Advertisement

Sulfate, Nitrate and Nitrite determinationReverse phase HPLC

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

10 posts Page 1 of 1
Hi,
I can determine Cl, NO3 and NO2 by using C18 column with the eluent n-Octylamine at 220 nm. However, I want to measure Sulfate additionally.

My methode is below

Column Eurosil Bioselect 300-5 C18, 120 x 4 mm
Eluent A 0.01 M n-Octylamine, set to pH 4
Flow rate 1.0 ml/min
Injection volume 5 µl
Column temperature 40 °C
Detection UV at 210 nm

Do you have an application to measure sulfate nitrate and nitrite together with HPLC-UV
Thanks
What else is in your mobile phase that you neglected to mention? Cl- is pretty much transparent at UV wavelengths, so you are *not* detecting it directly at 210 nm. Sulfate has the same issue: it does not absorb UV light.

Chloride and sulfate can be done via indirect UV (for example: http://www-user.uni-bremen.de/~koelling/so4.html) but it's much more straightforward with an ion-exchange column rather than ion-pair.
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
Hi Tom. Mobile phase is 0.01 M n-Octylamine at pH 4. But Once I checked Cl. I saw peak.
Hi Tom. Mobile phase is 0.01 M n-Octylamine at pH 4. But Once I checked Cl. I saw peak.
Probably the solvent front?
That size column would have an internal volume of around 1 mL. At 1 mL/min, that makes the dead time about 1 minute. Certainly anything eluting before 2 minutes is t0 noise. It's not good practice to quantitate any peak eluting with a k' less that 2 (in this case, retention time less than about 3 minutes).
-- Tom Jupille
LC Resources / Separation Science Associates
tjupille@lcresources.com
+ 1 (925) 297-5374
Ion exchange with conductivity detection is the simplest solution to this and an old conductivity detector should be easy enough to find and wouldn't cost much. I've run the same unit on and off for 20 years, have never done a thing to it, and it runs fine.

For $500, it'll be less expensive than spending half a day working on your RP method:

http://www.ebay.com/itm/Waters-431-Cond ... 3cd2df570d
http://the-ghetto-chromatographer.blogspot.com/
Hi Alpher,

I would have to concur with the previous comments above. Have you considered IC? Specifically designed for what you are trying to do...

http://www.dionex.com/en-us/products/io ... 72590.html

Thanks
Anthony

ThermoFisher Scientific
To add up an other IC player:

View Metrohm's IC
Dr. Markus Laeubli
Manager Marketing Support IC
(retired)
Metrohm AG
9101 Herisau
Switzerland
I've seen this done with a C18 "permanently coated" with some quaternary ammonium modifier with hydrophobic arms. The column never lasts long.

Friend, you could probably separate sulfate, nitrite, nitrate on a SAX guard column. Why re-invent wheel?
ELSD with one of our columns will address the issue:
http://www.sielc.com/Compound-Nitrate.html
http://www.sielc.com/Compound-Sulfate.html
Vlad Orlovsky
HELIX Chromatography
My opinions might be bias, but I have about 1000 examples to support them. Check our website for new science and applications
www.helixchrom.com
10 posts Page 1 of 1

Who is online

In total there are 20 users online :: 1 registered, 0 hidden and 19 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: Ahrefs [Bot] and 19 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