Bioactive Constituents in Cold-Pressed Plant Oils: Their Structure, Bioactivity and Chromatographic Analysis.

Cold-pressed oils are oils prepared from pressing plant materials with a screw or hydraulic press, yielding oils with little contamination of harmful chemicals and high content of nutrients and functional constituents. Cold-pressed oils have gained increasing recognition as food supplements for preventing and ameliorating body deterioration due to ageing and the progression of lifestyle diseases or non-communicable diseases. This article aimed to review their structure, bioactivity, and chromatographic analysis of the mostly found functional compounds in cold-pressed oils, including phytosterols, carotenoids, tocols (tocopherols and tocotrienols), phenolic compounds (flavonoids, phenolic acids, tannins, stilbenes, and lignans), and squalene.

Influence of black rice (Oryza sativa L.) bran derived anthocyanin-extract on growth rate, immunological response, and immune-antioxidant gene expression in Nile tilapia (Oreochromis niloticus) cultivated in a biofloc system.

This study investigated the effects of dietary supplementation with anthocyanin extracted from black rice bran (AR) on the growth rate, immunological response, and expression of immune and antioxidant genes in Nile tilapia raised in an indoor biofloc system. A total of 300 Nile tilapia fingerlings (15.14 ± 0.032 g) were maintained in 150 L tanks and acclimatized for two weeks. Five experimental AR diets (0, 1, 2, 4, and 8 g kg-1) with various anthocyanin doses were used to feed the fish. We observed that the growth and feed utilization of fish fed with different dietary AR levels increased significantly after eight weeks (p < 0.05). In addition, the serum immunity of fish fed AR diets was much greater than that of those fed non-AR diets (p < 0.05). However, there were little or no difference in between fish fed AR enriched diets and the control AR-free diet (p > 0.05). After eight weeks, fish fed AR-supplemented diets had significantly higher mRNA transcript levels in immune (interleukin [IL]-1, IL-8, and liposaccharide-binding protein [LBP]) and antioxidant (glutathione transferase-alpha [GST-α] and glutathione reductase [GSR]) genes compared to control fish fed the AR-free diet, with the greatest enhancement of mRNA transcript levels (in the case of IL-8 by up to about 5.8-fold) in the 4 g kg-1 AR diet. These findings suggest that dietary inclusion of AR extract from black rice bran at 4-8 g kg-1 could function as a herbal immunostimulant to enhance growth performance, feed consumption, and immunity in Nile tilapia.

Microwave treatment of rice bran and its effect on phytochemical content and antioxidant activity.

An alternative approach for rice bran stabilization is microwave treatment. However, the effects of the microwave treatment on the contents of bioactive compounds and antioxidant activities of the rice bran have rarely been reported in detail. In this study, microwave pretreatment (130-880 W for 0.5-5.0 min) of rice bran was proposed where the antioxidant activity, total flavonoids, and total phenolic contents were determined using UV-Vis spectrometry. Tocols, γ-oryzanols, squalene, phytosterols and phenolic compounds were quantified using high-performance liquid chromatography. The results showed an increase in the antioxidant activity (0.5 folds), total phenolic contents (1.3 folds), total flavonoid contents (0.9 folds), total tocols (2.6 folds), total γ-oryzanols (1.6 folds), and total phytosterols (1.4 folds). Phytochemicals were enhanced, especially trans-p-coumaric acid (10.3 folds) and kaempferol (8.6 folds). The microwave treatment at 440 W for 2.5 min provided the best contents of the bioactive compounds and antioxidant activity. This work revealed the microwave treatment as a potential tool for stabilizing rice bran and increasing the usability of its phytochemicals, which applies to several industries concerning the use of rice bran as an ingredient.

Comparative Studies on the Hepatoprotective Effect of White and Coloured Rice Bran Oil against Acetaminophen-Induced Oxidative Stress in Mice through Antioxidant- and Xenobiotic-Metabolizing Systems.

