Identification of a TBHQ-Interfering Peak in Crude Canola Oil Using AOCS Official Method Ce 6-86 and its Chromatographic Resolution.

AOCS Official Method Ce 6-86 "Antioxidants, Liquid Chromatographic Method" was originally developed to confirm the correct antioxidant was added at the specified concentration to refined oils. Today, this method is increasingly utilized to validate that antioxidants are absent from oil products. False positive results can have a significant impact on the ability to sell products in specific markets and can impart additional business expenditures for conclusive secondary analyses. In the current work, quantification of tert-butylhydroquinone (TBHQ) in crude canola/rapeseed oil using liquid chromatography (LC) with ultraviolet (UV) detection was compromised by an interfering peak. Analyses using liquid chromatography-mass spectrometry (GC-MS) and high-resolution accurate mass LC-MS identified the interferent as 2,6-dimethoxy-4-vinylphenol (canolol), an endogenous compound present in crude canola/rapeseed oil. Resolution of canolol and TBHQ using LC-UV can be achieved via minor modification of the chromatographic conditions.

Antioxidant (Tocopherol and Canolol) Content in Rapeseed Oil Obtained from Roasted Yellow-Seeded Brassica napus.

In this study, the effect of temperature (140, 160, 180 °C) and roasting time (5, 10, 15 min) on the bioactive compound content (canolol, tocopherol and plastochromanol-8) of cold-pressed oil from yellow-seeded rapeseed lines of different colors was investigated. Roasting increased the peroxide value in the seed oils compared to the oils from the control samples. However, roasting did not affect the acid values of the oils, which were 1.15-1.47 and 1.30-1.40 mg KOH/g, for line PN1 03/1i/14 (yellow seeds) and line PN1 563/1i/14 (brown seeds), respectively. In this study, the seeds of line PN1 03/1i/14 were characterized by different changes in canolol content during roasting than the seeds of PN1 563/1i/14. There was a 90-fold increase in canolol for the line PN1 03/1i/14 (768.26 µg/g) and a 46-fold increase for the line PN1 563/1i/14 (576.43 µg/g). Changes in tocopherol and PC-8 contents were also observed. There was an increase in the contents of γ-T and PC-8 in the oils obtained from the seeds roasted at 180 °C for 10 and 15 min. γ-T content increased by 17-18% after 15 min of roasting, whereas the PC-8 content increased twofold.

Defective ferritinophagy and imbalanced iron metabolism in PBDE-47-triggered neuronal ferroptosis and salvage by Canolol.

The brominated flame retardant 2,2',4,4'-tetrabromodiphenyl ether (PBDE-47) is a ubiquitous environmental pollutant that causes neurotoxicity. However, incomplete understanding of the underlying mechanisms has hampered the development of effective intervention strategies. Oxidative stress and related cell death are the modes of action for PBDE-47 neurotoxicity, which are also the characteristics of ferroptosis. Nonetheless, the role of ferroptosis in PBDE-47-induced neurotoxicity remains unclear. In the present study, we found that PBDE-47 triggered ferroptosis in neuron-like PC12 cells, as evidenced by intracellular iron overload, lipid peroxidation, and mitochondrial damage. This was confirmed by ferroptosis inhibitors including the lipid reactive oxygen species scavenger ferrostatin-1 and iron chelator deferoxamine mesylate. Mechanistically, PBDE-47 impaired ferritinophagy by disrupting nuclear receptor coactivator 4-mediated lysosomal degradation of the iron storage protein ferritin. Moreover, PBDE-47 disturbed iron metabolism by increasing cellular iron import via upregulation of transferrin receptor 1 and decreasing cellular iron export via downregulation of ferroportin 1 (FPN1). Intriguingly, rescuing lysosomal function by overexpressing cathepsin B (CatB) mitigated PBDE-47-induced ferroptosis by partially restoring dysfunctional ferritinophagy and enhancing iron excretion via the upregulation of FPN1. However, FPN1 knockdown reversed the beneficial effects of CatB overexpression on the PBDE-47-induced iron overload. Finally, network pharmacology integrated with experimental validation revealed that Canolol, the main phenolic compound in canola oil, protected against PBDE-47-evoked iron overload, resulting in ferroptosis by restoring defective ferritinophagy and improving abnormal iron metabolism via lowering iron uptake and facilitating iron excretion. Overall, these data suggest that ferroptosis is a novel mechanism of PBDE-47-induced neuronal death and that manipulation of ferritinophagy and iron metabolism via Canolol represents a promising therapeutic strategy.

