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ANSWER. Waters Magic Mix/AAA Wash is a general cleaning solution used to remove contaminants from internal HPLC and UPLC system components. The composition of the Magic Mix/AAA Wash is equal parts 25:25:25:25 MS/HPLC-grade H2O/ACN/MeOH/IPA plus 0.1 to 2% formic acid (typically).

5.0 PROCEDURE Change the mobile phase to filtered distilled water. ... Allow the water to flow through the column at the rate of 1ml / min. ... Change the mobile phase from water to HPLC grade 80% Acetonitrile. ... Set the instrument to flow rate 1 ml/ minute and wash the column for 30 minutes.

To wash the column usewater-miscible, nonaqueous solvents such as methanol, acetonitrile, or tetrahydrofuran. If you are using a buffered aqueous mobile phase, do not jump immediately to the strong solvent.

Cleaning and Regeneration An initial washing with deionised water is used to remove organic solvent and polar impurities, followed by a flush with a 0.5 M sodium chloride solution. Finally remove the salt solution with sufficient water and equilibrate the column with 80% (v/v) acetonitrile.

Regeneration Procedure for Reversed-Phase Silica Columns Disconnect the column and reconnect it to the system, with the flow through the column in the reversed direction. ... Flush out any salts/buffers with HPLC grade water. ... Flush column with 25 mL of isopropanol. Flush column with 25 mL of methylene chloride.

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:

A much better approach to washing the column is to first replace the buffer component of the mobile phase with water. For example, replace 60:40 acetonitrile–buffer with 60:40 acetonitrile–water. Wash the column with 5–10 mL of this — just enough to wash most of the buffer out.

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.

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:

0:04 1:32 How to Clean an HPLC Column - Reverse Flush Method - YouTube YouTube Start of suggested clip End of suggested clip Easiest way to remove. This is to reverse the flow on your column you can do this by taking out theMoreEasiest way to remove. This is to reverse the flow on your column you can do this by taking out the column. And seeing how the flow direction is currently.

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.

Using isopropanol as an intermediate solvent is recommended, followed by mobile phase without buffer, then finally with the starting mobile-phase composition. Tetrahydrofuran is another popular solvent that can be used for cleaning contaminated columns.