Chromatography Forum

Is there any HPLC column which can adequately retain histamine (and other biogenic amines) without derivatization, and possibly without using an ion-pair reagent?

Here are two methods for retention of histamine without IP reagent. One you can use for UV (TFA in the mobile phase) and another one is LC/MS (ammonium formate). For UV you can use TFA, sulfuric or phosphoric acid or corresponding buffers (Primesep 200 column):
http://www.sielc.com/compound_158.html

Histamine is retained by reverse phase and cation-exchange mechanisms.
Contact me directly if you have questions.

Regards,

Vlad

Histamine can be retained at pH 10 on a hybrid column such as XTerra or XBridge. Detection by UV.

In general, for the two analytes you specified both RP column and cation-exchange mixed-mode column should work. However, I would be interested in knowing what "other biogenic amines" are. If these unspecified analytes are hydrophilic, a RP column may not provide adequate retention and right selectivity even at pH10. Therefore, a cation-exchange mixed-mode column sounds more convenient. However, after checking the link provided above, the peak shape and efficiency on PrimeSep 200 seems a bit broad and low. I hope there are ways to improve them. To make the picture complete, you also could consider Ion Chromatography with the IonPac CS17 for analyzing some amines that would be difficult using toher means. Here is a link where you might find some useful information. http://www1.dionex.com/en-us/columns_ac ... s2727.html

The average plate count for Primesep columns is over 80K plates/meter. Methods in the link are less then 6 minutes with full base line resolution. For sharper peaks we suggest to use a longer column and play with pH and buffer concentration. For amines with double charge (histamine) pH and buffer is more critical as it interacts more strongly with the stationary phase.
With mixed mode chromatography you can achieve extremely sharp peaks by playing with pH. If you can "afford" gradient you will observe GC type peaks with double/triple gradients (pH buffer, concentration and ACN) due to focusing effect. This is not possible with RP or ion-exchange
http://www.sielc.com/application_122.html

"Plate count" (which is not an accurate definition in this case) is over 1.5M plates/meter. It would be interesting to see histamine on Dionex column (peak shape retention control, etc.)

P.S. There are many tricks to present chromatograms, if I create 40 minute scale for the one I cited above peak shape will be much much sharper, but this is a fast method to save time. Time is …very precious.

Any cation IC column is able to do that job.
Read Metrohm's information on biogenic amines in food or download the respective Poster.
Fluka (Sigma-Aldrich) offers a dedictated standard for these applications.