1. Active Substances ～ Contained in Familiar Plants ～ Mizuki Hashizume Kaede Horita Shiori Yoshida Members

2. Production of Natural Weed-killer The number of problems from using harmful weed-killer is increasing in recent years… We want to make natural weed-killer to solve food problems!!! SO

3. Characteristics of Red dead nettle ☺ grows very fast ☺ grows in colonies ☺ few other plants are around it

4. so we thought: Does it have a substance to inhibit sprouting and growth of other plants?

5. If we can extract the substance instead of using weed-killer, Can we solve food problems with weed-killer that is made from something in nature?

6. （ pigment found in tangerine peel ）

7. Characteristics ☺ Lower risks of skin cancer, large intestine cancer and lung cancer. ☺ Promotes the formation of human bone. Are there methods for extracting the effective substance efficiently?

8. Experiment 1 Extraction and refinement of active substances from Red dead nettle

9. Extraction of Substances which Inhibit Sprouting

10. Mash red dead nettle with methanol in a blender Do suction filtration and concentrate it to ¼ volume with low-pressure distillation ExperimentExperiment soak it in methanol and leave it for three weeks Add 200mL water and extract twice with 300mL chloroform extract with methanol extract with chloroform

11. Mash red dead nettle with methanol in a blender

12. Mash red dead nettle with methanol in a blender Do suction filtration and concentrate it to ¼ volume with low-pressure distillation ExperimentExperiment soak it in methanol and leave it for three weeks Add 200mL water and extract twice with 300mL chloroform extract with methanol extract with chloroform

13. Do suction filtration and, concentrate it to ¼ volume with low-pressure distillation

14. Mash red dead nettle with methanol in a blender Do suction filtration and concentrate it to ¼ volume with low-pressure distillation ExperimentExperiment soak it in methanol and leave it for three weeks Add 200mL water and extract twice with 300mL chloroform extract with methanol extract with chloroform

15. Separate and by column chromatography H01-1 H01-2

16. H01-2-1 H01-2-2 methanol 10ml chloroform 190ml two types Extracted liquid first Extracted liquid by methanol Extracted liquid H01-2 (2.55g) H01-1 (1.23g) Make

17. Sprouting Test

18. Experiment Experiment Left in our classroom for six days Cut and mashed clover leaves and roots with distilled water Soaked turnip, lettuce, and spinach’s seeds in it

19. Third dayFourth day Sixth day lettuce Distilled water ○ leaves ○ ○ roots ○ ○ turnip Distilled water ○ ○ ○ leaves ○○ roots ○ Spinach Distilled water ○ leaves ○ roots ○○

20. Extraction and purification of the dye contained in orange peel Experiment 2

21. Experiment Experiment soak it in 200ml hexane and leave it for a week M-01 M-02 Mash orange peel and mix with hexane Suction filtration Concentrated the extract to 1/5 volume by distillation under reduced pressure Added 50ml water and extracted with 200ml ethanol ethanol extract hexane extract

22. Crushed orange peel and mixed with hexane

23. Experiment Experiment soak it in 200ml hexane and leave it for a week M-01 M-02 Mash orange peel and mix with hexane Suction filtration Concentrated the extract to 1/5 volume by distillation under reduced pressure Added 50ml water and extracted with 200ml ethanol ethanol extract hexane extract

24. Suction filtration

25. Experiment Experiment soak it in 200ml hexane and leave it for a week M-01 M-02 Mash orange peel and mix with hexane Suction filtration Concentrated the extract to 1/5 volume by distillation under reduced pressure Added 50ml water and extracted with 200ml ethanol ethanol extract hexane extract

26. Concentrated the extract to 1/5 volume by distillation under reduced pressure

27. Experiment Experiment soak it in 200ml hexane and leave it for a week M-01 M-02 Mash orange peel and mix with hexane Suction filtration Concentrated the extract to 1/5 volume by distillation under reduced pressure Added 50ml water and extracted with 200ml ethanol ethanol extract hexane extract

28. Added 50ml water to the extract and extracted with 200ml ethanol

29. Experiment Experiment soak it in 200ml hexane and leave it for a week M-01 M-02 Mash orange peel and mix with hexane Suction filtration Concentrated the extract to 1/5 volume by distillation under reduced pressure Added 50ml water and extracted with 200ml ethanol ethanol extract hexane extract

30. M-02 Hexane extract Ethanol extract M-01 01 02

31. Test that this extracted substance is β-cryptoxanthin. If it is β-cryptoxanthin, refine it. Use our ingenuity in order to display its effect. Subjects in the future