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Anti  Salmonella typhi  activities of non aqueous  camelia sinensis  crude extract Tiurma PT Simanjuntak,STP M.Si Lecturer biochemistry
OUTLINE ,[object Object],[object Object],[object Object],[object Object]
Introductions ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Salmonella  Typhi ,[object Object],[object Object],[object Object],[object Object],[object Object]
Natural product Material Camelia   sinensis  sp. ,[object Object],[object Object],[object Object]
Drug discovery Acquisition  ( raw material, field colections, culture colections  ) Extractions Primary screens Isolation and Characterisation secondary screens structural elusidation pre-clinical development
On going research ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
Questions ,[object Object],[object Object]
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
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
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
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
Crude extract green tea with partitions by  n -heksan, benzen, diklorometan dan etil asetat
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
Diameter Inhibition area ,[object Object],[object Object],[object Object],[object Object],H B D E
Result from coloni counting plate
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  
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  
Summary ,[object Object],[object Object],[object Object],[object Object],[object Object]
Perkebunan teh desa mekar sari Subang  sources of green tea.
Thank you
Mekanisme pagositosis
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
 
 
Metode Agar Diffusion
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
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
Taksonomi  Camelia Sinensis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
 
 
Koloni hitung
Klasifikasi  Salmonella typhi ,[object Object],[object Object],[object Object],[object Object],[object Object]
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
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)
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
Kepolaran Pelarut Organik ,[object Object],[object Object],[object Object],[object Object],[object Object]
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
 
 

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My Biochemistry research

  • 1. Anti Salmonella typhi activities of non aqueous camelia sinensis crude extract Tiurma PT Simanjuntak,STP M.Si Lecturer biochemistry
  • 2.
  • 3.
  • 4.
  • 5.
  • 6. Drug discovery Acquisition ( raw material, field colections, culture colections ) Extractions Primary screens Isolation and Characterisation secondary screens structural elusidation pre-clinical development
  • 7.
  • 8.
  • 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
  • 15.
  • 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  
  • 19.
  • 20. Perkebunan teh desa mekar sari Subang sources of green tea.
  • 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
  • 24.  
  • 25.  
  • 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
  • 29.
  • 30.  
  • 31.  
  • 33.
  • 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
  • 37.
  • 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
  • 39.  
  • 40.