ABSTRACT
Simple thin-layer chromatographic procedures are outlined for the separation, isolation, and characterization of a complex of lipophilic naphthoquinones. Procedures are also described for the quantitative recovery of naphthoquinones from thin-layer plates. The general usefulness of the described methods is demonstrated by their application in the analysis of menaquinones from several microorganisms. The methods allow distinction between menaquinones varying in side-chain length, degree of saturation, and geometry, as well as in the presence or absence of ring methyl groups.
Subject(s)
Bacillus/analysis , Corynebacterium/analysis , Flavobacterium/analysis , Micrococcus/analysis , Mycobacterium/analysis , Streptomyces/analysis , Vitamin K/isolation & purification , Chromatography, Thin Layer , Evaluation Studies as Topic , Hydrogen-Ion Concentration , Light , Methods , Mycobacterium tuberculosis/analysis , Silver , Species Specificity , Spectrophotometry, Ultraviolet , Vitamin K/analysisSubject(s)
Naphthoquinones/analysis , Streptomyces/analysis , Vitamin K/analysis , Alkenes , Chemical Phenomena , Chemistry , Chromatography, Thin Layer , Fluorescent Dyes , Infrared Rays , Lipids/analysis , Magnetic Resonance Spectroscopy , Methods , Naphthoquinones/isolation & purification , Spectrophotometry , Spectrum Analysis , Ultraviolet Rays , Vitamin K/isolation & purificationABSTRACT
Synthetic phylloquinone was resolved into cis and trans isomers by thin-layer chromatography. The two isomers had identical ultraviolet spectra characteristic of vitamin K(1) and were differentiated by nuclear-magnetic-resonance spectroscopy on the basis of the displacement of the peak corresponding to the olefinic methyl group in the naphthoquinone side chain. Studies on the restoration of electron transport coupled to phosphorylation in irradiated preparations of Mycobacterium phlei showed that only the trans isomer was active with substrates linked to nicotinamide-adenine dinucleotide. The purified trans phylloquinone was enzymatically converted to the cis isomer. Under similar conditions, cis vitamin K(1) gave rise to the trans-naphthoquinone. The natural naphthoquinone of M. phlei vitamin MK(9)(II-H) was similarly resolved into cis and trans isomers.
Subject(s)
Mycobacterium/drug effects , Oxidative Phosphorylation/drug effects , Vitamin K 1/chemical synthesis , Chromatography, Thin Layer , Electron Transport , Magnetic Resonance Spectroscopy , NAD/pharmacology , Vitamin K 1/pharmacologyABSTRACT
1. delta-Tocotrienol (8-methyltocotrienol) was isolated from the latex of Hevea brasiliensis. This new member of the tocopherol family is a pale-yellow oil at room temperature. 2. The properties of delta-tocotrienol are very similar to those of delta-tocopherol and the small differences can be explained by the change in side chain. 3. The ultraviolet and infrared spectra of delta-tocotrienol were determined and a conversion factor for use with the Emmerie-Engel reaction was worked out. Details are given for the chromatography of delta-tocotrienol on thin layers (adsorption and partition) and reversed-phase paper, and the nitroso derivatives were formed. 4. An ethyl carbonate ester of delta-tocotrienol was prepared and compared with a similar ester of delta-tocopherol. 5. Hydroxymethylation of delta-tocotrienol followed by reduction gave beta-tocotrienol as a major product.
