Analysis of Alkylresorcinols in Wheat Germ Oil and Barley Germ Oil via HPLC and Fluorescence Detection: Cochromatography with Tocols

Alkylresorcinols are long‐chain phenolic compounds that have been reported to be localized in the outer layers of the kernels of wheat, rye, barley, and other grains. A sensitive HPLC method with fluorescence detection was recently reported for the quantitative analysis of alkylresorcinols in cereal grains and products. Using this new HPLC method we report for the first time that wheat germ oil contains moderate levels of alkylresorcinols, approximately 800–1,500 µg per gram of oil. We also found that commercial wheat germ oil and some experimental samples of wheat germ oil and barley germ oil also contained three unknown peaks. Upon further evaluation of these peaks it was found that the peaks appeared to be tocopherols (one peak of α‐tocopherol, one peak of δ‐tocopherol, and a peak with the combination of β‐ and γ‐tocopherol), even though the excitation and emission wavelengths for alkylresorcinols (excitation 274 nm and emission 300 nm) are different than those for tocols (excitation 294 nm and emission 326 nm). We also found that with this HPLC system one alkylresorcinol, AR17, had the same retention time (7.6 min) as δ‐tocotrienol and that another alkylresorcinol, AR19, had the same retention time (10.8 min) as α‐tocotrienol.     

Comparison of the chemical composition of extracts from Scutellaria lateriflora using accelerated solvent extraction and supercritical fluid extraction versus standard hot water or 70% ethanol extraction

The aqueous extract of American skullcap (Scutellaria lateriflora L. (S. lateriflora), Lamiaceae) has been traditionally used by North American Indians as a nerve tonic and for its sedative and diuretic properties. Recent reports stated that flavonoids and possibly amino acids are responsible for the anxiolytic activity. As a part of our search for environmentally friendly solvents to extract the active components from medicinal plants, we used S. lateriflora in a comparison of accelerated solvent extraction (ASE) using water, and supercritical fluid extraction (SFE) using CO2 and 10% EtOH as modifier, at different temperatures. Flavonoids and amino acids were quantified by HPLC-UV and HPLC-MS, respectively. The flavonoid content was compared with conventional extraction methods (hot water extraction and 70% ethanol). The use of ASE at 85 degrees C with water as solvent gave the best results for flavonoid glycosides and amino acids, whereas SFE gave higher yields of flavonoid aglycones. However, the results obtained for total flavonoids were not significatively superior to hot water extraction or 70% aqueous EtOH extract.     

Alkylresorcinols as markers of whole grain wheat and rye in cereal products

The alkylresorcinol (AR) content of Swedish wheat grain samples, as well as of cereal ingredients and cereal foods containing wheat and rye, was determined. The average total AR content in Swedish wheat was 412 microg/g (ranging between 227 and 639 microg/g), which is lower than that in Swedish rye analyzed in a previous study. The relative composition of AR homologues was consistent for wheat samples and differed markedly from that of rye. Notably, the ratio of the homologues C17:0/C21:0 was approximately 0.1 in wheat and approximately 1.0 in rye, indicating that it can be used to distinguish between those two cereals. The AR content in cereal foods commonly consumed in Sweden varied widely, from nondetectable levels in white wheat flour and products not containing the outer parts of wheat and/or rye to >900 microg/g in some whole grain rye products. AR content in cereal foods was calculated from their recipes using average AR values for the cereal ingredients determined in this study. As there was a good correlation between calculated and analyzed AR levels in cereal foods (R2 = 0.91), it is possible to estimate the proportion of whole grain wheat and/or rye in a given cereal product on the basis of AR content and C17:0/C21:0 ratio. ARs appear to be good markers of whole grain wheat and rye in foods, and their analysis may be an objective way to identify foods rich in whole grain wheat and/or rye or brans thereof.     

