Two new schemes for fractionation of complex lipid mixtures are presented. Their use for the study of lipids of beef brain, beef liver, and the sea anemone are described. Apparatus and techniques for working in an inert atmosphere, evaporation of solutions in the cold under nitrogen, use of infrared spectroscopy for examination of lipids and their hydrolysis products, preparation and clution of diethylaminoethyl (DEAE) cellulose and silicic acid‐silicate columns and general column combinations that can be used to fractionate complex lipid mixtures are considered in detail. The first scheme, employing DEAE cellulose columns followed by thin layer and paper chromatographic examination of the fractions, was applied to liver lipids. The many components, some of them new lipids not previously detected, are clearly seen with this technique but are not seen when paper or thin layer chromatography alone or silicic acid chromatography are used. The second scheme employing DEAE for initial fractionation, followed by complete separation on silicic acid and silicic acid‐silicate columns, was applied to lipids of the sea anemone and beef brain. Typical lecithin and phosphatidyl ethanolamine were isolated, but sphingomyelin was not found. A new sphingolipid, ceramide aminoethylphosphonate, with a free amino group and a direct carbon to phosphorus bond was isolated and characterized. The methods used for quantitative isolation, the infrared spectra, and the amounts of cholesterol, ceramide, cerebroside, galactosylglyceride, sulfatide, sphingomyelin, lecithin, phosphatidyl ethanolamine, phosphatidyl serine, phosphatidyl inositol, triphosphoinositide, phosphatidic acid, cardiolipin, and ganglioside of beef brain are presented. Finally, the types of lipid‐nonlipid interactions disclosed by column chromatography and their potential application to biological problems are discussed.

The described analytical method for histamine determination in foods requires sample homogenization in methanol, heating, centrifuging or filtering, several extractions, and fluorometric detection of histamine with o-phthalaldehyde. The method eliminates potential interference by other amines through a selective extraction step. Samples of 20 foods, including seafood products (fresh, frozen and canned), comminuted meats, cheeses and sauerkraut (canned), were analyzed for histamine content. Canned sauerkraut and tuna fish had the highest average histamine content among tested foods. This method can be used to detect histamine in food samples that contain as little as 0.02 mg histamine/ 1OOg of food.

A quantitative chromatographic procedure for the fractionation of complex lipid mixtures is described. The method utilizes diethylaminoethyl (DEAE) cellulose column chromatography followed by thin layer chromatography (TLC). Spots produced in TLC are charred with sulfuric acid‐potassium dichromate and heat and are then measured by quantitative densitometry. Results obtained with beef brain and beef heart mitochondrial lipids are presented, and the close correspondence between column isolation procedures and the new procedure is demonstrated. Methods utilizing only column chromatography, column chromatography and TLC, and one‐ and two‐dimensional TLC without column chromatography are compared.

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A total of 112 bacterial strains representing 38 species were tested for their potential to elicit food poisoning outbreaks via histamine formation in foods. Proteus morganii and Enterobacter aerogenes displayed a quantitative superiority in terms of histamine production on a trypticase‐soy broth‐histidine (TSBH) medium and a tuna fish infusion broth (TFIB). When bacteria were incubated under standardized conditions in TSBH medium, histamine accumulated to levels exceeding 50 nmoles/ml of media with a total of 23 strains, including 13 of 15 P. morganii strains, 3 of 3 E. aerogenes strains, 3 of 12 Hafnia alvei strains, 1 of 4 Providencia alcalifaciens strains, 1 of 5 Enterobacter cloacae strains, 1 of 1 Proteus rettgeri strains, and 1 of 1 Citrobacter diversus strains. However, only 8 of the 15 P. morganii strains and the 3 E. aerogenes strains were capable of generating histamine in excess of 200 nmoles/ml in the TSBH medium. Of the 23 strains capable of appreciable histamine production in TSBH medium, P. morganii and E. aerogenes were, by far, the most prolific histamine producers in TFIB. Of the organisms tested, only P. morganii and E. aerogenes would appear to have the capability of forming sufficient histamine in scombroid fish products to elicit food poisoning outbreaks.