Optimization of Ultrasound-Assisted Alkali Neutralization in the Refining of Safflower Oil to Minimize the Loss of Bioactive Compounds

In this study, ultrasound-assisted (UA) neutralization parameters are optimized using the response surface methodology to develop a novel alkali neutralization method based on the minimal refining concept. Sodium hydroxide (NaOH), magnesium oxide (MgO), and calcium hydroxide (Ca(OH)₂) are used in both the traditional (TR) and UA neutralizations. Optimum probe depth, duration, and intensity levels are calculated as 3.7 cm, 25 s, and 54.3%, respectively, for UA neutralization with NaOH, which is more successful at free fatty acid (FFA) reduction and total phenolic content (TPC) retention than MgO and Ca(OH)₂. Validation results of optimum conditions show that lowest average FFA content (0.29%) and highest average TPC (211.2 mg kg⁻¹) are determined for the UA-neutralized safflower oil samples. The comparison of all the neutralization experiments reveal that the UA neutralization under optimum conditions using NaOH reduced 82.8% of the FFA content, whereas the TR alkali neutralization reduced the FFA content at a maximum of only 47.8%. Practical Applications: From the results, it can be inferred that the UA neutralization exhibits good performance in FFA content reduction and bioactive compound retention while offering a good solution within the concept of minimal refining.     

Effect of minimal neutralization at optimal conditions on minor components and oxidation stability of sunflower oil

In this study, oxidatively stable minimal neutralized sunflower seed oils were produced using three chemicals (Ca(OH)₂, MgO, and Na₂SiO₃) under previously determined optimal process conditions. Lipid oxidation rates at these optimum conditions were compared to the oils neutralized with NaOH (0.20%, 40°C, 15 min). It was concluded that the oils neutralized by NaOH had the shortest hydroperoxide and hexanal lag phases, thus were the least stable oils. Oils neutralized by Ca(OH)₂, MgO, and Na₂SiO₃ had lower FFA and higher oxidative stability than oil neutralized by NaOH. The study focused on which weak alkaline has higher oxidation stability and minimum FFA content and maximum acceptable tocopherol content. The oil neutralized by Ca(OH)₂ had the lowest FFA value and highest total phenolics and α-tocopherol contents and it had better oxidative stability than oil neutralized by NaOH. It suggests that Ca(OH)₂ could be more effective in producing a high quality oil.     

Column Fractionation of Canola Oil Deodorizer Distillate Using Supercritical Carbon Dioxide

Semi-continuous column fractionation of canola oil deodorizer distillate using supercritical CO₂ (SCCO₂) was carried out to determine the feasibility of value-added processing of this feed material for the recovery of bioactive components such as sterols and tocopherols and to determine the effect of operating conditions [pressure (20, 25 MPa using a temperature gradient of 70–100 °C), temperature (70, 100 °C) and a linear temperature gradient (70–100 °C at 25 MPa)] on extract yield and separation efficiency. Total extract yield increased significantly (p ≤ 0.05) with pressure, whereas at isobaric conditions (25 MPa) the highest yield was obtained at the lowest temperature tested (70 °C). Fractionation efficiency was reflected in the composition of fractions and was affected by operating conditions. Residue composition was determined by extract yield in addition to selectivity. Use of the thermal gradient (70–100 °C) decreased the content of volatiles, free fatty acids and tocopherols while increasing sterol content significantly (p ≤ 0.05) to a level of 40% (GC area %) in the residue obtained at 25 MPa. The findings indicate the potential of canola oil deodorizer distillate as a source of sterols and warrant further research on the countercurrent column fractionation to improve the separation efficiency.     

Refining of rice bran oil by neutralization with calcium hydroxide

The applicability of calcium hydroxide (lime) in the neutralization of rice bran oil (RBO) was investigated. Crude RBO samples of three different free fatty acids (FFAs) (3.5–8.4 wt%) were degummed, dewaxed, bleached, and neutralized with lime and deodorized. The oils obtained thus were characterized by determining the color, peroxide value (PV), content of unsaponifiable matter (UM), and FFA. Conventionally practiced caustic soda neutralization (at 80–90°C) of FFA has in the present investigation been replaced by a high temperature (150–210°C) low pressure (2–4 mm Hg) reaction with lime. It was observed that neutralization with Ca(OH)₂ at high temperature (210°C) and under low pressure (2–4 mm Hg pressure) may substantially reduce the FFA content (0.8 wt%, after 2 h). The deodorized oil was found to be of acceptable color, PV, and content of UM and FFA. Neutralization of oil was also carried out by using NaHCO₃ and Na₂CO₃, nonconventional alkalies for neutralization, and the results were compared with NaOH and Ca(OH)₂. Overall recovery of oil in Ca(OH)₂ refining process (88.5 ± 0.6 wt%, for Sample 1 containing 8.4%‐wt FFA) was found to be more than other competitive processes studied.     

