Vitamin E isomers in grain amaranths (Amaranthus spp.)

Vitamin E isomers are important antioxidants, but their variation is poorly documented in pseduocereal grains such as amaranths. Using normal-phase, high-performance liquid chromatography with fluorescence detection, seeds of thirteen amaranth (Amaranthus cruentus L.,A. hypochondriacus L.) accessions were surveyed for the composition of tocols. The most common tocols found were α-tocopherol (2.97 to 15.65 mg/kg seed) and β-tocotrienol (5.92 to 11.47 mg/kg seed) and γ-tocotrienol (0.95 to 8.69 mg/kg seed), while someA. cruentus accessions contained δ-tocotrienol (0.01 to 0.42 mg/kg seed). This is the first report of tocotrienols in amaranths.Amaranthus cruentus grain-types of Mesoamerican origin had significantly (P≤0.01) greater levels of four tocols than didA. cruentus African vegetable-types. Unlike many cereal grains, amaranths have significant amounts of both β- and γ-tocotrienols; however, β-tocopherol was not detected in any of the amaranths. Using multiple linear regressions, α-tocopherol variation of both species and types was consistently explained by variation in tocols other than α-tocopherol. On the whole, fresh amaranth samples of both species tended to have higher levels of tocotrienols than samples stored for two years. Storage effects on amaranth tocol composition are suspected.     

Tocol-derived minor constituents in selected plant seed oils

Various crude and processed seed oils were analyzed for tocopherols (T) and tocotrienols (T₃) by reversed-phase HPLC with fluorescence detection (FL). The oils included processed canola oil, crude corn oil, crude milkweed oil, crude palm oil, crude/processed rice bran oils, crude/processed soybean oil, crude/processed sunflower oil, and related modified oil, crude/processed sunflower oil, and related modified oil varieties. The HPLC system consisted of a pentafluorophenylsilica (PFPS) column and a mobile phase of methanol and water. The results of comparative methodological studies with rice bran oils and milkweed oils indicated that the reversed-phase PEPS-HPLC method in conjunction with the use of less hazardous solvents proved to be superior and a viable alternative to the conventional normal-phase HPLC method. Unlike the traditional nonpolar octadecylsilica phase, which fails to resolve β-γ pairs of T and T₃, HPLC with the unique polar PFPS column enables separations of all compounds of interest. Except for palm oil, βT and γT were detected in all other crude oils. Although most milkweed oils contained moderale levels of βT and γT, the βT species was present in relatively low abundance in edible oils despite the observation of fairly high concentrations of γT in the latter oils. βT₃ and γT₃ were detected along with αT₃ and σT₃ only in palm and rice bran oils. Tocolderived antioxidant distribution data for zero-time processed oils provided potential utility in correlation studies of frying quality and stability. The variable distribution data for crude oils shed some light on market profitability of oilseeds with rich sources of vitamin E-related minor constituents.     

Some compositional properties of camelina (camelina sativa L. Crantz) seeds and oils

Fatty acid profiles (FAP), tocopherol (T), and tocotrienol (T3) contents, total lipid contents, and trypsin inhibitor activity were quantitated from thirteen accessions of camelina (Camelina sativa L. Crantz), a little-known oilseed. Camelina seeds of ten accessions were also assayed for ß-glucans. FAP (%) of camelina oils were: oleic (14.1 to 19.5), linoleic (18.8 to 24.0), linolenic (27.0 to 34.7), eicosenoic (12.0 to 14.9), erucic (0.0 to 4.0), all others (11.8 to 17.4). Camelina oil T and T3 contents (mg/100 g) were: αT (0.66 to 2.38), ßT (0.38 to 1.45), γT/ßt3 (4.37 to 18.68), δT (0.00 to 0.48), γT3 (0.00 to 0.79), γT3 (0.00), γT3 (0.00). Total tocols were higher in camelina than in canola, crambe, flax, soybean, and sunflower, with γT/ßT3 constituting 82% of total tocols. The oil content of camelina seeds ranged from 29.9 to 38.3%. Camelina seeds did not contain ß-glucans. Trypsin units inhibited ranged from 12 to 28 compared to 111 for raw soybean.     

