Organic Chemistry Lab.

SMJC 2701
Isolation of pigments form plant leave 22 May 2017
Note
1. Be careful with the treatment of diethyl ether, petroleum ether and acetone. These are
volatile, extremely flammable, harmful to skin.
2. Do not put chemical and organic waste into a sink. (solid waste→dust bin, liquid waste
→waste tank, glass → PET bottle)
3. Wash glassware (use detergent, rinse 10 times with tap water and 3 times with DW) and
wipe the experiment table. (Clean up and arrange equipment for next students)
4. Do not touch the surface of TLC plate.

Procedure
1. Obtain ca. 0.1 g grass leaf. and grind into powder with ca. 0.1 g silica gel using motar and
pestle until . If it remain moist add more silica gen.
2. Put the resulting powder in to a 1.5 mL sample tube, add ca. 0.5 mL diethyl ether, and
centrifuge it to obtain supernatant. (avoid the sample from light after the extraction of
pigment)
3. Put the developing solvent (petroleum ether : acetone = 7:3) into screw bottle ca. 5 mm
from the bottom, and close the cap. Stay for a while until the bottle is saturated by the
vapor of developing solvent
4. Put a weak mark with pencil at 1-2 cm from the bottom of TLC plate.
5. Apply the sample with capillary at the mark and dry well. If the color is weak, apply several
times.
6. Put the TLC plate into a bottle and close the cap.
7. When the front line of developing solvent reach to 1 cm from the top, remove the plate and
mark the front line.
8. Check the color and position of each spot.
9. Repeat 1 to 8 using Wakame (Undaria pinnatifida) instead of the plant leaf.

Important concepts
・Rf value
・Photosynthetic pigments
Questions
・Why the sample needs to avoid the light?
・Determine the Rf value and Identify the each pigment.
Organic Chemistry Lab. SMJC 2701
Isolation of pigments form plant leave 22 May 2017
For your junior
Please check your small lab kit II.
1. Put a silicon tube on the pump adaptor (small black piece) and check the pump.
2. Check the digital thermometer and remove a battery from the thermometer. Put the
removed battery it in a small plastic bag with the pump adaptor.
3. Clean glassware if those are dirty.
4. Add equipment if it those are lacking.