Chiaki Koizumi

High-pressure processing of fish and fish products

Abstract High hydrostatic pressure has recently been applied in food processing, and several commercial fruit and vegetable products have already been put on sale. High hydrostatic pressure results in protein denaturation, resulting in inhibition of some inherent enzymatic activities and of the biogenic activity of some microorganisms. However, high pressure also accelerates lipid oxidation in muscle tissues. Recent intensive research on the effects of high hydrostatic pressure on fish tissues has gradually revealed the benefits and defects of this novel processing technology.

Oxidative decomposition of cholesterol in fish products

Cholesterol oxides in fish products popular in Japan, including salted and dried, boiled and dried and smoked products, were qualitatively and quantitatively determined as trimethylsilyl ether derivatives by gas-liquid chromatography and mass spectrometry. The level of total cholesterol oxides ranged widely between 8.3 ppm in boiled and dried shrimp and 188.0 ppm in boiled and dried anchovy. 7β-Hydroxycholesterol and 7-ketocholesterol were the most prominent oxidative decomposition products of cholesterol. The levels of epimeric epoxides, cholestane triol and 25-hydroxycholesterol were relatively low. To elucidate a mechanism of cholesterol oxidation proceeding during fish processing and subsequent preservation, four model systems, consisting of a mixture of purified cod liver triglycerides plus cholesterol, of a mixture of authentic triolein plus cholesterol, of triolein alone and of cholesterol alone, were stored separately at 25°C in dry air for up to 104 d. The residual fatty acids of the triglycerides, and the cholesterol oxides produced, were recovered and determined. Oxygen uptake remained almost unchanged for the mixture of triolein plus cholesterol. No detectable amount of cholesterol oxides was produced, and the fatty acid content of the residual oleic acid, measured by an internal standard, remained almost unchanged. For the mixture of cod liver triglycerides plus cholesterol, a remarkable increase in oxygen uptake was observed. A continuous increase in the amount of cholesterol oxides was observed, accompanied by a remarkable concurrent decrease in polyunsaturated fatty acid residues, as well as of the oleic acid naturally present. These results strongly suggest that cholesterol oxidation in fish products proceeds in conjunction with oxidative decomposition of the coexisting polyunsaturated fatty acids of fish oils.

Oxidative stability of sardine and mackerel lipids with reference to synergism between phospholipids and α-tocopherol

Relationships between oxidative stability and the compositions of sardine and mackerel lipids were investigated in view of possible synergism between phospholipids andα-tocopherol (α-Toc). The total lipids extracted from viscera were highly susceptible to autoxidation, compared with lipids of white and red muscles and of skin. This seemed to be due to lower concentrations ofα-Toc and phosphatidylethanolamine (PE) in the tissue, but not to the level of polyunsaturated fatty acids. The synergistic effect of PE withα-Toc seemed to be slightly affected by the degree of unsaturation of its fatty acyl chains. The synergistic ability ofO-phosphoethanolamine, the base moiety of PE, was higher than that ofO-phosphoserine.O-Phosphocholine was only slightly effective. During the induction period of autoxidation, theα-Toc level decreased rapidly, and rapid lipid oxidation began only afterα-Toc was almost exhausted.

Influence of the position of unsaturated fatty acid esterified glycerol on the oxidation rate of triglyceride

The influence of the position of unsaturated fatty acid esterified glycerol on the oxidation rate of triglyceride was investigated at 50 C. Randomized triglycerides used were prepared by random interesterification between saturated and unsaturated monoacid triglycerides using sodium methoxide as catalyst. The monoacid triglycerides used were tripalmitin, tristearin, triolein and trilinolein. The molecular species of the randomized triglycerides were analyzed by high performance liquid chromatography (HPLC) in combination with gas liquid chromatography (GLC) and enzymatic hydrolysis. From the results of oxygen absorption measurement by GLC, the randomized triglycerides were more stable towards oxidation than the triglyceride mixtures which were prepared by mixing the equivalent quantities of the same monoacid triglycerides as used in the random interesterification. This may be due to the decrease in the contents of most unstable unsaturated monoacid triglycerides by random interesterification with saturated monoacid triglycerides. Furthermore, from the results obtained with the detailed analysis of the randomized triglycerides at different stages of oxidation, it became clear that the triglycerides having unsaturated fatty acids linked at the 2-position of glycerol are more stable towards oxidation than those linked at the 1(or 3)-position. The carbon chain length of saturated fatty acids has essentially no influence on the oxidation rates of unsaturated fatty acids esterified in the same glycerol.