High performance liquid chromatography of the tocols in corn grain

A sensitive and selective method was developed for analyzing the tocol isomers in corn grain by high performance liquid chromatography (HPLC) with fluorescence detection. The relative proportions and the total amounts of the tocol isomers (α-tocopherol, α-tocotrienol, γ-tocopherol and γ-tocortrienol) varied greatly among the 15 corn inbreds that were examined. Although γ-tocopherol has traditionally been considered to be the predominant vitamin E isomer in corn, inbreds with equal or higher levels of α-tocopherol have been discovered. No tocotrienols were found in corn germ oil, only α-and γ-tocopherols. Analysis of the tocopherols of the germ oils of inbreds and their reciprocal crosses indicated that the proportions of the α- and γ-isomers and the total amount of the tocopherols are heritable. Presented at the 74th AOCS annual meeting, Chicago, 1983.     

Further studies on sphingolipids in wheat grain

The principal molecular species of sphingolipids in wheat grain were confirmed to beN-2′-hydroxylignoceroyl-4-hydroxysphinganine for ceramide, and 1-O-β-glucosyl-, 1-O-[β-mannosyl(1→4)-O-β-glucosyl]-, 1-O-[β-mannosyl(1→4)-O-β-mannosyl(1→4)-O-β-glucosyl]-and 1-O[β-mannosyl(1→4)-O-β-mannosyl(1→4)-O-β-mannosyl(1→4)-O-β-glucosyl]-N-2′-hydroxypalmitoyl (or hydroxyarachidoyl)-cis-8-sphingenine for mono-, di-, tri- and tetraglycosylceramide, respectively. A novel glycolipid, cellobiosylceramide, was found as the minor diglycosylceramide; the major species was characterized to be 1-O-[β-glucosyl(1→4)-O-β-glucosyl]-N-2′-hydroxypalmitoyl (or hydroxyarachidoyl)-cis-8-sphingenine. It was observed in these sphingolipids that the dihydroxy bases were combined mainly with C₁₆ and C₂₀ acids, whereas the trihydroxy bases combined mostly with acids of chain length of 20 or more.     

Recovered oil from palm-pressed fiber: A good source of natural carotenoids,vitamin E,and sterols

Recovered fiber from pressed palm fruits, which is normally burned as fuel to provide energy for the palm oil mills, has now been found to be a rich source of carotenoids, vitamin E (tocopherol and tocotrienols), and sterols. Residual oil (5–6% on dry basis) extracted from palm press fibers contains a significant quantity of carotenoids (4000–6000 ppm), vitamin E (2400–3500 ppm), and sterols (4500–8500 ppm). The major identified carotenoids are α-carotene (19.5%), β-carotene (31.0%), lycopene (14.1%), and phytoene (11.9%). In terms of vitamin E, α-tocopherol constitutes about 61% of the total vitamin E present, the rest being tocotrienols (α-, γ-, and δ-). The major sterols present are β-sitosterol (47%), campesterol (24%), and stigmasterol (15%). The oil extracted from palm-pressed fiber is contaminated with about 30% of palm kernel oil. The quality of this fiber oil is slightly lower than that of crude palm oil in terms of the content of free fatty acids, peroxide value, and anisidine value.     

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.     

Assessment of olive oil adulteration by reversed-phase high-performance liquid chromatography/amperometric detection of tocopherols and tocotrienols

A method involving reversed-phase high-performance liquid chromatography with amperometric detection has been developed for the analysis of tocopherols and tocotrienols in vegetable oils. The sample preparation avoids saponification. Recoveries of α-tocotrienol and γ-tocotrienol in extra virgin olive oil were 97.0 and 102.0%, respectively. No tocotrienols were detected in olive, hazelnut, sunflower, and soybean oils, whether virgin or refined. However, relatively high levels of tocotrienols were found in palm and grapeseed oils. This method could detect small quantities (1–2%) of palm and grapeseed oils in olive oil or in any tocotrienol-free vegetable oil and might, therefore, help assess authenticity of vegetable oils.     

