Tris(2-carboxyethyl)phosphine hydrochloride Chemical Properties

Melting point:

177 °C

Density 

1.041 g/mL at 25 °C

refractive index 

n20/D 1.367

storage temp. 

2-8°C

solubility 

H₂O: 50 mg/mL

form 

powder

pka

7.66(at 25℃)

color 

white

PH

7.0(aqueous solution; pH was adjusted with ammonium hydroxide)

Water Solubility 

Soluble in water and methanol.

Sensitive 

Moisture Sensitive

λmax

λ: 260 nm Amax: 0.05
λ: 280 nm Amax: 0.03

Merck 

14,9085

BRN 

3724376

Stability:

Stable for 1 year from date of purchase as supplied. Solutions in distilled water may be stored at -20°C for up to 1 month.

InChIKey

PBVAJRFEEOIAGW-UHFFFAOYSA-N

CAS DataBase Reference

51805-45-9(CAS DataBase Reference)

Safety Information

Hazard Codes 

C,Xi

Risk Statements 

36/38-36/37/38-34

Safety Statements 

26-37/39-36/37/39-27-45

RIDADR 

UN 3261 8/PG 2

WGK Germany 

2

F 

10

TSCA 

No

HazardClass 

8

PackingGroup 

III

HS Code 

29319090

MSDS

• Language:English Provider:ACROS
• Language:English Provider:SigmaAldrich
• Language:English Provider:ALFA

Tris(2-carboxyethyl)phosphine hydrochloride Usage And Synthesis

Description

TCEP HCl (51805-45-9) is a reagent for selective reduction of disulfides in aqueous solutions1-3?with distinct advantages over DTT which are application specific3. For example, spin labels are more stable with TCEP than DTT, Ni2+?(leached from affinity columns) causes rapid oxidation of DTT with no effect on TCEP and for long term storage of proteins TCEP is more stable than DTT with no metal chelator (EGTA) present but DTT is more stable in the presence of metal chelators.3

Chemical Properties

White Crystalline Solid

Uses

TCEP (Tris(2-carboxyethyl)phosphine hydrochloride) is an alternative which is not pungent, more stable and works even at low pH. It is more selective, i.e. do not reduce metals and buried disulfides.

Uses

Reducing agent, specific for disulfidesTris(2-carboxyethyl)phosphine hydrochloride is widely used as a reducing agent to reduce disulfide bonds in proteins and trans-4,5-dihydroxy-1,2-dithiane. It is involved in the reduction of sulfoxides, sulfonyl chlorides, N-oxides, and azides. It plays an important role in the tissue homogenization process for RNA isolation. It acts as a chelating agent and forms complexes with various heavy metal ions such as Zn(II), Cd(II), Pb(II), and Ni(II). Furthermore, it is useful in the labeling of cysteine residues with maleimides.

Uses

Biochemical tool for selective reduction of disulfide bridges at low pH; reductant for redox assays.

Preparation

The preparation of Tris(2-carboxyethyl)phosphine hydrochloride is as follows:Trimethyl 3,3 ', 3 "-phosphine triphenylpropionate (92.5g, 0.28mol) prepared as described above was added to a 250 ml reaction flask under mechanical stirring conditions and a molar ratio of 12mol/(120ml, 1.44mol), the mixture was heated to 90°C, the solid gradually dissolved, continue to heat the reaction 4h, stirring and cooling with ice water, precipitation of white solid, filter, filter cake with distilled water to get white (2-carboxyethyl) phosphine hydrochloride, dried, weighing 61.3g, yield 82.4%

General Description

Tris (2-carboxyethyl) phosphine (TCEP) is very effective in cleaving disulfide bonds in aqueous solution. It dissolves in water and is odorless, unlike other trialkylphosphines (tributylphosphine). It is also less toxic than 2-mercaptoethanol. These advantages make it better than the other reducing agents.

Reactivity Profile

Nucleophilic Substitution by the Phosphorus Atom of TCEP. 1. The phosphorus atom attacks one sulfur atom along the S-S bond. 2. A thioalkoxyphosphonium cation and a sulfhydryl anion are formed. 3. A rapid hydrolysis releases the second sulfhydryl molecule and the phosphine oxide. There are two main steps in the breaking of disulfide bridges and forming a free sulfhydryl using TCEP: cleavage of the S-S bond; oxidation of the phosphine and release of sulfhydryl.

reaction suitability

reagent type: reductant

Biochem/physiol Actions

Tris(2-carboxyethyl)phosphine hydrochloride solution reduces the disulfide bonds and leaves other functional groups intact in proteins.

Synthesis

The general procedure for the synthesis of tris(2-carbonylethyl)phosphine hydrochloride from triethyl 3,3',3''-phosphinotriphenylpropionate was as follows: pre-prepared trimethyl 3,3',3''-phosphinotriphenylpropionate (92.5 g, 0.28 mol) was placed in a 250 ml reaction flask, and hydrochloric acid (120 ml, 1.44 mol) in a 12:1 molar ratio was added under mechanical stirring ). The reaction mixture was heated to 90°C until the solid was completely dissolved and the reaction was continued at this temperature for 4 hours. After completion of the reaction, it was cooled in an ice water bath and stirred to precipitate a white solid. The solid was separated by filtration and the filter cake was washed with distilled water to give white tris(2-carboxyethyl)phosphorus hydrochloride. The product was dried and weighed 61.3 g with a yield of 82.4%.

storage

Following reconstitution, aliquot and freeze (-20°C). TCEP retains stability when stored at an acidic or alkaline pH; avoid storage at pH 7.0-8.0. Also avoid exposure to high concentrations of phosphate ions (>150 mM). Stock solutions are stable for up to 3 months at -20°C.

References

[1] WOLFGANG H. FISCHER  A. G C  Jean E Rivier. In situ reduction suitable for matrix-assisted laser desorption/ionization and liquid secondary ionization using tris(2-carboxyethyl)phosphine[J]. Rapid Communications in Mass Spectrometry, 1993, 7 3: 225-228. DOI:10.1002/rcm.1290070312
[2] KIRLEY T L. Reduction and fluorescent labeling of cyst(e)ine-containing proteins for subsequent structural analyses[J]. Analytical biochemistry, 1989, 180 2: Pages 231-236. DOI:10.1016/0003-2697(89)90422-3
[3] ELISE BURMEISTER GETZ . A Comparison between the Sulfhydryl Reductants Tris(2-carboxyethyl)phosphine and Dithiothreitol for Use in Protein Biochemistry[J]. Analytical biochemistry, 1999, 273 1: Pages 73-80. DOI:10.1006/abio.1999.4203

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