比较四种提取技术分析沉积物中滴滴涕同系物

Four techniques, Soxhlet extraction （SOX）, ultrasonic extraction （USE）, fluidized-bed extraction （FBE） and accelerated solvent extraction （ASE） with different solvents （methanol, hexane/acetone and acetonitrile） were used for the extraction of DDT analogues in sediments. Results revealed that the four extraction techniques had high recoveries （〉 86.0%） with low standard deviations （〈 12.0%） for most of DDT analogues, meaning that they could all successfully extract DDT analogues in sediments. Accelerated solvent extraction using methanol and hexane/acetone （1：1）, fluidized-bed extraction using hexane/acetone （1：1） and the ultrasonic extraction using hexane/acetone （1：1） were comparable or better than Soxhlet extraction using hexane/acetone （1：1）. Considering solvent- and time-consumption, level of automation, and environmental risk, accelerated solvent extraction with hexane/acetone （1：1） was better than the other extraction techniques.     

土壤样品中多环芳烃分析方法研究进展

概述了国内外土壤样品中多环芳烃(PAHs)测定方法的研究状况,其中提取方法包括加速溶剂萃取方法、固相微萃取方法、超临界流体萃取方法、亚临界水萃取方法和流化床提取方法等,测定方法有HPLC法、GC法和免疫分析法等。重点介绍了PAHs的提取过程,同时总结了各种方法的优缺点。     

Alkylresorcinol content and homologue composition in durum wheat (Triticum durum) kernels and pasta products

The total alkylresorcinol (AR) content and relative homologue composition of 21 durum wheat (Triticum durum) kernel samples, as well as 5 pasta products and the corresponding flour mixtures, were determined. Durum wheat contained on average 455 microg/g ARs, and the average relative homologue composition was C17:0 (0.4%), C19:0 (14%), C21:0 (58%), C23:0 (21%), and C25:0 (6.5%). The homologue composition was found to be relatively consistent among samples, with durum wheat being different from common wheat by having a higher proportion of the longer homologues. No differences in content or homologue composition were observed in pasta products compared to flour ingredients, showing that alkylresorcinols are stable during pasta processing. The ratio of the homologues C17:0 to C21:0 was < or =0.02 for whole grain durum wheat products, which is different from those of common wheat (0.1) and rye (0.9).     

Alkylresorcinols in cereals and cereal products

The alkylresorcinol (AR) content of 8 commonly consumed cereals, 125 Triticum cultivars, milling fractions of wheat and rye, bread, and other cereal products was analyzed. ARs were found in wheat (489-1429 microgram/g), rye (720-761 microgram/g), triticale (439-647 microgram/g), and barley (42-51 microgram/g), but not in rice, oats, maize, sorghum, or millet. One durum wheat variety was found to have an exceptionally low level of ARs (54 microgram/g) compared to other durum wheat varieties (589-751 microgram/g) and Triticumspecies analyzed. The AR content of milling fractions closely followed the ash content and could be used as a marker of the presence of bran in flour. Using hot 1-propanol extraction, all ARs could be extracted from bread, contrary to previous studies which suggested that ARs were destroyed during baking. Cereal products varied greatly in AR content, with those containing wheat bran or whole rye having the highest content.     

Fat extraction from acid- and base-hydrolyzed food samples using accelerated solvent extraction

This paper describes a new in-cell method for pursuing accelerated solvent extraction (ASE) prior to lipid analysis from food samples. It is difficult to pursue direct ASE with acid- or base-hydrolyzed samples due to the corrosive nature of the reagents and material limitations. In this study ion exchange based materials were used to remove acid or base reagents in-cell without compromising the recovery of lipids. The performance data are presented here for the new methods for lipid extraction for a variety of food samples and compared to the Mojonnier method. NIST Standard Reference Materials (SRM-1546 and SRM-1849) were used to validate the ASE methods. Excellent fat recoveries were obtained for the ASE methods. The new methods presented here enhance the utility of ASE and eliminate labor intensive protocols.     

