A Guidebook to Plant Screening- Phytochemical and Biological

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A Guidebook to Plant ScreeningPhytochemical and Biological

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 GUIDEBOOK TO PLANT SCREENI

PHOCHEMICAL AND IOLOICA VD D 2005

BOTANY SECTION Alicia  Cast Rosie S Madlid PHYTOCHEMISTRY SECTION Aicia M Aginaldo Einda I Espeso Beatrice Q Gevara Maibe G Nonato You're Reading a Preview

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MICROBIOLOGY SECTION Download With Free Trial Edward A �into May Ann G Santos PH ARM ACOLOGY-TOXICOLOGY SECTION  Sign up to vote on this title Goia e CastoBenas  Useful  Not useful Rhodoa E Gonzaes Rosaina C de CastiloSoevila

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Published by: Research Center for the Natural Sciences Unversty of Santo Tomas Espaa Manla and UST Pubishing House You're Reading a Preview Beato Angelco Bldg Unlock full access with a free trial. Espaa Manla Download With Free Trial

Copyright© 2005 by the Unversity of Santo Tomas, Manila Philippnes A rghts reserved No part of ths book may be reproduced or copied in an wthout permisson n writng from the copyrght holder Sign up to vote on this title

Printed by: UST Publishng House

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FoREWORD

e Reseach Cente fo the Natal Scences (then Reseach Cente) o nvesty of Santo omas, n 980 pbshed a gdeboo to pant sceenng en Phytochemical, Microbiological and Pharmacological Screening  Medicinal P as a spplement to the Aca Manilana Ts was evsed n 985 Te boo conceptalzed and pepaed fom the 'handson dscpned eseach expeen faclty eseaches n the ed of natal podcts of the nvesty of Santo  a podct of the colaboatve eots of the botanst, chemsts, mcoboog phamacologsts The boo contaned the Herbarium Secion contbted by Aca L Ca the Pyocemisry Secion wth Afedo C Santos, Beatce Q Gevaa,  M Agnado, Mabe G Nonato and Belnda V Reco as contbtos Microbiology Secion had Patocno S Santos and Nmfa M Cha as contb whle the Parmacology Secion was contbted by Rosalnda C Soevl Lodes V Songco dtos of the boo wee B Q Gevaa and B V Reco Te pesent boo A Guidebook toReading Plant Screening: You're a Preview Phytochemical and Biol s made moe sefendy Ths boo povdes pdated 'doable pocedes  Unlock full access with a free trial. sceenng of pants fo boactve consttents, whethe they ae fo potenta soces o fo nttonal pposes Refeences gven Download With Free Trial at the end of each secto the lteate sed by the past and pesent eseaches n devsng the sce pocedes A panoamc ovevew ofthe ente gdeboo lstates at a glance the ana assays fo pant sceenng n the fo sectons of the gdeboo n ths ove the eseache s ed to the sees of the stdesSignnupode that the plant ma  to vote on this title fond pomsng n the sceenng pocedes, may eventay deveoped useful  Useful  Not be new podct, a natal podct fo medcnal o nttonal pposes he fo sectons of the pesent gdeboo ae the Boany Secion

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srongy recommend that hey conduct the stes unde he superso advsers It s hoped ha wth hs gudebook more scene stdents are enc pursue nerdscplnary sudes o unlock he secrets o pants or the w humanty Le he resuls of such studes serve as he engne of more concr conservng and propagang our rch plant resources n ther naural bo B Q

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PREACE o Phyochemical, Microbiological and Pharmacological creening of Medicinal Plants (Revsed EdonJune 985)

There has been n more recen mes an awakenng owards he use of drug her preparaons n a knd of"back-o naure movemen, nsead of he ca synhec compounds manufacured n advanced counres Whe he use ofsynhecs  s ofundoubed vaue, especay n advanced sa ness  s beeved ha he use ofherba medcnes of propery-esed ecacy w be of grea advanage n a deveopng counry ke ours, whch s s bessed boun pan resources; he dea shoud be o keep peope heahy by reang  a an eary sage nsead of resorng o reamen when aready a an advanced I s wh hs dea ha a manua for phyochemca, mcroboogca pharmacoogca screenng has been conceved o awaken he neres ofhe begn scens n esabshng scenc evdence n he use of herba medcne Ths m has been prepared speccay for he Reading use n he aboraory screenng of med You're a Preview pans for boogcay acve consuens Te mehods used have been so des Unlock full access with a free trial. as o provde a smpe and rapd mehod for screenng wh a mnmum of equpm reasonaby seecve for he knd of consuens under Download With Free Trialsudy wh provsons for quanave measure of evauaon he phyochemca screenng s conned o he deecon of mpo phyochemca consuens ha usuay exhb boogca acvy ke he aka saponns, cardenodes and bufadenodes, avonods, annns and poyphe compound, anhraqunones and cyanogycosdes Sign up to vote on this title  Te mcroboogca screenng s desgned o deec an-mcroba  Useful  Not useful acvty o pan exracs agans represenave es organsms of gram-posve, gram-neg and acd-fas groups ncudng yeas and fung