Fast quality screening of vegetable oils by HPLC-thermal lens spectrometric detection

Isocratic reversed-phase HPLC with thermal lens spectrometric (TLS) detection enabled identification of linseed, olive, sesame, and wheat germ vegetable oils to control the authenticity of the oils based on characteristic carotenoid/carotene profiles. Four characteristic regions of carotenoids (i.e., lutein, xanthophyll, carotene, and lycopene) have been identified in each type of oil. The concentrations of total β-carotene (BC) and α-carotene (AC), together with trans-to cis-isomers of β-carotene (TBC/CBC) and BC/AC ratios were shown to be reliable and useful indices for fast screening of oils for nutritional quality. The oil TBC/CBC ratio and the BC concentration (in μg/mL) should meet the following numerical criteria: linseed (≽2∶1, ≽1.7), olive (≽3∶1, ≽0.4), sesame (≽1∶1, ≽0.1), and wheat germ oil (≽1∶1, ≽1.7). Based on the above criteria, unsatisfactory olive oils differed significantly from the consumable ones. Likewise, the concentration of AC in consumable wheat germ and sesame oil should not be lower than 0.6 and 0.02 μg/mL, respectively. The AC level in safflower oil should not be higher than 0.04 μg/mL. The BC/AC ratios exceeding 3∶1, 6∶1, and 8∶1 should be used as an additional quality requirement for consumable wheat germ, sesame, and safflower oil, respectively.     

Effects of steeping conditions during wet-milling on the retentions of tocopherols and tocotrienols in corn

Vitamin E is a natural antioxidant that plays significant roles in food preservation and disease prevention. There are eight naturally occurring vitamin E isomers (tocols): α-, β-, γ-, and δ-tocopherols and α-, β-, γ-, and δ-tocotrienols. Corn oil is a major source of vitamin E. Most of the corn oil produced in the United States is a co-product of corn wet-milling. There is limited knowledge about the effects of corn wet-milling on the retention of these vitamin E isomers. A high-performance liquid chromatography method was developed for simultaneous determinations of tocols in steeped corn samples. Effects of steeping conditions (steeping time and SO₂ concentration) on retention of tocols in corn were investigated. α-Tocopherol, γ-tocopherol, α-tocotrienol, and γ-tocotrienol are the predominant vitamin E isomers in the corn variety used in the study. Steeping conditions had little effect on the concentration of α-tocopherol and α-tocotrienol. However, a higher concentration of SO₂ and a shorter steeping time gave a slightly higher γ-tocotrienol content and lower γ-tocopherol content. Corn kernels steeped in a vitamin C solution had a much higher concentration of the tocols than those steeped in SO₂ solution.     

Amaranth Oil Increased Fecal Excretion of Bile Acid but Had No Effect in Reducing Plasma Cholesterol in Hamsters

Hamsters were fed for 4 weeks on four different diets: control (C) (balanced diet containing 20 % corn oil as the lipid source), hypercholesterolemic (H) (identical to C but containing 12 % coconut oil, 8 % corn oil and 0.1 % cholesterol as the lipid source), amaranth oil (A) (identical to H without corn oil but with amaranth oil), and squalene (S) (identical to H but admixed with squalene in the ratio found in amaranth oil). There were no significant differences in lipid profile, and in the cholesterol excreted in the animals’ feces from amaranth oil (A) and squalene (S) groups. Fecal excretion of bile acids was greater in the amaranth oil (A) and squalene groups (S) as compared to the other groups. The scores of steatosis and parenchymal inflammation observed in the amaranth oil (A) and squalene groups (S) were superior to the ones observed in the other groups. Our findings demonstrated that amaranth oil, and its component squalene, increased the excretion of bile acids but did not have a hypocholesterolemic effect in hamsters fed on a diet containing high amounts of saturated fat and cholesterol.     