Rice bran oil (RBO) comprises various nutrients and phytochemicals which exhibit several health benefits. There are no studies regarding the functional effects of different colours of RBO. This study was aimed to compare the constituents and antioxidant activities of white rice bran oil (WRBO) and coloured rice bran oil (CRBO). Each RBO showed similar free fatty acid profiles. However, greater amounts of vitamin E, phytosterols, carotenoids, and chlorophylls were found in CRBO, which had lower γ-oryzanol content than WRBO. Oxidative stress was induced in male mice by an overdose of acetaminophen (APAP) at 300 mg/kg body weight. The mice were then fed with RBO at the equivalent dose to 100 mg/kg body weight of γ-oryzanol three hours later and sacrificed six hours after APAP treatment. The administration of 100 mg γ-oryzanol equivalent in CRBO ameliorated APAP-induced hepatotoxicity in mice more strongly than 100 mg γ-oryzanol equivalent in WRBO, as evidenced by the significant reduction of serum ALT, hepatocellular necrosis, and hepatic lipid peroxidation. CRBO could improve xenobiotic-metabolizing and antioxidant enzyme activities, including glutathione S-transferase, superoxide dismutase, glutathione peroxidase, and glutathione reductase, and also increase mRNA expression of various antioxidant-responsive genes. Vitamin E, phytosterols, carotenoids, and chlorophyll might be the protective compounds in CRBO that alleviate APAP-induced hepatotoxicity through the interruption of APAP metabolism and the activation of antioxidant systems at both transcriptional and enzymatic levels. These findings might provide a protective role of CRBO on oxidative stress associated with several degenerative diseases.

GC-MS and HPLC-DAD analysis of fatty acid profile and functional phytochemicals in fifty cold-pressed plant oils in Thailand.

Cold-pressed oil is one of the healthiest plant extracts, but its use is limited only in some kinds of plants. Therefore, we aimed to investigate some potential cold-pressed oils with attractive fatty acid profiles and high amounts of functional phytochemicals. Fifty cold-pressed plant oils were prepared from various plant materials in Thailand, in which some of them were from uncommon or unattended plant materials. The oils included were nut oils (n = 9), pseudo-cereal oils (n = 9), legume oils (n = 3), amaranth oils (n = 3), marrow seed oils (n = 8), cruciferous seed oils (n = 7), and leafy green seed oils (n = 11). Gas-chromatography mass-spectrometry (GC-MS) and high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) were employed to analyze fatty acid profile and five functional phytochemicals (e.g., phytosterols, cholecalciferol, and squalene). Saturated fatty acids were detected around 7.87-36.04%, monounsaturated fatty acids 10.17-80.25%, and polyunsaturated fatty acids nondetectable (ND)-78.25%, phytosterols 663-15123 µg g-1, squalene 265-5979 µg g-1, and cholecalciferol ND-1287.75 µg g-1. The study showed chemical characteristic of the analyzed oils: some contained good fatty acid composition and some were rich in functional phytochemical content. Among the obtained oils, marrow seed oils are a good source of phytosterol, cholecalciferol, and linoleic acid. Pseudo-cereal oils are rich in squalene and linolenic acid. Legume oils are rich in phytosterols and oleic acid. Besides, principal component analysis (PCA) was applied to identify the significance of oils that share compositional similarity (e.g., the samples from pseudo-cereal oil were found on the lower side of the PCA space, which separated them from marrow and leafy green seed oils distributed on the upper part of the plot). In summary, the qualitative and quantitative data would provide a good foundation for further application or selection of those plant oils for health purposes.

Cold plasma treatment to improve germination and enhance the bioactive phytochemical content of germinated brown rice.

The changes in the bioactive phytochemicals of six cultivars of Thai germinated brown rice (GBR) were monitored in parallel to those of cold plasma-treated GBR (PGBR). After treatment with the optimal plasma conditions, the germination percentage, root length, and seedling height measurements of the most sensitive rice cultivar increased by 84%, 57%, and 69%, respectively. For all of the rice cultivars, there were no significant differences in the antioxidant activities of the GBRs and PGBRs. Conversely, higher contents of γ-oryzanols were observed in the PGBR group than in the GBR group during the 2-day germination period. Certain cultivars in the PGBR group reached their maximum values for total phenolic compounds, total vitamin E, certain simple phenolics, phytosterols, triterpenoids, and anthocyanins one day earlier than the same values for GBR. In contrast, the concentrations of 2-acetyl-1-pyrroline in both the GBR and PGBR samples were reduced significantly with increased germination time.

Simultaneous determination of tocols, γ-oryzanols, phytosterols, squalene, cholecalciferol and phylloquinone in rice bran and vegetable oil samples.

In this study, a simultaneous analytical method of tocols, γ-oryzanols, phytosterols, squalene, cholecalciferol and phylloquinone were developed using HPLC-DAD-FLD. The developed method allowed the quantification of 18 compounds in 30 min. Method validation showed linearity of calibration curves (α = 0.05). RSD of intra-day, inter-day and inter-laboratory precision were less than 4.88%. The limit of detections (LODs) and limit of quantifications (LOQs) were low (0.009-2.166 µg g-1) with recoveries around 96.0-102.9%. Results derived from the established method demonstrated a wide variation of detected compounds in rice bran and vegetable oil samples (22.4-1774.6 µg g-1 tocols, ND-26484 µg g-1 γ-oryzanols, ND-12655 µg g-1 phytosterols, ND-3189 µg g-1 squalene, ND-105.3 µg g-1 cholecalciferol, and ND-54.4 µg g-1 phylloquinone). Thus, the developed HPLC-DAD-FLD method is a powerful analytical tool for the above mentioned compounds useful in food and pharmaceutical application.