Comparison of Three Extraction Systems for Determining Surface Lipid Content of Thickness‐Fractionated Milled Rice

The surface lipid content (SLC) of rice is often used to objectively measure the degree to which bran has been removed from rice kernels, commonly known as degree of milling (DOM). This study was conducted to evaluate new, rapid extraction technology for potential timesaving measurements of SLC of milled rice. The SLC of two long‐grain rice cultivars, Cypress and Drew, were determined using three extraction systems: Soxtec, accelerated solvent extraction (ASE), and supercritical fluid extraction (SFE). Before milling, rough rice was separated into three thickness fractions (<1.84, 1.84–1.98, and >1.98 mm) and samples from each thickness fraction were milled for durations of 10, 20, and 30 sec. Head rice collected from each milling duration was extracted using each of the three methods. Results showed that regardless of the extraction method, thinner kernels had lower SLC measurements than thicker fractions. In most cases, both the ASE and Soxtec produced SLC greater than that of the SFE. The ASE also showed SLC measurements at least as great as those from Soxtec extraction, suggesting that the ASE is as thorough in extracting lipids as commonly used methods.     

Synthesis and Inhibitory Activities against Colon Cancer Cell Growth and Proteasome of Alkylresorcinols

We have identified alkylresorcinols (ARs) as the major active components in wheat bran against human colon cancer cell growth (HCT-116 and HT-29) using a bioassay-guided approach. To further study the structure-activity relationships, 15 ARs and their intermediates (1-15) were synthesized expediently by the modified Wittig reaction in aqueous media, and six 5-alkylpyrogallols and their analogues (16-21) were prepared by the general Grignard reaction. The synthetic AR analogues were evaluated for activities against the growth of human colon cancer cells HCT-116 and HT-29 and the chymotrypsin-like activity of the human 20S proteasome. Our results found that (1) AR C13:0 and C15:0 (13 and 14) had the greatest inhibitory effects in human colon cancer cells HCT-116 and HT-29, while decreasing or increasing the side chain lengths diminished the activities; (2) two free meta-hydroxyl groups at C-1 and C-3 on the aromatic ring of the AR analogues greatly contributed to their antitumor activity; (3) the introduction of a third hydroxyl group at C-2 (20 and 21) into the aromatic ring of the AR analogues yielded no significant enhancement in activity against HCT-116 cells and decimated the effects against HT-29 cells, but dramatically increased the activity against the chymotrypsin-like activity of the human 20S proteasome; and (4) AR C11:0 (12) was found to have the greatest effect in a series of AR C9:0-C17:0 against the chymotrypsin-like activity of the human 20S proteasome.     

Analysis of zearalenone in cereal and Swine feed samples using an automated flow-through immunosensor

The development of a sensitive flow-though immunosensor for the analysis of the mycotoxin zearalenone in cereal samples is described. The sensor was completely automated and was based on a direct competitive immunosorbent assay and fluorescence detection. The mycotoxin competes with a horseradish-peroxidase-labeled derivative for the binding sites of a rabbit polyclonal antibody. Control pore glass covalently bound to Prot A was used for the oriented immobilization of the antibody-antigen immunocomplexes. The immunosensor shows an IC(50) value of 0.087 ng mL(-1) (RSD = 2.8%, n = 6) and a dynamic range from 0.019 to 0.422 ng mL(-1). The limit of detection (90% of blank signal) of 0.007 ng mL(-1) (RSD = 3.9%, n = 3) is lower than previously published methods. Corn, wheat, and swine feed samples have been analyzed with the device after extraction of the analyte using accelerated solvent extraction (ASE). The immunosensor has been validated using a corn certificate reference material and HPLC with fluorescence detection.     

Comparison of methods for the exhaustive extraction of hypericins, flavonoids, and hyperforin from Hypericum perforatum L

Renewed interest in plant-derived drugs has led to an increased need for efficient extraction methods. Hypericum perforatum L. contains several groups of bioactive compounds with noteworthy pharmacological activities. Direct sonication of H. perforatum was investigated and compared with conventional maceration, indirect sonication, Soxhlet extraction, and accelerated solvent extraction (ASE). Highly selective liquid chromatography/tandem mass spectrometry analysis showed that the content of six investigated active compounds (hypericin, pseudohypericin, hyperoside, rutin, quercitrin, and hyperforin) in extracts obtained by direct sonication was significantly higher than in extracts obtained by the other methods. The active compound contents increased on increasing the ultrasonic power from 40 to 60 W when using direct sonication. Conventional maceration gave the lowest amount of analyzed active compounds. Soxhlet extraction gave better results than ASE or indirect sonication.     