Evidence for multiple sterol methyl transferase pathways in <Emphasis Type="Italic">Pneumocystis carinii</Emphasis>

The sterol composition of Pneumocystis carinii, an opportunistic pathogen responsible for life-threatening pneumonia in immunocompromised patients, was determined. Our purpose was to identify pathway-specific enzymes to impair using sterol biosynthesis inhibitors. Prior to this study, cholesterol 15 (ca. 80% of total sterols), lanosterol 1, and several phytosterols common to plants (sitosterol 31, 24α-ethyl and campesterol, 24α-methyl 30) were demonstrated in the fungus. In this investigation, we isolated all the previous sterols and many new compounds from P. carinii by culturing the microorganism in steroid-immunosuppressed rats. Thirty-one sterols were identified from the fungus (total sterol=100 fg/cell), and seven sterols were identified from rat chow. Unusual sterols in the fungus not present in the diet included, 24(28)-methylenelanosterol 2; 24(28)E-ethylidene lanosterol 3; 24(28)Z-ethylidene lanosterol 4; 24β-ethyllanosta-25(27)-dienol 5; 24β-ethylcholest-7-enol 6; 24β-ethylcholesterol 7; 24β-ethylcholesta-5,25(27)-dienol 8; 24-methyllanosta-7-enol 9; 24-methyldesmosterol 10; 24(28)-methylenecholest-7-enol 11; 24β-methylcholest-7-enol 12; and 24β-methylcholesterol 13. The structural relationships of the 24-alkyl groups in the sterol side chain were demonstrated chromatographically relative to authentic specimens, by MS and high-resolution ¹H NMR. The hypothetical order of these compounds poses multiple phytosterol pathways that diverge from a common intermediate to generate 24β-methyl sterols: route 1, 1→2→11→12→13; route 2, 1→2→9→10→13; or 24β-ethyl sterols: route 3, 1→2→4→6→7; route 4, 1→2→5→8→7. Formation of 3 is considered to form an interrupted sterol pathway. Taken together, operation of distinct sterol methyl transferase (SMT) pathways that generate 24β-alkyl sterols in P. carinii with no counterpart in human biochemistry suggests a close taxonomic affinity with fungi and provides a basis for mechanism-based inactivation of SMI enzyme to treat Pneumocystis pneumonia.     

Identification and quantitation of γ-oryzanol components and simultaneous assessment of tocols in rice bran oil

A reverse-phase high-performance liquid chromatography method was developed for the simultaneous separation and quantitation of tocopherols, tocotrienols and oryzanols present in rice bran oil. Tocopherols and tocotrienols were quantitated by fluorescence detection and oryzanols (ferulic acid esters of sterols and triterpene alcohols) by photodiode array detection. Chemical ionization mass spectrometry was used to identify cycloartenyl ferulate, 24-methylene cycloartanyl ferulate, campesteryl ferulate, β-sitosteryl ferulate and cycloartanyl ferulate as the major oryzanols separated by this procedure. The levels of these nutritionally significant components were found to vary in fully processed, edible rice bran oils from different manufacturers.     

The chemistry and antioxidant properties of tocopherols and tocotrienols

This article is a review of the fundamental chemistry of the tocopherols and tocotrienols relevant to their antioxidant action. Despite the general agreement that α-tocopherol is the most efficient antioxidant and vitamin E homologuein vivo, there was always a considerable discrepancy in its “absolute” and “relative” antioxidant effectivenessin vitro, especially when compared to γ-tocopherol. Many chemical, physical, biochemical, physicochemical, and other factors seem responsible for the observed discrepancy between the relative antioxidant potencies of the tocopherolsin vivo andin vitro. This paper aims at highlighting some possible reasons for the observed differences between the tocopherols (α-, β-, γ-, and δ-) in relation to their interactions with the important chemical species involved in lipid peroxidation, specifically trace metal ions, singlet oxygen, nitrogen oxides, and antioxidant synergists. Although literature reports related to the chemistry of the tocotrienols are quite meager, they also were included in the discussion in virtue of their structural and functional resemblance to the tocopherols.     