1-O-alk-1′-enyl-2-acyl and 1-O-alkyl-2-acyl glycerophospholipids in white muscle of bonitoEuthynnus pelamis (Linnaeus)

The existence of ether-linked phospholipids, including 1-O-alk-1′-enyl-2-acyl and 1-O-alkyl-2-acyl-sn-glycero-3-phosphocholines and ethanolamines in bonitoEuthynnus pelamis (Linnaeus) white muscle, was investigated by gas chromatography and gas chromatography-mass spectrometry. Chemical ionization (iso-butane) mass spectrometry of trimethylsilyl ethers derived from the corresponding ether-linked glycerophospholipids proved effective not only for determining molecular weights but also for structural identification based on the ions [M−R]+, [M−RO]+ and [M+1]+. 1-O-Alk-1′-enyl-2-acyl-sn-glycero-3-phosphocholine and ethanolamine accounted for 3.0–6.0% and 3.6–7.6% of the total glycerophospholipids, respectively. 1-O-Alkyl-2-acyl-sn-glycero-3-phosphocholine and ethanolamine were also determined for one fish and accounted for 1.4% and 0.6% of the total glycerophospholipids, respectively. The predominant long chains in thesn-1 position of the glycerol moieties were 16∶0, 18∶0 and 18∶1 in the case of the alkenylacyl and alkylacyl components. Fatty acid distribution of individual glycerophospholipids was also determined.

Fatty chain compositon of ether and ester glycerophospholipids in the Japanese oysterCrassostrea gigas (Thunberg)

The fatty chain compositions of 1-O-alk-1′-enyl-2-acyl, 1-0-alkyl-2-acyl, and 1,2-diacyl glycerophospholipids of the Japanese oysterCrassostrea gigas (Thunberg) were investigated. Major fatty chains in thesn-1 position of 1-alk-1′-enyl-2-acyl ethanolamine phospholipids (EPL) were 18∶0 (64.7%) and 20∶1 (11.1%). Majorsn-1 chains of alkenylacyl choline phospholipids (CPL) were 18∶0 (63.3%) and 16∶0 (22.2%). In the case of 1-alkyl-2-acyl EPL, the predominant fatty chains in thesn-1 position were 18∶0 (51.5%), 16∶0 (16.0%) and 20∶1 (12.5%); in the case of 1-alkyl-2-acyl CPL, the majorsn-1 chains were 16∶0 (44.0%) and 14∶0 (23.4%). Saturated fatty chains were predominant in both EPL and CPL. Prominent fatty acids in thesn-2 position of the alkenylacyl EPL were 22∶6n−3 (29.0%), 20∶5n−3 (19.0%) and 22∶2 NMID (non-methylene interrupted dienes, 16.6%) contributing to about 65% of the total fatty acids, while alkenylacyl CPL was rich in the saturated acids 16∶0 (32.0%) and 18∶0 (9.2%). In the alkylacyl EPL, 16∶0, 18∶1n−9, 18∶0 and 16∶1n−7 were prominentsn-2 fatty acids and accounted for 30.6%, 10.0%, 9.8%, and 8.3%, respectively. Polyunsaturated fatty acids were detected, but were present at extremely low percentages. Majorsn-2 fatty acids in alkylacyl CPL were 16∶0 (25.4%), 22∶6n−3 (16.0%) and 20∶5n−3 (8.4%). The major fatty acids of diacyl EPL were 20∶5n−3 (22.3%), 16∶0 (17.9%), and 18∶0 (16.1%), and those of diacyl CPL were 16∶0 (30.4%), 20∶5n−3 (17.6%) and 18∶1n−7 (7.4%).