Effects of processing on the quality and stability of three unconventional Sudanese oils

In a refining experiment, on a laboratory scale, crude oils from Sclerocarya birrea (SCO), sorghum bugs (SBO), water‐extracted melon bugs (MBO H₂O) and solvent‐extracted melon bugs (MBO SOL) were processed by alkali refining. Quality changes were characterized by the determination of free fatty acids (FFA), peroxide value, tocopherols, sterols, phosphatides and stability against oxidation (Rancimat test). In addition, the fatty acid composition was determined. It is clear that the contents of phosphatides, peroxides, tocopherols, sterols as well as oxidative stability were reduced during processing, while FFA were nearly totally removed. The content of phosphorus was reduced in SCO, SBO, MBO H₂O and MBO SOL by 26, 19, 12, and 78%, respectively, while complete oil processing removed 95, 99, 96 and 99% of the FFA in crude oils, respectively. The level of total tocopherols decreased during processing by 38.7, 83.8, 100, and 33.3%, respectively. The color decreased through the processing steps up to bleaching; then, in the deodorization step, it darkened sharply in all samples. No change in the fatty acid composition was observed. The order of oxidation stability was crude > degummed > deodorized > neutralized > bleached, in SCO; and crude > degummed > neutralized > bleached = deodorized, in MBO H₂O; and crude > degummed > deodorized > neutralized > bleached in MBO SOL; while in SBO, the order of oxidative stability was deodorized > crude > degummed > neutralized = bleached. Total sterols decreased by 42–92% in the processed oils, compared with crude oils.     

Effect of Selected Polyhydric Alcohols on Refining Oil Loss in the Neutralization Step

Alkaline neutralization is a classical method for removal of free fatty acids (FFA) in crude oil. It is generally accompanied by neutral oil loss. Thus, reduction of refining losses associated with alkaline neutralization is very desirable. Refined, bleached and deodorized (RBD) palm oils with different FFA contents were used as oil models in this study. FFA in the oil models were neutralized with sodium hydroxide in polyhydric alcohols as neutralization media. Glycerol, propylene glycol and ethylene glycol in water were effective neutralization media. FFA in the oil models were totally removed in one step of neutralization, while percentages of refining losses were different. The losses were increased in the order of water > propylene glycol > ethylene glycol > glycerol used as neutralization media. Also, a higher concentration of polyhydric alcohol in the neutralizing media significantly reduced the percentage of refining loss (p < 0.05). Glycerol (90% in water) was the most effective neutralization media (p < 0.05). When neutralization was carried out on crude palm oil (containing 7.53% FFA), refining loss was reduced from 36.1% (in water) to 20.0% (in 90% glycerol in water).     

Comparative study on the stability of crude and refined rice bran oil during long‐term storage at room temperature

The effect of the full refining process on the stability of rice bran oil during storage at room temperature was studied. Crude and refined rice bran oil were kept in the dark and were exposed to light for 240 days, and every 10 days samples were drawn and analysed. The storage stability of crude and fully refined rice bran oil was determined and compared with respect to fatty acid composition, tocopherols, tocotrienols, sterols and γ‐oryzanol content. In addition, the oxidative status was evaluated by determining the concentration of polar compounds and the oil stability index (OSI). A good correlation between the decrease of total tocopherols and the OSI was found. α‐Tocopherol had the highest correlation coefficient (r² = 0.9653) in crude rice bran oil kept in the dark, and γ‐tocopherol showed the lowest in the refined sample (r² = 0.4722). The order of stability of tocopherols and tocotrienols in crude oil was completely different from that in refined oil. In comparison to tocopherols, sterols showed a better stability during the entire storage period. The exposure to daylight heavily affected the composition and the stability of both crude and refined rice bran oil.     

