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The 2009 ASHS Annual Conference

2950:
Separation and Detection of Antifungal Agents and Some Aflatoxins from Medicinal and Aromatic Plants Using Overpressured-Layer Chromatography and Bioautography Techniques

Tuesday, July 28, 2009: 3:00 PM
Laclede (Millennium Hotel St. Louis)
Nurhayat Tabanca, Natural Products Utilization Research Unit, USDA ARS, University, MS
David E. Wedge, Natural Products Utilization Research Unit, USDA ARS, University, MS
Emil Mincsovics, Natural Products Utilization Research Unit, USDA ARS, University, MS
Agnes M. Moricz, Natural Products Utilization Research Unit, USDA ARS, University, MS
Erno Tyihak, Natural Products Utilization Research Unit, USDA ARS, University, MS
Overpressured-layer chromatography (OPLC) is a forced flow planar layer liquid chromatographic technique which uses external pressure on the planar adsorbent layer and a pump system to introduce mobile phase into the pressurized adsorbent layer. OPLC provides better separation than conventional TLC and so the sample spots or bands are more compact and distinct. This technique is, therefore, especially suitable for sensitive biological detection. BioArena is a complex bioautographic system, which results in characteristic biological effects from the direct bioautography with OPLC separation using directed interactions in the adsorbent layer as well. Cinnamomum is one of the oldest known popular spices and remains an important mercantile commodity today with numerous uses in the food and cosmetic industries.  Ethnobotanical uses of Cinnamomum appear in Ayurveda and Traditional Chinese Medicine and seem to focus on its use for menstral problems and as antimicrobial agent in the treatment of wounds.  A tincture of the bark was useful in treating menorrhagia and for post-partum and other uterine hemmorages, Cinnamomum was one of the most prompt and efficient remedies in Material Medica. Two C. zeylanicum oil accessions, three C. cassia oil collections and one C. subavenium oil along with their major important compounds such as (E)-cinnamicaldehyde, o-methoxy cinamicaldehyde, eugenol, methyl eugenol, and methyl chavicol were separated using OPLC technique and biological detected by applying strawberry anthracnose-causing fungal plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides in the direct bioautographic system. We report on the suitability of bioautography using Pseudomonas savastanoi pv. Phaseolicola a pathogenic bacteria of bean for detection of aflatoxins after OPLC separation. Because it is hypothesized that the antibiotic-like activity of aflatoxins may be a result of the presence of formaldehyde (HCHO), we used L-arginine and glutathione as HCHO-capturing molecules to eliminate it from the chromatographic spots. BioArena system can be used to model interactions between mycotoxins, and other substance groups and diverse endogenous and exogenous substances in the culture medium in general. We also evaluated Cinnamomum oils and their major pure compounds for antimicrobial activity in BioArena system because according to recent observations the antimicrobial activities of different substance groups take place in general through HCHO and its reaction products.The results illustrated clearly the similarity of the mechanism of action of these two totally different substance groups as essential oils and mycotoxins. Detailed results will be presented in this study.