Advertisement

Mistaken the post-column mobile phase and pre-column phase

Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.

2 posts Page 1 of 1
Hi, I am Manik

I am a student who barely know anything about HPLC. I made a huge mistake in putting the pre-column and post column mobile phase. Anyway, my analytical condition as follows:

HPLC (Shimadzu, Japan), equipped with CDD-6A electroconductivity detector, double column of Shimpack SCR-102H (300×8 mm i.d., Shimadzu, Japan) and a guard column (50×8mm). Mobile phase containing 5 mM p-toluene sulfonic acid (pre-column) and post column detection was 5 mM p-toluene sulfonic acid, 100 μM EDTA and 20 mM Bis-Tris, the two were mixed (1:1, v/v).

One time during an analysis, I had mistaken 5 mM p-toluene sulfonic acid, 100 μM EDTA and 20 mM Bis-Tris as pre-column mobile phase and 5 mM p-toluene sulfonic acid as post column mobile phase. Since then the retention time delayed eventhough I had changed the mobile phase correctly.

So my question is:
Did 5 mM p-toluene sulfonic acid, 100 μM EDTA and 20 mM Bis-Tris damage the column (Shimpack SCR-102H ) in some or other way?
What is the influence of 5 mM p-toluene sulfonic acid, 100 μM EDTA and 20 mM Bis-Tris on the column described?
What should I do to make the retention time normal again?

Looking forward for your kind reply
Dear Manik

I assume the the column is not damaged by this wrong eluent unless the pH of that eluent is outside of the columns specs.
The BIB/TRIS sholud be washed out in a short time. EDTA may be removed with a few injections of a e.g. Ca standard.

Hope this helps
Dr. Markus Laeubli
Manager Marketing Support IC
(retired)
Metrohm AG
9101 Herisau
Switzerland
2 posts Page 1 of 1

Who is online

In total there are 15 users online :: 1 registered, 0 hidden and 14 guests (based on users active over the past 5 minutes)
Most users ever online was 4374 on Fri Oct 03, 2025 12:41 am

Users browsing this forum: Majestic-12 [Bot] and 14 guests

Latest Blog Posts from Separation Science

Separation Science offers free learning from the experts covering methods, applications, webinars, eSeminars, videos, tutorials for users of liquid chromatography, gas chromatography, mass spectrometry, sample preparation and related analytical techniques.

Subscribe to our eNewsletter with daily, weekly or monthly updates: Food & Beverage, Environmental, (Bio)Pharmaceutical, Bioclinical, Liquid Chromatography, Gas Chromatography and Mass Spectrometry.

Liquid Chromatography

Gas Chromatography

Mass Spectrometry