As the results of the previous studies on the lipids of micronektonic fishes and shrimps and zooplankton, it has been recognized that one of the most characteristic constituents of the midwater marineanimals' lipids is the wax esters. Moreover, a reversed relation betweon the triacylglycerol and wax ester contents has been observed, namely, the former is the main lipid class in the surface living animals, whereas the latter becomes the major one in the meso- and bathypelagic animals. In order to study the biosynthesis of wax ester by marine animal and microorganism the in vivo and in vitro experiments were carried out. By using intact zooplankters which were caught from different depths, 50 m, 400 m, 1000 m, by a net, the incorporation of hexadecyl alcohol [14CH2OH], -palmitic and -acetate into the wax esters was examined. In either cases, the higher radioactivity percentages were measured by the deeper zooplankters. Among the substrates used the highest incorporation was resulted from the use of hexadecyl alcohol. The inverse proportion between the triacylglycerol and wax ester formations with depth was recognized in the case of acetate incorporation. Furthermore, a limited interconversion of fatty acid and fatty alcohol moieties of the wax ester was demonstrated. The homogenate supernatants of myctophids, Diaphus glandulifer, decapods, Sergestes prehensilis, and copepods. Pareuchaeta elongata, synthesized the wax esters from [1-14C] hexadecyl alcohol, -palmitic and -oleic acids, -tripalmitoylglycerol and -acetate. The addition of cofactors, ATP and CoA, enhanced the incorporation of acids in to the wax esters as much as 2 to 3 times, but did not for the case of albumin complex of oleic acid [14COOH]. Marine microorganisms from different depths were collected on Millipore HA filters by filtrating 5 m, 50 m, 300 m, 1000 m layer waters. A tentative experiment on the biosynthesis and hydrolysis of wax esters by microorganisms was done on board. Unexpectedly high activities were detected in the classes of wax esters as well as of aldehydes and phospholipids. There were no clear relation of wax ester formation with depth. As the cofactors, ATP and CoA did not activate the synthesis of wax ester, but the addition of carnitine worked. Moreover, it was found that marine microorganisms also hydrolyzed oleyl [14COOH] palmitate used for substrate. As the conclusion from the results obtained here, all of these midwater animals and microorganisms biosynthesize the wax esters from the universally available precursors, acetate, fatty acids, as well as from long-chain alcohols. The data are discussed why wax esters are biosynthesized by them in midwater.
Boron trifluoride (BF3) -methanol complex is widely used for the preparation of methyl esters as the sample for gas chromatography. However, it is feared that short-chain fatty acids are lost, when esters are prepared from fats and oils containing these acids by the conventional BF3-methanol method. Collaborative studies were carried out for the purpose of establishing a standard method for the quantitative preparation of short-chain esters. Soybean oils containing definite amounts of glycerol trihexanoate and glycerol trioctanoate were prepared and used as the samples for the collaborative studies. GC analysis of the esters prepared from these samples by the conventional method revealed the appreciable loss of hexanoic- and octanoic acids through the procedure. Partial evaporation of the short chain esters and partial passage of them into aqueous sodium chloride layer were supposed to be responsible for the low recovery of these esters. Further collaborative studies were performed to examine the conditions for the prevention of the loss of short chain esters. In conclusion, the collaborative studies suggested that the recovery of the esters of short-chain acids (C6 and C8) could be improved by the following modification of the conventional method. a) Shaking the mixture after the addition of aqueous sodium chloride solution. b) Reextraction of esters from the aqueous layer after the separation of hexane (or heptane) layer. c) Immediate GC analysis after the esterification. It was concluded that 7 ml of 7%-BF3-methanol reagent was enough to esterify 300 mg of oil sample.
The five samples of N-n-alkyl-N-p-toluenesulfonylsulfanilates [3-n] (alkyl; n-butyl [3-4], n-hexyl [3-6], n-octyl [3-8], n-decyl [3-10], n-dodecyl [3-12]) were prepared from sodium N-p-toluenesulfonylsulfanilate and corresponding alkyl bromides or alkyl tosylates. They could well refined as anilinium salts. The physico-chemical studies on their aqueous solution were carried out. The results obtained were as follows. 1) It was found that [3-8] was the best of [3-n] in solubility for water and in the abilities for wettability and decrease of surface tension. 2) The effect of [3-10] for solubilization of Orange OT, was six times better than that of sodium N, N-dibenzylsulfanilate. 3) The aqueous solution of [3-12] showed high emulsificability.
