Chromatography Forum

Hello

I'm looking for a simple and convenient test mix for HILIC (Atlantis) columns for the purpose of performance check of the column over its lifetime.
(k' and N)

It should contain of a t0 marker and 2-3 substances which can isocratically separated.

Does anybody have something just by hand?

Thank you

Edit:
I'v googled a bit and found something at Alltech
Toluen, Hydantoin, Uracil, Caffein, Allantoin
ACN/NH4COOH pH3 95/5

sounds convenient to me and I have most of the substances in our lab.
I will give it a try.

As I've searched on the forum, toluen is a good choice for t0 marker isn't it? Or is naphthalen better?

We use Toluen (for t'0), uracil and cytosine.
Naphtalene is an equal good choice for t'0 but it smells....

At 80% acetonitrile, low ionic strength of buffer (5 mM overall) and neutral pH Uracil and cytosine elutes with retention factors between 0.5 and 2.0 on our ZIC®-HILIC phases, depending on pore and particle size.

These compounds are sufficiently stable (several months in solution in refridgerator and weeks at RT) and will be a simple and convenient test mix for column performance check (k' and N). They can also be used with different detection modes.

Thank you Patrik for your reply.

After some trials I used the following mix:

Toluene 1 µl/ml
Caffein 60µg/ml
Theobromine 60µg/ml
Adenosine 110 µg/ml
dissolved in H2O/MeOH/ACN 10/18/72

HPLC conditions
isocratic, premixed: 5 ml NH4COOH 200mM, pH 3.0 / 95 ml ACN
T Col: 30°C
Det: UV 270 nm
Inj Vol: 5 µl

With this I get the following retention factors on Waters Atlantis Hilic, 3µm 4.6x100
Toluene := 0
Caffein 0.6
Theobromine 0.9
Adenosine 3.3