Development of a Simultaneous Normal-Phase HPLC Analysis of Lignans, Tocopherols, Phytosterols, and Squalene in Sesame Oil Samples.

The objective of this study was to develop a simultaneous analytical method for the determination of lignans, tocols, phytosterols, and squalene using high-performance liquid chromatography coupled with a diode array and fluorescence detector (HPLC-DAD-FLD). The method employed a VertisepTM UPS silica HPLC column (4.6 × 250 mm, 5 µm) with a mobile phase mixture of n-hexane/tetrahydrofuran/2-propanol. This approach enabled the simultaneous analysis of ten compounds within 22 min. The linear correlation (R2) exceeded 0.9901. The limit of detection (LOD) and limit of quantitation (LOQ) were up to 0.43 µg mL-1 for lignans and tocopherols and up to 326.23 µg mL-1 for phytosterol and squalene. The precision and accuracy of the intra-day and inter-day variation were less than 1.09 and 3.32% relative standard deviations (RSDs). Furthermore, the developed method was applied for the analysis of targeted compounds in twenty-eight sesame oil samples (1775-8965 µg g-1 total lignans, 29.7-687.9 µg g-1 total tocopherols, 2640-9500 µg g-1 phytosterol, and 245-4030 µg g-1 squalene). The HPLC method that has been developed was proven to be a reliable and effective tool for the determination of those functional compounds among sesame oil samples.

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.

Mineral elements and their relation to anthocyanin content in pigmented rice plants using definitive screening design and self-organizing maps.

BACKGROUND: Mineral elements are nutrients required by an organism to perform functions necessary for survival. Stress-induced metabolism following nutritional stress has been reported to increase levels of anthocyanin. However, the role of mineral elements commonly found in soil and their contribution to the accumulation of anthocyanin content in rice plants is uncertain. RESULT: Amongst the ten mineral elements investigated, the cultivation of rice plants in clean sand showed that the Mg-, Se-, and Cu-treated plants had the highest accumulated anthocyanin content in the leaves, whereas B, Cr, and Se had the greatest effect on grains. Yield component data showed major positive effects from Mg, Cr, and B. The interaction of Zn*Se and Mg*Cu positively affected the anthocyanin content in grains. The self-organizing map indicated that the total anthocyanin content was relatively proportional to the concentration of Mn, B, and Cr, but disproportional to that of Se. However, rice plants with added Fe produced the smallest amount of total anthocyanin content, less than the control, in the four stages of rice growth. CONCLUSION: The appropriate concentrations of mineral elements in soil could promote the proliferation of anthocyanin content in rice plants and grains. © 2023 Society of Chemical Industry.

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.

Smartphone digital image colorimetric determination of the total monomeric anthocyanin content in black rice via the pH differential method.

In this research, the pH differential method was explored for the first time using a mobile phone as a detector, replacing UV-Vis spectrophotometry. A smartphone digital image colorimetric (SDIC) system was developed for the determination of the total monomeric anthocyanin content in black rice grains using colour values. The change in colour of anthocyanin cyanidin-3-glucoside (C3G) at pH 1.0 and pH 4.5 was investigated and described through colour model systems (RGB and CIELAB). Under optimum conditions, the quantitative method based on the chroma difference, was able to quantify the total amount of monomeric anthocyanins with a linear correlation (R2 = 0.989) to that of UV-Vis spectrophotometry and high performance liquid chromatography (HPLC) (reference method). The SDIC system displayed good precision (≤1.88% RSD) and a low detection limit (2.2 ± 0.1 µg g-1). The validated results demonstrated that the developed method was a cost-effective alternative for the quantitation of anthocyanins. In addition, the effect of six mineral elements on black rice cultivation was investigated. The results showed that the addition of the Ca fertiliser resulted in black rice grains with an anthocyanin content 2-fold higher than that of the control with a significant difference (p < 0.05).

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.

Effect of wall materials on some physicochemical properties and release characteristics of encapsulated black rice anthocyanin microcapsules.

Anthocyanin-rich extracts derived from the bran of a select Thai black rice cultivar, Leum Pua, were encapsulated by spray drying. In this microencapsulation method, twelve wall materials comprising maltodextrin (M), gum arabic (G), whey protein isolate (W), and their combinations were employed. For comparison, an ionic gelation method was employed using alginate-whey protein isolate (AL-W) as a wall material. Thirteen microcapsules were analysed by scanning electron microscopy, and their encapsulation efficiencies were almost 100%, except for that of the W (83%). M (88%) and its combination with W exhibited the best anthocyanin retention, while pure W contained the highest amount of total phenolic compounds and antioxidant activity. An in vitro digestion study using simulated gastrointestinal fluids revealed the highest released total phenolic content and antioxidant activity of the intestinal fluids when the microcapsule was spray-dried with pure W.

