What are the chemical properties of N, N-Bis (2-Hydroxyethyl) -2-Aminoethanesulphonic Acid?
N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid, often referred to as BES, is a commonly used zwitterion buffer in biochemical research. It has the following chemical properties:
1. ** pH characteristics **: The pKa value of BES is about 7.1-7.3 (25 ° C), which determines that it can exhibit excellent buffering ability in the pH range of about 6.7-7.7. In biochemical experiments, the pH required for many enzymatic reactions and protein stabilization systems is often in this range, so BES can effectively maintain the pH stability of the reaction system and prevent adverse effects on the structure and function of biomolecules due to large changes in pH.
2. ** Solubility **: BES is easily soluble in water to form a clear solution. This property makes it extremely convenient in various biochemical experiments using water as a solvent. It can quickly dissolve and prepare the required concentration buffer, and can be well mixed with other water-soluble reagents.
3. ** Chemical Stability **: Under normal storage and use conditions, BES is quite stable and is not easy to spontaneously decompose. However, although its stability is good, when exposed to strong oxidizing agents or reducing agents, structural changes may still occur due to reactions, affecting buffer performance. Therefore, it is necessary to avoid contact with such substances during storage.
4. ** Effects on biological systems **: BES has no significant toxicity and interference to most biological systems. Due to this, it is widely used in cell culture, protein purification, enzyme activity analysis, and many other experiments that need to maintain the activity of biomolecules, without significantly adverse effects on the normal function of biomolecules and cell physiological activities.
In which experiments are N, N-Bis (2-Hydroxyethyl) -2-Aminoethanesulphonic Acid commonly used?
N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid, commonly known as BES, is a commonly used amphoteric buffer in biochemical experiments. It is widely used in many biological and biochemical experiments.
In protein-related experiments, the stability and activity of proteins are highly susceptible to the influence of environmental pH. BES is often used in protein separation, purification and activity research because of its good buffering ability, which can effectively maintain a stable pH environment, prevent protein denaturation due to pH fluctuations, and thereby ensure the integrity of protein structure and function. For example, in the chromatographic separation of proteins, BES buffer can adjust the pH of the mobile phase to improve the separation effect of proteins.
Cell culture experiments are also common in BES. Cell growth is extremely sensitive to environmental pH, and small pH changes may cause cell growth inhibition or even death. The BES buffer system can precisely control the pH of the cell culture medium to create a suitable microenvironment for cell growth. For example, in animal cell culture, the culture medium containing BES can ensure the normal proliferation and metabolism of cells at a stable pH.
BES is also commonly used in enzyme activity research experiments. The catalytic activity of enzymes is closely related to pH, and the enzyme activity can be optimal at a specific pH. BES buffer can provide a stable pH condition, which is convenient for studying the change of enzyme activity under different conditions. For example, in some hydrolase activity measurement experiments, the BES buffer is used to maintain the stability of the pH of the reaction system, so as to accurately determine the enzyme activity.
How is N, N-Bis (2-Hydroxyethyl) -2-Aminoethanesulphonic Acid stored?
N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid, often referred to as BES, is a biological buffer. Its preservation method is extremely important, related to its quality and utility.
This material should be stored in a dry place. If the environment is humid, water vapor is easy to attach to it, or cause it to deliquescence, which in turn affects its chemical properties and buffering capacity. Therefore, a dry and well-ventilated place should be selected, away from water sources and moisture.
Temperature is also the key. It is suitable for storage in a cool environment, generally 2-8 ° C. If the temperature is too high, the thermal movement of the molecule will intensify, or it will cause its chemical reaction and cause it to deteriorate. If the temperature is too low, in some cases it may cause crystallization. Although it can be restored after heating up, it may also affect its performance.
And be careful to avoid light. Light may promote photochemical reactions to change its structure and properties. Can be placed in a brown bottle, or stored in a cabinet protected from light.
When storing, also ensure that the package is well sealed. To prevent excessive contact with air, so as not to be oxidized, or react with impurities in the air. After taking it, it should be sealed in time to maintain its stability. In this way, N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid must be properly stored so that it can play its due role in experiments or production.
