| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
Tetramethylfluoroformamidinium hexafluorophosphate does not target biological receptors. It is a chemical reagent used exclusively in organic synthesis for peptide bond formation. Its mechanism of action is chemical: it activates carboxylic acids by converting them to highly reactive acid fluorides in the presence of a base (e.g., DIPEA, TEA). The acid fluoride intermediate then reacts efficiently with the amine group of another amino acid to form an amide bond. TFFH is particularly useful for coupling hindered amino acids where traditional coupling agents (e.g., HOBt, HBTU) may be less effective. It is also used for the preparation of various carboxylic acid derivatives, including active esters and thioesters.
|
|---|---|
| ln Vitro |
The compound does not possess in vitro biological activity; it is strictly used as a chemical reagent. In peptide synthesis, the biological activity of the final peptide product, however, depends on the correct sequence and stereochemistry, which TFFH helps to achieve. By enabling efficient coupling of hindered amino acids, TFFH helps researchers synthesize biologically active peptides (e.g., peptide hormones, enzyme inhibitors, antimicrobial peptides) that may be difficult to produce with other coupling reagents. TFFH is effective even for challenging sequences, including those with N-methyl amino acids, C-terminal alpha,alpha-dialkyl amino acids, and other sterically hindered residues.
|
| ln Vivo |
The compound does not exhibit in vivo biological activity; it is not intended for in vivo use as a therapeutic agent. TFFH is a chemical reagent that is used for the synthesis of peptides and other compounds. The biological activity in vivo is exhibited by the final peptide product, not by the coupling reagent. When used in the synthesis of therapeutic peptides or peptide-based drugs (such as GLP-1 agonists, antibiotics, or peptide vaccines), TFFH can enable the efficient synthesis of these biologically active molecules. However, TFFH itself is not administered to animals or humans.
|
| Enzyme Assay |
A standard non-cellular (cell-free) protocol for using TFFH in solid-phase peptide synthesis (SPPS) involves the activation of an Fmoc-protected amino acid. The Fmoc-amino acid (1 mmol) is dissolved in 4 mL of DMF (N,N-dimethylformamide) or NMP (N-methyl-2-pyrrolidone). TFFH (1.1 mmol, 0.29 g) and DIPEA (N,N-diisopropylethylamine, 2-3 mmol, 0.35-0.52 mL) are added. The mixture is stirred for 5-10 minutes at room temperature to generate the reactive acid fluoride in situ. The activation solution is then added to the resin (substitution 0.6-0.8 mmol/g) containing the growing peptide chain with a free amine at the N-terminus. The coupling reaction is allowed to proceed for 30-60 minutes at room temperature with occasional mixing. The resin is drained and washed 3 times with DMF. Coupling efficiency can be monitored by the Kaiser test (ninhydrin test). If coupling is incomplete, a double coupling may be performed. After completion, Fmoc deprotection is carried out using 20% piperidine in DMF (2×10 min).
|
| Cell Assay |
Since TFFH is a chemical reagent used for organic synthesis and not a compound tested on live cells, there is no standard in vitro cell culture protocol for this compound. However, a protocol can be described for using a peptide synthesized with TFFH in cell culture experiments. After the peptide is synthesized using TFFH as a coupling reagent, it is cleaved from the resin (using TFA/triisopropylsilane/water (95:2.5:2.5)), precipitated with cold diethyl ether, purified by HPLC (reverse-phase C18 column), and lyophilized. The purified peptide is dissolved in a suitable solvent (e.g., DMSO, PBS, or cell culture medium) and tested in biological assays. For example, to assess the activity of a peptide hormone (e.g., a GLP-1 analogue), HEK293 cells expressing the GLP-1 receptor are seeded in 96-well plates at 1×10⁴ cells/well and cultured overnight. The next day, cells are treated with various concentrations of the peptide (0.1 pM to 1 microM) for 30 minutes, and cAMP levels are measured by a homogeneous time-resolved fluorescence (HTRF) cAMP assay. The EC50 of the peptide is calculated.
