Four fungi of the Aspergillusflavus group were differentiated to the species level and strain level by pyrolysis-gas-liquid chromotography. Comparisons of pyrochromatograms revealed more similarities than dissimilarities among both species and strains in the pyrolytic elution patterns. Quantitative analysis was made by comparing the number of peaks in which two strains or reference species agreed or disagreed, the degree of superimposability between the pyrolytic elution patterns of strains and reference species, and the presence or absence of peaks for strain pairs within each species. The accuracy and precision of these techniques suggest that pyrolysis-gas-liquid chromatography may have wide application in the detection, enumeration, and identification of fungi by nonmycologically trained personnel. Methods which have been used to characterize microorganisms include analysis of volatile products in head space (6), fatty acid composition (3, 8, 12), deoxyribonucleic acid base composition (5, 11), infrared spectroscopy (20), electrophoresis (19), and gas chromatography (1, 2). However, results from studies with these methods give an incomplete picture of the genotypic expression of microorganisms and are usually too tedious and time consuming for routine analysis. A comparatively new method for identifying and characterizing microorganisms combines pyrolytic degradation of cells or cell fragments with gas-liquid chromatographic analysis (15). Resulting pyrochromatograms show elution patterns characteristic of genus, species, and, in some instances, strains (16). Pyrolysis-gas-liquid chromatography (PGLC) has previously been used to characterize bacteria (14), fungi (13), and nucleotides or nucleosides (22). Differentiation of species or strains of fungi by PGLC has not been comprehensively described, although Oyama and Carle (13) examined one taxon from each of several fungal genera. Moreover, no workers, with the exception of Cone and Lechowich (4) who were unsuccessful in differ-957 Vol. 20, No. 6
AbslractStudies of the toxicity of 2,4,5,6-tetrachloroisophthalonitrile (TCIN) to cells of Stucluiromyce.s paslorianiis and ^'^•tlroHpora crasm showed that 2 to 4 [ig/ml of the tnxirant were required to inhihit giowth. Several thiol compounds reversed toxicity tn h tdticose oxidation was severely impaired in treated cells of both test organisms. The to.xicant at '2 or 4 ug/ml markedly reduced soluble thiol content of these cells. Unuiul thiol content was less affected in S. pastoritmus cells than soluble thiol content.I'ptake of toxicant by yeast cells was accompuniyd by lorination of derivatives, some of wliich ri'stMiihlcd tliosc formed by reaction of glutathionr with TCIN in vitro. Cot-nzynio A. glutathione, and 2-merca|)toethanol readily formed derivatives with TCIN in oitro. The nature of these derivatives was studied using 2-mercaptoethanol products as model suhstituents. Four derivatives were formed with '2-niercaptoethanol each with similar functional groups but showing dis.similar degree.s of mohility during silica gel chroniatogrii|)hy. Evidence indicaled tliat these are derivatives of I'CIN in which 1 to 4 of the halogens has been substituted by 2-mercaptoethanol.The mechanism of fungicidal action of TCIN is attributed to thiol inaclivalion.
InlroductionA iiilrile derivative, 2,4,5,(>tetrachloroisophlhalonitrile (TCIN). ha.s recently been introduced as a lunjiicide active aj,'ainst a broad .spectrum of phylopatho^'enic fungi (13). Chemically related to TCIN are tbe herbicide.s ;i5-dii()do-4-bydroxybenzonitrile. 2,6-dichiorobenzonitriIe (2,8,15), and 2,0dicbloro-3-bydroxybenzonitrile (10): an insecticide, p-cblorobenzonitrile (2); "Physiol. Plant., 21, 1068 [1249]
A high pressure liquid chromatographic isocratic procedure is described for determining and quantitating the 5 major alkaloids narcotine, papaverine, thebaine, codeine, and morphine in Papaver somniferum L. and thebaine in Papaver bracteatum Lindl. Other papaveraceous alkaloids, including salutaridine, oripavine, laudanosine, isothebaine, cryptopine, alpinigenine, narceine, protopine, and gnoscopine, were also quantitated. The values for morphine, codeine, and thebaine in P. somniferum were in agreement within 5—9% with values obtained by the United Nations Narcotics Laboratory by other methods. In contrast to previously reported procedures, the advantage of this method is that no precolumn or other purification other than solvent extraction of the capsular tissue is necessary. Isocratic chromatography alone on a single column resolved the 5 major alkaloids.
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