Selection of silica column for HILIC

[Kazimierz]

I would like determinate of ganciclovir in serum samples using HILIC.-HPLC.
In my experience some silica columns does not separate of ganciclovir and acyclovir (IS) f.e. Capital Silica. In literature autors used Intertsil or Inertsil,
but using HPLC/MS/MS. Actualy I plane to try with Zorbax Rx SIL 3.2x250 mm, 5 um (3um not available). Has anybody experience with ganciclovir, acyclovir, ribavirin or with similar drugs and compared differ brand of silica in hydrophilic interaction chromatography.

I was made some probes with zirconia and titania but such columns retained intensively ganciclovir causing very wide piks.

[Einar Ponten]

The zwitterionic ZIC®-HILIC column has successfully been used for similar compounds. I can not see why it should not be a proper choice in this case as well (i.e., for 2'-nor-2'-deoxyguanosine).


// I am new to this new forum, but I assume that ZIC®-HILIC will translate into a very happy smile //

[Kazimierz]

Dear Dr Pontén,
Thank you for your response.
I understand that your ZIC®-HILIC columns are dedicated to HILIC chromatography, but using HPLC-MS. The most their lenght is 10 cm only.
I'm using spectrofluorimetric detector and for this detection pH <2.2 is need.

I have obtained a reprint of the article that Zorbax Sil column separate
successfuly acyclovir in acetronitril:water(70:30) + 0.05 mM fosphoric acid.

Please send me materials and price data concerning your columns, if possible

[SIELC_Tech]

Dear Kazimierz,

After you try HILIC mode you might want to consider our mixed mode solution to this separation. Your compounds will retain on Primesep columns are similar to guanine. You can use veriety of additives to mobile phase (TFA, sulfuric, phosphoric acid) to help eluting your compounds:

http://allsep.com/makeCmp.php?cmp=Cmp_050

Contact us at mail@sielc.com if you have questions

[Kostas Petritis]

Dear Kazimierz,

Just courious if there is a specific reason for using HILIC chromatography for your application. All your compounds are very hydrophilic but most of them have been already been analysed with reversed phase LC and UV or amperometric detection. Furthermore, ganciclovir and acyclovir have been analysed simultaneously with RPLC in human plasma/serum.

Ribavirin has also been analysed by RPLC (although not in mixture with the above).

Does your compounds co-elute, or do you have problems of matrix interference when you are analysing these compounds by RPLC? If one of the above apply, or if you are using MS (which you do not) then I would use HILIC for these compounds -I can understand that due to the high polarity of these compounds you will need high aqueous mobile phases for their RPLC separation which are not the best conditions for high sensitivity with the MS-. Otherwise, as you are using UV, a RPLC seems more straightforward to me.

Here are some references about the use of RPLC for your compounds:

Title: A simple and simultaneous determination of acyclovir and ganciclovir in human plasma by high-performance liquid chromatography
Author(s): Teshima D, Otsubo K, Yoshida T, Itoh Y, Oishi R
Source: BIOMEDICAL CHROMATOGRAPHY 17, 8, 500-503 DEC 2003

Title: Liquid chromatographic method for the determination of ganciclovir and/or acyclovir in human plasma using pulsed amperometric detection
Author(s): Kishino S, Takekuma Y, Sugawara M, Shimamura T, Furukawa H, Todo S, Miyazaki K
Source: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 780 (2): 289-294 NOV 25 2002

R.H.A. Smith, B.E. Gilbert, J. Chromatgr. 414 (1987)
202.
G.G. Granich, D.J. Krogstad, J.D. Connor, K.L.
Desrochers, C. Sherwood, Antimicrob. Agents
Chemother. 33 (1989) 311.
M. Homma, A.L. Jayewardene, J. Gambertoglio, F.
Aweeka, Antimicrob. Agents Chemother. 43 (1999) 2716.
J.-O. Svensson, A. Bruchfeld, R. Schvarcz, L. Stahle,
Ther. Drug Monit. 22 (2000) 215.

Kostas

[Einar Ponten]

Dear Kazimierz,

In reply to your specific questions:
Our zwitterionic stationary phases (silica, polymeric) are designed for HILIC and availabel in practically any dimension from ID 75 µm to prep scale, both in PEEK and SS hardware.
Any detection mode can be used so you are not limited to MS, that is rather a feature, as well the wide pH range that may be used (2-11) depending on phase.