Effect of the full refining process on rice bran oil composition and its heat stability

The effects of the chemical refining process on the minor compounds of rice bran oil and its heat stability were investigated. After 8 h of heating, about 50% and 30% of total tocopherols remained in crude and refined rice bran oil, respectively. The individual tocopherols were differently affected by the refining process. The order of heat stability of tocopherols and tocotrienols in crude oil was found to be different from that in fully refined oil. A similar tendency was observed for sterols. After 8 h of heating, 65% and 72% of total sterols, and 14% and 46% of sterol esters, of crude or fully refined rice bran oil, respectively, disappeared. The heating process led to a 4% and 10.3% increase in polymer contents in crude and refined rice bran oil, respectively. Although refined rice bran oil showed good heat stability, when compared to crude oil its heat stability was decreased to some extent.     

Influence of the vegetable oil refining process on free and esterified sterols

The influence of the refining process on the distribution of free and esterified phytosterols in corn, palm, and soybean oil was studied. Water degumming did not affect the phytosterol content or its composition. A slight increase in the content of free sterols was observed during acid degumming and bleaching due to acid-catalyzed hydrolysis of steryl esters. A significant reduction in the content of total sterols during neutralization was observed, which was attributed to a reduction in the free sterol fraction. Free sterols probably form micelles with soaps and are transferred into the soapstock. The steryl ester content remained constant during all neutralization experiments, indicating that hydrolysis of steryl esters did not take place during neutralization. During deodorization, free sterols are distilled from the oil, resulting in a gradual reduction in the total sterol content as a function of the deodorization temperature (220–260°C). A considerable increase in the steryl ester fraction was found during physical refining, probably owing to a heat-promoted esterification reaction between free sterols and FA.     

The Effects of Refining on the Chemical Composition of Turkish Sunflower Seed Oil

For investigating the effects of degumming, neutralization, winterization and deodorization operations on the composition of sunflowerseed oil, samples were collected from each respective stage of a commercial oil refining factory in Tekirda?, Turkey. Based on laboratory analyses, it was concluded that there were no significant changes in specific gravities, refractive indices, saponification numbers. Iodine values and fatty acid compositions were altered slightly during winterization. There was a gradual decrease throughout each consecutive stage of refining, in free fatty acids, phospholipids, unsaponifiables, tocopherols, sterols and iron contents. Also, slight changes were observed in relative distribution pattern of individual tocopherol and sterol components. The overall oil quality, from points of oxidative stability and nutritional status, was not lower than the original crude sunflower seed oil.     

Supercritical fluid extraction of oil from millet bran

Proso millet bran [Panicum miliaceum (L.)], variety Dakota White, was extracted with supercritical carbon dioxide (SC-CO₂) to yield crude oil. The effects of operating parameters (pressure, temperature, and specific solvent flow) and of features of the raw material (moisture content and particle size) on oil extraction were investigated. Complete de-oiling of ground millet bran pellets was achieved under 300 bar at 40°C with a specific solvent flow of 2–10 h⁻¹ within 200 to 500 min. Solvent requirements were 20–30 kg CO₂/kg raw material. Composition of crude SC-CO₂ oil extracted under optimal conditions, i.e., fatty acid profile, amount of unsaponifiables, tocopherols, free fatty acids, sterols, sterol esters, waxes, hydrocarbons, and phospholipids, was compared to that of crude oil obtained by petroleum ether extraction. These two oils were similar in terms of fatty acid profile and amount of free fatty acids, unsaponifiables, peroxides, and tocopherols. They differed in respect to phospholipids (present in petroleum etherextracted oil and absent in SC-CO₂ extracted oil), metals, and waxes (lower levels in SC-CO₂ extracted oil). The effects of extraction procedures on oxidative stability of crude SC-CO₂ oil were studied. Ensuring that all pieces of the extractor in contact with the oil were in stainless steel; cleaning the separator, i.e., washing with KOH, rinsing, purging with N₂ and CO₂, and heating; performing a couple of extractions before the main extraction; and achieving the extraction without interruption all positively influenced the oxidative stability of the oil. Conversely, increasing CO₂ purity above 99.5% had no effect. Oxidative stability of the SC-CO₂ oil extracted under these conditions was only slightly lower than that of the oil extracted with petroleum ether.     

