Chemical characteristics and use of biological buffer HEPES (CAS 7365-45-9)

Biobuffering agent HEPES is a pH regulator commonly used in biochemical and cellular biology research. Although there are various types of biological buffering agents and HEPES is not irreplaceable, HEPES buffer is a zwitterionic buffering agent containing amino and carboxyl groups, with stable pH performance. So it is widely used in biochemical and cellular biology experiments that require precise control of pH.

Characteristics of HEPES

The molecular weight of HEPES is 247.32, and its molecular structure is stable. The pH buffer range is 6.8 to 8.2, showing neutrality and good thermal stability at room temperature. Due to its non ionic properties, it can still maintain high solubility in high ionic strength solutions.

Application Fields of HEPES

1. HEPES is commonly used in biochemical research to prepare enzyme reaction mixtures. In the presence of high concentrations of electrolytes, HEPES can also serve as an effective ion carrier. HEPES is often used as a buffer for cell culture media in cell culture;

2. HEPES is used as a pH regulator and moisturizer in the field of daily chemicals, commonly used in skincare products and shampoo, while maintaining the stability of product ingredients.

Although HEPES has a wide range of uses, it also has its limitations. In high concentration salt solutions or certain organic solvents, the buffering capacity of HEPES will decrease. In order not to affect the experimental results, please choose a biological buffer that meets the experimental requirements as appropriate to ensure the orderly progress of the experiment.

Preparation method of HEPES buffer solution

Taking the configuration of a HEPES solution with a pH of 7500 milliliters as an example:

1. 119g of HEPES powder, 500ml of deionized water, 0.5-1M sodium hydroxide solution, stirring rod, and large beaker need to be prepared;

2. Add 400ml of deionized water to 119g of HEPES and stir until dissolved;

3. Slowly add 0.5-1M sodium hydroxide and adjust to the required pH value of 7;

4. Add the remaining 100 milliliters of deionized water to a constant volume of 500ml;

Precautions for using HEPES

When preparing or using HEPES buffer, the following points need to be noted:

1. Avoid contact between HEPES and metals to prevent metal ions from reacting with the components in the buffer solution, which may affect the buffering effect;

2. Due to the potential toxicity of HEPES to cells at high concentrations, the concentration should be controlled during use and combined with the use of sodium carbonate to counteract changes in osmotic pressure;

3. When preparing HEPES solution, please wear protective masks and gloves, as contact between the reagent and the skin may cause discomfort;

4. The prepared buffer should be stored away from light to maintain its chemical stability.

The above listed mixing methods are relatively conventional and for reference only. When using them, flexible mixing should be carried out according to experimental requirements.

In summary, HEPES buffer has a wide range of uses, which can maintain pH stability in the experimental environment and protect the activity of biological samples from being affected. Understanding the characteristics, uses, and preparation methods of HEPES buffer is of great significance for biomedical researchers, as it can make their experiments smoother.

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