YMC-Pack Polymine II is highly effective in separation of a wide range of carbohydrates, including oligosaccharides and glycopeptides. It is used in normal phase separations with non-aqueous eluents or for the separation of ionic compounds by a combination of normal phase and weak anion exchange mode.
The example of analysis of ascorbic acid shown below, anions in the eluent and amino groups on the surfaces of the support material are in a state which allows formation of ion-pairs. If the ion-pairing has not yet reached ionic equilibrium, retention times of a sample can alter. YMC recommends that to enhance ion-pair formation and thereby avoid this problem, the preconditioning operation described below should be carried out prior to starting analysis.
Column preconditioning for ascorbic acid analysis (specifically for a 250 X 4.6mm I.D. column)
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1. |
Flush the column with water at the flow rate of 1.0 mL/min for 10 minutes. |
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2. |
Flush the column with 200mM aqueous solution of ammonium dihydrogen phosphate at the flow rate of 1.0mL/min for 40 minutes. |
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3. |
Flush the column with water at the flow rate of 1.0mL/min for 30 minutes. |
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4. |
Flush the column with the eluent for analytical use at the flow rate of 1.0mL/min for 60 minutes. |
However after using the eluents described above, sugar analysis using acetonitrile/water eluents can result in anomalies in peak shapes (such as peak splitting or broadening). Should this be a problem it is recommended that separate columns are used for the different analyses.
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1. Erythorbic acid
2. L-Asorbic acid |
| Column: |
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YMC-Pack Polyamine II
250x4.6 mm I.D. |
| Eluent: |
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50mM NH4H2PO4/acetonitrile(30/70) |
| Flow rate: |
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1.0 mL/min |
| Temperature: |
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30°C |
| Detector |
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UV at 250 nm |
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Start
