats are injected with iron dextran (e.g., 500 mg Fe/kg, IP) once weekly for 4 weeks. Control rats receive saline. After 4 weeks, the rats are euthanized, and the liver and spleen are excised. Tissue sections (5 um) are cut and incubated with 0.1 M potassium thiocyanate and a reducing agent (e.g., 10% hydroxylamine hydrochloride) for 30 minutes, then stained with 0.1% TPTZ in acetate buffer (pH 4.5) for 30 minutes. Iron deposits are visualized by light microscopy and appear as purple-blue granules. This protocol is used to assess iron accumulation in pathological conditions.
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| ADME/Pharmacokinetics |
TPTZ has a molecular formula of C1₈H12N₆ and a molecular weight of 312.33 g/mol. It is a yellow crystalline powder with a melting point of 247-249degC. It is soluble in methanol (100 mg/mL) and DMSO but only slightly soluble in water. It is stored at 2-8degC, protected from light. The compound is not intended for systemic use. It has a density of 1.276 g/cm3. TPTZ is a chromogenic reagent for iron detection.
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| Toxicity/Toxicokinetics |
TPTZ is classified as a mild skin and eye irritant. It may cause respiratory irritation if inhaled. It is not a drug. Standard laboratory safety practices should be followed (gloves, lab coat, goggles). Avoid creating dust. The compound is for research and manufacturing use (analytical chemistry).
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| References | |
| Additional Infomation |
2,4,6-Tris(2'-pyridyl)-1,3,5-triazine is the parent compound.
2,4,6-Tri-2-pyridinyl-1,3,5-triazine (TPTZ, CAS 3682-35-7) is the standard chromogenic reagent in the widely used Ferric Reducing Antioxidant Power (FRAP) assay for measuring the total antioxidant capacity of plasma, food extracts, and nutraceuticals. It is also used in clinical laboratories for the accurate determination of serum iron levels. The compound is a chelator, a colorimetric reagent, a fluorescent dye, and an iron detection probe. It is for research and analytical use only. |
| Molecular Formula |
C18H12N6
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|---|---|
| Molecular Weight |
312.33
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| Exact Mass |
312.112
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| CAS # |
3682-35-7
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| Related CAS # |
63451-30-9;monoperchlorate
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| PubChem CID |
77258
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| Appearance |
White to yellow solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
618.9±58.0 °C at 760 mmHg
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| Melting Point |
248-252ºC
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| Flash Point |
288.2±25.3 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.644
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| LogP |
2.3
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
323
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N1C(C2=C([H])C([H])=C([H])C([H])=N2)=NC(C2=C([H])C([H])=C([H])C([H])=N2)=NC=1C1=C([H])C([H])=C([H])C([H])=N1
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| InChi Key |
KMVWNDHKTPHDMT-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H12N6/c1-4-10-19-13(7-1)16-22-17(14-8-2-5-11-20-14)24-18(23-16)15-9-3-6-12-21-15/h1-12H
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| Chemical Name |
2,4,6-tripyridin-2-yl-1,3,5-triazine
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| Synonyms |
2,4,6-Tri-2-pyridyl-1,3,5-s-triazine; NSC 112125; 2,4,6-Tipyidyl-s-tiazie
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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) |
Methanol : ~50 mg/mL (~160.09 mM; with ultrasonication)
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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 | 3.2017 mL | 16.0087 mL | 32.0174 mL | |
| 5 mM | 0.6403 mL | 3.2017 mL | 6.4035 mL | |
| 10 mM | 0.3202 mL | 1.6009 mL | 3.2017 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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