Quality assessment and quantitative analysis of flavonoids from tea samples of different origins by HPLC-DAD-ESI-MS

Components of green tea ( Camellia sinensis) have been of considerable interest in recent years because of their potential utility as pharmaceutical agents, particularly for their antioxidant and anticarcinogenic activity. Responding to the increasing scientific validation of numerous health benefits of tea, a comprehensive approach was adopted to carry out analysis for the quality assessment of flavonoids in tea samples of different origins. For this purpose, extraction, separation, and mass spectrometric parameters were optimized. Extraction methods evaluated include reflux extraction, a modified accelerated solvent extraction (ASE), namely, Aquasolv extraction, and microwave-assisted extraction (MAE) using different percentages of solvents. Separation was performed by a specifically developed reversed phase high-performance liquid chromatography (RP-HPLC) method using different C18 and C8 stationary phases. Optimization of extraction techniques clearly proved the performance of MAE, which delivered highest yields in a very short time. Additionally, the comparison with Aquasolv extraction provided new insights, as variations in quantified amounts of target compounds between the extracts could be explained on the basis of thermal degradation and epimerization phenomena. Especially the epimerization phenomenon for catechin/epicatechin oligomers, that is, of procyanidins P 2 and P 3, was observed for the first time. Finally, an optimized extraction and separation system was used for qualitative and quantitative investigations of compounds from different green tea samples from Ceylon (cultivated under biologically controlled conditions), Japan, India, and China as well as from one black tea sample from India.     

Improved Method for n-Alkanes and n-Methyl-Ketones Determination in Litters and Soils

Eurasian Soil Science - A new technique to extract lipid biomarkers (n-alkanes and n-methyl-ketones) from forest litters and soils using the accelerated solvent extraction (ASE) method was...     

Extraction of Antioxidants from Wheat Bran Using Conventional Solvent and Microwave-Assisted Methods

Total phenolic and tocopherol contents and free radical scavenging capability of wheat bran extracted using conventional and microwave-assisted solvent extraction methods were studied. Three different solvents (methanol, acetone, and hexane) were used in the conventional solvent extraction. Methanol was the most effective solvent, producing higher extraction yield (4.86%), total phenolic compound content (241.3 μg of catechin equivalent/g of wheat bran), and free radical scavenging capability (0.042 μmol of trolox equivalent/g of wheat bran) than either acetone or hexane. However, there was no significant difference in the total tocopherol contents (13.6–14.8 μg/g of wheat bran) among the three different solvent extraction methods. Microwave-assisted solvent extraction using methanol significantly increased the total phenolic compound content to 467.5 and 489.5 μg of catechin equivalent; total tocopherol content to 18.7 and 19.5 μg; and free radical scavenging capability to 0.064 and 0.072 μmol of trolox equivalent/g of wheat bran at extraction temperatures of 100 and 120°C, respectively. However, extraction yields of conventional methanol solvent and microwave-assisted methanol extractions at different temperatures were not significantly different.     

Alkylresorcinols in wheat varieties in the HEALTHGRAIN Diversity Screen

The contents of alkylresorcinols (AR) were analyzed in 131 winter wheats, 20 spring wheats, 10 durum wheats, 5 spelt wheats, and 10 early cultivated forms of wheat (5 diploid einkorn and 5 tetraploid emmer), which are part of the HEALTHGRAIN diversity screen. AR were analyzed by gas chromatography (GC), which provides both total contents and relative homologue compositions, as well as with a Fast Blue colorimetric method that provides only total contents but which is fast and easily screens a large number of samples. There was considerable variation in the total AR content analyzed with GC: winter wheat (220-652 microg/g of dm), spring wheat (254-537 microg/g of dm), durum wheat (194-531 microg/g of dm), spelt (490-741 microg/g of dm), einkorn (545-654 microg/g of dm), and emmer wheat (531-714 microg/g of dm). The relative AR homologue composition was different for different types of wheat, with a C17:0 to C21:0 ratio of 0.1 for winter, spring, and spelt wheats, 0.04 for einkorn and emmer wheat, and 0.01 for durum wheat. The total AR content analyzed with the Fast Blue method was lower than that analyzed with GC but there was a good correlation between the two methods (R(2) = 0.76).     

