Comparison of oil and phytosterol levels in germplasm accessions of corn, teosinte, and Job's tears

Seeds of 49 accessions of corn (Zea mays ssp. mays), 9 accessions of teosinte (Zea species that are thought to be ancestors and probable progenitors to corn), and 3 accessions of Job's tears (Coix lacryma), obtained from a germplasm repository, were ground and extracted with hexane. Whole kernel oil yields and levels of four phytonutrients (free phytosterols, fatty acyl phytosterol esters, ferulate phytosterol esters, and gamma-tocopherol) in the oils were measured. Among the seeds tested, oil yields ranged from 2.19 to 4.83 wt %, the levels of ferulate phytosterol esters in the oil ranged from 0.047 to 0.839 wt %, the levels of free phytosterols in the oil ranged from 0.54 to 1.28 wt %, the levels of phytosterol fatty acyl esters in the oil ranged from 0.76 to 3.09 wt %, the levels of total phytosterols in the oil ranged from 1.40 to 4.38 wt %, and the levels of gamma-tocopherol in the oil ranged from 0.023 to 0.127 wt %. In general, higher levels of all three phytosterol classes were observed in seed oils from accessions of Zea mays ssp. mays than in seed oils from accessions of the other taxonomic groups. The highest levels of gamma-tocopherol were observed in teosinte accessions.     

Reinvestigation of the effect of heat pretreatment of corn fiber and corn germ on the levels of extractable tocopherols and tocotrienols

We previously reported that heat pretreatment of corn fiber (150 degrees C, 1 h) caused a tenfold increase in the levels of extractable gamma-tocopherol. The current study was a reinvestigation of the previous effect, using improved methods (HPLC with fluorescence detection, diode-array UV detection, and mass spectrometry) for tocol analysis. Heat pretreatment did not cause an increase in the levels of any of the tocopherols or tocotrienols in corn fiber oil, but lowered the levels of three of the tocols and had no effect on the levels of the other two tocols. Heat pretreatment of corn germ had a similar effect. UV and mass spectra indicated that the peak that we had identified as gamma-tocopherol in our previous report was probably a mixture of oxidation products of triacylglycerols. Thus, heat treatment of corn germ or other corn-oil containing fractions at high temperatures leads to decreases in gamma-tocopherol, gamma-tocotrienol, and delta-tocotrienol and to the production of triacylglycerol oxidation products.     

Yield and Phytosterol Composition of Oil Extracted from Grain Sorghum and Its Wet‐Milled Fractions

Corn fiber contains an oil with high levels of three potential cholesterol‐lowering phytosterol compounds. Little information is available about the levels and types of phytosterols in sorghum. In this study, phytosterols were evaluated in grain sorhgum and its wet‐milled fractions and were compared with the phytosterols in corn. The study showed that sorghum kernels can provide a significant source of two phytosterol classes, free phytosterols (St) and fatty acyl phytosterol esters (St:E). Most of these phytosterols are concentrated in the wet‐milled fiber fraction followed by the germ fraction. In addition to phytosterols, other lipid classes such as wax esters and an aldehyde (50% C28 and 50% C30) are also present in the sorghum oil. Comparison of sorghum and corn kernels show that corn has 72–93% more phytosterols than sorghum.     

Comparison of Yield and Composition of Oil Extracted from Corn Fiber and Corn Bran

We recently reported that corn fiber oil contains high levels of three potential cholesterol-lowering phytosterol components: ferulate-phytosterol esters (FPE) (3–6 wt%), free phytosterols (1–2 wt%), and phytosterol-fatty acyl esters (7–9 wt%). A previous study also indicated that corn bran oil contained less phytosterol components than corn fiber oil. The current study was undertaken to attempt to confirm this preliminary observation using more defined conditions. Accordingly, oil was extracted from corn fiber and corn bran prepared under controlled laboratory conditions, using the same sample of corn hybrid kernels for each, and using recognized bench-scale wet-milling, and dry-milling procedures, respectively. After extraction, the chemical composition of the phytosterol components in the oil were measured. This study confirmed our previous observation—that FPE levels were higher in corn fiber oil than in corn bran oil. During industrial wet-milling, almost all of the FPE are recovered in the fiber fraction (which contains both fine and coarse fiber). During laboratory-scale wet-milling, ≈60–70% of the FPE are recovered in the coarse fiber (pericarp) and 30–40% are recovered in the fine fiber. During laboratory-scale dry-milling, <20% of the FPE are recovered in the bran (pericarp), and the rest in the grits. The recoveries of the other two phytosterol components (free phytosterols and phytosterol-fatty acyl esters) revealed a more complex distribution, with significant levels found in several of the dry- and wet-milled products.     

