1. University of Health Sciences Proposition of monograph of Terminalia nigrovenulosa Pierre ex. Prepared by: Channeth Meng 1, Sothea-Kim 1-2, Philippe-Bessioud 2-3, Davy Theng, Chhommonica Pech 2, Mathieu Leti 2-3, Bernard Fabre 2-3.

2. University of Health Sciences Outline I.Introduction II.Project’s purpose III.Research methodology IV.Result V.Conclusion

3. University of Health Sciences I.Introduction: Traditional medicine (TM) still plays an important role in healthcare system in Cambodia, especially in rural area. There are many types of medicinal plants that are used. Among those Terminalia nigrovenulosa is frequently used as anti-diarrhea and dysentery. Terminalia nigrovenulosa’s stem bark is used in treatment of aphtha, generally in Asian country (weerapeeyakul, 2012).

4. University of Health Sciences II.Project’s purpose : To propose the monograph of Terminalia nigrovenulosa in Cambodian pharmacopeia.

5. University of Health Sciences III.Research methodology: 1.Plant collection 2.Microscopic study 3.Colored reaction 4.Thin Layer chromatographic (TLC) Test 5.Determine the quantity of total polyphenol

6. University of Health Sciences 1.Plant collection 10 samples testing 2.Microscopic study Identify the micro-element In powder stem’s bark

7. University of Health Sciences 3.Colored reaction Identify the tannin: 5ml extract + 1ml solution FeCl3 Black color/green black Identify the tannin catechic: 10ml extract + 5ml stiasny Precipitation (2) Identify tannin gallic: Filtrate (2) + C 2 H 3 00Na + 1ml FeCl 3 Blue-black precipitation

8. University of Health Sciences 4.TLC Test: Control reagent: Gallic acid and Rutin Mobile phase: Water/ Formic acid/ Ethyl acetate (8/8/80) Revealer : blue salt solid B

9. University of Health Sciences 5.Determine the quantity of total polyphenol UV/Visible Spectrophotometry Control reagent : gallic acid Stem bark extract : Ethanol-water 50% extract

10. University of Health Sciences IV.Result 1.Plant collection: Collection dateCodePlant partCollect region 7/18/2013CL1448 (003)Stem barkKampot 11/16/2013CL1470Stem barkPhnom Penh 11/18/2013MPM284Stem barkPhnom Penh 11/30/2013MPM286Stem barkPhnom Penh 12/1/2013MPM288Stem barkPhnom Penh 12/2/2013MPM289Stem barkPhnom Penh 12/2/2013MPM291Stem barkSeim Reap 12/3/2013MPM302Stem barkPhnom Penh 1/14/2014CL1486 Stem bark Kep 3/23/2014CL1492 Stem bark Mondulkiri

11. University of Health Sciences 2.Identify the micro-element of stem bark: Type of micro-elementPhoto of fragments Fragment of parenchyma 10x Sclerotic cells of the outer seed coat assie 10x Calcium oxalate cristal 10x

12. University of Health Sciences 2.Identify the micro-element of stem bark: Fiber bundle Associates with liberoligneur 10x Mesocarp sclerotic cells 10x Debris suber in yellow colored walls 10x

13. University of Health Sciences 3.Colored reaction: Reaction T. chebula (CL 1411) T. catapa (CL1482) T. nigrovenulosa (CL1486) T. nigrovenulosa (MPM302) Colored reaction Tannins ++++ Tannins catechic ++++ Tannins gallic ++++

14. University of Health Sciences 4.TLC Test: Top of plat Gallic acid : red band (Rf ~ 0,90) Rutin : yellow band (Rf ~ 0,16) 1 Red band low intensity (Rf ~ 0,65) 2 Red band(Rf ~ 0,5, 0,46) 2 Red band with low intensity (Rf ~ 0,27, 0,17) One red band (Rf ~ 0.11) Control reagent Stem bark extract

15. University of Health Sciences 5.Determine the quantity of total polyphenol: Plant codeAverage concentration (%)SDVariance(%) CL144814.750.503.42 CL147016.480.382.29 CL148614.980.110.74 CL149212.380.252.02 MPM28413.381.047.77 MPM28614.740.453.06 MPM28812.410.403.20 MPM28913.560.634.64 MPM29115.290.593.84 MPM30215.280.241.58

16. University of Health Sciences 5.Determine the quantity of total polyphenol

17. University of Health Sciences V.Conclusion The simple quality controls’ methods of Terminalia nigrovenulosa are developed and proposed.