User:Dhea Patel/Notebook/Phosphorylation/2012/06/25

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A silica gel column was prepared to''' purify the crude ruthenium complex'''. Glass wool was used to plug the bottom of the column (with a little bit of water). After the water had been drained, about 1 cm of sand was poured into the flask so that the glass wool was completely covered in sand. 300mL silica gel was mixed with ~400mL acetonitrile-sodium nitrate saturated solution. After the mixture had reacted and settled, the gel mixture was poured into the column. The column material was allowed to settle. When no significant change had occurred, the stopcock was opened and air (pressure) was added to help the packing settle. The liquid was drained so that the top was dry. The sample was then added to the column followed by about 1 cm of sand. The column was filled with acetonitrile (saturated with sodium nitrate) solution. The acetonitrile solution was used until the large band appeared to have stopped moving. Pressure (air) was used to run the column efficiently while also using a little less liquid. Then, 90% acetonitrile solution was used until the large dark-orange band was collected. The column was then dried by draining all the liquid and adding pressure (air) for a few hours. The ruthenium band that was collected was rotovapped.
A silica gel column was prepared to''' purify the crude ruthenium complex'''. Glass wool was used to plug the bottom of the column (with a little bit of water). After the water had been drained, about 1 cm of sand was poured into the flask so that the glass wool was completely covered in sand. 300mL silica gel was mixed with ~400mL acetonitrile-sodium nitrate saturated solution. After the mixture had reacted and settled, the gel mixture was poured into the column. The column material was allowed to settle. When no significant change had occurred, the stopcock was opened and air (pressure) was added to help the packing settle. The liquid was drained so that the top was dry. The sample was then added to the column followed by about 1 cm of sand. The column was filled with acetonitrile (saturated with sodium nitrate) solution. The acetonitrile solution was used until the large band appeared to have stopped moving. Pressure (air) was used to run the column efficiently while also using a little less liquid. Then, 90% acetonitrile solution was used until the large dark-orange band was collected. The column was then dried by draining all the liquid and adding pressure (air) for a few hours. The ruthenium band that was collected was rotovapped.
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Notes:
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#The ATP was on the side of the flask after being rotovapped. When the DMF is added, it needs to be mixed such that the product is reacted fully.
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#200mL of silica gel was too great a quantity for the ruthenium purification column. Next time use about 200mL of the silica gel.
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#When pouring the silica gel mixture into the column, be careful that the sand is not disturbed.
==Data==
==Data==

Revision as of 14:44, 25 July 2012

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Contents

Objective

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Description

  1. 0.0245g ATP (disodium ATP) was measured out. 10mL of 0.1M TEAB was added and the ATP dissolved in the solution.
  2. This solution was run through a column equilibrated to 0.1M TEAB.
  3. Eleven 5mL fractions were collected and spotted on a TLC plate. Using UV, it was determined that fractions 3, 4, and 5 contained ATP.
  4. These fractions were combined and rotovapped.
  5. A white powder formed as residue. 10mL mehtanol was added to the flask and rotovapped three times. [The residue was on the side of the flask, so when DMF is added, it needs to be mixed such that the product is reacted fully.]

A silica gel column was prepared to purify the crude ruthenium complex. Glass wool was used to plug the bottom of the column (with a little bit of water). After the water had been drained, about 1 cm of sand was poured into the flask so that the glass wool was completely covered in sand. 300mL silica gel was mixed with ~400mL acetonitrile-sodium nitrate saturated solution. After the mixture had reacted and settled, the gel mixture was poured into the column. The column material was allowed to settle. When no significant change had occurred, the stopcock was opened and air (pressure) was added to help the packing settle. The liquid was drained so that the top was dry. The sample was then added to the column followed by about 1 cm of sand. The column was filled with acetonitrile (saturated with sodium nitrate) solution. The acetonitrile solution was used until the large band appeared to have stopped moving. Pressure (air) was used to run the column efficiently while also using a little less liquid. Then, 90% acetonitrile solution was used until the large dark-orange band was collected. The column was then dried by draining all the liquid and adding pressure (air) for a few hours. The ruthenium band that was collected was rotovapped.


Notes:

  1. The ATP was on the side of the flask after being rotovapped. When the DMF is added, it needs to be mixed such that the product is reacted fully.
  2. 200mL of silica gel was too great a quantity for the ruthenium purification column. Next time use about 200mL of the silica gel.
  3. When pouring the silica gel mixture into the column, be careful that the sand is not disturbed.

Data

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Notes

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