Effect of Various Acids and Sulfites in Steep Solution on Yields and Composition of Corn Fiber and Corn Fiber Oil

The addition of six acids (organic and inorganic) and four sulfite compounds (including gaseous SO₂) during the conventional corn wet‐milling steeping process of two yellow dent corn hybrids were evaluated for the effect on corn fiber yield, corn fiber oil yield, and the composition of three phytosterol compounds (ferulate phytosterol esters [FPE], free phytosterols [St], and phytosterol fatty acyl esters [St:E]) in the corn fiber oil. No significant effect of different sulfite compounds and acids were observed on corn fiber yields. However, a significant effect was observed on corn fiber oil yield and the composition of corn fiber oil for phytosterol compounds. Three of the sulfite compounds (including gaseous SO₂) caused very little effect on the levels of phytosterol compounds compared with the control sample (corn steeped with sodium metabisulfite and lactic acid). However, for one hybrid, ammonium sulfite gave a significantly higher yield of FPE and St:E and had no effect on the yield of St. For the other hybrid, it gave a significantly higher yield of FPE and had no effect on the yield of St and St:E compared with the control sample. This indicates that the effect of these sulfite compounds on yields of these phytosterol compounds in corn fiber oil is probably hybrid‐dependent. No significant effect of acids was observed on corn fiber yields, but significant effects were observed on corn fiber oil yields and yields of phytosterol compounds in the corn fiber oil. The effect also seems to be hybrid‐dependent because different acids affected the two hybrids differently. Overall, it seems that weak acids have a positive effect on increasing the individual phytosterol compounds in the corn fiber. When comparing the effect of experimental acids and sulfites on the two hybrids, acids have a more positive effect than sulfites in increasing the yield of phytosterol compounds in corn fiber oil.     

Effect of Alternative Milling Techniques on the Yield and Composition of Corn Germ Oil and Corn Fiber Oil

The effects of alternative corn wet‐milling (intermittent milling and dynamic steeping (IMDS), gaseous SO₂ and alkali wet‐milling) and dry grind ethanol (quick germ and quick fiber with chemicals) production technologies were evaluated on the yield and phytosterol composition (ferulate phytosterol esters, free phytosterols, and fatty acyl phytosterol esters) of corn germ and fiber oil and compared with the conventional wet‐milling process. Small but statistically significant effects were observed on the yield and composition of corn germ and fiber oil with these alternative milling technologies. The results showed that the germ and fiber fractions from two of the alternative wet‐milling technologies (the gaseous SO₂ and the IMDS) had, for almost all of the individual phytosterol compounds, either comparable or signficantly higher yields compared with the conventional wet‐milling process. Also, both of the modified dry grind ethanol processes (the quick germ and quick fiber) with chemicals (SO₂ and lactic acid) can be used as a new source of corn germ and fiber and can produce oils with high yields of phytosterols. The alkali wet‐milling process showed significantly lower yields of phytosterols compounds in germ but showed significantly higher yield of free phytosterols, fatty acyl phytosterol esters and total phytosterols in the fiber fraction.     

Hybrid Variability and Effect of Growth Location on Corn Fiber Yields and Corn Fiber Oil Composition

The variability in commercial corn hybrids for corn fiber yields, amounts of extractable oil, and levels of individual and total phytosterol components in corn fiber oil was determined. Also, the effect of growth location on fiber yields, fiber oil content, and the levels of individual and total phytosterol compounds was determined. Significant variation was observed in the commercial hybrids for fiber yield (13.2–16.6%) and fiber oil yield (0.9–2.4%). No significant correlation was observed between fiber and oil yields. Significant variations in the commercial corn hybrids were also observed in the individual phytosterol compounds in corn fiber oil: 2.9–9.2% for ferulate phytosterol esters (FPE); 1.9–4.3% for free phytosterols (St); and 6.5–9.5% for phytosterol fatty acyl esters (St:E). Positive correlations were observed among the three phytosterol compounds in the corn fiber oil (R = 0.75 for FPE and St:E; 0.48 for St:E and St; and 0.68 for FPE and St). The effect of location on dependent variables was also significant. The same hybrids grown at different locations showed a variation (range) of 4.0–17.5% for FPE, 4.9–12.2% for St:E, and 1.95–4.45% for St. Relative ranking of hybrids with respect to phytosterol composition was consistent for almost all of the growth locations.     

