Search PubMed⌕ Search

Biomedical subjects

R G Ackman

Publications and source records attributed to R G Ackman.

At least 19 recordsLinked to original sources

Different tocopherols and the relationship between two methods for determination of primary oxidation products in fish oil.

The effectiveness of 2.32 mmol/kg (approximately 1000 ppm) of alpha-, gamma-, or delta-tocopherol (TOH), as well as different levels of alpha TOH, on the formation of hydroperoxides in fish oil was studied by monitoring the peroxide value (POV) and the formation of conjugated dienes (CD) during storage at 30 degrees C. The same order of antioxidant activity was observed by both methods. Linear regression of POV on CD showed that these data were strongly correlated (r(2) > or = 0.98). The value of the slope of the regression lines, however, differed substantially and decreased with increasing hydrogen-donating ability of the different tocopherols and with increasing alpha TOH concentration. It is suggested that this is due, at least in part, to the contribution from hydroxy compounds to the CD measurements and a greater contribution from hydroperoxy epidioxides (two peroxide groups per conjugated diene unit) to the POV than to the CD value. The degrees of formation of both these groups of oxidation products are expected to be influenced by the rate of scavenging of lipid peroxyl and alkoxyl radicals by tocopherol (alpha TOH > gamma TOH > delta TOH).

Antioxidants↗

Purification of ethoxyquin and its two oxidation products.

2,6-Dihydro-2,2,4-trimethyl-6-quinolone (QI) and 1,8'-bis(1, 2-dihydro-6-ethoxy-2,2,4-trimethylquinoline) (DM) are two oxidation products of 1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline (ethoxyquin, EQ). This paper describes several methods for the purification of technical grade EQ and for the production of pure QI and DM as standards with the purity required (>99%) for calibration of quantitative determination methods. EQ of high purity was obtained through vacuum distillation followed by a quick column chromatographic purification on silica gel. Preparative scale purity DM could be obtained through recrystallization from methanol, but QI could be purified only by a high-pressure liquid chromatographic method.

Chromatography, High Pressure Liquid↗

Separation of paralytic shellfish poisoning toxins on Chromarods-SIII by thin-layer chromatography with the Iatroscan (mark 5) and flame thermionic detection.

Thin-layer chromatography (TLC) on Chromarods-SIII with the Iatroscan (Mark-5) and a flame thermionic detector (FTID) was used to develop a rapid method for the detection of paralytic shellfish poisoning (PSP) toxins. The effect of variation in hydrogen (H2) flow, air flow, scan time and detector current on the FTID peak response for both phosphatidylcholine (PC) and PSP were studied in order to define optimum detection conditions. A combination of hydrogen and air flow-rates of 50 ml/min and 1.5-2.0 l/min respectively, along with a scan time of 40 s/rod and detector current of 3.0 A (ampere) or above were found to yield the best results for the detection of PSP compounds. Increasing the detector current level to as high as 3.3 A gave about 130 times more FTID response than did flame ionization detection (FID), for PSP components. Quantities of standards as small as 1 ng neosaxitoxin (NEO), 5 ng saxitoxin (STX), 5 ng B1-toxins (B1), 2 ng gonyautoxin (GTX) 2/3, 6 ng GTX 1/4 and 6 ng C-toxins (C1/C2) could be detected with the FTID. The method detection limits for toxic shellfish tissues using the FTID were 0.4, 2.1, 0.8 and 2.5 micrograms per g tissue for GTX 2/3, STX, NEO and C toxins, respectively. The FTID response increased with increasing detector current and with increasing the scan time. Increasing hydrogen and air flow-rates resulted in decreasing sensitivity within defined limits. Numerous solvent systems were tested, and, solvent consisting of chloroform: methanol-water-acetic acid (30:50:8:2) could separate C toxins from GTX, which eluted ahead of NEO and STX. Accordingly, TLC/FTID with the Iatroscan (Mark-5) seems to be a promising, relatively inexpensive and rapid method of screening plant and animal tissues for PSP toxins.

Calibration↗

Dietary menhaden, seal, and corn oils differentially affect lipid and ex vivo eicosanoid and thiobarbituric acid-reactive substances generation in the guinea pig.