Quantitative Determination of Butylated Hydroxyanisole, Butylated Hydroxytoluene, and tert-Butyl Hydroquinone in Oils, Foods, and Biological Fluids by High-Performance Liquid Chromatography with Fluorometric Detection

Concentrations of synthetic antioxidants butylated hydroxyanisole, butylated hydroxytoluene, and tert-butyl hydroquinone were quantified using a high-performance liquid chromatograph with spectrofluorometric detector. The antioxidants were separated and eluted on a reversed-phase column by gradient of a mixture of H2O/acetonitrile/acetic acid (66.5: 28.5∶5, by vol) and a mixture of acetonitrile/acetic acid (95∶5, vol/vol). The eluants were monitored at emission and excitation wavelengths of 310 and 280 nm, respectively. Calibration curves obtained using peak areas against concentration showed ligh coefficients of multiple determination (R2>0.99) for all antioxidants. Known concentrations of added antioxidant standards were recoverable within 98–99% from oils and over 93% from mouse blood. This method requires minimum sample extraction and purification before analysis and provides a relatively high percentage recovery. The method has been applied successfully for the measurement of antioxidant concentrations in oils, dried foods, and biological fluids.

Application of selective ion monitoring to the analysis of molecular species of vegetable oil triacylglycerols separated by open-tubular column GLC on a methylphenylsilicone phase at high temperature

Analyses of the molecular species of authentic triacylglycerols and of soybean triacylglycerols, which were separated on a fused silica open-tubular column coated with a methylphenylsilicone gum stationary phase, were carried out by selective ion monitoring mass spectrometry. It was determined that peak assignments could be made by selecting certain characteristic ions with the same retention time on the SIM profile, that is, three ions of the type RCO+ corresponding to the fatty acyl residues (R1CO+, R2CO+ and R3CO+) and the corresponding three M-acyl ions ([M-OCOR1]+, [M-OCOR2]+ and [M-OCOR3]+ instead of the molecular ion. The yield of [RCO-1]+ and [RCO-2]+ and relating these to the unsaturated fatty acyl residues is inconvenient for an exact peak assignment. The SIM method was further applied to the peak assignment of the triacylglycerols from palm, cottonseed and safflower oils and certain new fatty acid combinations are proposed.

Effect of Dietary Fatty Acids on Ca2+-ATPase Activity of the Sarcoplasmic Reticulum of Rainbow Trout Skeletal Muscle

Abstract Effects of different dietary lipids, corn oil, cod liver oil, and beef tallow, on lipid compositions and Ca 2+ -ATPase activity of the rainbow trout Salmo gairdneri sarcoplasmic reticulum (SR) were examined. The levels of phosphatidylethanolamine (PE) of the muscle and the SR were lower in fish fed for 2 months on diets rich in n -6 fatty acids or deficient in polyunsaturated fatty acids (PUFA) than in those fed n-3 fatty acid rich diet. Fatty acid compositions of muscle well reflected those of the diets. On the other hand, the SRs contained higher levels of polyunsaturated fatty acids, such as 22:6 n -3, than muscle. In particular, PE fractions of each SR exhibited very high contents of polyunsaturated fatty acids and feeding PUFA-deficient diet for 3 months failed to reduce markedly the level of 22:6 n -3 in PEs. Fatty acid compositions of phosphatidylcholine (PC) fractions of the SRs were altered by dietary fatty acids more easily than those of PEs. n -3 Fatty acid rich diet feeding caused reduction of SR Ca 2+ -ATPase activities. There was hardly any difference in peptide mapping patterns of Ca 2+ -ATPase proteins through the experimental stages, suggesting that there was practically no change of the Ca 2+ -ATPase moiety itself in the primary structure. These results suggested that dietary fatty acid compositions would alter lipid class and fatty acid compositions of the rainbow trout SRs and consequently lipid conformation changes. It was concluded that such physiological changes in lipids might affect the SR Ca 2+ -ATPase activities.

New developments in surimi technology

Abstract Surimi - minced and washed fish meat - has primarily been produced from white-muscled fish species, since the desirable odor, light color and gel-forming characteristics of the resultant surimi are very important to enable further processing to heat-induced gel products such as crab leg analog. The production of lowfat surimi with desirable gel-forming characteristics from sardine and mackerel by conventional processing methods has had only limited success; however, recent developments have opened the way to the utilization of red-meat fish for surimi production. During surimi processing, the loss of water-soluble proteins into the wash water leads not only to the waste of certain valuable nutrients but also to pollution of the water; new methods for the recovery of water-soluble proteins from the wash water are described. The role of cryoprotectants in improving frozen surimi storage is explained by the glass transition concept.