A New Phenolic Acid Decarboxylase from the Brown-Rot Fungus Neolentinus lepideus Natively Decarboxylates Biosourced Sinapic Acid into Canolol, a Bioactive Phenolic Compound.

Rapeseed meal (RSM) is a cheap, abundant and renewable feedstock, whose biorefinery is a current challenge for the sustainability of the oilseed sector. RSM is rich in sinapic acid (SA), a p-hydroxycinnamic acid that can be decarboxylated into canolol (2,6-dimethoxy-4-vinylphenol), a valuable bioactive compound. Microbial phenolic acid decarboxylases (PADs), mainly described for the non-oxidative decarboxylation of ferulic and p-coumaric acids, remain very poorly documented to date, for SA decarboxylation. The species Neolentinus lepideus has previously been shown to biotransform SA into canolol in vivo, but the enzyme responsible for bioconversion of the acid has never been characterized. In this study, we purified and characterized a new PAD from the canolol-overproducing strain N. lepideus BRFM15. Proteomic analysis highlighted a sole PAD-type protein sequence in the intracellular proteome of the strain. The native enzyme (NlePAD) displayed an unusual outstanding activity for decarboxylating SA (Vmax of 600 U.mg-1, kcat of 6.3 s-1 and kcat/KM of 1.6 s-1.mM-1). We showed that NlePAD (a homodimer of 2 × 22 kDa) is fully active in a pH range of 5.5-7.5 and a temperature range of 30-55 °C, with optima of pH 6-6.5 and 37-45 °C, and is highly stable at 4 °C and pH 6-8. Relative ratios of specific activities on ferulic, sinapic, p-coumaric and caffeic acids, respectively, were 100:24.9:13.4:3.9. The enzyme demonstrated in vitro effectiveness as a biocatalyst for the synthesis of canolol in aqueous medium from commercial SA, with a molar yield of 92%. Then, we developed processes to biotransform naturally-occurring SA from RSM into canolol by combining the complementary potentialities of an Aspergillus niger feruloyl esterase type-A, which is able to release free SA from the raw meal by hydrolyzing its conjugated forms, and NlePAD, in aqueous medium and mild conditions. NlePAD decarboxylation of biobased SA led to an overall yield of 1.6-3.8 mg canolol per gram of initial meal. Besides being the first characterization of a fungal PAD able to decarboxylate SA, this report shows that NlePAD is very promising as new biotechnological tool to generate biobased vinylphenols of industrial interest (especially canolol) as valuable platform chemicals for health, nutrition, cosmetics and green chemistry.

Effects of High-Canolol Phenolic Extracts on Fragrant Rapeseed Oil Quality and Flavor Compounds during Frying.

Fragrant rapeseed oil (FRO) is a frying oil widely loved by consumers, but its quality deteriorates with increasing frying time. In this study, the effect of high-canolol phenolic extracts (HCP) on the physicochemical properties and flavor of FRO during frying was investigated. During frying, HCP significantly inhibited the increase in peroxide, acid, p-anisidine, and carbonyl values, as well as total polar compounds and degradation of unsaturated fatty acids. A total of 16 volatile flavor compounds that significantly contributed to the overall flavor of FRO were identified. HCP was effective in reducing the generation of off-flavors (hexanoic acid, nonanoic acid, etc.) and increased the level of pleasant deep-fried flavors (such as (E,E)-2,4-decadienal). Therefore, the application of HCP has a positive effect on protecting the quality and prolonging the usability of FRO.

Optimization of Canolol Production from Canola Meal Using Microwave Digestion as a Pre-Treatment Method.

