Protocol for Lactose Separation using Chromatography

[smsaufi]

Hi,
I try to separate lactose using chromatography system. Does anyone can help me to get the protocol for that?

Thanks in advance.
Saufi

[Uwe Neue]

Separate from what?

Here is a possible method: Waters Carbohydrate Analysis column with 75% acetonitrile and 25% water.

[smsaufi]

Actually I used standard lactose in water. I try to develop new affinity ligand that can bind a lactose. I plan to check the static and dynamic binding capacity for my new ligand. But, I'm not very sure about the suitable mobile phase for that binding experiment.

[Uwe Neue]

The chromatographic system that I proposed can be used for assaying the lactose concentration.

I can not advise you how to do the binding to your affinity ligand, since I do not know what it is.

[smsaufi]

The ligand used is peptide. Anyway thanks for your help:)

[tom jupille]

If all you need to see is the single peak, there are numerous carbohydrate analysis columns available, designed to run with different mobile phases:
- water/ACN (per Uwe's suggestion),
- pure water (e.g., HPX-87 series columns from Bio-Rad), or
- basic buffers (e.g., columns designed for use with the PAD detector from Dionex).