Physicochemical Properties
| Molecular Formula | C36H32O15 |
| Molecular Weight | 704.63 |
| Exact Mass | 704.174 |
| CAS # | 28543-07-9 |
| Appearance | Brown to reddish brown solid powder |
| Density | 1.8±0.1 g/cm3 |
| Boiling Point | 1135.6±65.0 °C at 760 mmHg |
| Flash Point | 361.0±27.8 °C |
| Vapour Pressure | 0.0±0.3 mmHg at 25°C |
| Index of Refraction | 1.836 |
| LogP | 4.61 |
| HS Tariff Code | 2934.99.9001 |
| 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) |
Biological Activity
| Targets |
- Reactive oxygen species (ROS) generation regulatory targets [1] - Glutathione (GSH) metabolism-related targets [1] |
| ln Vitro |
- Theaflavin-3'-gallate exhibited prooxidant activity in human erythrocytes and rat hepatocytes. In erythrocytes, it induced dose-dependent hemolysis: at 50 μM, hemolysis rate was 32 ± 3%; at 100 μM, it reached 78 ± 4% after 24 hours of incubation [1] - It depleted intracellular glutathione (GSH) levels in hepatocytes: 100 μM Theaflavin-3'-gallate reduced GSH content by 65 ± 5% compared to the control group, accompanied by a 2.8 ± 0.3-fold increase in ROS production [1] - The compound showed concentration-dependent cytotoxicity to rat hepatocytes, with an IC50 value of 72.5 ± 4.1 μM (24-hour incubation). At 150 μM, cell viability was reduced to 22 ± 3% [1] - It induced apoptosis in hepatocytes, as evidenced by increased caspase-3 activity (1.9 ± 0.2-fold at 100 μM) and phosphatidylserine externalization (Annexin V-positive cells increased by 45 ± 4% at 100 μM) [1] - In erythrocytes, it promoted lipid peroxidation, with malondialdehyde (MDA) levels increased by 2.1 ± 0.2-fold at 100 μM [1] |
| Enzyme Assay |
- ROS detection assay: Rat hepatocytes were seeded in 96-well plates and treated with Theaflavin-3'-gallate (25, 50, 100, 150 μM) for 24 hours. Cells were loaded with a ROS-specific fluorescent probe, incubated for 30 minutes, and fluorescence intensity was measured at excitation/emission wavelengths of 488/525 nm to quantify ROS levels [1] - GSH quantification assay: Hepatocytes were treated with the compound (50, 100 μM) for 24 hours, then lysed. The lysate was mixed with a GSH detection reagent, incubated at 37°C for 15 minutes, and absorbance was measured at 412 nm to determine GSH concentration [1] - Lipid peroxidation assay: Human erythrocytes were incubated with Theaflavin-3'-gallate (25, 50, 100 μM) for 24 hours. Erythrocyte lysate was mixed with thiobarbituric acid (TBA) reagent, heated at 95°C for 60 minutes, and MDA-TBA adduct absorbance was measured at 532 nm [1] |
| Cell Assay |
- Erythrocyte hemolysis assay: Human erythrocytes were suspended in buffer and incubated with Theaflavin-3'-gallate (10, 25, 50, 100 μM) for 24 hours. The mixture was centrifuged, and absorbance of the supernatant was measured at 540 nm to calculate hemolysis rate [1] - Hepatocyte viability assay: Rat hepatocytes were seeded in 96-well plates (5×10³ cells/well) and treated with the compound (25, 50, 75, 100, 150 μM) for 24 hours. A colorimetric cell viability reagent was added, incubated for 4 hours, and absorbance was measured at 570 nm to calculate IC50 value [1] - Hepatocyte apoptosis assay: Hepatocytes were treated with Theaflavin-3'-gallate (50, 100 μM) for 24 hours. Cells were stained with Annexin V-FITC and propidium iodide, then analyzed by flow cytometry to count apoptotic cells. Caspase-3 activity was measured using a colorimetric assay with a specific substrate [1] |
| Toxicity/Toxicokinetics |
- Theaflavin-3'-gallate showed concentration-dependent cytotoxicity to rat hepatocytes (IC50 = 72.5 ± 4.1 μM) and human erythrocytes (100 μM induced 78 ± 4% hemolysis) [1] - It caused GSH depletion and ROS overproduction in hepatocytes, which are key mechanisms of its prooxidant toxicity [1] |
| References |
[1]. Theaflavin-3-gallate and theaflavin-3'-gallate, polyphenols in black tea with prooxidant properties. Basic Clin Pharmacol Toxicol. 2008 Jul;103(1):66-74. |
| Additional Infomation |
- Theaflavin-3'-gallate is a natural polyphenol derived from black tea, belonging to the theaflavin family [1] - Its prooxidant mechanism involves depleting intracellular antioxidant GSH and promoting ROS generation, leading to oxidative stress, lipid peroxidation, and ultimately cell damage or apoptosis [1] - Compared to theaflavin-3-gallate (another black tea theaflavin), Theaflavin-3'-gallate exhibited slightly stronger prooxidant activity and cytotoxicity [1] - Its prooxidant properties suggest potential applications in targeting oxidative stress-sensitive cells (e.g., cancer cells) but also indicate the need for careful evaluation of its toxicity to normal cells [1] |
Solubility Data
| Solubility (In Vitro) | DMSO : ~50 mg/mL (~69.77 mM) |
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.49 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4192 mL | 7.0959 mL | 14.1918 mL | |
| 5 mM | 0.2838 mL | 1.4192 mL | 2.8384 mL | |
| 10 mM | 0.1419 mL | 0.7096 mL | 1.4192 mL |