This investigation was carried out to characterize the effects of specific dietary marine oils on tissue and plasma fatty acids and their capacity to generate metabolites (prostanoids, lipid peroxides). Young male guinea pigs were fed nonpurified diet (NP), or NP supplemented (10%, w/w) with menhaden fish oil (MO), harp seal oil (SLO), or corn oil (CO, control diet) for 23 to 28 d. Only the plasma showed significant n-3 polyunsaturated fatty acid (PUFA)-induced reductions in triacylglycerol (TAG) or total cholesterol concentration. Proportions of total n-3 PUFA in organs and plasma were elevated significantly in both MO and SLO dietary groups (relative to CO), and in all TAG fractions levels were significantly higher in MO- than SLO-fed animals. The two marine oil groups differed in their patterns of incorporation of eicosapentaenoic acid (EPA). In guinea pigs fed MO, the highest levels of EPA were in the plasma TAG, whereas in SLO-fed animals, maximal incorporation of EPA was in the heart polar lipids (PL). In both marine oil groups, the greatest increases in both docosahexaenoic acid (22:6n-3, DHA) and docosapentaenoic acid (22:5n-3, DPA), relative to the CO group, were in plasma TAG, although the highest proportions of DHA and DPA were in liver PL and heart TAG, respectively. In comparing the MO and SLO groups, the greatest difference in levels of DHA was in heart TAG (MO > SLO, P < 0.005), and in levels of DPA was in heart PL (SLO > MO, P < 0.0001). The only significant reduction in proportions of the major n-6 PUFA, arachidonic acid (AA), was in the heart PL of the SLO group (SLO > MO = CO, P < 0.005). Marine oil feeding altered ex vivo generation of several prostanoid metabolites of AA, significantly decreasing thromboxane A2 synthesis in homogenates of hearts and livers of guinea pigs fed MO and SLO, respectively (P < 0.04 for both, relative to CO). Lipid peroxides were elevated to similar levels in MO- and SLO-fed animals in plasma, liver, and adipose tissue, but not in heart preparations. This study has shown that guinea pigs respond to dietary marine oils with increased organ and plasma n-3 PUFA, and changes in potential synthesis of metabolites. They also appear to respond to n-3 PUFA-enriched diets in a manner that is different from that of rats.

Adipose Tissue↗

Distribution of alpha- and gamma-tocopherols in Atlantic salmon (Salmo salar) tissues.

Groups of Atlantic salmon parr (mean initial weight 9.5 g) were fed three diets, the first containing no tocopherol supplement, the others supplemented with either all-rac-alpha-tocopherol (A-T) or RRR-gamma-tocopherol (G-T). Tocopherol concentrations in the liver, serum, testes, kidney, brain, gill, muscle, and perivisceral fat were measured after 36 wk. Despite a higher dietary intake of G-T, compared to A-T, deposition of gamma-tocopherol (gammaT) was less efficient than of alpha-tocopherol (alphaT) in most tissues except in the perivisceral fat, an adipose tissue. In fish fed the G-T diet, the gammaT/alphaT ratio was highest in the perivisceral fat and lowest in the liver, indicating that the liver is the most discriminatory organ for retaining alphaT as compared to gammaT, and the perivisceral fat is more suitable for the storage of gammaT. A negative correlation (P < 0.01) was observed between the gammaT/alphaT ratio and the corresponding tissue phospholipid content, suggesting that gammaT is less efficiently deposited compared to alphaT in the phospholipid-rich membranes which are presumed to be the functional site for lipid antioxidants in vivo. During restricted intake of alphaT, the liver and muscle exhibited the greatest reduction of this tocopherol among the tissues analyzed. The presence of minimal alphaT in the muscle from fish fed the tocopherol-unsupplemented diet led to greater susceptibility to lipid peroxidation after frozen storage than was the case for muscle containing higher concentrations of either alphaT or gammaT. However, both alphaT and gammaT were effective stabilizers of salmon muscle lipids during frozen storage.

Animals↗

Digestion of the 1-O-alkyl diacylglycerol ethers of Atlantic dogfish liver oils by Atlantic salmon Salmo salar.

