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Change the mobile phase to a mixture of 0.01 N H2SO4: ACN (85: 15). ... Allow the mixture to flow through the column at the rate of 0.6 ml/min for 2 hours. ... Change the mobile phase from (H2SO4, ACN) to water. ... Set the instrument to flow rate 0.6 ml/ min. ... Ensure that the column contains water.

How to Clean an HPLC Column in Daily & Deeper Cleaning - YouTube YouTube Start of suggested clip End of suggested clip Around. And you can flow into a container. So you don't dirty your detector. The method for reverseMoreAround. And you can flow into a container. So you don't dirty your detector. The method for reverse flush would be to start with 95. Percent water. And 5 methanol for about 10 column. Volumes.

Wash the column with the current mobile phase without buffer, salt or acid for 10-15 min. If the ratio of organic solvent and water is changed, salt may precipitate. Wash impurities in a column to achieve a stable base line: Use a solvent with strong elution properties in which the sample dissolves well.

HPLC column washing method for normal phase mode. - YouTube YouTube Start of suggested clip End of suggested clip Group. In normal phase mode columns flushed with water or methanol may change retention. Usually itMoreGroup. In normal phase mode columns flushed with water or methanol may change retention. Usually it becomes irreversibly faster it may stabilize with a faster retention.

Wash with 2 column volumes of 6 M guanidine hydrochloride (Table 21 for recommended flow). Wash immediately with at least 5 column volumes of buffer at pH 7?8 (Table 21 for recommended flow). Rinse with at least 2 column volumes of distilled water (same flow as step 2) until the UV-baseline and eluent pH are stable.

The following procedure should be satisfactory to remove common contaminants: Wash with at least 2 column volumes of 2 M NaCl (Table 21 for recommended flow). Wash with at least 4 column volumes of 1 M NaOH (same flow as step 1). Wash with at least 2 column volumes of 2 M NaCl (same flow as step 1).

In your case wash the column with 70% water; 15% methanol; 15% acetonitrile. Divert the column eluent to waste not to contaminate your detector(s). Wash the column slowly over to 100% methanol and wash for at least 15 minutes. Wash the column over to 100% acetonitrile and wash for at least 15 minutes.

Typical Regeneration Procedure for Reversed Phase Columns: Disconnect column from detector and run wash solvents into a beaker. Start with your mobile phase without buffer salts (water/organic). ... Next, use 100% organic (methanol or acetonitrile). Check pressure to see if it has returned to normal; if not, then: