Materials Required

 

• 0.1M phosphate buffer

 

• Screw cap tube

 

• Blender

 

• 0.2M p- nitrophenyl acetate

 

• Potassim permanganate solution in reagent spray bottle

 

• Acetone

 

• TLC plate

 

• TLC chamber

 

• Ferric chloride – potassium ferricyanide solution in reagent bottle

 

• Vaccum filter

 

• TLC solvent (70:30 petroleum ether and ethyl acetate)

 

• Orange peel

 

• Shaker

 

Procedure

 

1. The orange peel is collected from orange with a citrus zester

 

2. 40ml of 0.1M phosphate buffer is poured into it and the resulting mixture is homogenized via blender

 

3. The mixture is vacuum filtered over a thin layer of celite 545.

 

4. The blender was rinsed with 5- 10 ml of buffer and filtrate the material

 

5. The clear orange filtrate is collected in a beaker. The extract is stable for at least 72 hour at 2-8°C.

 

6. Two 5ml of screw topped vial is taken and the following two reactions are set up:

 

a) 6 drops pNPA solution + phosphate buffer, pH 7.0 for VT = 3ml

 

b) 6 drops pNPA solution + Orange peel filtrate for VT = 3ml

 

7. The two reaction mixtures are capped and allowed to stir

 

8. The reactions are monitored by TLC every 5 minutes

 

 

 

Help:

 

                    Unfortunately, this Virtual Lab requires Adobe Flash player.  Please see additional information if this does not work on your computer

• For Google Chrome - https://support.google.com/chrome/answer/6258784?co=GENIE.Platform%3DDesktop&hl=en

• For Microsoft Edge - https://support.microsoft.com/en-in/help/4532571/microsoft-edge-turn-on-flash 

• For Mozilla Firefox - https://support.mozilla.org/en-US/kb/install-flash-plugin-view-videos-animations-games

 

On GNU/Linux machines, please see appropriate online help. For example, 

 

• on Ubuntu - https://help.ubuntu.com/stable/ubuntu-help/net-install-flash.html.en 

• on Fedora - https://docs.fedoraproject.org/en-US/quick-docs/using-adobe-flash/