Dogfish (Squalus acanthias) liver poses a waste disposal problem in Canada because it is not utilized for any commercial purpose. The liver of Atlantic dogfish, which is often up to 20% of the weight of the fish, contains 40-70% oil. The oil contains about 30-40% 1-O-alkyl diacylglycerol ethers (DAGE) which render it unacceptable for human use, and it has also not been considered satisfactory for animal feed use. Polyunsaturated fatty acids (20:5n-3 and 22:6n-3) are present in dogfish liver oils at levels comparable to those in herring oil. Dogfish liver oil could be a source of essential fatty acids for Atlantic salmon (Salmo salar), but their ability to hydrolyze DAGE from dogfish oil has not been examined. Experiments were designed to measure the digestibility of fatty acids of DAGE in salmon. The fatty acid moieties were liberated by the digestive enzymes of the fish and made readily available as a source of energy. The 1-O-alkylglycerol ether moiety was absorbed to a small extent but should not constitute a health problem in either the fish or the human fish consumer. The long-chain polyunsaturated fatty acids were particularly well absorbed, with an apparent digestibility in salmon of 87-95% when feeding on dogfish liver oil. The total fatty acids and other lipids were in fact both absorbed to the extent of approximately 85%.

Animals↗

Diets enriched in menhaden fish oil, seal oil, or shark liver oil have distinct effects on the lipid and fatty-acid composition of guinea pig heart.

The purpose of this investigation was to determine whether diets supplemented with oils from three different marine sources, all of which contain high proportions of long-chain n-3 polyunsaturated fatty acids (PUFA), result in qualitatively distinct lipid and fatty acid profiles in guinea pig heart. Albino guinea pigs (14 days old) were fed standard, nonpurified guinea pig diets (NP) or NP supplemented with menhaden fish oil (MO), harp seal oil (SLO) or porbeagle shark liver oil (PLO) (10%, w/w) for 4-5 weeks. An n-6 PUFA control group was fed NP supplemented with corn oil (CO). All animals appeared healthy, with weight gains marginally lower in animals fed the marine oils. Comparison of relative organ weights indicated that only the livers responded to the diets, and that they were heavier only in the marine-oil fed guinea pigs. Heart total cholesterol levels were unaffected by supplementing NP with any of the oils, whereas all increased the triacylglycerol (TAG) content. The fatty-acid profiles of total phospholipid (TPL), TAG and free fatty acid (FFA) fractions of heart lipids showed that feeding n-3 PUFA significantly altered the proportions of specific fatty-acid classes. For example, all marine-oil-rich diets were associated with increases in total monounsaturated fatty acids in TPL (p < 0.05), and with decreases in total saturates in TAG (p < 0.05). Predictably, the n-3 PUFA enriched regimens significantly increased the cardiac content of n-3 PUFA and decreased that of n-6 PUFA, although the extent varied among the diets. As a result, n-6/n-3 ratios were significantly lower in all myocardial lipid classes of marine-oil-fed guinea pigs. Analyses of the profiles of individual PUFA indicated that quantitatively, the fatty acids of the three marine oils were metabolized and/or incorporated into TPL, TAG and FFA in a diet-specific manner. In animals fed MO-enriched diets in which eicosapentaenoic acid (EPA) > docosahexacnoic acid (DHA), ratios of DHA/EPA in the hearts were 1.2, 2.2 and 1.5 in TPL, TAG and FFA, respectively. In SLO-fed guinea pigs in which dietary EPA approximately DHA, ratios of DHA/EPA were 0.9, 3.4 and 2.1 in TPL, TAG and FFA, respectively. Feeding NP + PLO (DHA/EPA = 4.8), resulted in values for DHA/EPA in cardiac tissue of 2.1, 10.6 and 2.9 in TPL, TAG and FFA, respectively. In the TAG and FFA, proportions of n-3 docosapentaenoic acid (n-3 DPA) were equal to or higher than EPA in the SLO- and PLO-fed animals. The latter group exhibited the greatest difference between the DHA/n-3 DPA ratio in the diet and in cardiac TAG and FFA fractions (7, 3.4 and 3.1, respectively). Quantitative analysis indicated that > or = 85% of the n-3 PUFA were in TPL, 7-11% were in TAG, and 2-6% were FFA. Specific patterns of distribution of EPA, DPA and DHA depended on the dietary oil. Both the qualitative and quantitative results of this study demonstrated that in guinea pigs, n-3 PUFA in different marine oils are metabolized and/or incorporated into cardiac lipids in distinct manners. In support of the concept that the diet-induced alterations reflect changes specifically in cardiomyocytes, we observed that direct supplementation of cultured guinea pig myocytes for 2-3 weeks with EPA or DHA produced changes in the PUFA profiles of their TPL that were qualitatively similar to those observed in tissue from the dietary study. The factors that regulate specific deposition of n-3 PUFA from either dietary oils or individual PUFA are not yet known, however the differences that we observed could in some manner be related to cardiac function and thus their relative potentials as health-promoting dietary fats.