Accelerated solvent extraction of ochratoxin a from rice samples

In this paper, accelerated solvent extraction (ASE) for the analysis of ochratoxin A (OTA) is applied for the first time. Optimization of the method is given for the extraction of OTA from rice samples. Several parameters such as type of solvent, temperature, pressure, static time, and cell size were investigated thoroughly to find the optimal extraction conditions. The optimum ASE operating conditions were methanol as extraction solvent, 1500 psi, 40 degrees C, 5 min of static time, 50% flush volume, 60 s of purge, 1 cycle, and 11 mL cell size. The total extraction time was approximately 15 min. OTA was determined by liquid chromatography coupled with a fluorescence detector and confirmed by methyl ester derivatization. The analytical performance of the method was monitored with quality control parameters. Finally, the optimized method was used to evaluate 12 rice samples, 1 of which was positive with an OTA content of 4.17 ng/g. The proposed method offers the possibility of a fast and simple process to obtain a quantitative extraction of OTA.     

Determination of trifloxystrobin and its metabolites in Hawaii soils by ASE-LC-MS/MS

Analytical methods for the determination of trifloxystrobin and four of its metabolites were developed in a leaching study conducted in Hawaii. To duplicate plots at each of five locations representing various agricultural areas in Hawaii, trifloxystrobin was applied at label rates and allowed to leach under normal rain and irrigation conditions. Soil samples were collected at weekly to monthly intervals and the residual concentrations of trifloxystrobin and metabolites measured. A quantitative analytical method for their determination in various soil samples was developed using accelerated solvent extraction (ASE), coupled with liquid chromatography-tandem mass spectrometry. Extraction solvent with various ratios of methanol to water, addition of EDTANa2 to the extract solvent, and ASE cell temperature were adjusted to improve recovery. Deuterated (E, E)-trifloxystrobin was chosen as the internal standard of the analytical method. The limit of quantitation was 2.5 ppb in the soil for trifloxystrobin and its metabolites. Laboratory aerobic degradation studies with the soils from the five sites were also conducted to measure the same compounds.     

From Field to Table: A Review on the Microbiological Quality and Safety of Wheat‐Based Products

Throughout history, wheat‐based foods have been considered among the safest of all foods produced for human consumption. In part, this claim reflects both the low risk profile of low‐moisture foods and the thermal processes used to produce the finished product. Nevertheless, raw flour contains a number of potential hazards, which, if not properly managed, may have the potential to result in serious public health consequences. These hazards are mostly microbiological in origin and arise mainly during production and distribution through the wheat supply chain. The physical processes carried out during milling have minimal impact on the level of contamination present on grain; therefore, the initial microbiological quality of wheat grain has a strong influence on the ultimate quality and safety of milling end products. Although most flour‐based foods are processed and consumed in forms that are less likely to be contaminated with pathogens, refrigerated dough products have the potential to be a safety hazard to consumer health because they could be consumed raw or undercooked. The risk for exposure to pathogenic microorganisms present in raw flour by eating uncooked baking mixture has been demonstrated by several outbreaks. Such food safety incidents have led to heightened manufacturer and consumer awareness about safety related to flour‐containing foods.     

Development of a new method for the complete extraction of carotenoids from cereals with special reference to durum wheat (Triticum durum Desf.)

Due to the growing interest in the role of carotenoids in human health, their qualitative and quantitative analysis in foods is becoming more and more important. High-performance liquid chromatography has become the method of choice for the determination of these phytochemicals. A crucial step prior to the chromatographic separation is the quantitative extraction from the food matrix which was proven to be impeded in durum wheat. To optimize the extraction procedure, several factors with influence on extractability of carotenoids were investigated. Finally, it was shown that soaking of samples in water for 5 min prior to extraction with organic solvents had the strongest impact on extraction yield and led to the most rapid and gentle method. Contents of carotenoids in the extracts of several durum wheat and corn samples were doubled by soaking in water before extracting with methanol/tetrahydrofuran (1/1, v/v). In light of these findings, literature data on contents of carotenoids in cereal grains have to be viewed critically regarding the extraction procedures employed.