How is Acetone Used in Chromatography Calculations?

Click For Summary
SUMMARY

The discussion centers on the use of acetone in chromatography calculations, specifically in determining the volume accessible to solvent (Vg) in a column. The Pharmacia Superose 6 column is referenced, where acetone is utilized to measure the accessible volume, yielding a value of 19.5 mL. The total column volume (Vt) is calculated as 24.4 mL, leading to a Vg of 4.9 mL. A participant questions the validity of using acetone to determine both the void volume (Vo) and the internal volume (Vi) in the context of the equation Vt = Vi + Vg + Vo.

PREREQUISITES
  • Understanding of chromatography principles
  • Familiarity with volume calculations in chromatography
  • Knowledge of UV detection methods
  • Experience with Pharmacia Superose 6 column specifications
NEXT STEPS
  • Research the role of small molecules in chromatography, focusing on acetone
  • Study the calculation of void volume (Vo) and internal volume (Vi) in chromatography
  • Learn about UV detection techniques in liquid chromatography
  • Examine the specifications and applications of the Pharmacia Superose 6 column
USEFUL FOR

This discussion is beneficial for chromatography researchers, laboratory technicians, and students studying separation techniques who seek to understand the calculations involved in determining solvent accessibility and volume in chromatography systems.

indoubt
Messages
28
Reaction score
0
chromatography...using acetone!

the protocol:

Vg = volume not accessable to solvent (volume of the gel matrix)

Vg cannot be measured directly. However, it should apparent that:
Vg = Vt - volume accessable to solvent

We can measure the volume accessable to the solvent, using a small molecule such as acetone which can be easily visualized by a UV detector.

example:
Pharmacia Superose 6 column - volume accessable to solvent = 19.5 mL (measured with acetone)
Vg = Vt - 19.5 mL = 24.4 - 19.5 mL = 4.9 mL

---------------------------------------------------------------------------------------

hi guys!
i just find this calculation very unreasonable, since as we know Vt=Vi+Vg+Vo,
where Vt =total column volum, Vo= void volum due to EXTREMELY huge substances, Vg= volum of matrix and Vi=volum inside the beads due to EXTREMELY small substances.

my question is, how come we use acetone (small molecule) to determine both Vo and Vi, since Vg= Vt- (Vo+Vi)?


hopes for replies!

thanks!
 
Chemistry news on Phys.org
by the way here is the link; http://itsa.ucsf.edu/%7Ehdeacon/Stokesradius.html
 
Last edited by a moderator:

Similar threads

Protocol and solvent help needed for gel-permeation chromatography
  • · Replies 11 ·
Replies
11
Views
7K
How can I improve molecule elution in thin layer chromatography?
  • · Replies 15 ·
Replies
15
Views
7K
Chemistry How to Calculate Equivalent Gas Volumes Using Avogadro's Law?
  • · Replies 3 ·
Replies
3
Views
3K
Undergrad Some questions about Cosmic Inflation
  • · Replies 4 ·
Replies
4
Views
4K