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cKNOWLEDGEMENT

O heafel gade o D Fonao S evlla III fo challengng  and pgade he Aca Manlana spplemen enled Phyohemia Miob and Phamaologial eening fMediinal Plans whch was pblshed n  hank he conbos o hs spplemen who have moved on o ohe c o have joned o Ceao o he nvesy ahoes fo he knd and whole heaed sp eseach and s exenson sevce o he commny, o he UNESCO N fo he Chemsy of Naal Podcs n Soheas Asa and o he A adjnc Newok fo he Chemsy of Bologcally Impoan Naal Po senghenng he compeences and capables of naal podcs ese US Reseach Cene fo he Naal Scences, o hem goes he whole gade of he conbos o hs gdebook We gaeflly acknowledge he nd help of D Ma Naala R Dm he compe geneaed gaph of he mcobologcal assay daa shown  M You're Reading a Preview e nal sage of hs gdebook wold no have been possble who Unlock full access with a free trial. of yong alens, M James R Jane, who dd he lne skeches n hs g and Ms Rolln S dela Cz, Download who paenly and ably anscbed and fom With Free Trial encoded ex and scanned ges no he eleconc fom eady fo he  of he book Of cose, we ae gael fo he nd sppo of he oce Reseach Cene, Ms Chamane M Caga, Ms Ma Ada A Caalee Macelo A Gazo  Fo We gaeflly acknowledge he US Reseach Endowmen Sign up toand vote on this title Inc fo he nal nancal sppo n he pepaaon pdaed g Nothe useful  Useful of and o he Phlppne Concl fo Healh Reseach and Developmen, Dep of Scence and echnology he gan fo he s pnng of he gdeboo

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Phytochemical and iological A OVERVIEW F THE GUIDEBK

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Ethnbtanial  pharmatial - pharmalgial - mdial

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* Invro

srning - against batria

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 Plant idntiati dmnta


hytochmstry Scto  hytochm Download Withfr Free Trial Studs * Srning sndary *

Mcrobology Sco

Fild lltin f spimn

plant mtablits Srning fr anti xidants

 Fratina f pr m strtr l & idnti 

Sign up to vote on this title harmacoogy-Toxcology  Useful  Not useful Sco * In-vro srning - harmacolo

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Research Center for the Natural Sciences, University of

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TAE O CONNS Foreword Preface Acknowledgement Overview of the Guidebook Drug Deelopment of pomoea murca L.Jacq, "Tonn

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BT SEi  Overew

Inoducon to Plant Studes 

Plant Samples needed for Botnc Idencaon

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Prepaon for Field Collecn   

Objecves and econs om e Tp Preparaons needed for e Botanc Collecon Equpment and Supples 21 ds o the plant collecon 22 Preparation for the herbarum sme 2 ds n the ·dumentaton o the plans colected You're Reading a Preview

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e Feld Collecon

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Prelmna Preparaons of te Plant Specmens n e Feld . .


    

Pressng e Plant Speme Download With Free Trial Dng e Presse-t pecen n e Feld Preseng te Pressed Plant Speen   n e Feld

Fnal Preparaon of eHerbaru/oucher Speen n the Laborato Fnl Preseaon of e Plant Speen 

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HTCHEMITR SECTI n Ovrw Inoducon  Prang  Plant Matral for Stu

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 Praraon of Stock Plant Ec  Phytochcal Scrnng for Scondary Plant Mtabolts

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alod  211 eld e for loid e Clvenorgerald mehod 212 borory e be mehod for lkloid nalyi 2121 relminry y 2122 Conor e 212 Te for qernry be nd/or mine oide 21 hinlyer hromogrph analyi for loid e FrnworhEuler meod 211 Eion of he hloroform olble fron 212 riion nd nlyi of he hloroformolble fri 21 Eion nd nlyi of he werolble frion

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Strods: Cardnolds and Bufanolds  221 Te be reon mehod You're Readinge a Preview 2211e KellerKilini for 2deogr 2212e iebermnnBuhrd e for nured erod Unlock full access with a free trial. 221e Kedde e for nred lone Download ph Trial for unured lone 222nlyer homo meod & With Free

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rnods 21 reprion f he pln er 22 nlyer hromogrphi nlyi

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nthranons  21 Te be eening meod Sign up to vote on this title  11e Bornrger e  Useful  Not useful 212he modied Bornrger e 22 Sreenng by hinlyer hromogrphi mehod



