天然植物甾醇的分离纯化

天然植物甾醇具有多种重要生理功能，目前广泛应用于医药、食品及化妆品等行业。随着这些行业对天然植物甾醇需求量的增加，如何制备高纯度的天然植物甾醇逐渐受到研究者的关注。由此，对国内外天然植物甾醇的分离与提纯方面研究进展做了概述。     

Effect of dry‐heat treatments on the nutritional value of maize germ

Maize germ is a by‐product of the maize milling process that is characterised by a high nutritional value. Currently, heat treatments are employed to prevent full‐fat maize germ from spoilage. The aim of this research was to study the effect of five dry‐heat treatments on the nutritional value of full‐fat maize germ. The results confirmed that after each dry‐heat treatment, the lipase activity decreases but the use of high temperatures could be detrimental for phytosterol and thiamine concentrations. The main negative effects have been observed after treatments at 140 °C for 30 min and 160 °C for 10 min. No significant difference has been observed for protein, ash or fatty acid contents. The treatment at 140 °C for 20 min resulted an optimal combination between temperature and heating time to inactivate lipase without altering deeply the nutritional value and the colour of maize germ.     

Fatty acid and phytosterol accumulation during seed development in three oilseed species

This study was carried out to compare the kinetic accumulation of bioactive lipids during seed development in three oil crops rapeseed, sunflower and woad. Field experiments were conducted under organic conditions during 3 years. After flowering, seeds were collected each 4–5 days until harvest. The three species differed by the quantity and quality of both fatty acids and sterols. Higher levels of phytosterols and fatty acids were reached between 20 and 40 days after flowering (DAF) in sunflower, 40 and 60 DAF in rapeseed and 25 and 45 DAF for woad. The modification of lipid composition during the grain filling depends on species. Knowledge of the composition and accumulation of fatty acids and phytosterols in sunflower, rapeseed and woad seeds would assist in efforts to achieve industrial applications. These seeds may give an interesting source of bioactive lipids.     

Variability of Phytosterols in Jatropha curcas Germplasm

Jatropha curcas L. has great potential for biofuel and phytosterol production. The objective of this research was to evaluate G × E variability for kernel phytosterol content and composition in Jatropha germplasm. Freshly matured seeds from 21 accessions grown in Málaga, Spain were collected at two stages of development. Significant genetic variation was detected for total kernel phytosterol content, which ranged from 2,246 to 2,883 mg kg^?1; and stigmasterol concentration, which ranged from 7.6 to 11.5 % of the total phytosterols. An accession with 9.2 % Δ⁵‐avenasterol was also identified. The coefficient of variation for kernel phytosterol content and stigmasterol concentration was 6.2 and 14.0 % respectively between accessions and 7.2 and 10.2 % respectively within accessions. Accordingly, evaluation of plant to plant variation is advisable. The existence of variability for kernel phytosterol content and composition in Jatropha will enable breeding for enhanced levels of these compounds.     

Analysis of sterols from various food matrices

Two methods suitable for routine phytostanol/phytosterol analysis of various sterol‐enriched food matrices and phytostanyl/phytosteryl fatty acid ester ingredients are introduced. A method based on hot saponification of a sample with ethanolic potassium hydroxide in the presence of an internal standard (5β‐cholestan‐3α‐ol) is adequate for most matrices, such as spread, milk and yoghurt. Some matrices, like pasta, require acid hydrolysis in order to release matrix‐incorporated bound sterols or sterols from steryl glycosides before the saponification step. After saponification, the unsaponifiable material containing phytostanols and phytosterols is extracted into an organic solvent (e.g. heptane), followed by evaporation of the solvent to dryness. Sterols are separated as their trimethylsilyl ether derivatives with a gas‐liquid chromatograph (GC), on a column coated with 5% phenyl/95% dimethylpolysiloxane, and detected with a flame ionization detector. The GC conditions applied provide efficient separation of the most abundant phytostanols/phytosterols in 15 min, a wide linear range of stanols/sterols without the need of defining sterol response factors. The methods are repeatable and accurate, as shown with standard addition trials. These methods were applied to determine phytostanol/phytosterol contents of several sterol‐enriched functional food products, and the analyzed amounts were in good accordance with the information provided on the packaging labels.     

