Validation of an Isocratic HPLC Method for Simultaneous Estimation of Major Phytosterols in Prunus spinosa L. extracts.

This study aimed to develop a rapid method for separation of stigmasterol, campesterol and ß-sitosterol in Prunus spinosa L. (sloe) fruit extracts by High Performance Liquid Chromatography system. Samples were prepared by Soxhlet extraction method and separated on a high strength silica C18 column using acetonitrile-methanol mobile phase and Photodiode Array Detector. The optimized method resulted in a linear calibration curve ranging from 1.7 ng mL-1 to 130 ng mL-1 for all three phytosterols. Analyses of internal and external phytosterol standards showed good linearity (R2 of 0.998 to 0.999); LOD and LOQ were determined to be 2.33×10-7-2.18×10-4 and 7.07×10-7-6.60×10-4 mg mL-1, respectively. Repeatability and reproducibility precision analyses showed acceptable values of RSD %. ß-sitosterol was the predominant phytosterol (51.53-81.03 % of total) among all samples. Method validation parameters indicated that this analytical method can be applied for accurate and precise determination of campesterol, stigmasterol and ß-sitosterol, in selected extracts.

Characterization of Silybum marianum and Silybum eburneum seed oils: Phytochemical profiles and antioxidant properties supporting important nutritional interests.

Milk thistle seed oil is still not a well-known edible oil. Silybum marianum (milk thistle), is present in several countries and is the only known representative of the genus Silybum. However, Silybum eburneum, which is an endemic plant in Spain, Kenya, Morocco, Algeria, and Tunisia, is considered a marginalized species. The present work is the first report that gives information on the lipid and phenolic profiles of Tunisian S. eburneum seed oil compared to those of Tunisian S. marianum seed oil. In addition, the antioxidant properties of these oils were determined with DPPH, FRAP, and KRL assays, and their ability to prevent oxidative stress was determined on human monocytic THP-1 cells. These oils are characterized by high amounts of unsaturated fatty acids; linoleic acid and oleic acid are the most abundant. Campesterol, sitosterol, stigmasterol, and ß-amyrin were the major phytosterols identified. α-tocopherol was the predominant tocopherol found. These oils also contain significant amounts of phenolic compounds. The diversity and richness of Silybum marianum and Silybum eburneum seed oils in unsaturated fatty acids, phenolic compounds, and tocopherols are associated with high antioxidant activities revealed by the DPPH, FRAP, and KRL assays. In addition, on THP-1 cells, these oils powerfully reduced the oxidative stress induced by 7-ketocholesterol and 7ß-hydroxycholesterol, two strongly pro-oxidant oxysterols often present at increased levels in patients with age-related diseases. Silybum marianum and Silybum eburneum seed oils are therefore important sources of bioactive molecules with nutritional interest that prevent age-related diseases, the frequency of which is increasing in all countries due to the length of life expectancy.

MOF-derived nanozyme CuOx@C and its application for cascade colorimetric detection of phytosterols.

Phytosterols (PSs), a class of naturally occurring bioactive lipid compounds, have been found to possess a significant cholesterol-lowering effect. In developing countries, the consumption of rapeseed oil is the primary pathway of PS intake for the general population. However, developing low-cost, real-time, and high-throughput screening techniques for PSs remains a challenge. Here, a Cu-based nanocomposite CuOx@C was synthesized via a simple method of the calcination of HKUST-1 and systematically characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The CuOx@C demonstrated excellent peroxidase-like (POD-like) activity, functioning as a peroxidase mimic to facilitate the catalysis of 3,3',5,5'-tetramethylbenzidine (TMB) into its oxidized form (oxTMB), thereby initiating a discernible color response. On the basis of this discovery, a CuOx@C-based colorimetric method for detecting total sterols in rapeseed was successfully constructed via cascade reactions. After optimizing the conditions, the high-throughput screening of total sterols in rapeseed could be completed in only 21 min, which significantly facilitated the sensing of PSs. A linear range of 0.6-6 mg/g was achieved for the detection of total sterols in rapeseed samples, thereby satisfying the requirements for detection. In addition, due to the high stability of CuOx@C and the specificity of cholesterol oxidase, the developed method had excellent stability and selectivity toward PSs, indicating that this work has huge prospects for commercial application. This innovative work overcomes the limitation of the instrumental method and provides a portable and reliable tool for total sterols detection. It can also facilitate the development of oilseeds with a high content of PSs.