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2.6.2 Semi-quantitative assay for saponins (O, 1998) 2.6.2. Preliminary test 2.6.2.2 Main test 2.6. Liebermann-Burchard test for unsaturated sterols nd ritrenes 2.6. in-layerhromatographi (TLC test for saognins'

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...................... Tannns and Polyphe'nols 2.7. Sreening methods for tannins 2.7.1.he test tube method 2.7..2 Spot test for tannins 2.7.2 Sreening method for polhenols by dry disllation

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Cyanogenc Glycosdes 2.8.he Guignard Test

nlaye Choatogaphc (TLC) Seeg of Boacve Copounds n Plants 

Pepaaon of e Plant ts

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Sceenng by nlaye Choatogaphy

Anoxdants n Plants 



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Basi Tehniques in in-layer Chromatography 1.0 Preparaon of the Plates ..  .... ...... .... 2.0 Preparation of the Silia Gel G Layer with . ........... .... the Use of a Glass Rod .0 Saturaon of the Developing Chamber ..............  ...  Sign up to vote on this title . ..... .. 4.0 Sample Appliaon ......... useful............  Useful  Not....... 5.0 Developmen of the Chromatogram: One-dimensiona Chromatography 6.0 Visualization of the Chromaogram .

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 2 Ppaatin f th tst gansm  inclm  2   Bactia: Gam psitv  Gam ngativ 22 Filamnts ngi  2  3 Yasts

13 he Microbiological Assay Methods 3 Ppaatin f th assa plats 3h aga-cp/agawll disin mthd 3 2h pap disc disin mthd

1.4 Reading the Assay Plates and Documenting the Results 15 Cleaning up Microbiological Wastes 2.0 Determination of the Minimal Inhibitory Concentraon (MIC) and Minimal Bactericidal Concentration (MBC) of Plant Extracts 21 Preparation of the Plant Extract for Assay 22 Preliminary Preparations 22 Clt mdia 222 Ppaatin f th tst ganisms 223 Antibitic standads

23 Methods of Analysis

You're Reading a Preview 23 Analsis f th plant xtact against bactia 23 Dtminatin f Minimal Unlock full access with a free trial. Inhibit Cncntat 232 Dtminatn f Minmal Bactcida Cncntati

Download Free against Trial lamnts ngi 23 2 Analsis f th plantWith xtact 2 3 2  Dtmnatin f Minimal Inhibit Cncntati 2 322 Dtminatin f Mnimal Fngcidal Cncntatn

233 Analsis f th plant xtact aganst ast 2 32  Dtminatin f Minmal Inhibity Cncntatin 2 322 Dtminatin f Minma Fngicidal Cncntatin

24 Reading and Documentation


 Not useful 3.0 Comparison of Antimicrobi ActivityofUseful the Plant Extract with a Standard Antibiotic

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50 Bioautography

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 9

51 Materias Needed



5.2 Preiminary Preparations 5 2 h lt mdia 5 22 h tst ganism 5 23  hmatgam



53 Preparation o the Base and Seeded Layers in the Assay Pates



54 Bioautographic Assay ndirect Mehod

 

5.5 Documentation o Results Reerences



POLOGY-OXOLOGY ON An Oeriew Introduction 10 Generl Considerations 11Test Drug Materias 1.2 Experimentl Anims 2.0 ToxicityTests


2.1 Determination o the Appromate Letha Dose Download With Free Trial (ALD) by single dose method 2.2 Determination o Median Lethl Dose LD by singe dose method 23 he Brine Shrimp Assay 23. Pant tst xtat ppaatin 23.2 Hahing th shimp Sign up to vote on this title 233 Biassay useful  Useful  Not 2 3 4 Dtminatin  th Mdian Ltha Cnntatin (LC) 30 SensitiityTest to Local andTopical Applications

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4.6 Subjeciveess 47 Auopsy 5.0 Specic Pharmacologicaless 51 Anagesic Propery 511 Ati aid writhing aay 5.2 Anipyreic Propery 5.3 Aniinammaory Propery 53 In-vv rning mthd 53 Carragnan-indd rat paw dma 53..2 Carragnanindd plriy 53..3 Crtn il drmatiti mthd

5.4 Anispasmodic Propery 60 Muageniciy and lasogeniciyess 61 Ameses 6 Gnral nidratinDirt mtagn aay 6. Prparatin f th plant xtrat and pitiv ntrl 61  2 Prparatin f tp agar 6.1.13 Prparatin f th ba agar 614 Prparatin f ntrint brth ltr You're Reading a Preview 6.2 Aay prpr Unlock full access with a free trial. 6.3 Gnral nidratin-Indirt mtagn aay 6.13.1 Eipmnt and gawar Download With Free Trial 63 2 Chmial 613 3 Tt anima