Facile purification of tocopherols from soybean oil deodorizer distillate in high yield using lipase

Tocopherols have been purified from deodorizer distillate produced in the final deodorization step of vegetable oil refining by a process including molecular distillation. Deodorizer distillate contains mainly tocopherols, sterols, and free fatty acids (FFA); the presence of sterols hinders tocopherol purification in good yield. We found that Candida rugosa lipase recognized sterols as substrates but not tocopherols, and that esterification of sterols with FFA could be effected with negligible influence of water content. Enzymatic esterification of sterols with FFA was thus used as a step in tocopherol purification. High boiling point substances including steryl esters were removed from soybean oil deodorizer distillate by distillation, and the resulting distillate (soybean oil deodorizer distillate tocopherol concentrate; SODDTC) was used as a starting material for tocopherol purification. Several factors affecting esterification of sterols were investigated, and the reaction conditions were determined as follows: A mixture of SODDTC and water (4∶1, w/w) was stirred at 35°C for 24 h with 200 U of Candida lipase per 1 g of the reaction mixture. Under these conditions, approximately 80% of sterols was esterified, but tocopherols were not esterified. After the reaction, tocopherols and FFA were recovered as a distillate by molecular distillation of the oil layer. To enhance further removal of the remaining sterols, the lipase-catalyzed reaction was repeated on the distillate under the same reaction conditions. As a result, more than 95% of the sterols was esterified in total. The resulting reaction mixture was fractionated to four distillates and one residue. The main distillate fraction contained 65 wt% tocopherols with low contents of FFA and sterols. In addition, the residue fraction contained high-purity steryl esters. Because the process presented in this study includes only organic solvent-free enzymatic reaction and molecular distillation, it is feasible as a new industrial purification method of tocopherols. This work was presented at the Biocatalysis symposium in April 2000, held at the 91st Annual Meeting and Expo of the American Oil Chemists Society, San Diego, CA.     

Effect of Extraction Solvent on Oil and Bioactives Composition of Commercial Indian Niger (Guizotia abyssinica (L.f.) Cass.) Seed

Commercially available niger (Guizotia abyssinica (L.f.) Cass.) seed was investigated to evaluate the effect of extraction solvent on oil and bioactives composition. For this purpose, niger seeds were subjected to solvent extraction using solvents of different polarity, viz., hexane, petroleum ether, chloroform, acetone, methanol and ethanol. The oil content of niger seeds obtained after extraction with solvents of different polarities was in the range of 31.8–41.3 g/100 g. The extracted oil was characterized by the following parameters: color (40.0–95.0 Lovibond units), free fatty acids (3.6–12.3 g/100 g), peroxide value (3.2–7.8 mequiv O₂/kg), iodine value (137.6–140.3 cg I₂/g), saponification value (177.3–185.9 mg KOH/g) and unsaponifiable matter (1.3–4.3 g/100 g). Among fatty acids, linoleic acid (69.4–73.2 %) was the major fatty acid and trilinolein (31.2–33.4 %) was the major triacylglycerol. The composition of bioactive molecules was 171.9–345.8 ppm of total tocopherols; 247.1–2,647.7 ppm of total phenolics; 1,249.6–6,309.3 ppm of total sterols and 18.9–181.0 ppm of total carotenoids. Among the tocopherols, α-tocopherol was the major component with 154–276 ppm. Of the total phenolics, vanillic acid with 176–1,709 ppm was the major phenolic compound in the oil extracted using different solvents. Ethanol-extracted oil showed a 13.9-fold better oxidative stability and a higher radical scavenging activity (IC₅₀ value of 9.2 mg/mL) compared to hexane-extracted oil (IC₅₀ value of 40.3 mg/mL). This is probably the first report of its kind on solvent extractability of bioactives of niger seed.     

