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ARTICLES, APPNOTES, FAQ (9292)
- AppNotes Library (1888)
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Changing retention times of amino acids in biological extracts using ammonium acetate in HPLC methods. – Troubleshooting.

KB Home/ARTICLES, APPNOTES, FAQ /ARTICLES, APPNOTES, FAQ / HPLC Column Information / TYPE-C Column Information / Cogent Diamond Hydride Information / ARTICLES, APPNOTES, FAQ / HPLC Column Information / ARTICLES, APPNOTES, FAQ / HPLC Suggestions and Information / Trouble Shooting Suggestions / ARTICLES, APPNOTES, FAQ / HPLC Column Information / TYPE-C Column Information / Changing retention times of amino acids in biological extracts using ammonium acetate in HPLC methods. – Troubleshooting.

June 25, 2013
Bill Ciccone

KB Home

ARTICLES, APPNOTES, FAQ / Cogent Diamond Hydride Information / HPLC Column Information / Trouble Shooting Suggestions / TYPE-C Column Information

If you observe changing retention times of amino acids using a Cogent Diamond Hydride™ HPLC column, consider the following.

Suggestions, tips and tricks
1. Acid in the Mobile Phase; Formic or acidic acid is generally used when analyzing amino acids, so the use of ammonium acetate (or formate) might be a problem.
2. Contamination from the biological samples. Try our conditioning protocol for biological samples.
3. Suggested Mobile phase: 50% methanol or 50% isopropanol +0.1 % formic acid or 0.2% acetic acid would be suggested to use with biological samples and amino acid analysis when using the Cogent TYPE-C™ Silica columns.
4. How long is your gradient’s re-equilibration time? Follow our protocol on choosing a proper post time.
5. Is the column thermostated? If not, the variation may be due to changes in lab temperature.

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