Interference of polar lipids with the alkalimetric determination of free fatty acid in fish lipids

An examination of the suitability of an alkalimetric method for the determination of free fatty acid (FFA) contents in fats, oils, and lipid extracts was conducted by comparing AOCS method Ca 5a-40 with a method based on a Chromarod-latroscan thin-layer chromatography-flame-ionization detector (TLC-FID) system. The FFA contents determined by the alkalimetric method were consistently higher than the genuine FFA contents obtained by the latroscan TLC-FID method. Phospholipids were found to be the major components that contributed to the alkali-titratable, nongenuine FFA in the total FFA determined alkalimetrically. Contributions from other polar lipid components were smaller, but they dominated as the proportion of phospholipids fell. The other alkali-titratable polar components may include oxidized lipids and their by-products bound to protein fragments. The accurate determination of FFA contents by alkalimetric methods may only be applicable to those commercially refined fats and oils that contain negligible amounts of phospholipids. Corrections for the alkalimetrically determined FFA contents should be made for those fats and oils with relatively high phospholipid contents by correlating the nongenuine FFA contents and the phospholipid contents.     

Lipid analysis of Greek broad bean oil: Preparation of liposomes and physicochemical characterization

Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba L. (syn. Fabae calabaricae)] oil and their physical properties were studied. The method of preparation was the hydration of the thin lipid film, while the probe sonication methodology was used for reducing the size of the vesicles. The seeds of the broad bean were collected in two different periods of maturity and extracted by the Bligh‐Dyer method, and the lipid classes were studied by HPTLC/FID. The oils were found to be rich in polar lipids (63.1% and 60.2% of total lipids) and low in neutral lipids (36.9% and 39.8% of total lipids) for the immature and mature seed oils, respectively. The neutral lipid fraction consisted mainly of triacylglycerides (34.2% and 32.3%) whereas the polar lipids mainly consisted of phospholipids (60.2% and 54.2%) for the mature and immature seed oils, respectively. Sphingolipids (8.9%) were identified only in the immature seed oil. The overall goal of this study was the preparation of a new liposomal formulation with physicochemical properties such as unique lipid composition, size and ζ‐potential, which are important factors influencing drug delivery to the target tissues.     

Impact of Frying on Changes in Clam (Ruditapes philippinarum) Lipids and Frying Oils: Compositional Changes and Oxidative Deterioration

The current study shows the compositional changes and oxidation development of clam (Ruditapes philippinarum, R. philippinarum) lipids and frying oils when subjected to different processing conditions. Parameters measured include acid value, peroxide value (POV), thiobarbituric acid-reactive substances (TBARS), total oxidation (TOTOX), lipid classes, fatty acid composition, phosphatidylcholine (PC), and phosphatidylethanolamine (PE) contents together with major glycerophospholipid (GP) molecular species. Deep-fat frying increased triacylglycerol (TAG) content and decreased the contents of PC, PE, and GP molecular species in clam in a time-dependent manner. Meanwhile, minor amounts of free fatty acids, diacylglycerols, monoacylglycerols, and polar lipids were detected in frying oils, indicating lipid migration between the clam and frying oils. The time-dependent increase of POV, TBARS, and TOTOX in fried clams and frying oils with concurrent reduction of docosahexenoic acid and eicosapentaenoic acid indicates extensive oxidative degradation of clam lipids. Moreover, the moisture-rich clam aggravated the deterioration of frying oils. Consequently, deep-fat frying significantly altered the lipid profile and decreased the nutritional value of clams.     

Influence of Commercial Dietary Oils on Lipid Composition and Testosterone Production in Interstitial Cells Isolated from Rat Testis

The aim of this study was to examine the influence of dietary fat on lipid composition, as well as on the steroidogenic function of interstitial cells isolated from Wistar rats that had been fed semi-synthetic diets containing four different commercial oils (S soybean, O olive, C coconut, and G grape seed). Steroidogenic enzyme activities, lipid composition, and androgen production were measured in testicular interstitial cells. Lipid analysis included measurement of the contents of major lipid subclasses (neutral lipids, polar lipids, free and esterified cholesterol), as well as principal polar and neutral lipid fatty acyl compositions. Significant differences in lipid composition were observed among the groups, most of them reflecting the specific fatty acyl composition of the diet tested. Testosterone concentration was higher in O and C groups compared with S or G. In agreement with this observation, the activity of both key enzymes involved in testosterone biosynthesis (3-β-HSD and 17-β-HSD) was higher in O and C groups with significant differences between them (O > C). A significant negative correlation was found between cellular testosterone production and cellular cholesterol ester content. Additionally, testosterone concentration directly correlated with cholesterol levels. We conclude that dietary oils qualitatively and quantitatively modified the lipid composition of interstitial cells, producing either a direct or indirect regulatory effect on testicular steroidogenic function.     