Comparison of Supercritical Fluid and Hexane Extraction Methods in Extracting Kenaf (<Emphasis Type="Italic">Hibiscus cannabinus</Emphasis>) Seed Oil Lipids

The objective of this study was to investigate and compare fatty acids, tocopherols and sterols of kenaf seed oil extracted by supercritical carbon dioxide and traditional solvent methods. Fatty acids, tocopherols and sterols were determined in the extracted oils as functions of the pressure (400 bar, 600 bar), temperature (40 °C, 80 °C) and CO₂ flow rate (25 g/min) using a 1-L extraction vessel. Gas chromatography was used to characterize fatty acids and sterols of the obtained oils while tocopherols were quantified by HPLC. No differences were found in the fatty acid compositions of the various oil extracts and the main components were found to be linoleic (38%), oleic (35%), palmitic (20%) and stearic acid (3%). Extraction of tocopherols using high pressure (600 bar/40 °C, 600 bar/80 °C) gave higher total tocopherols (88.20 and 85.57 mg/100 g oil, respectively) when compared with hexane extraction which gave yield of 62.38 mg/100 g oil. Extraction of kenaf seed oil using supercritical fluid extraction at high temperature (80 °C) gave higher amounts of sterols when compared with hexane extraction.     

Minor constituents of vegetable oils during industrial processing

We report the effects of individual steps of industrial refining, carried out in Brazil, on the alteration of selected minor constituents of oils, such as corn, soybean, and rapeseed oils. Total sterols, determined by capillary gas chromatography (GC), decreased by 18–36% in the fully refined oils, compared with the crude oils. The total steradienes, dehydration products of sterols, were determinedvia a simple clean-up on a short silica gel column, followed by high-performance liquid chromatography (HPLC) with ultraviolet detection. The level of steradienes, normally not present in crude oils, increased after each refining step, especially after deodorization. Thus, the content of steradienes increased after deodorization by about 15- to 20-fold in corn and soybean oils, and by about 2-fold in rapeseed oil. The total steryl esters were also determinedvia clean-up on a short silica gel column, followed by HPLC with evaporative light scattering mass detection. A minor decrease in the level of steryl esters was observed after complete refining. The individual tocopherols and tocotrienols were determined by HPLC with a fluorescence detector. The level of total tocopherols and tocotrienols decreased by about 2-fold after complete refining of corn oil and by about 1.5-fold in soybean and rapeseed oils. In all three cases, maximum reduction of tocopherols was observed after the deodorization step. The level of polymeric glycerides, determinedvia clean-up on a short silica gel column followed by size-exclusion HPLC, increased to some extent (0.4–1%) during refining. The level oftrans fatty acids, determined by capillary GC, also increased to a substantial extent (1–4%) after refining. Part of doctoral thesis of Roseli Ap. Ferrari to be submitted to Faculdade de Engenharia de Alimentos, Universidade de Campinas, Campinas, Brazil.     

Isolation of tocopherol and sterol concentrate from sunflower oil deodorizer distillate

The isolation of tocopherols and sterols together as a concentrate from sunflower oil deodorizer distillate was investigated. The sunflower oil deodorizer distillate was composed of 24.9% unsaponifiable matter with 4.8% tocopherols and 9.7% sterols, 28.8% free fatty acid (FFA) and 46.3% neutral glycerides. The isolation technology included process steps such as biohydrolysis, bioesterification and fractional distillation. The neutral glycerides of the deodorizer distillates were hydrolyzed byCandida cylindracea lipase. The total fatty acids (initial FFA plus FFA from neutral glycerides) were converted into butyl esters withMucor miehei lipase. The esterified product was then fractionally distilled in a Claisen-vigreux flask. The first fraction, which was collected at 180–230°C at 1.00 mm of Hg for 45 min, contained mainly butyl esters, hydrocarbons, oxidized products and some amount of free fatty acids. The fraction collected at 230–260°C at 1.00 mm Hg for 15 min was rich in tocopherols (about 30%) and sterols (about 36%). The overall recovery of tocopherols and sterols after hydrolysis, esterification and distillation were around 70% and 42%, respectively, of the original content in sunflower oil deodorizer distillate.     