Canola meal, the by-product of canola oil refining, is a rich source of phenolic compounds and protein. The meal, however, is primarily utilized as animal feed but represents an invaluable source of nutraceuticals. Of particular interest are the sinapates, sinapine and sinapic acid, with the decarboxylation of the latter to form canolol. Extracting these phenolics has been carried out using a variety of different methods, although there is an urgent need for environmentally safe and sustainable methods. Microwave-assisted solvent extraction (MAE), as a green extraction method, is receiving considerable interest. Its ease of use makes MAE one of the best methods for studying multiple solvents. The formation of canolol, from sinapine and sinapic acid, is primarily dependent on temperature, which favors the decarboxylation reaction. The application of MAE, using the MultiwaveTM 500 microwave system with green extractants, was undertaken to assess its ability to enhance the yield of sinapates and canolol. This study examined the effects of different pre-treatment temperature-time combinations of 140, 150, 160, and 170 °C for 5, 10, 15, 20, and 30 min on the extraction of canolol and other canola endogenous phenolic compounds. Total phenolic content (TPC), total flavonoid content (TFC), as well as metal ion chelation (MIC) and DPPH radical activity of the different extracts were assessed. The results confirmed that extractability of canolol was optimized with methanol at 151 °C and with ethanol at 170 °C with pre-treatment times of 15.43 min and 19.31 min, respectively. Furthermore, there was a strong positive correlation between TPC and TFC (p < 0.05) and a negative correlation between TFC and DPPH radical activity. Interestingly, no significant correlation was observed between MIC and DPPH. These results confirmed the effectiveness of MAE, using the novel MultiwaveTM 500 microwave instrument, to enhance the yield of canolol. This was accompanied by substantial improvements in the antioxidant activity of the different extracts and further established the efficacy of the current MAE method for isolating important natural phenolic derivatives for utilization by the nutraceutical industry.

Challenges and advances in biotechnological approaches for the synthesis of canolol and other vinylphenols from biobased p-hydroxycinnamic acids: a review.

p-Hydroxycinnamic acids, such as sinapic, ferulic, p-coumaric and caffeic acids, are among the most abundant phenolic compounds found in plant biomass and agro-industrial by-products (e.g. cereal brans, sugar-beet and coffee pulps, oilseed meals). These p-hydroxycinnamic acids, and their resulting decarboxylation products named vinylphenols (canolol, 4-vinylguaiacol, 4-vinylphenol, 4-vinylcatechol), are bioactive molecules with many properties including antioxidant, anti-inflammatory and antimicrobial activities, and potential applications in food, cosmetic or pharmaceutical industries. They were also shown to be suitable precursors of new sustainable polymers and biobased substitutes for fine chemicals such as bisphenol A diglycidyl ethers. Non-oxidative microbial decarboxylation of p-hydroxycinnamic acids into vinylphenols involves cofactor-free and metal-independent phenolic acid decarboxylases (EC 4.1.1 carboxyl lyase family). Historically purified from bacteria (Bacillus, Lactobacillus, Pseudomonas, Enterobacter genera) and some yeasts (e.g. Brettanomyces or Candida), these enzymes were described for the decarboxylation of ferulic and p-coumaric acids into 4-vinylguaiacol and 4-vinylphenol, respectively. The catalytic mechanism comprised a first step involving p-hydroxycinnamic acid conversion into a semi-quinone that then decarboxylated spontaneously into the corresponding vinyl compound, in a second step. Bioconversion processes for synthesizing 4-vinylguaiacol and 4-vinylphenol by microbial decarboxylation of ferulic and p-coumaric acids historically attracted the most research using bacterial recombinant phenolic acid decarboxylases (especially Bacillus enzymes) and the processes developed to date included mono- or biphasic systems, and the use of free- or immobilized cells. More recently, filamentous fungi of the Neolentinus lepideus species were shown to natively produce a more versatile phenolic acid decarboxylase with high activity on sinapic acid in addition to the others p-hydroxycinnamic acids, opening the way to the production of canolol by biotechnological processes applied to rapeseed meal. Few studies have described the further microbial/enzymatic bioconversion of these vinylphenols into valuable compounds: (i) synthesis of flavours such as vanillin, 4-ethylguaiacol and 4-ethylphenol from 4-vinylguaiacol and 4-vinylphenol, (ii) laccase-mediated polymer synthesis from canolol, 4-vinylguaiacol and 4-vinylphenol.

Canolol and its derivatives: A novel bioactive with antioxidant and anticancer properties.

The health and safety concerns associated with synthetic antioxidants has resulted in an urgent search for natural sources of antioxidants. Such antioxidants are not only convenient but may also have important therapeutic properties. Oilseed crops, for example, are rich in phenolic compounds some of which exhibit powerful antioxidant properties that have broad applications in both the food and feed industry. Often, the concentration of these phenolic compounds is affected by many processing conditions including temperature, pressure, pH, and extracting solvents. Hence it is important to optimize processing conditions to obtain maximum levels of those antioxidants with superior antioxidant activity. Oilseeds, such as canola and mustard, are rich sources of sinapates and kaempferol derivatives. When subjected to different processing conditions, including pressurized temperature, sinapates are converted to vinyl phenol derivatives, of which the major one is canolol. This chapter will focus on the nature of canolol and its applications in food and medicine.