Tocotrienols Suppress Proinflammatory Markers and Cyclooxygenase-2 Expression in RAW264.7 Macrophages

Tocotrienols are powerful chain breaking antioxidant. Moreover, they are now known to exhibit various non-antioxidant properties such as anti-cancer, neuroprotective and hypocholesterolemic functions. This study was undertaken to investigate the anti-inflammatory effects of tocotrienol-rich fraction (TRF) and individual tocotrienol isoforms namely δ-, γ-, and α-tocotrienol on lipopolysaccharide-stimulated RAW264.7 macrophages. The widely studied vitamin E form, α-tocopherol, was used as comparison. Stimulation of RAW264.7 with lipopolysaccharide induced the release of various inflammatory markers. 10 μg/ml of TRF and all tocotrienol isoforms significantly inhibited the production of interleukin-6 and nitric oxide. However, only α-tocotrienol demonstrated a significant effect in lowering tumor necrosis factor-α production. Besides, TRF and all tocotrienol isoforms except γ-tocotrienol reduced prostaglandin E₂ release. It was accompanied by the down-regulation of cyclooxygenase-2 gene expression by all vitamin E forms except α-tocopherol. Collectively, the data suggested that tocotrienols are better anti-inflammatory agents than α-tocopherol and the most effective form is δ-tocotrienol.     

Phytosterols,triterpene alcohols,and phospholipids in seed oil from white lupin

This study was conducted to determine effects of genotypes and growing environment on phytosterols, triterpene alcohols, and phospholipids (PL) in lupin (Lupinus albus L.) oil from seven genotypes grown in Maine and Virginia. The unsaponifiable lipid (UNSAP) and phospholipid (PL) fractions ranged from 2.1 to 2.8% and from 2.6 to 2.8% of oil, respectively. UNSAP in lupin oil contained 19.9 to 28.7% sterols and 17.3 to 22.0% triterpene alcohols. Growing location significantly affected contents of total PL, PS, phosphatidylglycerol, β-sitosterol, campesterol, and β-amyrin. Genotypic effects were significant for stigmasterol. PC (32.6 to 46.3% of PL), PE (21.6 to 32% of PL), and PS (11.2 to 17.9% of PL) were the major PL in lupin oil. The concentration of PL classes in lupin oil were in the following descending order: PC>PE>PS>PI>phosphatidic acid > lysophosphatidylcholine > phosphatidylglycerol > diphosphatidylglycerol. In descending order of abundance, the sterols present in lupin oil were: β-sitosterol > campesterol > stigmasterol > Δ⁵-avenasterol > Δ⁷-stigmastenol Lupeol was the most prominent triterpene alcohol in lupin seed oil. In general, growing environment had a much greater influence on lupin oil characteristics than the genotypes.     

The Antioxidant Functions of Tocopherol and Tocotrienol Homologues in Oils,Fats, and Food Systems

This review paper is focused on the relative antioxidant activities of tocopherols and tocotrienols in oils and fats and certain food systems. α-Tocopherol generally showed better antioxidant activity than γ-tocopherol in fats and oils, but at higher concentrations γ-tocopherol was found to be a more active antioxidant. The results of studies on the optimum antioxidant concentrations of tocopherols in oils and fats indicated that the optimal level for α-tocopherol is usually lower than other tocopherols, meaning less α-tocopherol is needed for maximum antioxidant protection. There are comparatively very few studies related to the antioxidant activities of tocotrienols in oils and fats. It has been stated that generally γ-tocotrienol has higher antioxidant effect than α-tocotrienol, and tocotrienols may be better antioxidants than their corresponding tocopherols in certain oils and fats systems. Studies on the antioxidant activity of various tocopherols in food systems are varied and cannot be uniformly evaluated because experiments have generally focused on different foods and used various methods for the detection of antioxidant activities. Depending on the food system, in certain cases tocopherols were better antioxidants than synthetic antioxidants such as butylhydroxy toluene (BHT) or butylhydroxy anisole (BHA). However, in certain other food systems the synthetic antioxidants were more effective to increase the shelf life and the stability of foods than those containing tocopherols.     