Minor constituents of palm oil

Crude palm oil contains about 1% of minor components including carotenoids, tocopherols, sterols, triterpene alcohols, phospholipids, glycolipids and terpenic and paraffinic hydrocarbons. The nutritionally important components such as carotenes and tocopherols also improve stability of the oil. Although a highly valued product, carotene unfortunately is bleached or destroyed in refining because suitable recovery technology is not available. Thermal degradation of carotene, previously suspected of giving rise to undesirable chemicals, now is known to furnish mainly harmless hydrocarbons, most of which are removed by the deodorization step in refining. Tocopherols, being natural antioxidants, need to be carefully preserved during milling, refining, fractionation and modification of palm oils. The accumulation of tocopherols in the palm fatty acid distillate promises to provide a new source for the recovery of this valuable substance. The role played by phospholipids is frequently misunderstood because they can act in two ways, i.e. as an antioxidant synergist and a surface active agent to disperse impurities in oil. In crude palm oil the phospholipid content is small, because most of it is removed during milling; the phosphorus content is due mainly to inorganic phosphorus. Among the sterols, cholesterol constitutes too small a percentage to be of much concern. Sterols, triterpenoids and terpenoid hydrocarbons are also potentially useful side products should recovery technology become available. Other newly characterized minor and trace components also are discussed.     

Impacts of Refining and Antioxidants on the Physico-Chemical Characteristics and Oxidative Stability of Watermelon Seed Oil

Compositional analyses of seeds from two cultivars (Mateera and Sugar baby) was performed to evaluate their suitability as oilseeds. Watermelon seeds and kernels contained 21.9–25.5 % and 38.9–46.9 % oil of exceptionally high quality. The crude oil was expelled with a screw press and then refined to obtain a odor free and colorless oil. The moisture content, unsaponifiable matter content, refractive index, and specific gravity were within the narrow ranges. Refining influenced the color, acid value, saponification value, peroxide value, and free fatty acid contents. Linoleic acid (C18:2) was the principal fatty acid constituting 64.5–67.2 % of the total fatty acids. Oxidative stability increased with the addition of tocopherols, butylated hydroxyl anisole (BHA), and tert-butyl hydroxyl quinine (TBHQ). The high amount of polyunsaturated fatty acids (PUFA) along with physicochemical properties were similar to soybeans, sunflower and other common vegetable oils, suggesting the suitabilty of watermelon seed oil for industrial production.     

Quality of Rapeseed Oil Produced by Conditioning Seeds at Modest Temperatures

Conditioning rapeseed can significantly increase the amount of bioactive compounds in the crude oil, but if the conditioning temperatures are too high, they can cause unwanted side effects such as darker color and sensory defects. Modest conditioning temperatures may be more suitable, but little is known about the effects on the quality and bioactive composition of the resulting oil. Oil was recovered from five rapeseed cultivars by cold pressing (CP) or by pressing seeds conditioned at 80 °C for 30 min (HP). Conditioning rapeseed increased oil yield without changing fatty acid composition and increased the amount of total sterols by 16 %, total tocopherols by 20 %, and the levels of polyphenols. Levels of the polyphenol canolol were up to 55-fold higher in HP oil than in CP oil. These higher levels of bioactive compounds gave HP oil higher radical scavenging activity. Although HP oil also had higher free fatty acid contents, peroxide levels, and specific UV extinctions (K values). The quality parameters of HP and CP oils were within codex limits indicating high quality. Modest conditioning temperatures can be used to produce rapeseed oil with high quality and radical scavenging activity.     

Effect of heat treatments on canola press oils. I. Non- triglyceride components

Increasing heat treatment given to canola seed prior to pressing resulted in press oils with progressively increasing contents of non-triglyceride components. Phosphorus and chlorophyll contents ranged from 13 ppm and 7 ppm, respectively, in cold press oil to 64 ppm and 68 ppm, respectively, in oil from heated seeds. Refining reduced the amount of these components to 19 ppm and 60 ppm, respectively, in degummed oil and to 4 ppm and 11 ppm, respectively, in bleached oil. Oil with the lowest amount of non-triglyceride material was obtained by cold pressing and/or bleaching. The major sterols wereβ-sitosterol (55%), campesterol (35%) and brassicasterol (10%), and the major tocopherols were y (60%), α (30%) and δ (10%). The content of sterols and tocopherols ranged from 620 to 773 mg/100 g and from 47 to 64 mg/100 g, respectively, in the press oils. The total content of sterols was reduced by 15% and a further 1% on degumming and bleaching, respectively. The total tocopherol content was reduced by 20% and 60% on degumming and on subsequent bleaching. Refining had no effect on the sterol isomer ratio, but there was a significant relative loss ofα-tocopherol on bleaching.