Air frying pretreatment and the recovery of lipophilic sinapates from the oil fraction of mustard samples.

Roasting of mustard seeds prior to oil extraction is a well-documented unit operation essential to produce canolol and other lipophilic sinapates. This study investigated the effectiveness of air frying as a seed roasting treatment operation for enhancing the recovery of lipophilic sinapates from various mustard samples and fractions/products. Air frying of seeds, powder, cake, bran, and flour from different mustard varieties was carried out at temperature-time combinations of 160, 170, and 180°C for 5, 10, 15, and 20 min, respectively. Oil was extracted using the Soxtec method. Lipophilic sinapates were extracted from the oil using equal volumes of hexane to methanol 70% (v/v) and quantified by high performance liquid chromatography-diode array detection (HPLC-DAD). The total phenolic content (TPC) and antioxidant activity of the oils were also evaluated. The results showed a time-temperature dependency for the recovery of major oil-soluble sinapates in all mustard samples and fractions. The optimum air frying condition 180°C for 15 min produced the maximum yield of canolol as well as other unidentified oil-soluble sinapates (retention time (RT)-7.7, RT-11.50, RT-14.95, and RT-16.24 min). The oil from lower grade yellow mustard seeds (LGYMS) roasted at 180°C for 20 mins specifically had the highest TPC (3402.22 ± 58.79 mg GAE/g oil), while LGYMS oils generally showed better antioxidant activities (2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion reducing antioxidant power (FRAP), and inhibition of linoleic acid oxidation) but were lower in metal ion chelating capacity. This information would be beneficial to the oil industry because air frying generated valuable canolol and other antioxidant lipophilic sinapates from mustard varieties and their fractions. PRACTICAL APPLICATION: A major limitation in the application of natural extracts in vegetable oils is the poor lipophilic nature of phenolic compounds. This study employed a new thermal treatment (air frying) in the recovery of canolol and other lipophilic antioxidants. Such treatments can enrich mustard-based ingredients with canolol and other lipophilic antioxidants for domestic and industrial applications.

Rapidoxy® 100: A Solvent-Free Pre-treatment for Production of Canolol.

RapidOxy® 100 is an automated instrument originally designed for measuring the oxidative stability of both solid and liquid samples. The compact and portable design of RapidOxy® 100, and its built-in pressurized heating chamber, provides a suitable environment for studying processing conditions. The feasibility of using oxygen or an inert atmosphere provides the ideal environment to study the effect of dry heat pre-treatment on canola antioxidants. The current study used RapidOxy® 100 to examine the impact of pressurized dry heat pre-treatment, under nitrogen, on the ultrasonic extraction of phenolic compounds. The effect of different pre-treatment temperature-time combinations of 120, 140, 160, and 180°C for 2, 5, 10, 15, and 20 min on the subsequent extraction of canola phenolic compounds was examined. The major sinapates identified by HPLC were sinapine, sinapic acid, and canolol. The optimum RapidOxy® condition for the maximum recovery of canolol was 160°C for 10 min. RapidOxy® 100 proved to be a novel and versatile instrument for enhancing the extraction of phenolic compounds.

Understanding of the Role of Pretreatment Methods on Rapeseed Oil from the Perspective of Phenolic Compounds.

The thermal pretreatment of oilseed prior to oil extraction could increase the oil yield and improve the oil quality. Phenolic compounds are important antioxidants in rapeseed oil. In this study, we investigated the impact of thermal pretreatment method on the rapeseed oil based on phenolic compound levels. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) analysis showed that the phenolic compound contents in the microwave-pretreated oil were higher than those in the oven- and infrared-treated oils. Sinapic acid (SA) and canolol (CA), which are the top two phenolic compounds in rapeseed oil, exerted well 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity with IC50 values of 8.45 and 8.80 µmol/L. The cell experiment uncovered that SA and CA have significant biological activities related to rapeseed oil quality, including increase of antioxidant enzymes superoxide dismutase (SOD), alleviation of reactive oxygen species (ROS), and cytotoxicity of HepG2 cells after the intake of excessive oleic acid. Further investigation indicated that SA and CA reduced cell apoptosis rate through Bax-Bcl-2-caspase-3 and p53-Bax-Bcl-2-caspase-3, respectively. Taken together, our findings suggest that microwave pretreatment is the best method to improve the content of phenolic compounds in rapeseed oil compared with oven and infrared pretreatments.