Modified ferrous oxidation‐xylenol orange method to determine lipid hydroperoxides in fried snacks

A modified ferrous oxidation‐xylenol orange (FOX) method was adapted to measure lipid hydroperoxides (LHP) in lipid extracts from snack foods fried in vegetable oils. First, a methanol‐based FOX reaction medium was assayed, but this became turbid upon addition of the lipid extracts dissolved in ethanol. To avoid the precipitation of lipids, the polarity of the reaction medium was reduced by lowering its water content and by replacing the methanol as the basis of the medium for less polar solvents. Some of the solvents used instead of methanol yielded a lower FOX reaction response. Of the reaction media assayed, the one based on dichloromethane/ethanol (3:2, vol/vol) was not turbid at high lipid extract concentrations (assayed at up to 25 mg of lipid extract/mL reaction medium) and provided the same response level as the methanol‐based medium. Thus, this FOX method shows high sensitivity and is particularly useful for lipid extracts with low LHP content. This method was also successfully applied to edible oils. Solvents such as 2‐propanol, ethyl acetate and butanol were discarded, because they easily produce hydroperoxides, which interfere in the FOX reaction. Xylenol orange preparations from a number of suppliers were tested, and some differences affecting the sensitivity of the reaction were observed.     

Lipid class composition during embryonic and early larval development in Atlantic herring (Clupea harengus,L.)

The lipid class compositions of Atlantic herring eggs and larvae were determined immediately before fertilization, after fertilization and at various times during subsequent embryonic and early larval development. Total lipid constituted 15% of the dry wt of ripe eggs, 70% of the total lipid being polar lipid with phosphatidylcholine (PC) accounting for almost 90% of the polar lipid. In general, the total lipid content decreased gradually during embryogenesis and in particular during larval development. Within 3 hr after fertilization the relative percentage of neutral lipid decreased slightly. This was followed by a general decrease in polar lipid which, by the stage of yolk sac absorption, was reduced to 52% of the total lipid. The decreased percentage of polar lipid was due entirely to a decrease in PC, which was reduced to 66% of the polar lipids at the stage of yolk sac absorption. The accompanying increase in the percentage of neutral lipids was mainly due to increased percentages of triacylglycerols (TAG) up to yolk sac absorption and cholesterol esters in the larval stages. During the first 4 days after hatching, phospholipids and to a lesser extent cholesterol were preferentially depleted in the yolk sacs, which also had higher levels of free fatty acids. The results are discussed in relation to possible roles of different lipids during embryonic and early larval development.     

A methodology study to evaluate quality of soybeans stored at different moisture levels

The quality of soybeans and oil extracted from seeds stored at different moisture contents was evaluated by static headspace gas chromatography, near-infrared spectrometry, fluorescence measurements, and silicic acid chromatography. Headspace gas chromatographic analysis of both ground beans and crude oils provided a sensitive measure of oxidative deterioration based on hexanal and total volatiles. Near-infrared analyses at 2260 nm showed a correlation coefficient of 0.864 with titratable free fatty acids. Fluorescence measurements on chloroform-methanol extracts were much less sensitive and showed an increase only in the most damaged samples. Silicic acid chromatography of crude oils showed a significant decrease of polar lipids and increase of less polar lipids with storage at high moisture levels, in agreement with the decrease in phosphorus observed. Among the methods tested, headspace gas chromatography is most sensitive to evaluate oxidative deterioration, and near-infrared analysis is most suitable and rapid to evaluate hydrolytic deterioration in stored soybeans. This methodology can be used to evaluate factors affecting the food quality of soybeans for domestic and foreign markets.     

Fluorescence screening of antioxidant capacity in pumpkin seed oils and other natural oils

Plant oils provide a rich source of dietary polyunsaturated fatty acids (PUFAs) and mostly lipophilic antioxidants. PUFAs are both in their free form and as components of glycerolipids preferred targets of free radical‐induced oxidation, leading to the formation of highly atherogenic compounds. Thus, stabilization of polyunsaturated lipids by radical scavengers in the oils is important in order to avoid pathophysiological side effects of these essential components of our diet. To determine lipid oxidizability and its inhibition by endogenous antioxidants, we developed a simple fluorescence technique. It is based on solubilisation of the oils in aqueous buffer, labeling of the resulting emulsions with a suitable reporter fluorophore, which reflects lipid oxidation, and continuous monitoring of the decomposition process. Using this method, we found that oxidizability of the oils depended only to a limited extent on the content of lipophilic antioxidants. In addition, a smaller fraction of polar (phenolic) compounds showed comparable protective effects, especially in pumpkin seed oil, which is a non‐refined product therefore containing antioxidative components that are removed from most other edible oils during processing. Therefore, the contribution of these “minor” compounds has to be taken into account when potential biological effects of plant oils are evaluated.     