FooDOxS: a database of oxidized sterols content in foods.

Dietary oxidized sterols (DOxS) are cholesterol-like molecules known to exert pro-inflammatory, pro-oxidant, and pro-apoptotic effects, among others. We present the FooDOxS database, a comprehensive compilation of DOxS content in over 1680 food items from 120 publications across 25 countries, augmented by data generated by our group. This database reports DOxS content in foods classified under the NOVA and What We Eat in America (WWEIA) systems, allowing a comprehensive and statistically robust summary of DOxS content in foods. Notably, we evaluated the efficacy of using NOVA and WWEIA classifications in capturing DOxS variations across food categories. Our findings provide insights into the strengths and limitations of these classification systems, enhancing their utility for assessing dietary components. This research contributes to the understanding of DOxS in food processing and suggests refinements for classification systems, holding promise for improved food safety and public health assessments.

Evaluation of sterols as markers of fungal spoilage in red pepper powder.

Red pepper powder (RPP) made from ground dried red pepper (Capsicum annuum L.) is prone to adulteration with fungal-spoiled RPP to gain unfair profits in Korea. This study aimed to investigate the effects of fungal infection on the ergosterol and phytosterol content of RPP and evaluate the potential of the sterol content as a marker for identifying fungal-spoiled RPP. Ergosterol was detected only in fungal-spoiled RPP and not in unspoiled RPP [

Comprehensive gas chromatography with mass spectrometry analysis of sterols in red goji berries (Lycium sp.).

Gas chromatography with mass spectrometry (GC/MS) and fractionation steps were used to determine the sterol patterns of red goji berries in detail. Twenty-five sterols were detected in fresh berries of two species (Lycium barbarum and L. chinense) from bushes grown in the botanical garden of the University of Hohenheim, and 20 sterols were identified. The rarely occurring campesta-5,24(25)-dienol, ß-sitosterol, Δ5-avenasterol, campesterol, and cycloartenol represented >60 % of the total sterol content. Maturity and drying of fresh red goji berries caused small changes but did not affect the characteristic sterol pattern. This was confirmed by analyzing various commercial dried red goji berry samples from different sources. Separated flesh and seed samples revealed pronounced differences in the sterol pattern. A new method of merging GC/MS chromatograms showed that ∼75 % of the sterols were present in seeds and ∼25 % in flesh. The unique sterol profile may be exploited to authenticate red goji berries.

Optimizing niger seed (Guizotia abyssinica) oil quality: A comprehensive analysis of infrared-heat induced changes in bioactive profile, physiochemical attributes, and oxidative stability.

Infrared heating (IRH) at 140, 160, and 180°C for varying durations (5, 10, and 15 min) was employed for improving the niger (Guizotia abyssinica) seed oil (NSO) quality for diverse food applications. The study explored changes in phenolic profile, oxidative stability index (OSI), tocopherols, phytosterols, fatty acid profiles, and physicochemical attributes of NSO. Upon IRH at 180°C for 10 min, the oil yield, total phenolic, and flavonoid contents increased from 33.09% to 40.56%, 6.67 to 173.62 mg GAE/kg, and 24.76 to 120.64 mg QE/kg, respectively. The viscosity, chlorophylls, carotenoids, radical scavenging activity, OSI, caffeic, protocatechuic, vanillic, and syringic acids were highest upon IRH at 180°C for 15 min. The tocopherols and phytosterols initially augmented while decremented upon raising IRH conditions. The infrared spectra indicated no adverse impact of IRH on NSO quality. The appropriate IRH conditions can be considered for improving NSO quality and making it valuable for various edible products.

Supercritical CO2 Extraction of Fatty Acids, Phytosterols, and Volatiles from Myrtle (Myrtus communis L.) Fruit.

