Understanding Flowthrough, Wash and Eluate Fractions

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Discussion Overview

The discussion revolves around the differences between flowthrough, wash, and eluate fractions in chromatography. Participants explore the definitions and roles of these fractions within the context of column chromatography, with some references to related techniques like thin layer chromatography.

Discussion Character

  • Exploratory
  • Technical explanation
  • Conceptual clarification
  • Homework-related

Main Points Raised

  • One participant expresses familiarity with the eluate fraction but seeks clarification on flowthrough and wash fractions.
  • Another participant suggests that discussing what the eluate fraction is could help others understand the topic better.
  • A participant mentions that chromatography relates to the affinity of substances in a mixture to the stationary phase, although the context of this statement is unclear.
  • One participant outlines the steps in chromatography, indicating that washing removes non-binding molecules while elution involves high-affinity molecules being washed out.
  • There is a clarification that the discussion is about column chromatography rather than thin layer chromatography, highlighting the differences in techniques and applications.

Areas of Agreement / Disagreement

Participants do not reach a consensus on the definitions of the fractions, and multiple interpretations and clarifications are presented without resolution.

Contextual Notes

Some assumptions about the definitions of flowthrough, wash, and eluate fractions are not explicitly stated, and there may be dependencies on specific contexts or applications of chromatography that remain unresolved.

Who May Find This Useful

Students and individuals studying chromatography or related laboratory techniques may find this discussion beneficial for understanding the terminology and processes involved.

indoubt
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chromatography:

we have to collect the fractions from the column. i wonder what is the difference between;

flowthrough fraction, wash fraction and eluate fraction?

i think i know what it means with eluate fraction, but what about the other two?

hope for replies!
thanks a bunch!
 
Last edited:
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It seems there are a few folks posting here working on similar topics in class (i.e., chromatography). Why don't you try to help each other out a bit? You'll learn it better to try answering each other's questions (it's like practicing for a quiz or test).

Please start out by telling us what you think the eluate fraction is, since that's the one you know.
 
I'm not sure quite what you mean, but I know chromatography has to do with the substances in a mixture's affinity to the paper. I hope this helps you out a little.

Paden Roder
 
indoubt, just think about the different steps you go through during chromotography when you apply your sample to the column. You apply your sample, you wash it, you elute it. Washing takes away non-binding or low-affinity binding molecules, eluting is a wash step where the high-affinity molecules binding to the column will lower their affinity and wash out.
 
Last edited:
PRodQuanta said:
I'm not sure quite what you mean, but I know chromatography has to do with the substances in a mixture's affinity to the paper. I hope this helps you out a little.

Paden Roder

You're thinking of thin layer chromatography (TLC). Indoubt is asking about column chromatography. The basic principles are similar in terms of adsorption of a compound to a matrix, but column chromatography takes it a further, allowing you to not only separate your sample, but collect it in a more purified form so you can then use it for something else.

Here's a website with some basic column chromatography methods and pictures of the procedure. At the end, it demonstrates using TLC to confirm successful separation of the components of the original mixture. http://www.wfu.edu/academics/chemistry/courses/CC/
 
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