Accumulation of phospholipids and glycolipids in seed kernels of different sunflower mutants (Helianthus annuus)

Phospholipids are essential components of the oil bodies present in seeds, and they are also the main components of the commercial seed lecithins used in many food formulas. In the present study, we analyzed the characteristics of the polar lipid fraction of seeds from different sunflower FA mutants. In sunflower seeds the accumulation of polar lipids reaches a maximum 25 d after anthesis before diminishing during the final stages of maturation and desiccation. We have developed an HPLC method, using ELSD, that produces optimal separation of all polar seed lipids. This method improves the results that could be obtained with previous HPLC methods and hence, we have used it to analyze the polar lipid fraction of sunflower seeds. We show that this fraction comprises phospholipids and glycolipids, of which PC is the most abundant species. Moreover, we found that the relative polar lipid content in control and mutant seeds is similar, suggesting that the mutant traits do not affect polar lipid synthesis. The degradation of polar lipids in isolated seeds was also examined and we found that the PC and PE present in developing sunflower seed kernels were rapidly degraded owing to the activity of D-type phospholipases.     

A new test method for determination of wax content in crude oils,residues and bitumens

A new method for determining wax content in petroleum materials is developed. It is based on thin layer chromatography with flame ionization detection (TLC-FID) and involves two-step development with two solvents. The principle of the test method is first to separate saturates from other more polar components based on good solubility of saturates in n-heptane and weak strength of interaction with an adsorbent (silica). Waxes are then separated from the saturate fraction using a poor solvent methyl-ethyl ketone (MEK) at such a low temperature (typically ?20 °C) that waxes are in solid state. The separated fractions are quantified with FID. The test method is verified using various model compounds including n-alkanes of different molecular weight, isoalkane, as well as commercial waxes. Results indicate that the TLC-FID method detects the waxes mainly composed of n-alkanes ranging from C20 to C40, and large isoalkanes and cycloalkanes which are soluble in n-heptane. The method has been satisfactorily applied to a variety of samples of crude oils, residues, and bitumens. It is simple, quick, and reliable. By changing MEK temperature in the development chamber, waxes may be further characterized.     

A Rapid Method for Determining the Oxidative Stability of Oils Suitable for Breeder Size Samples

A method utilizing thin-layer chromatography with a flame ionization detector (TLC-FID) was developed for assessing the stability of breeder’s oil seed samples based on the formation of polar compounds. The results showed a linear relationship between peroxide value (PV) and the content of polar material in the oxidized oil. Oil samples oxidized very readily on chromarods, even at low temperature, which is a particular advantage for antioxidant screening. At 45 °C, the oil oxidation rate was relatively low, but the relationship between the content of polar material and reaction time was linear. At 65 °C, if the content of polar material was below 50 %, the above relationship was still linear. At different temperatures, the action of tocopherol appeared to vary slightly. For example, at 65 °C, the oxidative stability of the oil sample was determined by the content of tocopherol, especially γ-tocopherol. At 45 and 55 °C, the oxidative stability was determined by both the content of tocopherol and polyunsaturated fatty acids. Of the tocopherol isomers, γ-tocopherol exhibited the highest antioxidant potency, consistent with the published literature. These results suggest that chromarods provide good media for monitoring oil oxidation for antioxidant screening. A particular advantage is the use of very small oil samples, usually 1–2 μL, and the ability to analyze multiple samples at the same time.     

Quantitative isolation of lipids of partially defatted and whole peanuts by a dry column method

A dry column method of lipid extraction was found to be applicable to peanuts—partially defatted as well as whole. Total lipid was obtained from the peanut/sodium sulfate/Celite 545 columns by isocratic elution with dichloromethane/methanol 9:1. Moreover, neutral lipids were obtained by sequential elution that were completely free from polar lipids. First, dichloromethane eluted the neutral lipids, then the 9:1 solvent eluted the polar lipids—at times containing small quantities of neutral lipids. Total lipid values obtained by the column extraction method were slightly higher than those obtained by the standard Soxhlet extraction procedure. This was due in part to the more complete polar lipid isolation produced by the column method. In partially defatted peanuts produced by mechanical pressing, 99% of the polar lipids remained in the retained oil, and these were shown to be slightly less unsaturated than the neutral lipids.     