Animals↗

Positional isomerization of trans-3-hexadecenoic acid employing 2-amino-2-methyl-propanol as a derivatizing agent for ethylenic bond location by gas chromatography/mass spectrometry.

The effect of derivatization with 2-amino-2-methyl-propanol on trans-3-hexadecenoic acid was investigated as part of the identification of the trans-3-hexadecenoic acid in two Nova Scotian seaweeds. After the extraction of the total fatty acids and their methylation, the monoenoic trans fraction was isolated by thin-layer chromatography on silica gels impregnated with silver nitrate. This fraction was first analyzed by gas chromatography and showed the presence of the trans-3-hexadecenoic acid; other fatty acids were not present. The isolated fraction was derivatized with 2-amino-2-methyl-propanol prior to analysis by gas chromatography/mass spectrometry. The chromatogram obtained showed the presence of a positional isomer formed during the derivatization of the trans-3-hexadecenoic acid. The mass spectrum showed a prominent [M+H] and diagnostic ions for the identification of the unknown isomer, corresponding to the 4,4-dimethyloxazoline (DMOX) derivative of a presumed 2-hexadecenoic acid. Definitive confirmation of the ethylenic bond position was obtained by oxidative ozonolysis of the DMOX derivatives of the fatty acids under investigation. Infrared spectroscopy showed that the artifact formed during the DMOX derivatization of trans-3-hexadecenoic acid was the DMOX derivative of cis-2-hexadecenoic acid.

Gas Chromatography-Mass Spectrometry↗

Very long chain polyunsaturated fatty acids in the blubber of ringed seals (Phoca hispida sp.) from Lake Saimaa, Lake Ladoga, the Baltic Sea, and Spitsbergen.

Blubbers of four ringed seal subspecies from Lake Saimaa, Lake Ladoga, the Baltic Sea, and Spitsbergen were analyzed for very long chain polyunsaturated fatty acids (VLCPUFA; > C22) using gas-liquid chromatography and gas chromatography/mass spectrometry. The VLCPUFA of the blubber oils were mainly n-3 polyunsaturated fatty acids--23:5n-3, 24:3n-3, 24:4n-3, 24:5n-3, 24:6n-3, 26:5n-3, 26:6n-3, and 28:7n-3. The largest VLCPUFA components in all populations were 24:5n-3 (0.1-0.2 wt% of total fatty acids) and 24:6n-3 (0.1%), but 24:4n-3 (0.1%) was also prominent in the Baltic specimens. The blubber oils of the freshwater species contained considerably more 24:4n-6 and 24:5n-6 than the blubbers of the marine species. The differences among the VLCPUFA in these subspecies appear to be mainly due to different dietary VLCPUFA.

Adipose Tissue↗

Marine and freshwater fish oil varying in arachidonic, eicosapentaenoic and docosahexaenoic acids differ in their effects on organ lipids and fatty acids in growing rats.

Arachidonic acid [20:4(n-6)] and docosahexaenoic acid [22:6(n-3)] are important to normal neurodevelopment and visual function. Infants fed formula often have low blood lipid 20:4(n-6) and 22:6(n-3). Consumption of fish oils high in eicosapentaenoic acid [20:5(n-3)] and 22:6(n-3) with no 20:4(n-6) increases tissue 20:5(n-3) and 22:6(n-3) but decreases 20:4(n-6). Some freshwater fish oils contain higher 20:4(n-6) and lower 20:5(n-3) than usual marine fish oils, but their effects on tissue fatty acids are not well known. Therefore, the effects of feeding weaning rats 30 d with 12% (wt/wt) soybean oil [0.0% 20:4(n-6), 20:5(n-3) and 22:6(n-3)], 2% safflower oil with 10% marine fish oil [0.9% 20:4(n-6), 15.1% 20:5(n-3), 7.3% 22:6(n-3)] or 10% freshwater fish oil [3.3% 20:4(n-6), 5.9% 20:5(n-3), 8.0% 22:6(n-3)] on plasma, tissue and brain fatty acids was determined. Levels (g/100 g) of 20:4(n-6) were significantly higher and 20:5(n-3) lower in plasma, liver, kidney and brain of rats fed freshwater fish oil rather than marine fish oil. Marine fish oil, but not freshwater fish oil resulted in a higher brain 20:5(n-3) and 22:6(n-3), and lower 20:4(n-6) than soybean oil. Plasma and liver triglyceride concentrations were significantly lower in rats fed marine fish oil, but not in rats fed soybean oil when compared with those fed freshwater fish oil. The results indicate dietary 20:4(n-6) prevents the decline in plasma and tissue 20:4(n-6) caused by dietary 20:5(n-3) and/or 22:6(n-3). Oils with 20:4(n-6) may affect cholesterol and triglyceride metabolism differently than usual fish oils.