Cold-pressed rapeseed (Brassica napus) oil: Chemistry and functionality.

Rapeseed oil is the second most abundant produced edible oil in the world with low erucic acid and low glucosinolate. Thus, the quality of rapeseed oil had attracted global attention. Cold-pressed rapeseed oil appeared to be a preferred choice than refined oil as no solvent and less processing involved in the cold-pressing. The methods of cold-pressing and microwave pre-treatment on the extraction yield and bioactive compounds of rapeseed oil have been reviewed in this paper. Cold-pressed rapeseed oil offers health benefits due to its preserved fatty acid profile and bioactive compounds. High phenolic compounds, tocopherols, phytosterols, and carotenoids contents in the cold-pressed rapeseed oil offer health benefits like regulating blood lipid profile, insulin sensitivity, and glycemic control, as well as offer antioxidant and cytotoxic activity. Besides using as edible oil, cold-pressed rapeseed oil find applications in animal feed, chemical, and fuel.

Preparation of rapeseed oil with superhigh canolol content and superior quality characteristics by steam explosion pretreatment technology.

In this study, rapeseed was pretreated by steam explosion pretreatment technology and subsequently pressed to prepare rapeseed oil. GC, UPLC, and HPLC techniques were employed to analyze the quality characteristics of the rapeseed oil, including the canolol content and other quality characteristics. Additionally, the effect of steam explosion pretreatment technology on the canolol content of rapeseed oil was studied and the formation mechanism of canolol elucidated. The results revealed that when the steam explosion pressure reached 1.0 MPa, the canolol content of the tested oil increased from 41.21 to 2,168.69 mg/kg (52.63-fold increase) and that sinapic acid played a significant role in the conversion of canolol. Thus, the sinapine was converted into the intermediate (sinapic acid) by hydrolysis, which in turn was transformed into canolol through decarboxylation. The instantaneous high-energy environment generated by steam explosion pretreatment could intensify the hydrolysis and decarboxylation reactions of sinapine and sinapinic acid, thereby significantly increasing the canolol content of the oil. To prove the superiority of steam explosion pretreatment, we compared it with other pretreatment technologies, including traditional high-temperature roasting and popular microwave pretreatment. The results revealed that rapeseed oil prepared by steam explosion pretreatment displayed the best quality characteristics. This study can be a reference for the preparation process of rapeseed oil with superhigh canolol content and superior quality characteristics using steam explosion pretreatment.

Canolol Dimer, a Biologically Active Phenolic Compound of Edible Rapeseed Oil.

Rapeseed and its oil are the source of many biologically active substances. From crude rapeseed oil, canolol is isolated and from edible oil its dimer. Herein, we tested the biological activity of those two compounds isolated from the oils by determining their antioxidant capacity and in vitro cytotoxicity on human tumor cell lines. Canolol and its dimer showed antiproliferative activity on both cell lines with IC50 values of 46.45 µM in HeLa, and 51.19 µM in MCF7 cells, respectively. Evaluation of cell death was also done, while the oxygen radical absorbance capacity method confirmed that the canolol dimer has higher antioxidant potential than canolol. Stability of canolol and its dimer under different storage conditions showed that for a longer period of time both compounds should be stored in a freezer, but also that the dimer is more stable against degradation than canolol. Presented results indicate possible applications of canolol and its dimer in the food and pharmaceutical industry as a natural antioxidant and an anticancer agent, respectively.

Dietary canolol induces apoptosis in human cervical carcinoma HeLa cells through ROS-MAPK mediated mitochondrial signaling pathway: In vitro and in vivo.

Canolol (4-vinylsyringol), extracted form crude canola oil, is the promising drug toward cancer prevention and treatment. The current studies focus on the role of COX-2 signaling pathway in canolol-induced apoptosis in cancer cells. It is still unknown whether mitochondria and MAPK signaling pathways are involved. To elucidate the roles of above signaling pathways in canolol-induced apoptosis in cancer cells, human cervical carcinoma cell line HeLa and HeLa xenograft tumor model are adopted. Canolol induced apoptosis of HeLa cells and inhibited tumor growth with low systemic adverse effect, accompanying with excess generation of intracellular ROS and lysosome rupture. The results in vitro and in vivo confirmed that MAPK signaling pathways mediated mitochondrial signaling pathway activation were involved in canolol-induced apoptosis. In conclusion, these data showed that canolol induced apoptosis in HeLa cells through ROS-MAPK mediated mitochondrial signaling pathway, providing a view of the potential application of canolol as an anticancer agent.

