What is the elution volume and how does it relate to protein purification?

[mountain]

hi guys!

i have difficulty in understanding this sentence, since English is not my language. here it is : "The elution volume (Ve) is the volume of buffer before which the peak of protein elutes." hallo! yes, i don't get it. does it mean Ve of that protein is the volume of the buffer before the UV-absorbance of that protein or does it mean that the volume of the buffer to the middle of the peak?

p.s! UV-absorbance and peak are different things here.

thanks for helping!

 

[chem_tr]

Hello,

As I'm not a native English speaker, I think I found the logic behind this sentence

You use a buffer solution to "elute" some material onto a chromatographic column. What I mean with "elute" is to persuade a mixture through a column, causing any impurities or simply "different" substances to be separated one another. I am quite sure you know these, but I wanted to stress them anyway.

Well, what you're asking is closer to the second alternative in my opinion. But if we are talking about HPLC with UV detection, I presume that they are supporting each other. Peak centers are considered to identify peaks, so your second suggestion fits better. I have poor knowledge about UV detection, but if it is calibrated according to that kind of protein before analysis by recording the full spectrum and monitoring the absorption maxima, it points to the same analyte, I think.

 

[ravenprp]

Hi there! I can understand the confusion with this sentence, as it can be a bit technical and may require some background knowledge in chemistry or biology. Let me try to explain it in simpler terms.

The elution volume (Ve) is a term used in chromatography, which is a technique used to separate and purify different substances. In this case, we are talking about proteins. The elution volume refers to the volume of the buffer (a solution used in chromatography) where the protein peak appears. The protein peak is the point on the graph where the protein is most concentrated and can be measured through its UV-absorbance.

So, to answer your question, the Ve of the protein is the volume of the buffer before the protein peak appears. It is not referring to the volume of the buffer to the middle of the peak. I hope this helps clarify things for you. Let me know if you have any other questions!