Animals↗

Confirmation by gas chromatography/mass spectrometry of two unusual trans-3-monoethylenic fatty acids from the Nova Scotian seaweeds Palmaria palmata and Chondrus crispus.

The structures of two unusual fatty acids, the known trans-3-hexadecenoic acid and a novel trans-3-tetradecenoic acid, both isolated from the Nova Scotian seaweeds Palmaria palmata and Chondrus crispus, were positively identified. After the extraction of the total fatty acids by saponification, followed by methylation, the monoenoic trans fractions were isolated by thin-layer chromatography on silica gel impregnated with silver nitrate. The monoenoic trans fractions were derivatized with 2-amino-2-methyl-propanol prior to analysis by gas chromatography/mass spectrometry. The mass spectra showed prominent [M + 1]+ ions but lacked the 12 amu interval useful for identifying the double bond position. Hence, alternative diagnostic peaks were used to confirm the position of the double bond in these two fatty acids. As the trans-3-hexadecenoic acid is found in the photosynthetic tissue of all plants, it may also be present in ruminant fats and, presumably, in human adipose tissue.

Buffers↗

The absorption of fish oils and concentrates.

Both preventive and curative therapies have created a considerable demand for eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. The most common sources for omega 3 fatty acids are fish oil. The concentrations of EPA and DHA in commercial oils, after modest enrichment, reach about 300 mg/g; alternative technologies can produce reasonably priced fish oils containing 400 or even 500 mg/g of omega 3 acids. When the acids are liberated from the glycerides, concentrates of ethyl esters or free acids with 65 to 70% total omega 3 fatty acids (at least 50% EPA + DHA) are readily prepared. Difficulties have arisen because most clinical trials have used fish oils of unspecified composition, and some trials are now based on either ethyl esters or free acids. There are at least three different, but not mutually exclusive, absorption routes in humans, namely the preduodenal route, the lymphatic route via chylomicrons, and the route via the portal vein to the liver. This makes it difficult to compare results. The difficulty in obtaining dose-related clinical data may in part be due to the form in which the omega 3 acids are offered and due in part to the natural presence of these fatty acids in the body. The nontriglyceride forms, especially the free acids, have been advocated for standardization of trials to facilitate interlaboratory comparisons.

Absorption↗

Cholestane as a digestibility marker in the absorption of polyunsaturated fatty acid ethyl esters in Atlantic salmon.

Salmonid fish require long-chain n-3 fatty acids in their diet. The digestibility of different chemical forms of fish oil fatty acids, fed as triacylglycerols, free fatty acids or ethyl esters, was examined in 300 g farmed Atlantic salmon (Salmo salar) using cholestane as an indicator of fat absorption in lieu of the chromium oxide (Cr2O3) which is commonly used as a marker in digestibility studies. It was established that the two digestibility markers gave similar results. Conveniently, cholestane does not require a separate analysis if fatty acids are to be determined by appropriate gas-liquid chromatography. The long-chain polyunsaturated fatty acids were particularly well absorbed, the apparent digestibility being 90-98% when feeding triacylglycerols or free fatty acids. However, the digestibility of monounsaturated fatty acids (75-94%) was lower, and lower still for saturated fatty acids (50-80%). Ethyl esters of fatty acids were significantly less well absorbed (P less than 0.05) than were the corresponding fatty acids in free acid or triacylglycerol form. Irrespective of dietary fat type, only free fatty acids were identified in feces, indicating total hydrolysis of triacylglycerols and ethyl esters.

Animals↗