|
| Animal Protocol |
No in vivo animal protocol exists for TFFH itself, as it is not intended for in vivo administration. However, a protocol can be described for using a peptide synthesized with TFFH in animal studies. After the peptide is synthesized and purified, it is formulated in a suitable vehicle (e.g., saline, PBS, or 5% DMSO in saline). For an efficacy study, male C57BL/6 mice (6-8 weeks old) are administered the peptide via intraperitoneal (ip) injection (e.g., 0.1-10 mg/kg). Blood samples are collected at various time points for pharmacokinetic analysis (ELISA or LC-MS/MS to measure peptide concentration) or pharmacodynamic readouts (e.g., blood glucose measurement for a GLP-1 analog). Control groups receive vehicle only. For toxicology studies, animals are treated with the peptide for up to 14-28 days, and body weight, food consumption, clinical signs, and organ weights are monitored. The investigator can then attribute any biological activity to the peptide itself, not the synthetic reagent (TFFH) used in its production.
|
| ADME/Pharmacokinetics |
Tetramethylfluoroformamidinium hexafluorophosphate is a chemical reagent; as such, it is not characterized by typical pharmacokinetic parameters (Cmax, Tmax, AUC, half-life) because it is not administered as a drug. When used in peptide synthesis, TFFH is not present in the final product after purification. Any residual TFFH in the final peptide product would be considered an impurity and must be removed during purification. The reagent itself is unstable in aqueous solutions and degrades rapidly, which is another reason why it is not suitable as a therapeutic agent. Researchers should ensure that final peptide products are purified to remove traces of TFFH and other synthetic reagents before biological testing.
|
| Toxicity/Toxicokinetics |
Tetramethylfluoroformamidinium hexafluorophosphate is a chemical reagent and should be handled with appropriate safety precautions. The compound is corrosive; it causes severe skin burns and eye damage. It is harmful if swallowed, inhaled, or absorbed through the skin. Use only in a well-ventilated area (e.g., a chemical fume hood). Wear appropriate personal protective equipment: chemical-resistant gloves (e.g., nitrile), safety goggles, and a lab coat. In case of contact, rinse skin or eyes immediately with plenty of water for at least 15 minutes and seek medical attention. TFFH is not intended for human or animal use. The compound should be stored in a tightly sealed container at 2-8degC (refrigerated), protected from light and moisture. Decomposition may occur upon prolonged exposure to heat or humidity.
|
| Additional Infomation |
Structure in the first source
Tetramethylfluoroformamidinium hexafluorophosphate (TFFH, CAS 164298-23-1) is a fluorouronium-based coupling reagent widely used in solid-phase and solution-phase peptide synthesis. It is also known as Fluoro-N,N,N‘,N‘-tetramethylformamidinium hexafluorophosphate and is available from commercial sources with a purity of ≥97% (typical). TFFH is particularly valuable for coupling sterically hindered amino acids, such as alpha,alpha-disubstituted amino acids, N-methylated amino acids, and residues with bulky protecting groups. Unlike some other coupling reagents (e.g., HBTU, HATU), TFFH generates a less reactive O-acylisourea intermediate that minimizes the risk of racemization. TFFH is also used for the synthesis of acid fluorides and as a reagent for the preparation of various carboxylic acid derivatives (e.g., esters, thioesters, amides). This product is for research use only and is not intended for clinical or therapeutic applications. |
| Molecular Formula |
C5H12F7N2P
|
|---|---|
| Molecular Weight |
264.12
|
| Exact Mass |
264.063
|
| CAS # |
164298-23-1
|
| PubChem CID |
2774761
|
| Appearance |
White to off-white solid powder
|
| Melting Point |
104-109 °C(lit.)
|
| LogP |
3.528
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
8
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
15
|
| Complexity |
153
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
[P-](F)(F)(F)(F)(F)F.F/C(/N(C([H])([H])[H])C([H])([H])[H])=[N+](/C([H])([H])[H])\C([H])([H])[H]
|
| InChi Key |
ZAVXOOLKAGPJPI-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C5H12FN2.F6P/c1-7(2)5(6)8(3)4;1-7(2,3,4,5)6/h1-4H3;/q+1;-1
|
| Chemical Name |
[dimethylamino(fluoro)methylidene]-dimethylazanium;hexafluorophosphate
|
| 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 |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| 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 | 3.7862 mL | 18.9308 mL | 37.8616 mL | |
| 5 mM | 0.7572 mL | 3.7862 mL | 7.5723 mL | |
| 10 mM | 0.3786 mL | 1.8931 mL | 3.7862 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