61 4 Indtin f rat livr nzym fr aringni ativatin 6.15 Prparatin flivr hmgnat fratin ( S9 ) 6.151 Prparatin fS9 mx

6.2 Micronuceuses


62 Gnral nidratin  Useful  Not useful 6. 2 1 Eipmnt and glawar 62.12 Chmial

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70 Angiogenic Studies Chicken Chorioaantoic Membrane as a Mode System 7.1 Genera Considerations 71.1 Experimental animal 71.2 Materials and equipment

72 Assay Proper 73 Visua Assessment and Photography References

...................... 13

Appendx Poliies on Researh involving Animals 1. Experimental Animals used in Biologial Studies 1.1 Ethia Considerations 1.2 Researh Design 13 Suppy of Animals 14 Training on Handling and Care of Animals 15 Animal Handing and Restraint 15 1 Handling smal mammals 1.5 11 Handling the mouse 15 .12 Handling the guinea pig 1. 5.1 3 Handling theYou're rat Reading a Preview 1. 5.1 Handling the rabbit Unlock full access with a free trial. 1.6 Speiations for Experimental Animals 16.1 Swiss mie Download With Free Trial 1.62 Guinea pigs 1.63 Rats 16 Rabbits 17 Method of Drug Administration 1.8 Animal Disposal

References

LIST OF FIGURES

...................... 14


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Figure BO. Mounting labels a and b Figure B Mounted sheet with eld label, herbarium label and paket for detahed plant parts

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............. .............

gure P. igure P2 Figure P3. Figure P4. gure P. Figure P6 Figure P7.

  .  ..  .. Set-up for reux Set-up for ltration through anhydrous sodium sulfate .  ........  ........... Set-up for ltration  .... .... Set-up for reux     ..  .. Test ube setup for the Guignard test e spotted plate in the equilibrated hamber .  ........ Fabriated glass sprayer pumped by a rubber aspirator .  ...........

Figure M. gure M2.

Flowhart of the mirobiologial sreening methods Adjusting inoulum turbidity against a arland barium sulfate standard Pouring agar medium into a Petri dish Swabbed agar plate Position of the agar ups/wells in the plae Preparing the agar well with a ork borer Plaing paper diss onto agar plate Flowhart for determining Minimal Inhibitory You're Reading a the Preview Conentration  MIC) of a plant extrat Unlock full access with a free trial. Flowhart for the omparison of the antimirobial ativity of the plant extrat with a standard antibioti Download With Free Trial Arrangement of the agarups in the assay plate Graph of the data obtaied in the bioassay of peniillin G against Bacllus subtlis Sterile lens paper laid evenly on the agar plate Chromatogram faed downward with the silia oated side laid on the lens paper

gure M3 Figure M4 Figure M Figure M6. Figure M7. Figure M. Figure M9 gure MO. gure M Figure M12. gure M13 .


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Figure PT. Lateral position to the spinal groove of a guinea pig ............. gure PT2 Measurement of paw edema by the plethysmometer ............

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Figure PT Mouse handling gure PT2. Mouse restraint Figure PT3 . Guinea pig handling Figure PT14. Guinea pig restraint gure PT15 Rat handling gure PT6  Rat restraint Figure PT7. More rat restraint igure PT8. Rabbit handling, grasping the loose sin of the sru and plaing hand underneath the belly gure PT19. Rabbit restraints, a and b gure PT20. Rabbit handling when returning the rabbit to its age positioned to fae one of he side walls Figure PT2. Subutaneous injetion into the ank of the rat Figure PT22. Subutaneous injetion into the bak of the shoulder of the rat gure PT23  Intramusular injetion of the rat Figure PT24. Intraperitoneal injetion of the rat Figure PT2. Restraint of rabbit for intravenous injetion gure PT26. Intravenous injetion site into the marginal ear vein of the rabbit Fgure PT27. Intravenous injetion site into the tail vein of the mouse Figure PT28 Intraoral administration into the mouse

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 14  14  143  143  1  14  14  14

 145  146

 147  147

 147  147  14  14  1  1


 OFAL



Table P. Table P2 Table P3 Table P4 Table P5.

 Serial dilution of the plant extrat with phosphate buer  Serial dilution of the plant extrat, undiluted or diluted with phosphate buer  Groups of plant onstituents that would possibly  Sign up to vote on this title be distributed in the dierent extrats useful  Useful  Not  Developing solvent system for TLC of Solutions A B or C  List of plant onstituents, their visualizing

44

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5

5

5

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Table PT.

Table PT2

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

Site ofinjetion, needle size and maximum normally aepted volume to administer for eah speies of test animals Gavae equipment reommended for eah speies of test animals

.




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A Guidebook to Plant Screening- Phytochemical and Biological

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