BACKGROUND: Myrtle (Myrtus communis L.) is a coastal Mediterranean aromatic medicinal plant rich in essential oil components, flavonoids, and phenolic acids. Studies highlight the potential health benefits of myrtle bioactive compounds with antioxidant and antiproliferative properties. Since limited research exists on myrtle fruit's lipid fraction, the aim of this study was to apply supercritical CO2 extraction to obtain bioactive compounds from myrtle berries focusing on the fatty acids, sterols, and essential oils. METHODS: The optimization of the supercritical CO2 extraction of myrtle fruit using CO2 as solvent was carried out using the response surface methodology with Box-Behnken experimental design. The following conditions were tested: temperature (40, 50, and 60 °C), pressure (200, 300, and 400 bar), and flow rate (20, 30, and 40 g min-1) on the yield of lipid extract as well as on the yield of fatty acids, phytosterols, and volatiles present in the extract and constituting its bioactive potential. RESULTS: In the extracts examined, 36 fatty acids, 7 phytosterols, and 13 volatiles were identified. The average yield of the extract was 5.20%, the most abundant identified fatty acid was essential cis-linolenic acid (76.83%), almost 90% of the total phytosterols were ß-sitosterol (12,465 mg kg-1), while myrtenyl acetate (4297 mg kg-1) was the most represented volatile compound. The optimal process conditions obtained allow the formulation of extracts with specific compositions.

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.

Resolving phytosterols in microalgae using offline two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography coupled with quadrupole time-of-flight mass spectrometry.

Sterols are unsaponifiable lipids resulting from plant metabolism that exhibit interesting bioactive properties. Microalgae are a major source of specific phytosterols, most of which are still not fully characterized. The similarity in sterol structures and the existence of positional isomers make the separation of phytosterols challenging. A method was developed based on an offline two-dimensional (2D) system, reversed-phase liquid chromatography (RPLC)-supercritical fluid chromatography (SFC)/quadrupole time-of-flight (Q-ToF) mass spectrometry, for the identification of sterols in microalgae. Subsequent positive-mode MS/MS was used to confirm the identified phytosterols. The 2D chromatogram exhibited a pattern related to the positions of the double bonds, which were confirmed by standard injection, enabling structural elucidation. The analysis of the unsaponifiable fraction of two algae, namely Scenedesmus obliquus, a freshwater microalgae, and Padina pavonica, a marine macroalgae, highlighted the ability of the method to distinguish a large number of sterol isomers.

Deciphering the effect of phytosterols on Alzheimer's disease and Parkinson's disease: the mediating role of lipid profiles.

BACKGROUND: Studies have suggested that blood circulating phytosterols, plant-derived sterols analogous to cholesterol, were associated with blood lipid levels and the risk of Alzheimer's disease (AD) and Parkinson's disease (PD). This Mendelian randomization (MR) study is performed to determine the causal effect of circulating phytosterols on AD and PD and evaluate the mediation effect of blood lipids. METHODS: Leveraging genome-wide association studies summary-level data for phytosterols, blood lipids, AD, and PD, univariable and multivariable MR (MVMR) analyses were conducted. Four types of phytosterols (brassicasterol, campesterol, sitosterol, and stigmasterol), three blood lipids parameters (high-density lipoprotein cholesterol [HDL-C], non-HDL-C, and triglyceride), two datasets for AD and PD were used. Inverse-variance weighted method was applied as the primary analysis, and false discovery rate method was used for adjustment of multiple comparisons. RESULTS: Using the largest AD dataset, genetically proxied higher levels of stigmasterol (OR = 0.593, 95%CI = 0.431-0.817, P = 0.004) and sitosterol (OR = 0.864, 95%CI = 0.791-0.943, P = 0.004) significantly correlated with a lower risk of AD. No significant associations were observed between all four types of phytosterols levels and PD. MVMR estimates showed that the above causal associations were missing after integrating the blood lipids as exposures. Sensitivity analyses confirmed the robustness of these associations, with no evidence of pleiotropy and heterogeneity. CONCLUSION: The study supports a potential beneficial role of blood stigmasterol and sitosterol in reducing the risk of AD, but not PD, which is dependent on modulating blood lipids. These insights highlight circulating stigmasterol and sitosterol as possible biomarkers and therapeutic targets for AD.

Negative Association of Lignan and Phytosterol Intake with Stress Perception during the COVID-19 Pandemic-A Polish Study on Young Adults.

