The isolation, extraction, and spectrophotometric determination of acetylcholine from Lactobacillus plantarum ATCC 10241 is described. Acetylcholine was extracted with a mixture of sodium tetraphenylboron-butylethylketone-acetonitrile and was measured enzymatically at 340 nm.
Skazin's observation that maple flavor is not present as such in sugar maple sap but develops at boiling temperatures (100–104 °C.) has been confirmed. Levorotatory glucosides cannot be detected in sap concentrated to 1/150 its volume, and are therefore probably not present in amounts exceeding 1 gm. (calculated as coniferin) per 50 litres. In maple wood, pyrocatechol tannins were detected but no alcohol-soluble glucosides. The sap contains succinic acid, a glucosidase, an unsaponifiable oil, a water-soluble substance melting at 191.5 °C. and having the composition C11H21O9, and an acetone-soluble substance giving lignin-like color tests. Maple flavor could not be developed in any individual constituent of the sap.
A method of fractionating the chloroform soluble constituents of maple syrup has been devised. Marked differences were observed among Quebec syrups of different years. Fat constituted half the weight of the extract of the 1935 syrup but was completely absent in the 1936, and present in very small quantity in the 1937, product. Crystals of carbonyl compounds having vanillin odour were isolated from bisulphite fractions of the 1935 and 1936 extracts, but the crystals from the two years' syrups differed from each other in melting point and chemical behaviour. Vanillin was not found. An odourless fraction constituting 35% in 1935 and 65% in 1936 had a composition and a behaviour similar to those of lignin. The substance chiefly responsible for maple odour is indicated to be an enolic viscous oil, volatile at 0.03 mm., and present in the 1936 and 1937 syrups in the proportions of 0.6 gm. per 100 gal. of syrup, or about 1 p.p.m.
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