Lipids in wheat from various classes and varieties

Lipids were extracted with petroleum ether and with water-saturated n-butanol from 8 hard red winter, 5 hard red spring, and one each from soft red, durum, and club wheat varieties from 2 harvests. The butanol-extracted lipids were fractionated into nonpolar and polar lipids by silicic acid column chromatography, and the two major fractions were subfractionated by thin-layer chromatography. Durum wheats contained the highest lipid contents, and the highest concentration of nonpolar lipids. The breadmaking wheat varieties had a lipid content which was consistent for the 2 years examined. The total and nonpolar lipid contents of hard red spring wheats were higher than of hard red winter wheats. The polar lipid contents of wheats from the two classes were essentially equal. Total lipid contents were substantially higher in wheats than in flours milled from the wheats. Nonpolar lipids constituted about one-half of the flour lipids and two-thirds of the wheat lipids. Concentrations of triglycerides were higher in wheat than in flour nonpolar lipids. Glycolipids were present in comparable concentrations in wheat and in flour polar lipids; concentration in polar lipids of phosphatidyl choline was higher and of other phospholipids was lower in wheat than in flour polar lipids. No. 547, Kansas Agrieultural Experiment Station, Manhattan. Done in part under contract with the U.S. Department of Agriculture and authorized by the Research and Marketing Act of 1946. Supervised by the Western Utilization Research and Development Div., ARS.     

Oxidation at elevated temperatures: competition between α‐tocopherol and unsaturated triacylglycerols

The competitive oxidation between α‐tocopherol and unsaturated fatty acyls at thermoxidation conditions (180 and 240 °C) was evaluated using purified triacylglycerols from nine fats and oils (refined coconut, palm, tallow, olive, high oleic sunflower, sunflower, corn, soybean, and flaxseed oil). α‐Tocopherol degraded faster in less unsaturated lipids and a linear correlation between the iodine value (x) and the residual tocopherol content (y) was obtained after 2 h of heating at 240 °C (y = 3.72x + 137.5, R² = 0.9463). The formation of polar oxidation products was established and the results were explained by a non‐selective oxidation of unsaturated fatty acyls and α‐tocopherol by highly reactive alkoxyl and hydroxyl radicals generated by decomposition of hydroperoxides.     

Fatty acids in lipids of maturing wheat

Two varieties of hard red winter wheat were sampled at various stages of maturity. The lipids in those samples were fractionated into free polar, free nonpolar and bound lipids. Fatty acids of those fractions were determined. Major acids present were palmitic, oleic, linoleic and linolenic. Both wheat samples showed similar qualitative, but not quantitative patterns in distribution of fatty acids during maturation. In the free polar lipid fraction, the palmitic acid content decreased with maturation while the linoleic acid content increased. The free nonpolar fractions showed a slight increase in linoleic acid; the concentration of other acids decreased slightly as the wheat matured. The bound lipid fraction showed a marked increase in linoleic acid, accompanied by decreases in the other major fatty acids, especially linolenic. Cooperative investigations of Kansas Agricultural Experiment Station and Crops Research Div., ARS, USDA.     

Comparative evaluation of solvent extraction methods for the determination of neutral and polar lipids in beef

Samples of lean (< 5% fat), medium (13–15%) and high-fat (> 20%) ground beef were extracted for total lipid by 4 methods of wet extraction employing chloroform/methanol (CM), n-hexane/iso-propanol (HIP) and ethyl alcohol/ethyl ether (AE), and by 3 methods of soxhlet extraction of freeze-dried material by petroleum ether (PE) or eithyl ether (EE), CM and methylene chloride/ methanol (MM). The purified lipid was fractionated into neutral and polar lipid fractions by silicic acid chromatography and the frac-tions were analyzed for fatty acid distribution by gas liquid chroma-tography (GLC). The soxhlet procedure employing either PE or EE extracted less than 75% of total lipid, 89% of triglycérides and 15% of polar lipids from lean beef as compared to other methods, and as the fat content increased from 3 to 20%, extracted amounts of polar lipid which increased to 40% of that extracted by other methods. The fatty acid distribution of the fractionated triglycerides and polar lipids was generally within experimental error for each frac-tion, irrespective of the method of extraction. The percentages of 16:0 and 18:1 were significantly less in polar lipids than in trigly-cerides. In addition to significantly higher percentage of 18:2, the polar lipids contained up to 20% of long-chain fatty acids not detected in triglycerides. The soxhlet procedures with CM or MM were as effective as wet extraction procedures in extracting neutral and polar lipids. Presented at the 73rd AOCS annual meeting, Toronto, 1982. Contribution No. 512, Food Research Institute, Agriculture Canada.     