Phytosterol and tocopherol components in extracts of corn distiller's dried grain

As the ethanol industry continues to grow, it will become very important to develop value-added markets for its coproducts in order for the industry to remain profitable. Corn distiller's dried grain (DDG) is a major coproduct of ethanol fermentation from corn processed by dry-milling and is primarily sold as livestock feed. The objective of this research was to determine if valuable phytochemicals found in corn oil and corn fiber oil, such as phytosterols and their saturated equivalents, phytostanols, ferulate phytosterol esters (FPE), tocopherols, and tocotrienols, are retained in DDG. Hexane and supercritical carbon dioxide (CO2) extracts of DDG were similar in their concentrations of total phytosterols (15.8-17.3 mg/g of extract), FPE (3.75-3.99 mg/g of extract), and tocols (1.7-1.8 mg/g of extract). Ethanol extracts were slightly lower in concentration of phytosterols (8.9-11.4 mg/g of extract), FPE (1.62-1.98 mg/g of extract), and tocols (0.73-0.76 mg/g of extract).     

Effect of garlic oil on neutrophil infiltration in the small intestine of endotoxin-injected rats and its association with levels of soluble and cellular adhesion molecules

Garlic ( Allium sativum ) possesses anti-inflammatory effects. This study investigated the effects of garlic oil on endotoxin-induced neutrophil infiltration in the small intestine. Wistar rats received by gavage 10, 50, or 100 mg/kg body wt garlic oil (GO) or the vehicle (corn oil; 2 mL/kg body wt) every other day for 2 weeks before being injected with endotoxin (ip, 5 mg/kg body wt). Control rats were administered corn oil and injected with sterile saline. Blood samples for the measurement of soluble adhesion molecules were collected at various time points after injection, and all other samples were collected 18 h after injection. The 10 and 50 mg/kg doses suppressed endotoxin-induced neutrophilia, serum levels of sL-selectin and sICAM-1, cellular CD11b on neutrophils, intestinal ICAM-1 content, and neutrophil infiltration (P < 0.05). The 100 mg/kg dose significantly lowered local ICAM-1 and cellular CD11b on neutrophils (P < 0.05) but did not have a beneficial effect on neutrophil infiltration. In addition, 100 mg/kg of GO worsened the elevation of the local TNF-α level and neutrophilia. Appropriate doses of garlic oil have a preventive effect on endotoxin-induced neutrophil infiltration and damage to the small intestine.     

Simultaneous analysis of free phytosterols/phytostanols and intact phytosteryl/phytostanyl fatty acid and phenolic acid esters in cereals

An approach based on solid-phase extraction for the effective separation of free phytosterols/phytostanols and phytosteryl/phytostanyl fatty acid and phenolic acid esters from cereal lipids was developed. The ester conjugates were analyzed in their intact form by means of capillary gas chromatography. Besides free sterols and stanols, up to 33 different fatty acid and phenolic acid esters were identified in four different cereal grains via gas chromatography-mass spectrometry. The majority (52-57%) of the sterols and stanols were present as fatty acid esters. The highest levels of all three sterol and stanol classes based on dry matter of ground kernels were determined in corn, whereas the oil extract of rye was 1.7 and 1.6 times richer in fatty acid esters and free sterols/stanols than the corn oil. The results showed that there are considerable differences in the sterols/stanols and their ester profiles and contents obtained from corn compared to rye, wheat, and spelt. The proposed method is useful for the quantification of a wide range of free phytosterols/phytostanols and intact phytosteryl/phytostanyl esters to characterize different types of grain.     

Effect of Corn Milling Practices on Aleurone Layer Cells and Their Unique Phytosterols

Coarse and fine fiber fractions obtained from the corn wet‐milling processes, with and without steeping chemicals (SO₂ and lactic acid), were evaluated microscopically for structure and analytically for recovery of phytosterol compounds from the fiber oil. Microscopic results showed that wet milling, with and without chemicals during steeping, changed the line of fracture between pericarp and endosperm and therefore affected the recovery of the aleurone layer in coarse (pericarp) and fine (endosperm cellular structure) fiber. Analytical results showed that most of the phytosterols and mainly phytostanols in corn fiber are contributed by the aleurone layer. Hand‐dissection studies were performed to separate the two layers that comprise the wet‐milled coarse fiber, the aleurone, and pericarp layer. Analyses revealed that the aleurone contained 8× more phytosterols than the pericarp.     