The effects of the physico-chemical properties of the aqueous solution of 38 carboxylate builders (Table-1) on detergencies have been studied. The correlation between the building activity and molecular structure has been discussed. The detergency tests were carried out on naturally soiled cotton fablics and the detergency powers were evaluated by Scheffe's method. Measurements were made on the surface tension, the critical micelle concentration, the emulsifying and solubilization capacities of the builder solution with DBS. The buffering and dispersing capacities for carbon black and manganese dioxide of the aqueous builder solution, and the chelate stability constants of the builders with alkaline earth metals were determined. These results indicate that the washing efficiencies of formulations containing the carboxylate builder correlate well with their dispersing capacities for manganese dioxide (Fig.-7) and chelate stability constants with calcium ions (Fig.-8). Other physico-chemical properties of the builder solutions have no relation to their washing efficiencies.
2-Formylpyridineketoxime, 2- acetylpyridineketoxime and 2-benzoylpyridineketoxime were found to coordinate to iron (II) ion in aqueous solution to give intensely colored complexes. Their molar extinction coefficients at the maximum wavelength were about 104. The complexes were fairly stable even in a highly alkaline solution (ca. pH 12.0). The conjugation over the ligand and the chelate ring of the present complexes was considered to play an important role on these characteristic color reactions. The absorption spectra of the complexes were examined at marious pH ranges. These organic ligands were found to be applicable to the colorimetric determinatin of iron (II) ion.
Two types of new cyclic ketones were prepared from conjugated hydrocarbons by the combination of Diels-Alder reaction, Grignard reaction and cyclization. (1) New cyclohexenyl alkenyl ketones were obtained by the reaction of various methyl cyclohexenyl carboxylates and vinylmagnesium chloride; for example, 1- (4'-methyl-3'-cyclohexen-1'-yl) -4-penten-1-one was produced from methyl 4-methyl-3-cyclohexen-1-carboxylate and vinylmagnesium chloride. (2) Several hydroindene derivatives were prepared from 2, 7-dimethyl-1, 3, 7-octatriene via Diels Alder reaction followed by acidic cyclization ; for example, a mixture of 2-acetyl-5, 7, 7-trimethylbicyclo [4, 3, 0] non -1 (6) -ene, 2-acetyl-5, 7, 7-trimethylbicyclo [4, 3, 0] non-1-ene and 2-isopropyl-3, 6-dimethylindene was obtained by the cyclization of 4-acetyl-1-methyl-3- (3'-methyl-3'-butenyl) -1-cyclohexene.
The effect of ionic properties of poly (L-Glu) [poly (L-Glutamic acid)] and poly (L-Lys) [poly (L-Lysine)] on metal-catalyzed fatty acid oxidation were studied as a model of metal-fatty acid-protein interaction in aqueous solution. In the poly (L-Glu) system, most of added Fe3+ combined with poly (L-Glu), and the resultant Fe3+ poly (L-Glu) complex caused the inhibition of the oxidant activity of added Fe3+. On the other hand, about 70% of added Cu2+ was remained in ionic state in the poly (L-Glu) system, and the oxidant activity of added Cu2+ was observed. In the poly (L-Lys) system, the insoluble fatty acid-poly (L-Lys) complex was formed, and most of added metal ion was contained in the complex. The rate of metal catalyzed fatty acid oxidation increased with an increase in poly (L-Lys) concentration and witn a rise in temperature under the formation of the insoluble fatty acid-poly (L-Lys) complex.
Oxygen content in fats and oils was investigated by a modification of the Schutze-Unterzaucher method. Oxygen in soybean oil and colza oil heated at 180°C in air was determined periodically. The oxygen contents of soybean oil and colza oil were 9.9% and 9.2% initially and 8.2% and 9.1% after 30 hours, respectively, therefore the change in the oxygen contents was very little in this case. Oxygen was passed through linseed oil heated at 100°C and 200°C, and the oxygen content in linseed oil determined perisdically. The oxygen content in linseed oil was initially 11.0%, that in gelatinized linseed oil heated at 100°C for 10 hours was 17.8%, and that in gelatinized one heated at 200°C for 5 hours was 15.0%. Then, oxygen content in perilla oil was similarly determined. The oxygen content in linseed oil was determined in the process of drying test by using cobalt naphthenate as a drier. The oxygen content in linseed oil was 8.5% initially and 21.6% after 6 hours. To linseed oii, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) or isopentyl gallate was added, each mixture heated at 100°C or 200°C under the passage of oxygen, and the oxygen content was determined. This determination of oxygen content in fats and oils affected by their aging seems to be effective for clarifying the process of their oxidation and degradation, in addition to other usual instrumental analyses.