BACKGROUND: There has been an increasing global prevalence of depression and other psychiatric diseases in recent years. Perceived stress has been proven to be associated with psychiatric and somatic symptoms. Some animal and human studies have suggested that consuming foods abundant in lignans and phytosterols may be associated with lower levels of stress, depression, and anxiety. Still, the evidence is not yet strong enough to draw firm conclusions. Thus, we investigated the association between dietary intake of these phytochemicals and the level of stress experienced by adult individuals. METHODS: Diet was assessed using self-reported 7-day dietary records. The intakes of lignans and phytosterols were estimated using databases with their content in various food products. The Perceived Stress Scale (PSS) was implemented to measure the level of perceived stress. A logistic regression analysis was used to test for associations. RESULTS: The odds of elevated PSS were negatively associated with dietary intake of total phytosterols, stigmasterol, and ß-sitosterol, with evidence of a decreasing trend across tertiles of phytochemicals. The analysis for doubling the intake reinforced the aforementioned relationships and found protective effects against PSS for total lignans, pinoresinol, and campesterol. CONCLUSIONS: Habitual inclusion of lignans and phytosterols in the diet may play a role in psychological health. To address the global outbreak of depression and other mental health issues triggered by stress, it is important to take a holistic approach. There is a need to develop effective strategies for prevention and treatment, among which certain dietary interventions such as consumption of products abundant in lignans and phytosterols may play a substantial role.

Phytosterol and phytostanol-mediated epigenetic changes in cancer and other non-communicable diseases: a systematic review.

Phytosterols/phytostanols are bioactive compounds found in vegetable oils, nuts and seeds and added to a range of commercial food products. Consumption of phytosterols/phytostanols reduces levels of circulating LDL-cholesterol, a causative biomarker of CVD, and is linked to a reduced risk of some cancers. Individuals who consume phytosterols/phytostanols in their diet may do so for many years as part of a non-pharmacological route to lower cholesterol or as part of a healthy diet. However, the impact of long term or high intakes of dietary phytosterols/phytostanols has not been on whole-body epigenetic changes before. The aim of this systematic review was to identify all publications that have evaluated changes to epigenetic mechanisms (post-translation modification of histones, DNA methylation and miRNA expression) in response to phytosterols/phytostanols. A systematic search was performed that returned 226 records, of which eleven were eligible for full-text analysis. Multiple phytosterols were found to inhibit expression of histone deacetylase (HDAC) enzymes and were also predicted to directly bind and impair HDAC activity. Phytosterols were found to inhibit the expression and activity of DNA methyl transferase enzyme 1 and reverse cancer-associated gene silencing. Finally, phytosterols have been shown to regulate over 200 miRNA, although only five of these were reported in multiple publications. Five tissue types (breast, prostate, macrophage, aortic epithelia and lung) were represented across the studies, and although phytosterols/phytostanols alter the molecular mechanisms of epigenetic inheritance in these mammalian cells, studies exploring meiotic or transgenerational inheritance were not found.

Phytosterol intake and risk of coronary artery disease: Results from 3 prospective cohort studies.

BACKGROUND: Phytosterols are structurally similar to cholesterol and partially inhibit intestinal absorption of cholesterol, although their impact on coronary artery disease (CAD) risk remains to be elucidated. OBJECTIVES: This study aimed to prospectively assess the associations between total and individual phytosterol intake and CAD risk in United States health professionals. METHODS: The analysis included 213,992 participants from 3 prospective cohorts-the Nurses' Health Study (NHS), NHSII, and Health Professionals Follow-Up Study-without cardiovascular disease or cancer at baseline. Diet was assessed using a validated food frequency questionnaire every 2-4 y since baseline. Associations between phytosterol intake and the risk of CAD, such as nonfatal myocardial infarction and fatal CAD, were evaluated using Cox proportional hazards regression models. RESULTS: More than 5,517,993 person-years, 8725 cases with CAD were documented. Comparing extreme quintiles, pooled hazard ratios (95% CIs) of CAD were 0.93 (0.86, 1.01; P-trend = 0.16) for total phytosterols, 0.89 (0.82, 0.96; P-trend = 0.05) for campesterol, 0.95 (0.88, 1.02; P-trend = 0.10) for stigmasterol, and 0.92 (0.85, 1.00; P-trend = 0.09) for ß-sitosterol. Nonlinear associations were observed for total phytosterols, campesterol, and ß-sitosterol: the risk reduction plateaued at intakes above ∼180, 30, and 130 mg/d, respectively (P-nonlinearity < 0.001). In a subset of participants (N range between 11,983 and 22,039), phytosterol intake was inversely associated with plasma concentrations of total cholesterol, triglycerides, high-density lipoprotein cholesterol, and IL-6 and positively associated with adiponectin, whereas no significant associations were observed for low-density lipoprotein cholesterol or C-reactive protein concentrations. CONCLUSIONS: Higher long-term intake of total and major subtypes of phytosterols may be associated with a modest reduction in CAD risk, displaying a nonlinear relationship that plateau at moderate intake levels. The role of phytosterols in preventing CAD warrants further investigation.

