ABSTRACT
Cell-Based Assays........................................................................................................... 391 10.3.1 Immortal Cell Lines ........................................................................................................ 392
10.3.1.1 Hep G2 (Human Hepatocellular Carcinoma).................................................. 392 10.3.1.2 MDA-MB-231 (Human Mammary Adenocarcinoma).................................... 393 10.3.1.3 PC-3 (Human Prostatic Carcinoma)................................................................ 393 10.3.1.4 5637 (Primary Bladder Carcinoma) ................................................................ 393
10.3.2 Primary Tissue Culture ................................................................................................... 393 10.3.2.1 Primary Rat Hepatocytes ................................................................................. 393
10.3.3 Antiviral Assays .............................................................................................................. 394 10.3.3.1 Anti-HSV ......................................................................................................... 394
10.4 Invertebrate-Based Assays............................................................................................................ 395 10.4.1
Artemia salina
(Brine Shrimp) ....................................................................................... 395 10.4.2
Drosophila melanogaster
(Fruit Fly).............................................................................. 395 10.4.3
Solenopsis invicta
(Red Imported Fire Ant) ................................................................... 396 10.4.4
Meloidogyne incognita
(Root-Knot Nematode) Nematocidal Assay............................. 396 The Test Procedure.......................................................................................................... 397
10.4.5
Caenorhabditis elegans
Anthelmintic Assay.................................................................. 397 The Test Procedure.......................................................................................................... 398
10.4.6
Epilachna varivestis
(Mexican Bean Beetle) Antifeedant Assay................................... 398 10.4.6.1 Dual-Choice Antifeedant Assay....................................................................... 398 10.4.6.2 Contact Toxicity ............................................................................................... 399 10.4.6.3 No-Choice Feeding Test for Insect Growth Regulator ................................... 399
10.4.7
Biomphalaria glabrata
(Freshwater Snail) Molluscicidal Assay................................... 399 The Test Procedure.......................................................................................................... 401
10.5 Biochemical Screens .................................................................................................................... 401 10.5.1 Based on Spectrophotometric Detection ........................................................................ 401
10.5.1.1 Papain Inhibition Assay ................................................................................... 402 10.5.1.2 Trypanothione Reductase Inhibition................................................................ 402
10.5.2 Based on Gel Electrophoresis......................................................................................... 402 10.5.2.1 Topoisomerase II Inhibition............................................................................. 403
10.6 Evaluation of Structural Interactions via NMR and MS Methods ............................................. 403 10.6.1 NMR-Based Methods...................................................................................................... 404
10.6.1.1 Ligand-Based Screening Methods................................................................... 404 10.6.1.2 Diffusion Spectroscopy-Based Methods ......................................................... 404 10.6.1.3 SHAPES........................................................................................................... 406 10.6.1.4 Receptor-Based Screening Methods................................................................ 406 10.6.1.5 SAR by NMR .................................................................................................. 407
10.6.2 MS Methods .................................................................................................................... 408
10.7 Conclusions .................................................................................................................................. 408 References .............................................................................................................................................. 409
The driving force behind much phytochemical research is the discovery of new biologically active compounds for medicinal or agricultural uses. Biological assays, then, must be carried out in order to identify promising plant extracts, to guide the separation and isolation, and to evaluate lead compounds. In this chapter, we present some bioassays that are routinely used in our laboratories. These include
in vitro
assays for antimicrobial or cytotoxic activities, assays with invertebrates such as brine shrimp and insects, and some biochemical assays. We do not include biological assays using vertebrate animals or human clinical trials.