Stability of sterols in phytosterol-enriched milk under different heating conditions

Commercially available phytosterol-enriched milk was subjected to usual and drastic heating conditions to evaluate the stability of the sterols at different treatments. Products showed 422.2 mg of phytosterols/100 g of milk and 132 microg of sterol oxidation products (SOPs)/g of fat (277 microg of SOPs/100 g of milk). Schaal oven conditions (24 h/65 degrees C, equivalent to 1 month of storage at room temperature) reduced the phytosterol content by only 4%. Drastic heating treatments (2 min of microwave heating at 900 W or 15 min of electrical heating at 90 degrees C) led to a 60% decrease of total phytosterol content, with a significant increase of TBARs. The oxysterol amount under those conditions (which was higher in microwave-treated samples) was lower than expected, probably because of the degradation of the oxidation products. Usual heating conditions (1.5 min of microwaves) maintained phytosterol content on physiologically active values (301 mg/100 g of milk) with oxidation percentages around 0.12-0.40% for phytosterols and 1.13% for cholesterol.     

Composition of Functional Lipids in Hulled and Hulless Barley in Fractions Obtained by Scarification and in Barley Oil

Two cultivars of hulled barley (Thoroughbred and Nomini) and two cultivars of hulless barley (Doyce and Merlin) were scarified to abrade the outer layers of hull and pericarp. The resulting scarification fines fractions were evaluated as potential sources of functional lipids (phyto‐sterols, tocopherols, and tocotrienols). The levels of total phytosterols and total tocotrienols in the barley scarification fine fractions were probably not high enough to justify their use as functional foods. However, the levels of total phytosterols and total tocotrienols in the oils extracted from both whole kernels and scarified fines were both sufficiently high to make it reasonable to consider their potential use as new functional oils. Indeed, the levels of total tocotrienols in barley oils (2,911–6,126 mg/kg of oil) are several‐fold higher than those reported in two other oils that are being marketed as high in tocotrienols: palm oil (530 mg/kg) and rice bran oil (770 mg/kg). The levels of total phytosterols in barley oils range from 1.20 to 9.60 g/100 g of oil.     

Recovery of Fiber in the Corn Dry-Grind Ethanol Process: A Feedstock for Valuable Coproducts

A new process was developed to recover corn fiber from the mash before fermentation in dry-grind ethanol production. In this process, corn is soaked in water (no chemicals) for a short period of time and then degermed using conventional degermination mills. In the remaining slurry, corn coarse fiber is floated by increasing the density of the slurry and then separated using density differences. The fiber recovered is called quick fiber to distinguish it from the conventional wet-milled fiber. This study evaluated the percent of quick fiber recovery for a normal yellow dent and high oil corn hybrid. The quick fiber was analyzed for levels of corn fiber oil, levels of ferulate phytosterol esters (FPE) and other valuable phytosterol components in the oil and compared with conventional wet-milled corn coarse and fine fiber samples. Fiber samples were also analyzed and compared for yields of potentially valuable corn fiber gum (CFG, hemicellulose B). Comparisons were made between the quick fiber samples obtained with and without chemicals in the soakwater. An average quick fiber yield of 6–7% was recovered from the two hybrids and represented 46–60% of the total fiber (fine and coarse) that could be recovered by wet-milling these hybrids. Adding steep chemicals (SO₂ and lactic acid) to the soakwater increased the quick fiber yields, percent of FPE recoveries, and total percent of phytosterol components to levels either comparable to (for the dent corn hybrid) or higher than (for the high oil corn hybrid) those recovered from the total conventional wet-milled fiber samples. CFG yields in the quick fiber samples were comparable to those from the wet-milled fiber samples. CFG yields in the quick fiber samples were not significantly affected by the addition of chemicals (SO₂ and lactic acid) to the soakwater.     