Chemical conversions of free phytosterols during the bleaching of corn oil.

Phytosterols have health benefits; however, they are partially removed during the bleaching of corn oil. We evaluated the chemical conversion of free phytosterols (FPs) during bleaching. FP degradation accelerated with increased time and temperature, following a first-order kinetic model. In the n-heptane system, air and activated clay promoted the chemical conversion of the FPs. Sterenes formation was analysed under different conditions using a zero-order kinetic model. The apparent activation energies revealed sterene formation decreasing in the following order: campesta-3,5-diene ≈ stigmasta-3,5,22-triene > stigmasta-3,5-diene. Isomers of the above were not detected, indicating that these sterenes were the only primary products of FPs. The desorption test indicated that the FP loss from corn oil was not only due to FPs being adsorbed the activated clay, but also FPs adsorbed at acidic activated sites being degraded. This study presents a vital scientific foundation for retaining FPs to develop healthier and more nutritious oils.

Age-related reference intervals for serum phytosterols in children by gas chromatography-mass spectrometry and its application in diagnosing sitosterolemia.

BACKGROUND AND AIMS: Serum phytosterol profiles are essential for the diagnosis and management of sitosterolemia. However, pediatric reference interval (RI) studies are scarce and various mass spectrometry (MS) approaches for phytosterol analysis still face multiple limitations. Therefore, an optimized gas chromatography (GC)-MS assay and age-related RIs in children are both required. MATERIALS AND METHODS: Cholesterol and phytosterols (sitosterol, campesterol, cholestanol, stigmasterol, and sitostanol) were simultaneously determined by optimized GC-MS and performance was verified by the lower limit of quantification (LLOQ), linearity, precision, recovery, matrix effects, and method comparison. Healthy children (247 males and 263 females) were recruited, sex and age dependence were assessed using quantile regression (2.5th percentile and 97.5th percentile), and RIs were established according to Clinical and Laboratory Standards Association guideline C28-A3. These RIs were validated in 19 patients with sitosterolemia and 23 patients with hypercholesterolemia. RESULTS: The optimized method shortened the sample processing time by approximately 60 min. Among the five phytosterols, all precision, recoveries (ranging from 89.97% to 104.94%), and relative matrix effects (%CV: ranging from 0.08% to 13.88%) met the specifications. GC-MS showed good agreement with lower cholesterol concentrations compared to conventional enzymatic methods. No significant differences between males and females were observed for all phytosterols, but age dependency was found and age-related RIs were established accordingly. Five phytosterols were significantly higher than RIs in patients with sitosterolemia. CONCLUSION: We established age-related RIs for five phytosterols in children based on an optimized GC-MS assay, providing a screening tool for the diagnosis of sitosterolemia in children.

Effects of different deodorization methods on the oxidation of sterol components in rice bran oil.

Rice bran oil contains a significant quantity of phytosterols that have various active functions and are natural active substances beneficial to humans. It is well known that deodorization during refining affects the quality of rice bran oil. However, changes in phytosterols fraction caused by stripping with nitrogen compared to water vapor remain unexplored. We measured phytosterols in rice bran oil after deodorization with nitrogen and water vapor. The variations in sitosterol fraction, which accounts for the highest percentage of phytosterols in rice bran oil, were analyzed by Gas chromatography (GC), Fourier transform infrared spectrometer (FTIR), and Nuclear magnetic resonance (NMR). Results showed that using nitrogen as the stripping gas was more suitable for deodorization. It promoted the formation of phytosterol esters, reduced the production of phytosterol oxidation products and improved the oil quality. This study provides a theoretical basis for improving the industrial production quality of rice bran oil.

Evaluation of urban pollution in a tropical lacustrine ecosystem by using n-alkanes and sterols as biomarkers.