Salivary lipid profiles of the leech (Hirudo medicinalis)

Saliva was collected from sixteen leeches (Hirudo medicinalis). The saliva was analyzed for its total lipid content (3.26±0.31 mg of total lipids per 100 mL saliva). The lipids were separated into polar and nonpolar by chromatographic techniques. The neutral fraction was approximately 2/3 of the total, and the polar fraction was approximately 1/3 of the total lipids. Thin-layer chromatography was used to obtain the individual profiles of the polar and nonpolar lipids. Of the identified lipids, phosphatidic acids and free fatty acids represented the largest percentage. These results suggest that the leech contains a unique lipid distribution, and that some of these components may be potent phospholipases and lipases that probably are present in its saliva for the purpose of preventing plugging or healing of the wound in the attacked organism.     

Lipids from Wastewater-Activated Sludge Cultivated on Acetic Acid as Potential Alternatives to High-Value Oils and Fats

This study focuses on new interpretations of the published literature by statistically evaluating the potential of microbial lipids from activated sludge (AS) as alternatives to high-value oils and fats. There are two data analysis stages involved in this study after compilation and organization of fatty acid profiles from the literature databases: (1) comparison of fatty acid profiles of the cultivated AS lipids with that of oils and fats found in the literature databases, and (2) hierarchical cluster analysis of the fatty acids of the combined dataset of literature oils and fats, and the AS lipids. Results show that fatty acid profiles of lipids from cultivated AS were similar to the fatty acid profiles of some oils and fats of plant, animal, single-microbial cultures, and algal origins; hence, lipids from AS could be potential alternatives to specialty oils and fats. The cultivation conditions of AS during lipid content enhancement may influence lipid application.     

Oxidative stability of black cumin (Nigella sativa L.), coriander (Coriandrum sativum L.) and niger (Guizotia abyssinica Cass.) crude seed oils upon stripping

Information on stability of edible oils is important for predicting the quality deterioration of the oil during storage and marketing. Stripping of crude oils removes most of non‐triacylglycerol components, including polar lipids and phenolics. Oxidative stability of black cumin (Nigella sativa L.), coriander (Coriandrum sativum L.) and niger (Guizotia abyssinica Cass.) crude and stripped seed oils was investigated and compared. The factors influencing the oxidative stability of different seed oils were also discussed. Oil samples were stored under accelerated oxidation conditions for 21 d. The progress of oxidation at 60 °C was followed by recording the ultraviolet absorptivity and measuring the formation of oxidative products (peroxide and p‐anisidine values). Inverse relationships were noted between peroxide values and oxidative stabilities and also between secondary oxidation products, measured by p‐anisidine value and stabilities at termination of the storage. Absorptivity at 232 nm and 270 nm increased gradually with the increase in time, due to the formation of conjugated dienes and polyenes. In general, oxidative stabilities of crude oils were stronger than their stripped counterparts and the order of oxidative stability was as follows: coriander > black cumin > niger seed. Levels of polar lipids in crude oils correlated with oxidative stability. Thus, the major factor that may contribute to the better oxidative stability of crude oils was the carry‐over of their polar lipids.     

Determination of nonvolatile components of heated soybean oils separated with high-efficiency mixed-bed polystyrene/divinylbenzene columns

Whole heated soybean oils and their polar fractions were analyzed for nonvolatile components by high-performance size-exclusion chromatography (HPSEC) with evaporative light scattering detection (ELSD). High molecular-weight (MW) polymer compounds with MW ≥ trimer were efficiently separated with new 3-μm mixed-bed styrene/divinylbenzene copolymer columns. Peaks of high MW polymer components in the new column system appeared to be sharper and more symmetrical than those obtained with other columns. In the model systems studied, continuous addition of water to partially simulate frying conditions resulted in a significant increase (up to 30%) in the polar lipid content of the heated oils evaluated. Due to relatively high concentrations of monomeric triglycerides (84.6–93.5%) present in the whole unfractionated oils, small but erratic variations in the compositional distribution of components were observed in oils containing different amounts of added water. On the other hand, HPSEC-ELSD analyses of the polar fractions (monomeric triglycerides, 25.4–62.6%) showed significant changes in the content and composition of nonvolatile components with the amount of water added. In general, prolonged heating with increasing amounts of water accelerated hydrolysis and polymerization of heated soybean oils. Discrepancies in total polymerization of heated soybean oils. Discrepancies in total polymeric materials obtained from HPSEC composition data for whole oils and polar fractions are discussed in terms of nonuniformity in sample matrices, detection limitations for minor components, and a nonlinear ELSD response rationale.