degassed buffer（Degassing Buffer The Importance of Removing Dissolved Gases for Accurate Results）

Degassing Buffer: The Importance of Removing Dissolved Gases for Accurate Results

Degassing buffer, also known as degassed buffer, refers to buffers that have had their dissolved gases removed. In many life science applications, accurately measuring pH, conductivity, and other properties of solutions is crucial for obtaining reliable results. This is why researchers often use degassing buffers to ensure that their measurements are not skewed by the presence of dissolved gases. In this article, we will discuss the importance of using degassing buffers and how they can be beneficial in various applications.

The Significance of Dissolved Gases in Buffers

When preparing buffers, it's common for dissolved gases such as oxygen, nitrogen, and carbon dioxide to be present in the solution. However, these gases can influence the pH and other properties of the buffer, causing errors in experimental results. Oxygen, for example, can react with components in the buffer, leading to oxidation or reduction reactions that can affect the pH. Carbon dioxide can form carbonic acid, which can influence pH measurements. Additionally, dissolved gases can also cause changes in viscosity, which may impede or delay mixing. By using degassing buffers, the eliminated dissolved gases help guarantee the stability of buffer pH and improve overall experimental accuracy.

The Benefits of Degassing Buffer in Biological Studies

Degassing buffers can be especially vital in biological research, where even small changes in pH can affect enzyme activity, protein folding, or cell physiology. For example, if a biological buffer like Tris-HCl contains dissolved oxygen, the oxidation of Tris (a component that maintains buffer pH) can occur, and cause inaccurate pH readings. The presence of dissolved gas can also be detrimental to delicate proteins, which is why degassing buffers are commonly used in protein purification and analysis. Similarly, if the buffer is used in cell culture, the presence of dissolved gases can impact cell growth or other processes. Degassing buffers can reduce variables that affect cell physiology or metabolism, contributing to more consistent experimental results.

Methods of Degassing Buffer Preparation

There are several ways to degas buffers, such as using a vacuum pump, a degassing chamber, or a membrane filter. The choice depends on the lab equipment available, the sample volume, and the buffer components. One common way is aspirating the buffer into a vacuum chamber and removing the dissolved gas by applying a vacuum. To enhance degassing efficiency, the solution can be heated and stirred under vacuum for a more extended period. Another method is membrane filtration. The buffer is passed through a specialized membrane, which allows gases to diffuse through it; thus removing the dissolved gases. With both methods, it's important not to agitate the buffer to prevent reintroducing air into the solution after degassing.

In summary, removing dissolved gases is crucial for obtaining accurate results and consistent experimental outcomes. By using degassing buffers, researchers can avoid errors caused by dissolved gases in buffers, reduce variables that may affect physiological processes or reaction rates, and ultimately produce more reliable data.