Quality evaluation of rapeseed oil in Chinese traditional stir-frying.

Canolol is a potential antioxidation ingredient in rapeseed oil. Rapeseed oil with two levels of canolol (528.9 vs. 250.5 mg/kg) was used for stir-frying different foods (potatoes, tofu, and vegetables). Comprehensive evaluations indicated that the canolol content in high canolol rapeseed oil (HCR) and low canolol rapeseed oil (LCR) after stir-frying were in the range of 187.8-237.7 and 45.6-96.4 mg/kg, respectively. The degradation rate of total phenol was 58.4% and 80.3% in HCR and LCR, respectively. The loss rates of α- and γ-tocopherol were 24.5% and 47.6%, respectively. Phytosterol concentration decreased by 20% and trans-fatty acid was not detected in either rapeseed oil. In addition, the peroxide value, anisidine value, and malondialdehyde content in HCR were lower than those in LCR. The oxidative stability index in HCR was longer, showing lower extent of deterioration. Rapeseed oil with high canolol content displayed good oxidation resistance due to significant positive correlation with oxidation induction time (p < .01).

Dietary canolol protects the heart against the deleterious effects induced by the association of rapeseed oil, vitamin E and coenzyme Q10 in the context of a high-fat diet.

BACKGROUND: Obesity progressively leads to cardiac failure. Omega-3 polyunsaturated fatty acids (PUFA) have been shown to have cardio-protective effects in numerous pathological situations. It is not known whether rapeseed oil, which contains α-linolenic acid (ALA), has a similar protective effect. Omega-3 PUFAs are sensitive to attack by reactive oxygen species (ROS), and lipid peroxidation products could damage cardiac cells. We thus tested whether dietary refined rapeseed oil (RSO) associated with or without different antioxidants (vitamin E, coenzyme Q10 and canolol) is cardio-protective in a situation of abdominal obesity. METHODS: Sixty male Wistar rats were subdivided into 5 groups. Each group was fed a specific diet for 11 weeks: a low-fat diet (3% of lipids, C diet) with compositionally-balanced PUFAs; a high-fat diet rich in palm oil (30% of lipids, PS diet); the PS diet in which 40% of lipids were replaced by RSO (R diet); the R diet supplemented with coenzyme Q10 (CoQ10) and vitamin E (RTC diet); and the RTC diet supplemented with canolol (RTCC diet). At the end of the diet period, the rats were sacrificed and the heart was collected and immediately frozen. Fatty acid composition of cardiac phospholipids was then determined. Several features of cardiac function (fibrosis, inflammation, oxidative stress, apoptosis, metabolism, mitochondrial biogenesis) were also estimated. RESULTS: Abdominal obesity reduced cardiac oxidative stress and apoptosis rate by increasing the proportion of arachidonic acid (AA) in membrane phospholipids. Dietary RSO had the same effect, though it normalized the proportion of AA. Adding vitamin E and CoQ10 in the RSO-rich high fat diet had a deleterious effect, increasing fibrosis by increasing angiotensin-2 receptor-1b (Ag2R-1b) mRNA expression. Overexpression of these receptors triggers coronary vasoconstriction, which probably induced ischemia. Canolol supplementation counteracted this deleterious effect by reducing coronary vasoconstriction. CONCLUSION: Canolol was found to counteract the fibrotic effects of vitamin E + CoQ10 on cardiac fibrosis in the context of a high-fat diet enriched with RSO. This effect occurred through a restoration of cardiac Ag2R-1b mRNA expression and decreased ischemia.

Cold-pressed and hot-pressed rapeseed oil: The effects of roasting and seed moisture on the antioxi- dant activity, canolol, and tocopherol level.