The Jacarepaguá Lagoon System (JLS) receives industrial and domestic waste in an urban area with high population density and intense economic activity. The hydrography of the lagoons favours the sedimentation of particulate material transferred from the drainage basin. Water engineering, such as channel dredging and subsea outfall, did not satisfactorily mitigate pollution effects. Therefore, the environment is highly eutrophic, presents frequent blooms of algae and generates high emissions of greenhouse gases. There is no record in the literature on the analysis of organic compounds in the water compartment. The present work applies sterols as biomarkers to quantify the degree of pollution caused by biogenic compounds in riverine and lacustrine water of the JLS. n-Alkanes were applied to estimate the fractions of petrogenic contaminants. The sums of n-alkanes and sterols analysed had average concentrations of 21 ± 20 µg L-1 and 10 ± 8 µg L-1, respectively, in the river samples and 235 ± 156 µg L-1 and 30 ± 28 µg L-1, respectively, in the lagoon samples. The work also showed that the organic compounds inside the lagoons are evenly distributed, and approximately 7% of them are transferred to the marine ecosystem. Biogenic biomarkers and the absolute concentrations of sterols showed that sewage contaminants transferred by the rivers are partially decomposed in the lagoons. The correlations between indices and physicochemical parameters indicated that the degradation of organic compounds in the lagoons occurs mainly in the sediment compartment under anoxic conditions. The indices for sewage indicate that the ecosystem has exceeded its carrying capacity. The indices based on n-alkanes reported strong contamination at all sampling stations and inferred that 75-100% of these compounds were derived from petrogenic sources. These indices did not show any difference between rivers and the lagoon, which demonstrates the resilience of these compounds in the ecosystem.

Effects of Extraction Methods on the Bioactivities and Nutritional Value of Virginia and Valencia-Type Peanut Oil.

This study aimed to evaluate the effects of peanut varieties cultivated in Morocco (Virginia and Valencia) and extraction methods (cold press, CP; Soxhlet, Sox and maceration, and Mac) on the fatty acid profile, phytosterol, and tocopherol contents, quality characteristics, and antioxidant potential of peanut seed oil. The DPPH method was used to determine the antioxidant activity of the oils. The results revealed that fatty acid content was slightly affected by the extraction technique. However, the CP method was shown to be an excellent approach for extracting oil with desirable quality features compared to the Sox and Mac methods. Furthermore, the peanut oil extracted via CP carried a higher amount of bioactive compounds and exhibited remarkable antioxidant activities. The findings also revealed higher oleic acid levels from the Virginia oil, ranging from 56.46% to 56.99%. Besides, a higher total phytosterol and tocopherol content and DPPH scavenging capacity were obtained from the Valencia oil. Analyzing the study, it can be inferred that extraction method and variety both affect the composition of the peanut oil's bioactive compounds and antioxidant activity. This information is relevant for extracting peanut oil with a greater level of compounds of industrial interest.

Analysis of Changes in the Amount of Phytosterols after the Bleaching Process of Hemp Oils.

Unrefined vegetable oils from niche oilseeds are now sought after by consumers because of their unique nutritional properties and taste qualities. The color and flavor intensity of niche oils is a big problem, and their refining is not industrially feasible due to the small production scale. The study undertaken aimed analyze the effect of changing the amount of phytosterols (PSs) after the bleaching process of hemp oils of the 'Finola', 'Earlina 8FC' and 'Secuieni Jubileu' varieties. Cold-pressed (C) and hot-pressed (H) crude vegetable oils were bleached with selected bleaching earth (BE) at two concentrations. The post-process BE was extracted with methanol. The amount of PSs in the crude oils and the extract after washing the BE with methanol was analyzed by GC (gas chromatography). The study shows that the bleaching process did not significantly affect the depletion of PSs in the oils. Trace amounts of PSs remain on the BE and, due to methanol extraction, can be extracted from the oil. The conclusion of the performed research is that the bleaching of hemp oil does not cause depletion of the oil, and it significantly improves organoleptic properties. The oil bleaching process results in an oil loss of less than 2% by weight of the bleached oil, while the loss depends on the type of BE used. The study shows that the loss of phytosterols after the bleaching process averages 2.69 ± 0.69%, and depends on the type of BE used and the oil extracted from different varieties of hemp seeds.