The document summarizes research on the anti-Salmonella typhi activities of crude extracts from Camellia sinensis (green tea). Key findings include: - Crude extracts from green tea showed inhibition of S. typhi in agar diffusion tests, with diameter inhibition areas of 33.3-40% for ethanol extracts and 39.5-44.7% for water extracts. - Further fractionation of the crude extract in non-aqueous solvents like n-hexane, benzene, dichloromethane, and ethyl acetate showed reduced inhibition compared to the water fraction. - Colony counting plate experiments confirmed a decrease in bacterial colonies with the crude water extract, providing

1. Anti Salmonella typhi activities of non aqueous camelia sinensis crude extract Tiurma PT Simanjuntak,STP M.Si Lecturer biochemistry

6. Drug discovery Acquisition ( raw material, field colections, culture colections ) Extractions Primary screens Isolation and Characterisation secondary screens structural elusidation pre-clinical development

9. Methodology 50 g green tea + 175 mL water as solvent Extract green tea ( brown ) agar difussion method Crude extract green tea (dry) Crude extract green tea (liquid) With n -heksan agar difussion method extraction with soklet Drying with vacuum konsentrator on 30 C Re-solvent again screens extractions

10. Fasa n -heksan Fasa water Extractions with benzen Activity antibacterial Fasa water Fasa benzen Extractions with diklorometan Fasa water Fasa diklorometan Fasa water Fasa etil asetat Extractions with etil asetat Activity antibacterial Activity antibacterial Activity antibacterial 200 uL dried (fraksi water 1 ) Drying and resolvent again Drying and resolvent 200 uL dried (fraksi water 2) 200 uL dried (fraksi water 3 ) 200 uL dried (fraksi water 4 ) Drying and resolvent Dried and resolvent again

11. Result and analysis green tea (50 g) Soxletation 175 with water as solvent for10 hours Crude extract (50 ml) Crude extract dried (2,186 g) Liquid Crude extract Dried 60 °C, 2 days Resolvent again with water Green tea crude extract (200  l) Liquid Crude extract (800  l) Partitions n-heksan (1:1) …………… ... dried

12. continue Fraksi water (0,051 g) Fraksi heksan (0,002 g) Partitions with 600 ml benzen p.a (1:1) Fraksi water (0,038 g) Fraksi benzen (0,004 g) Fraksi water (0,077 g) Fraksi water (0,077 g) Partitions with 400 ml diklorometan (1:1) Partitions with 200 ml etil asetat (1:1) Fraksi diklorometan (0,003 g) Fraksi etil asetat (0,003 g) Screening with agar diffusion method and coloni counting plate method

13. Crude extract green tea with partitions by n -heksan, benzen, diklorometan dan etil asetat

14. Result from activity anti S.thyphi by Agar Difussion methods B A C D A : 10 ul crude extract green tea 54,65mg/ ml B : 10 ul liquid extract green tea C : 10 ul aquadest D: 10 ul ampisilin 100 mg/ml Diameter inhibitions area of extract green tea. Ampicilin as positive control Aquadest as negative control

16. Result from coloni counting plate

17. Analysis 54.65 54,65 54,65 54,65 54,65 0,5 1 2 1 2 83 420 550 320 940 Control 1140 Result and analysis from crude extract green tea Crude extract green tea Consentration (mg/ml) Diameter inhibitions (mm) Total colonies Fractions water Fractions water 1 Fractions water 2 Fractions water 3 Fractions water 4

18. Analysis 54,65 54,65 54,65 54,65 0 1 0 1 500 0 2 0 Analysis and result from green tea non aqueous solvent Extractions Consentrations (mg/ml) Diameter inhibi tions (mm) Total coloni N-heksan benzena diklorometan Etil asetat

20. Perkebunan teh desa mekar sari Subang sources of green tea.

21. Thank you

22. Mekanisme pagositosis

23. DNA amplifications from chromosome S.typhi 1419 517 396 214 75 65 Dna kromosom s.typhi kontrol positif DNA kromosom S typhi yang hidup dalam ekstrak 0.4 kb pb PUC/HinfI

26. Metode Agar Diffusion

27. Metoda koloni hitung Media luria bertani padat Media LB padat + ekstrak teh 20 ul Media LB + ekstrak teh hasil partisi benzena 20 ul dan diklorometan + 30 ul bakteri + 30 ul bakteri + 30 ul bakteri Masing-masing di spread dan di inkubasi 37OC Hitung jumlah koloni

28. Tabel.Berat kering ekstrak yang diperoleh dalam pelarut selain air. Tabel. Berat kering ekstrak yang diperoleh dalam pelarut air 0,077 200  L Fraksi air 4 0,077 200  L Fraksi air 3 0,038 200  L Fraksi air 2 0,051 200  L Fraksi air 1 0,075 200  L Fraksi air Berat kering yang diperoleh (gr) Volume yang dikeringkan suhu 30 o C Ekstrak teh 0,003 150  L Fraksi etil asetat 0,003 300  L Fraksi diklorometan 0,004 550  L Fraksi benzen 0,002 750  L Fraksi n-heksan Berat kering yang diperoleh (gr) Volume yang dikeringkan suhu 30 o C Ekstrak pelarut teh

32. Koloni hitung

34. Analisis porsentase koloni hitung pada pelarut tanpa ekstrak 100 1140 KONTROL 0,87 10 Etilasetat 3,5 40 diklorometan 12,63 144 Benzen 80,52 918 n-heksan % koloni hidup Jumlah koloni hidup Ekstrak teh

35. Analisis porsentase koloni hitung pada ampicilin dalam berbagai konsentrasi 100 450 0 66,6 300 25 40 180 50 28,57 120 75 7,1 32 100 % koloni hidup Jumlah koloni hidup Konsentrasi (mg/ml)

36. Analisis porsentase koloni hitung pada pelarut 100 1140 Kontrol 0 0 Etilasetat 0,17 2 Diklorometan 0 0 Benzen 43,85 500 N-heksan % koloni hidup Jumlah koloni hidup Ekstrak pelarut

38. Analisis porsentase koloni hitung pada fraksi air 82,45 940 Fraksi air4 28,07 320 Fraksi air3 48,24 550 Fraksi air2 36,84 420 Fraksi air1 7,2 83 Fraksi air % koloni hidup Jumlah koloni hidup Ekstrak teh