BACKGROUND: The paper looks at the levels of canolol, tocopherols and antioxidant activity in cold-pressed and hot-pressed rapeseed oils produced from seeds of various moisture levels (5%, 7.5%, and 10%). The paper also considers the effects of seed roasting on the levels of these compounds. METHODS: The material used for the tests was rapeseed cv. Adrianna. The quality of the oils obtained is determined using peroxide and acid values. The levels of canolol and tocopherols are analyzed using HPLC. The DPPH radical-scavenging activity method for oil samples and phenolic extract from oils was used. RESULTS: It has been demonstrated that the oils produced from rapeseeds with a 5% moisture content, and   in particular from cold-pressed oils, were characterized by the lowest peroxide values. Cold-pressed oils produced from rapeseeds with a 5% moisture content were characterized by higher levels of tocopherols and plastochromanol-8. In the case of hot-pressed oils, the highest levels of tocopherols were found in oils pro- duced from seeds with a 7.5% moisture content, and the greatest amount of PC-8 (more than 4 mg/100 g) was found in oils produced from seeds with a 10% moisture content. Hot-pressed oils have been shown to have higher levels of these compounds than cold-pressed oils. Both roasting and hot pressing led to an increase in the amount of canolol in the oils investigated. When analysing the antioxidant activity of the oils and phenolic extracts it was shown that phenolic compounds are responsible for approx. 10% of total antioxidant activity. CONCLUSIONS: Various levels of biologically active compounds were shown to be present in the rapeseed oil obtained from raw materials of a varying moisture content. The type of pressing process (cold-pressing or hot-pressing) and whether the seeds have undergone roasting has also been shown to affect the resulting oil and the level of native antioxidants it contains.

The effect of microwave pretreatment of seeds on the stability and degradation kinetics of phenolic compounds in rapeseed oil during long-term storage.

Storage stability and degradation kinetics of phenolic compounds in rapeseed oil pressed from microwave treated seeds (0, 2, 4, 6, 8, 10min, 800W) during long-term storage (12months) at a temperature of 20°C was discussed in the current study. The dominant phenolic compound detected in rapeseed oil was canolol, followed by minor amounts of free phenolic acids and sinapine. The most pronounced effect of seeds microwaving was noted for canolol formation - after 10-min exposure the quantity of this compound was approximately 63-fold higher than in control oil. The degradation of phenolics during storage displayed pseudo first-order kinetics. Differences in the initial degradation rate (r0) demonstrated significant impact of the period of seeds microwave exposure on the degradation rates of phenolic compounds. Results of the half-life calculation (t1/2) showed that the storage stability of phenolic compounds was higher in oils produced from microwave treated rapeseeds than in control oil.

A Two-Step Bioconversion Process for Canolol Production from Rapeseed Meal Combining an Aspergillus niger Feruloyl Esterase and the Fungus Neolentinus lepideus.

Rapeseed meal is a cheap and abundant raw material, particularly rich in phenolic compounds of biotechnological interest. In this study, we developed a two-step bioconversion process of naturally occurring sinapic acid (4-hydroxy-3,5-dimethoxycinnamic acid) from rapeseed meal into canolol by combining the complementary potentialities of two filamentous fungi, the micromycete Aspergillus niger and the basidiomycete Neolentinus lepideus. Canolol could display numerous industrial applications because of its high antioxidant, antimutagenic and anticarcinogenic properties. In the first step of the process, the use of the enzyme feruloyl esterase type-A (named AnFaeA) produced with the recombinant strain A. niger BRFM451 made it possible to release free sinapic acid from the raw meal by hydrolysing the conjugated forms of sinapic acid in the meal (mainly sinapine and glucopyranosyl sinapate). An amount of 39 nkat AnFaeA per gram of raw meal, at 55 °C and pH 5, led to the recovery of 6.6 to 7.4 mg of free sinapic acid per gram raw meal, which corresponded to a global hydrolysis yield of 68 to 76% and a 100% hydrolysis of sinapine. Then, the XAD2 adsorbent (a styrene and divinylbenzene copolymer resin), used at pH 4, enabled the efficient recovery of the released sinapic acid, and its concentration after elution with ethanol. In the second step, 3-day-old submerged cultures of the strain N. lepideus BRFM15 were supplied with the recovered sinapic acid as the substrate of bioconversion into canolol by a non-oxidative decarboxylation pathway. Canolol production reached 1.3 g/L with a molar yield of bioconversion of 80% and a productivity of 100 mg/L day. The same XAD2 resin, when used at pH 7, allowed the recovery and purification of canolol from the culture broth of N. lepideus. The two-step process used mild conditions compatible with green chemistry.