Tocopherols and tocotrienols in Finnish foods: Oils and fats

The tocopherols and tocotrienols of vegetable oils, cod liver oil, margarines, butter and Voimariini dairy spread were analyzed by HPLC. The total tocopherol content varied from 4 (coconut oil) to 242 mg/100 g (wheat germ oil). α-tocopherol equivalents varied from 2 (coconut oil) to 225 mg/100 g (wheat germ oil). Semisoft and soft margarines had an average total tocopherol of 53 and 61 mg, and an average α-tocopherol equivalent of 17 and 27 mg/100 g, respectively. Hard margarines averaged 29 mg total tocopherol and 9 mg α-tocopherol equivalent/100 g. The average tocopherol content of butter and Voimariini was 2 and 15 mg/100 g, respectively, and the average α-tocopherol equivalent 2 and 6 mg/100 g.     

Rapid method for the quantitative determination of individual tocopherols in oils and fats

Oils and fats are frozen two times from acetone solution at -80 C under protection by ascorbyl palmitate. Tocopherols (T) and tocotrienols (T₃) present in the filtered extract are separated into their homologues by one-dimensional thin layer chromatography on precoated silica gel plates in the n-hexane/ethyl acetate system 92.5:7.5. Complete separation of the positional isomers β-T and γ-T is accomplished as well, whereas β-T₃ and γ-T are assumed to form identical bands. Suitable spray- and detection systems including new found coloring ones are described for the qualitative estimation of the chromatograms. The content of individual tocopherols (T and T₃) of 24 commercial vegetable oils is quantitatively determined using the Emmerie-Engel procedure.     

Fatty acid and triacylglycerol compositions of seed oils of five Amaranthus accessions and their comparison to other oils

This paper reports the fatty acid and triacylglycerol (TAG) compositions of five Amaranthus accessions (RRC1011, R149, A.K343, A.K432, and A. K433) representing two species and a cross between one of these and a third species. Seed oils of these were analyzed by gas chromatography and reversed-phase high-performance liquid chromatography, and their compositional properties compared with buck-wheat (Fagopyrum esculentum), corn (Zea mays), rice bran (Oryza sativa), soybean (Glycine max L. Merr.), sesame (Sesamum indicum), quinoa (Chenopodium quinoa), and cottonseed (Gossypium hirsutum) oils. All Amaranthus accessions were relatively high in palmitic (21.4–23.8%) and low in oleic (22.8–31.5%) and linolenic (0.65–0.93%) acids when compared to most of the grain and seed oils. The fatty acid composition of Amaranthus accessions K343, K433, and K432 (group I) were different from R149 and RRC1011 (group II) in mono and polyunsaturated fatty acids, but the saturate/unsaturate (S/U) ratios were very similar. All Amaranthus accessions were similar in TAG type, but showed slight differences in percentage. High similarities in UUU, UUS, and USS composition were observed among Amaranthus K343, K433 and K432, and between R149 and RRC1011. The fatty acid compositions of Amaranthus oil (group I) and cottonseed oil were similar, but their TAG compositions were different. The grain and oilseed oils were different from each other and from the Amaranthus accessions oils in terms of fatty acid composition, S/U, and TAG ratios. The UUU, UUS, and USS percentages were very diverse in grain and seed oils. The percentages of squalene in the TAG sample from the Amaranthus accessions were 8.05% in K343, 11.10% in K433, 11.19% in K432, 9.96% in R149, and 9.16% in RRC1011. Squalene was also tentatively identified in quinoa and ricebran oils at levels of 3.39 and 3.10%, respectively.