Extracting Biologically active substances from Plants Haruna Kobayashi Yohei Nosaka Yuta Fukushima Eriko Minamiya Koshi High School

We listened to our seniors’ study “Making chemical-free fertilizer with plant components” and we will extract component from plants to find good applications for our life. ~Motive~

 Cinnamon  Cork Tree  Bracket fungus  Chaga  Rooibos ~Research Material~

・ Spice ・ We say it has hyperthermic action. Cinnamon Cork tree ・ Rutaceae Rooibos ・ we say it is a medicine of perpetual youth and long life ・ It has antioxidant action

・ Shelf-type mushrooms ・ It is born in the stem of natural trees, fallen trees, and dead trees Bracket fugus ・ Cold-resistant mushroom ・ It is born in cold areas Chaga

Bracket fugus and Chaga are relatives. But both of them are different in trees. Do they have different components!?

Crush material with ethanol using a machine. Leave it for three weeks. Concentrate it. Add water to it, to get n-Hexane Extracts. ～ First Experiment ～

① Make 10ml of solution which is 90% Chloroform and 10% Methanol (we will just call it “the solution” after this), and make a sample with the solution. ～ Method ～ Vacuum Filtration Concentration ～ Thin Layer Chromatography(TLC) ～

② Spot the bottom of the TLC plate with the concentrate from step ①. Then, we put 5ml of the solution into a glass case and we put the TLC into the case.

③ After the solution rises enough, check if you can see component spots with UV light.

… Spots of components （ A thicker mark means the component is more concentrated. ） … The first spot ～ Results of Experiment ～ Ⅰ Cinnamon Ⅱ Rooibos Ⅲ Bracket fugus Ⅳ Cork tree, Chaga The line of rising solution

～ Column Chromatography ～ ① Make 200ml of the solution ② Put Silica gel and the solution into an Erlenmeyer flask and mix them. ～ Method ～～ Method ～ Chloroform Methanol Silica gel

③ Put ② into a column and pour in a little solution. After that we put 5ml of the solution mixed with a sample into the Column.

④ Put a little Silica gel on ③ and pour little by little another 200ml of the solution. → Silica gel → Sample → Solution → Expand Mixed Silica gel and 5ml solution

⑤ Divide the liquid into 2 equal parts, put in a Flask and concentrate them.

Result of Experiment ColorFragrance C－０C－０ strong red C－１C－１ YellowWeak C-0 smell C－２C－２ YellowStrong C-0 smell C－３C－３ Reddish brown Ⅰ :Cinnamon

Ⅱ :Rooibos Ｒ－ ０ Ｒ－ １ Ｒ－２Ｒ－２ ColorFragrance R－０R－０ Strong yellow-green R－１R－１ Weak yellow-green R－２R－２ Weak Yellow-green

Ⅲ： Bracket fungus Ｓ－ １ Ｓ－ ２ Ｓ－ ３ Ｓ－ ４ Ｓ－ ５ ※ There is not S-0 picture. ColorFlagrance S－０S－０ Yellow-green S－１S－１ Strong yellow S－２S－２ Weak yellowS-0 smell S－３S－３ Weak yellowSour smell S－４S－４ Weak yellowSour smell S－５S－５ Weak yellow

ⅠⅡⅢ … Spot of components （ a thicker mark means the spot is more concentrated ） …… The first spot C －１ C －２ C －３ R－１R－１ R－２R－２ C －０ R －０ S －１ S－０,S－０, ２～５

～ Consideration ～ ・ C-2 has a strong C-0 smell ⇒ We think C-2 has fragrance components. ・ C-3 has C-0 color ⇒ We think C-3 has color components. ・ R-2 is like C-0 color ⇒ We think R-2 has color components. Cinnamon Rooibos Bracket fugus ・ S-2 is like S-0 smell ⇒ We think S-2 has fragrance components.

～ Future Activities ～ ・ Try to do other experiments. ・ Try to extract component of other plants. ・ Conduct experiments to test the effects of components. We will examine whether Rooibos have inhibits UV light and so on.

Thank you f r listening listening o