Some Strategies for Utilization of Rice Bran Functional Lipids and Phytochemicals.

Rice bran contains a great amount of functional lipids and phytochemicals including γ-oryzanols, tocotrienols, and tocopherols. However, utilization of those compounds is limited and needs some proven guidelines for better implementation. We introduce some effective strategies for the utilization of rice functional lipids, including an introduction of pigmented rice varieties for better bioactive compounds, biofortification of rice tocotrienols, plasma technology for improving rice phytochemicals, supercritical CO2 extraction of high quality rice bran oil, and an example on the development of tocotrienol-fortified foods.

Supercritical CO2 Extraction of Rice Bran Oil -the Technology, Manufacture, and Applications.

Rice bran is a good source of nutrients that have large amounts of phytochemicals and antioxidants. Conventional rice bran oil production requires many processes that may deteriorate and degrade these valuable substances. Supercritical CO2 extraction is a green alternative method for producing rice bran oil. This work reviews production of rice bran oil by supercritical carbon dioxide (SC-CO2) extraction. In addition, the usefulness and advantages of SC-CO2 extracted rice bran oil for edible oil and health purpose is also described.

Effect of post-harvest treatment on bioactive phytochemicals of Thai black rice.

Because black rice is rich in antioxidants, appropriate methods of post-harvest treatment are necessary for maintaining these bioactive phytochemicals. Drying methods, storage temperatures, storage duration, and packaging methods affected the contents of some bioactive compounds in the two varieties of Thai black rice used in this research. Sun drying reduces the loss of anthocyanins and γ-oryzanols more than does hot air drying. Glutinous black rice stored as paddy at cool room temperature retains more anthocyanins, γ-oryzanols, and vitamin E than does paddy stored at room temperature. Nylon/LLDPE pouches containing N2 are the most suitable packaging for preserving the key aroma compound 2-acetyl-1-pyrroline (2AP), total phenolic, and anthocyanin contents of unpolished aromatic black rice. These pouches also retard the formation of some common off-flavor compounds.

Simultaneous quantification of vitamin E, γ-oryzanols and xanthophylls from rice bran essences extracted by supercritical CO2.

Since the nutrition value of rice is diminished during rice processing, technology that can preserve and sustain functional compounds is necessary. In this study, supercritical carbon dioxide (SC-CO2) extraction was optimized for operational conditions (time, temperature, pressure and modifier) to extract vitamin E, γ-oryzanols and xanthophylls from rice bran. The simultaneous quantification of the compounds was developed using high-performance liquid chromatography with diode array and fluorescence detectors. Central composite design and respond surface methodology were applied to achieve optimum extraction conditions. The optimized conditions were 60min, 43°C, 5420psi with 10% ethanol as a modifier. Pigmented rice bran extracts contained greater amounts of functional phytochemicals than non-pigmented rice bran extracts (0.68, 1410, and non-detectable µg/g compared with 16.65, 2480, and 0.10µg/g of vitamin E, γ-oryzanols and xanthophylls in pigmented and non-pigmented ones, respectively). SC-CO2 extraction with modifier would be promising for preparation of phytochemical essences for therapeutic purpose.

A novel gelatin crosslinking method retards release of mulberry 1-deoxynojirimycin providing a prolonged hypoglycaemic effect.

Mulberry 1-deoxynojirimycin (DNJ), a potent α-glycosidase inhibitor, has therapeutic potency in the suppression of postprandial blood glucose levels thereby possibly preventing diabetes mellitus. However, DNJ has a relatively short half-life in vivo (about 2 h). Therefore, several doses of mulberry DNJ are required to achieve optimal therapeutic results. This study aimed to delay the release of mulberry DNJ with biodegradable matrices to maintain the intestinal DNJ concentration and prolong the hypoglycaemic effect in vivo. A novel, simple, and commercially viable method was adopted to develop DNJ-entrapped microspheres (DNJ-MSs). A higher extent of crosslinking and the larger sized DNJ-MS decreased the rate of mulberry DNJ release in vitro. Consequently, an in vivo study was performed in Wistar rats over a 6 h period. The area under curve (AUC) of rats with DNJ-MS was significantly increased, compared to animals dosed with mulberry powder (control). DNJ-MS suppressed postprandial glucose from sucrose administration at the initial and 3 h time points indicating a prolonged hypoglycaemic effect.