Phase transitions, solubility, and crystallization kinetics of phytosterols and phytosterol-oil blends

The thermal properties, solubility characteristics, and crystallization kinetics of four commercial phytosterol preparations (soy and wood sterols and stanols) and their blends with corn oil were examined. Differential scanning calorimetry (DSC) revealed narrow melting peaks between 138 and 145 degrees C for all phytosterol samples, reversible on rescan. Broader and less symmetrical melting transitions at lower temperatures with increasing oil content were observed for two samples of phytosterol-oil admixtures. The estimated, from the solubility law, deltaH values (34.7 and 70.7 mJ/mg for wood sterols and stanols, respectively), were similar to the DSC experimental data. Fatty acid esters of soy stanols differing in the chain length of the acyl groups (C2-C12) exhibited suppression of the melting point and increase of the fusion enthalpy with increasing chain length of the acyl group; the propionate ester exhibited the highest melting point (Tm: 151 degrees C) among all stanol-fatty acid esters. Solubility of phytosterols in corn oil was low (2-3% w/w at 25 degrees C) and increased slightly with a temperature rise. Plant sterols appeared more soluble than stanols with higher critical concentrations at saturation. The induction time for recrystallization of sterol-oil liquid blends, as determined by spectrophotometry, depended on the supersaturation ratio. The calculated interfacial free energies between crystalline sediments and oil were smaller for sterol samples (3.80 and 3.85 mJ/m2) than stanol mixtures (5.95 and 6.07 mJ/m2), in accord with the higher solubility of the sterol crystals in corn oil. The XRD patterns and light microscopy revealed some differences in the characteristics among the native and recrystallized in oil phytosterol preparations.     

Enrichment of tocopherols and phytosterols in canola oil during seed germination

The effect of canola (Brassica napus L.) seed germination under illuminated and dark environments on the total concentration and the composition of tocopherols and phytosterols in seedlings and extracted oil were investigated. During the first 10 days of germination, a decrease in gamma-tocopherol was offset by an increase in alpha-tocopherol, indicating the interconversion of these isomers. From day 10 to day 20 under illumination, there was a net increase in alpha-tocopherol and total tocopherols suggesting the synthesis of new tocopherols, whereas there was no net increase in tocopherols in dark. Tocopherols were mainly concentrated in the leafy seedling tops rather than in the non-photosynthesizing bottoms, whereas phytosterols were equally distributed across both sections. The total tocopherol content of oil extracted from 20-day-old seedlings was 4.3- to 6.5-fold higher than that of intact seeds. On a dry seedling basis, the content and composition of phytosterols did not change significantly (p > 0.05) over the sprouting period, but the concentration of total phytosterols in the oil fraction increased 4.2- to 5.2-fold. The concentration of these valuable phytochemicals in the oil fraction is largely due to the depletion of oil reserves during germination, as well as the de novo synthesis of new alpha-tocopherol stimulated by the presence of light. Germination may represent a viable means to naturally concentrate these high-value constituents in canola oil, offering improvements in oil quality based on the nutritional value and oxidative protection offered by tocopherols and the health benefits provided by both tocopherols and phytosterols.     

Pretreatment of Wet‐Milled Corn Fiber to Improve Recovery of Corn Fiber Oil and Phytosterols

The phytosterol‐containing oil in the corn fiber (corn fiber oil) has potential use as a natural low‐density lipoprotein (LDL) lowering nutraceutical but its low concentration (1–3%) makes it difficult and expensive to extract. Pretreatment of corn fiber with dilute acid or glucosidases removed nonlipid components of fiber, producing oil‐enriched fractions that should be more amenable to efficient and inexpensive oil extraction. Acid, as well as enzymes, significantly increased the content of corn fiber oil and its phytosterol compounds by hydrolyzing (and removing) the starch and nonstarch (cell wall) polysaccharides from the wet‐milled corn fiber. Dual treatment of the fiber with acid and enzyme greatly increased the concentrations of corn fiber oil and its phytosterol components, compared with acid or enzyme treatments alone. Depending on the treatment, the oil concentration in the residual solids increased from 0.3 to 10.8% (21–771% increase in conc.) and the total phytosterol concentration increased from 19.8 to 1256.2 mg/g of fiber (11–710% increase in conc.) compared with untreated fiber.     

Relationship between fatty acid profile and vitamin E content in maize hybrids (Zea mays L.)