What are the preparation methods of N, N-Bis (2-Hydroxyethyl) -2-Aminoethanesulphonic Acid?
N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid, often abbreviated as BES, is prepared as follows:
** 1. Sodium 2-chloroethanesulfonate and diethanolamine are used as raw materials **
Sodium 2-chloroethanesulfonate is placed in the reactor, and diethanolamine is slowly added according to a certain molar ratio, and the nucleophilic substitution reaction occurs between the two. The reaction process needs to strictly control the temperature, generally maintained at 50-70 ° C. Due to high temperature, side reactions are easy to occur, resulting in lower product purity. At the same time, the reaction system needs to maintain an alkaline environment, and an appropriate amount of sodium carbonate or sodium hydroxide can be added to adjust the pH value to 8-9 This alkaline condition is conducive to the nucleophilic attack of nitrogen atoms in diethanolamine against chlorine atoms in sodium 2-chloroethanesulfonate. After the reaction lasted for several hours, the reaction progress was monitored by thin-layer chromatography. After that, the reaction liquid was cooled to room temperature, and an appropriate amount of organic solvent, such as ethanol or acetone, was added to promote the crystallization of the product. Pure N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid can be obtained by suction filtration, washing and drying.
** Second, ethanolamine and sodium 2-bromoethanesulfonate are used as raw materials **
In the reaction vessel, ethanolamine and sodium 2-bromoethanesulfonate are added, and the molar ratio of the two should also be controlled. The reaction temperature is set at 40-60 ° C, and the appropriate temperature can ensure the reaction rate and product quality. To promote the forward reaction, an appropriate amount of acid binding agent, such as triethylamine, can be added to neutralize the hydrogen bromide generated by the reaction in time. As the reaction proceeds, regular sampling and testing are carried out. After the reaction is completed, the reaction mixture is treated, the solvent and low-boiling impurities are removed by vacuum distillation, and then the column chromatography or recrystallization is used to further purify it to obtain high-purity target products.
The two methods have their own advantages and disadvantages. The former has relatively common raw materials and low cost, but the reaction conditions are more demanding; the latter has relatively mild reaction and high product purity, but the price of raw material 2-bromoethanesulfonate may be higher. In actual preparation, it is necessary to choose reasonably according to specific needs and conditions.
What are the uses of N, N-Bis (2-Hydroxyethyl) -2-Aminoethanesulphonic Acid?
N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid, often referred to as BES, is a commonly used zwitterion buffer in biochemical research. It has a wide range of uses and plays a key role in many fields.
In biochemistry and molecular biology experiments, BES is essential. Many enzymatic reactions are extremely sensitive to environmental pH, and BES can create and maintain a specific and stable pH environment, ensuring that enzymes can exhibit optimal activity and stability. For example, in the separation, purification, and activity measurement experiments of proteins, the BES buffer system can make the protein at an appropriate pH level, prevent it from being denatured due to pH fluctuations, and ensure the accuracy and reproducibility of the experimental results.
BES is also commonly found in the field of cell culture. When cells are cultured in vitro, they have strict pH requirements for the environment they are in. BES buffer can effectively maintain the pH stability of the culture medium, providing a good environment for cell growth and promoting normal cell metabolism and proliferation. Like mammalian cell culture, BES plays an indispensable role in precisely regulating the pH to an appropriate range.
In addition, in immunoassays, such as enzyme-linked immunosorbent assays (ELISA), BES buffers can optimize reaction conditions. By maintaining an appropriate pH, the specific binding of antigens and antibodies is enhanced, the sensitivity and reliability of detection are improved, and the experimental results are more accurate.
In short, N, N-bis (2-hydroxyethyl) -2-aminoethanesulfonic acid is an important reagent for ensuring the smooth development of experiments and accurate and reliable results in many life science research branches, such as biochemistry, cell biology, and immunoassay. It is of great significance in the process of life science research.