Alternate oils in fish feeds

Nearly half of the fish consumed as food worldwide are raised on fish farms rather than caught in the wild, as shown by FAO statistics. The increasing aquaculture of predatory carnivorous fish demands new sources of feed constituents, particularly oils at the moment. Common terrestrial plant oils contain only traces of the long‐chain polyunsaturated fatty acids. In connection with fish feed, especially the lack of n‐3 “marine fatty acids” is obvious. Recommendations on the required amounts of the fatty acids DHA (22:6n‐3) and EPA (20:5n‐3) exist from 1994. When plant oil‐based diets are fed during the growing phase and replaced by a fish oil‐based diet during a period prior to slaughter, most of the beneficial lipid composition of fish in terms of human dietary recommendations is restored. Little attention has been focused on the fish welfare in connection to substitution of dietary oil sources, and studies are still scarce. New fish diets will rely heavily on the use of alternate ingredients such as plant oils also for carnivorous cold‐temperate water fish species. In the future, an addition of synthetic or GMO‐produced “marine” fatty acids is a possible scenario. The aim of this review is to highlight some plant oils used in fish feeds, with special emphasis on compounds other than fatty acids. We also include some results from an ongoing study, where the effect of dietary soy oil on gonad maturation in Atlantic cod (Gadus morhua) is indicated.     

Phytochemical Characterization of Veronica officinalis L., V. teucrium L. and V. orchidea Crantz from Romania and Their Antioxidant and Antimicrobial Properties

Aerial parts of Veronica species are used in Romanian traditional medicine for the treatment of various conditions like kidney diseases, cough, and catarrh, and are known for their wound-healing properties. In the present study, the phenolic and sterolic content and the antioxidant and antimicrobial activities of three Veronica species (Plantaginaceae), V. officinalis L., V. teucrium L. and V. orchidea Crantz, were studied. The identification and quantification of several phenolic compounds and phytosterols were performed using LC/MS techniques and the main components were p-coumaric acid, ferulic acid, luteoline, hispidulin and β-sitosterol. More than that, hispidulin, eupatorin and eupatilin were detected for the first time in the Veronica genus. Nevertheless, representatives of the Veronica genus were never investigated in terms of their phytosterol content. The antioxidant potential investigated by Trolox equivelents antioxidant capacity (TEAC) and EPR spectroscopy revealed that V. officinalis and V. orchidea extracts presented similar antioxidant capacities, whilst the values registered for V. teucrium extract are lower. Regarding the antimicrobial activity of the investigated species, Staphylococcus aureus, Listeria monocytogenes and Listeria ivanovii were the most sensitive strains with MIC values between 3.9 and 15.62 mg/mL. The results obtained by this study may serve to promote better use of representatives from the genus Veronica as antioxidant and antimicrobial agents.     

Simultaneous Analysis of Tocopherols, Cholesterol, and Phytosterols Using Gas Chromatography

ABSTRACT: A method for the simultaneous analysis of tocopherols and sterols was developed. Lipids were extracted with the Folch method, saponified by KOH-ethanol, and then tocopherols, cholesterol, and phytosterols were extracted with hexane. The extracted samples were dried under a nitrogen stream, derivatized using trimethylsilyl compounds, and then subjected to a gas chromatography. The recovery rates for cholesterol, stigmasterol, and sitosterol were about 100%, but recovery rates for tocopherols were low (25% forδtocopherol and 66% for γ-tocopherol) and varied according to compound structures. However, the recovery rates for δ- and γ-tocopherols increased to about 100% when the amounts of water and hexane were increased to 15 mL at the extraction step after saponification.     

植物甾醇硬脂酸酯的合成研究

采用植物甾醇与硬脂酸直接酯化时方法合成了甾醇硬脂酸酯．通过正交实验法，筛选出酯化反应最优工艺条件为n(甾醇)：n(硬脂酸)=1：1．2(mol／mol)，反应温度135℃，反应时间7h，酯化率可以达到87．8％．研究了甾醇硬脂酸酯的重结晶精制工艺．     

Determination of tocopherols and phytosterols in sunflower seeds by NIR spectrometry

The objective of this work was to develop a near‐infrared reflectance spectrometry (NIRS) calibration estimating the tocopherol and phytosterol contents in sunflower seeds. Approximately 1000 samples of grinded sunflower kernels were scanned by NIRS at 2‐nm intervals from 400 to 2500 nm. For each sample, standard measurements of tocopherol and phytosterol contents were performed. The total tocopherol content was obtained by high‐performance liquid chromatography coupled with a fluorescence detector, while the total phytosterol content was assessed by gas chromatography. For tocopherol, the calibration data set ranged from 175 to 1005 mg/kg oil (mean value around 510 ± 140 mg/kg oil), whereas for the phytosterol content, the calibration data set ranged from 180 to 470 mg/100 g oil (mean value of 320 ± 50 mg/100 g oil). The NIRS calibration showed a relatively good correlation (R² = 0.64) between predicted by NIRS and real values for the total tocopherol content but a poor correlation for the total phytosterol content (R² = 0.27). These results indicate that NIRS could be useful to classify samples with high and low tocopherol content. In contrast, the estimation of phytosterol contents by NIRS needs further investigation. Moreover, in this study, calibration was obtained by a modified partial least‐squares method; the use of other mathematical treatments can be suitable, particularly for total phytosterol content estimation.