The balance between the vitamin E (tocochromanols) and polyunsaturated fatty acid (PUFA) contents mainly determines the susceptibility to lipid peroxidation and the storage stability of corn oil. In 1997, field experiments were conducted at two different locations to evaluate a collection of 30 corn hybrids for fatty acid profiles and tocochromanol contents. Hybrids differed significantly (p < 0.01) for major fatty acids, as well as for tocochromanol contents and composition. The major fatty acids were palmitic, oleic, and linoleic acids, whose contents were in the ranges 9.2-12.1%, 19.5-30.5%, and 53.0-65.3%, respectively. The tocopherol contents ranged as follows: alpha-tocopherol, 67-276 mg (kg of oil)(-1); beta-tocopherol, 0-20 mg (kg of oil)(-1); gamma-tocopherol, 583-1048 mg (kg of oil)(-1); delta-tocopherol, 12-71 mg (kg of oil)(-1); total tocopherol, 767-1344 mg (kg of oil)(-1). gamma-Tocopherol was the predominant derivative among all tocopherols. The tocotrienol contents were in the ranges 46-89, 53-164, and 99-230 mg (kg of oil)(-1) for alpha-, gamma-, and total tocotrienol contents, respectively. The tocotrienol profile was not characterized by the predominance of any tocotrienol homologue. alpha-Tocopherol was positively correlated with PUFA (r = 0.41) and with the vitamin E equivalent (vit E equiv) (r = 0.84), and it was not correlated with gamma-tocopherol. gamma-Tocopherol was highly correlated with total tocopherol and tocochromanol contents (r = 0.93 and r = 0.90, respectively), indicating that the contribution of this vitamer to the total tocochromanol content is the most important among all tocochromanols. The high positive correlation found between the vit E/PUFA ratio and the vit E equiv, as well as the absence of correlation between this ratio and PUFA indicates that a higher vit E/PUFA ratio can be easier achieved be increasing the vitamin E content than by modifying fatty acid profile in corn oil.     

Evaluation of a commercial enzyme-based serum cholesterol test kit for analysis of phytosterol and phytostanol products

Plant sterols (phytosterols) have been shown to possess serum cholesterol-lowering properties. In recent years, several phytosterol-enriched functional food products have been developed and marketed. Some phytosterol products contain common unsaturated sterols and some contain a subset of phytosterols called phytostanols (saturated sterols, also called plant stanols). Current methods for the quantitative analysis of plant sterols are labor intensive and require sophisticated gas or liquid chromatographs. In this study, a popular commercial spectrophotometric serum cholesterol test kit was evaluated for the analysis of plant sterols. The results indicate that the method could be modified to analyze phytosterols and phytostanols by increasing the incubation time. Both free phytosterols and fatty acyl phytosteryl esters were quantitatively analyzed, but ferulate phytosteryl esters, such as those that are found in corn and other cereals, were not hydrolyzed by the enzymes in the test kit and therefore were not detected.     

Influence of environmental stresses on stability of O/W emulsions containing cationic droplets stabilized by SDS-fish gelatin membranes

Oil-in-water (O/W) emulsions containing small oil droplets (d32 approximately 0.22 microm) stabilized by sodium dodecyl sulfate (SDS)-fish gelatin (FG) membranes were produced by an electrostatic deposition technique. A primary emulsion containing anionic SDS-coated droplets (zeta approximately -40 mV) was prepared by homogenizing oil and emulsifier solution using a high-pressure valve homogenizer (20 wt % corn oil, 0.46 wt % SDS, 100 mM acetic acid, pH 3.0). A secondary emulsion containing cationic SDS-FG-coated droplets (zeta approximately +30 mV) was formed by diluting the primary emulsion with an aqueous fish gelatin solution (10 wt % corn oil, 0.23 wt % SDS, 100 mM acetic acid, 2.00 wt % fish gelatin, pH 3.0). The stabilities of primary and secondary emulsions with the same oil concentration to thermal processing, ionic strength, and pH were assessed by measuring particle size distribution, zeta potential, microstructure, destabilized oil, and creaming stability. The droplets in secondary emulsions had good stability to droplet aggregation at holding temperatures from 30 to 90 degrees C for 30 min, [NaCl] < or = 100 mM, and pH values from 3 to 8. This study shows that the ability to generate emulsions containing droplets stabilized by multilayer interfacial membranes comprised of two or more types of emulsifiers, rather than a single interfacial layer comprised of one type of emulsifier, may lead to the development of food products with improved stability to environmental stresses.