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.

Phosphatidylcholine hydroperoxide promotes VEGF-induced angiogenesis in endothelial cells and rat aorta ring cultures.

BACKGROUND: Phosphatidylcholine hydroperoxide (PCOOH) is a primary oxidation product of PC, and is markedly accumulated in blood plasma and arterial walls in atherosclerotic animals and humans. The role of PCOOH in the induction of angiogenesis is unknown. METHODS: In this study, we investigated whether PCOOH stimulated angiogenic responses (e.g., vascular endothelial growth factor (VEGF)-induced cell proliferation, migration, and tube formation, and angiogenesis-related gene/protein expression) in human umbilical vein endothelial cells (HUVEC) and in an ex vivo rat aorta model. RESULTS: VEGF induced proliferation, migration, and tube formation of HUVEC, and these angiogenic responses were all enhanced by PCOOH but not by native (nonoxidized) PC. The angiogenic effects of PCOOH are considered to be mediated via generation of reactive oxygen species and activation of both PI3K/AKT and MAPK pathways. The angiogenic activities of PCOOH were also confirmed by the rat aortic ring assay. CONCLUSIONS: These results indicate that PCOOH can elicit several angiogenic responses. GENERAL SIGNIFICANCE: The present study implies an important role of PCOOH in atherosclerosis progression and plaque instability.

Amadori-glycated phosphatidylethanolamine, a potential marker for hyperglycemia, in streptozotocin-induced diabetic rats.

It has been demonstrated in vivo that lipid glycation products such as Amadori-glycated phosphatidylethanolamine (Amadori-PE) accumulate in the plasma of diabetic humans and animals, but how lipid glycation products are formed under hyperglycemic conditions are not clear. We sought to clarify the occurrence of lipid glycation and its relationships with lipid peroxidation and protein glycation during the development of hyperglycemia using the streptozotocin (STZ)-induced diabetic rat model. A significant increase in Amadori-PE was observed in STZ rats 7 days after STZ treatment, and Amadori-PE (especially 18:0-20:4 Amadori-PE) was found at high levels in the blood and in organs that are strongly affected by diabetes, such as the kidney. Significant changes in Amadori-PE appeared to occur prior to changes in levels of oxidized lipids, which increased after 21-28 days. In addition, accumulation of Nε-(carboxymethyl)lysine (CML), a protein glycation product, proceeded somewhat more slowly and moderately than that of Amadori-PE, suggesting that Amadori-PE and CML are early and advanced glycation products, respectively. Our results suggest that Amadori-PE may be a useful predictive marker for hyperglycemia, particularly in the early stages of diabetes. Similar speculations have been made from previous human studies, but this study provides a direct evidence to support the speculations in rat study.

Amyloid ß-induced erythrocytic damage and its attenuation by carotenoids.

The presence of amyloid ß-peptide (Aß) in human blood has recently been established, and it has been hypothesized that Aß readily contacts red blood cells (RBC) and oxidatively impairs RBC functions. In this study, we conducted in vitro and in vivo studies, which provide evidence that Aß induces oxidative injury to RBC by binding to them, causing RBC phospholipid peroxidation and diminishing RBC endogenous carotenoids, especially xanthophylls. This type of damage is likely to injure the vasculature, potentially reducing oxygen delivery to the brain and facilitating Alzheimer's disease (AD). As a preventive strategy, because the Aß-induced RBC damage could be attenuated by treatment of RBC with xanthophylls, we suggest that xanthophylls may contribute to the prevention of AD.

gamma-Tocotrienol reduces squalene hydroperoxide-induced inflammatory responses in HaCaT keratinocytes.

Squalene hydroperoxide (SQ-OOH), the primary peroxidation product of squalene (SQ), accumulates at the surface of sunlight-exposed human skin. There are however only a few studies on the pathogenic actions (i.e., inflammatory stimuli) of SQ-OOH. Here, we evaluated whether SQ-OOH induced inflammatory responses in immortalized human keratinocytes (HaCaT). We found that SQ-OOH caused an increase in the expression of inflammatory genes such as the interleukins as well as cyclooxygenase-2 (COX-2). In concordance with the upregulation of COX-2 mRNA, SQ-OOH enhanced reactive oxygen species generation, nuclear factor kappa B activation, COX-2 protein expression, and prostaglandin E2 production. Therefore, the pro-inflammatory effects of SQ-OOH may be mediated in part via COX-2. On the other hand, gamma-tocotrienol (gamma-T3, an unsaturated form of vitamin E) was found to ameliorate the SQ-OOH actions. These results suggest that SQ-OOH induces inflammatory responses in HaCaT, implying that SQ-OOH plays an important role in inflammatory skin disorders. As a preventive strategy, inflammation could be reduced via the use of gamma-T3.