Effects of co-administration of tea epigallocatechin-3-gallate (EGCG) and caffeine on absorption and metabolism of EGCG in humans.

Based on the ratios of (-)-epigallocatechin-3-gallate (EGCG) and caffeine (CAF) levels found in commercial tea drinks, EGCG and CAF were co-administered to human volunteers at various EGCG/CAF ratios, and plasma EGCG was determined by high performance liquid chromatography with chemiluminescence detection. As for the results, in plasma taken after ingestion of a beverage containing 95 mg of EGCG alone, the area under the plasma EGCG concentration-time curve (AUC) was 857 ngxh/ml. A higher AUC (1,370 ngxh/ml) was observed when subjects ingested a beverage containing EGCG (95 mg) and a low amount of CAF (40 mg). In the case of ingestion of a beverage containing EGCG (95 mg) and a high amount of CAF (180 mg), the AUC tended to be somewhat higher (1,165 ngxh/ml), but not significantly so, compared with the beverage with EGCG alone. These findings (modulation of plasma EGCG level by CAF) provide ideas for modulating the bioavailability of tea catechins, which can be applied to tea-related drinks and foods.

Antioxidant effect of lutein towards phospholipid hydroperoxidation in human erythrocytes.

Peroxidised phospholipid-mediated cytotoxity is involved in the pathophysiology of many diseases; for example, phospholipid hydroperoxides (PLOOH) are abnormally increased in erythrocytes of dementia patients. Dietary carotenoids (especially xanthophylls, polar carotenoids such as lutein) have gained attention as potent inhibitors against erythrocyte phospholipid hydroperoxidation, thereby making them plausible candidates for preventing diseases (i.e. dementia). To evaluate these points, we investigated whether orally administered lutein is distributed to human erythrocytes, and inhibits erythrocyte PLOOH formation. Six healthy subjects took one capsule of food-grade lutein (9.67 mg lutein per capsule) once per d for 4 weeks. Before and during the supplementation period, carotenoids and PLOOH in erythrocytes and plasma were determined by our developed HPLC technique. The administered lutein was incorporated into human erythrocytes, and erythrocyte PLOOH level decreased after the ingestion for 2 and 4 weeks. The antioxidative effect of lutein was confirmed on erythrocyte membranes, but not in plasma. These results suggest that lutein has the potential to act as an important antioxidant molecule in erythrocytes, and it thereby may contribute to the prevention of dementia. Therefore future biological and clinical studies will be required to evaluate the efficacy as well as safety of lutein in models of dementia with a realistic prospect of its use in human therapy.

Tocotrienol content in hen eggs: its fortification by supplementing the feed with rice bran scum oil.

Tocotrienol (T3) is an unsaturated vitamin E having health benefits (e.g., anti-angiogenesis). We measured T3 in commercial eggs, and developed T3-fortified eggs by adding rice bran scum oil (RBO, containing 1.3% T3) to the feed. Commercial eggs contained about 0.11 mg of T3/egg, while the T3 content was improved to 0.62 mg/egg after RBO supplementation to the feed of hens for 7 d.

Tumor anti-angiogenic effect and mechanism of action of delta-tocotrienol.

Anti-angiogenic therapy mediated by drugs and food components is an established strategy for cancer prevention. Our previous cell-culture studies identified a food-derived anti-angiogenic compound, tocotrienol (T3, an unsaturated vitamin E), as a potential angiogenic inhibitor. Among T3 isomers, delta-T3 is considered as the most potent compound. The purpose of this study was therefore to evaluate the inhibitory effect of delta-T3 on tumor angiogenesis. As growth factors (e.g., vascular endothelial growth factor and fibroblast growth factor) play critical roles in tumor angiogenesis, a conditioned medium rich in these growth factors from human colorectal adenocarcinoma cells (DLD-1-CM) was used as an angiogenic stimulus. Delta-T3 (2.5-5 microM) significantly suppressed DLD-1-CM-induced tube formation, migration, and adhesion on human umbilical vein endothelial cells. These effects were partly associated with reactive oxygen species generation by delta-T3. Western blot analysis revealed that the anti-angiogenic effect of delta-T3 is attributable to regulation of growth factor-dependent phosphatidylinositol-3 kinase (PI3K)/phosphoinositide-dependent protein kinase (PDK)/Akt signaling as well as to induction stress response in endothelial cells. Moreover, we conducted an in vivo mouse Matrigel plug angiogenesis assay, and found that delta-T3 (10-20 microg) exhibits dose-dependent inhibition of DLD-1-induced vessel formation. These results suggest that T3 has potential use as a therapeutic dietary supplement for minimizing tumor angiogenesis.