Direct Solvent Extraction Research Articles

2-methyl-3-buten-2-ol: A Pheromone Component of Conifer Bark Beetles Found in the Bark of Nonhost Deciduous Trees

Volatiles from bark of aspen,Populus tremulaL. and two species of birch: silver birch (Betula pendulaRoth.) and common birch (B. pubescensEhrh.), were collected by direct solvent extraction and aeration of both newly cut bark chips and undamaged stems in June 1998 and subjected to GC-MS analysis. The results showed the presence of 2-methyl-3-buten-2-ol (MB), one of the two principal aggregation pheromone components of the spruce bark beetle,Ips typographus, in bark extraction samples of all the three deciduous tree species tested. In addition, one more oxygenated hemiterpene, 3-methyl-3-buten-2-one, and (E)-3-penten-2-ol were also found in the bark extracts. Only trace amounts of MB were detected in some aeration samples of the fresh bark chips, and no MB was found from the aeration samples of undamaged stems at detectable levels. The occurrence of this compound was also confirmed in the bark of four exotic birch species:B. albosinensisSchneid.,B. ermaniiCham.,B. jacquemontiiSpach, andB. maximowiczianaRegel, but not yet in the European pines/spruces and the common yeasts. Our results raise major questions regarding the evolution, the tropospheric chemistry, and the ecological role of this hemiterpene alcohol. They also suggest that comparative studies on the biosynthetic pathways for MB in different sources would be of considerably evolutionary interest.

Identification of Irradiated Prawn (Penaeus monodon) Using Thermoluminescence and 2-Alkylcyclobutanone Analyses

Thermoluminescence (TL) and 2-alkylcyclobutanone (2-ACB) analyses were performed to identify irradiated prawns ( Penaeus monodon ). With the TL method, minerals were extracted from prawns using acid hydrolysis. The experimental results satisfied the evaluation criteria of European Norm (EN) 1788, even after low-dose irradiation (0.5 kGy) and a 60 day storage at -20 °C. With the 2-ACB method, 2-dodecylcyclobutanone (2-DCB) and 2-tetradecylcyclobutanone (2-TCB) were successfully extracted from prawns by direct solvent extraction with purification using a conventional silica column and a sulfoxide column, which was used for 2-ACB for the first time. Both 2-ACB derivatives were absent from the non-irradiated samples but were identified in all irradiated samples by gas chromatography-mass spectrometry. Moreover, 2-DCB and 2-TCB production correlated with the applied dose (2.5-10 kGy), and the correlation did not diminish after 60 days of storage at -20 °C for any dose. Therefore, these two techniques provide rapid, simple, and promising methods for routine investigation of frozen prawns.

Comparative study of two extraction techniques to obtain representative aroma extracts for being analysed by gas chromatography–olfactometry: Application to roasted pistachio aroma

This research paper presents a comparative study of two different extraction and concentration techniques to obtain representative pistachio aroma extracts: the traditional direct solvent extraction (DSE) followed by high-vacuum transfer (HVT) and the headspace solid-phase microextraction (HS-SPME). The results showed that, although both techniques provide accurate information about the aromatic composition that will be perceived by the consumer, the precision in terms of within-day repeatability and between-days repeatability (intermediate precision) of the chromatographic areas presented better values for HS-SPME than for DSE-HVT. Moreover the solvent-free HS-SPME allows the extraction of more odour-active regions, requires very little sample handling and shorter time for sampling.

Effects of co-existed proteins on measurement of pesticide residues in blood by gas chromatography–mass spectrometry

Accurate measurement of pesticides in biological fluids such as blood is important for quantifying environmental exposures. Beyond sample enrichment and separation, the method presented here is focused on studies of interactions between pesticides and co-existed proteins. It was experimentally demonstrated that entrapped or adsorbed pesticide residues within the folded native structures of proteins were poorly recovered using direct solvent extraction solely. We described here an effective approach termed Enzymatic Digestion-Organic Solvent Extraction (eDOSE) that utilizes the enzymatic approach to disrupt the folded structures of proteins and release entrapped or adsorbed pesticide residues. In this approach, samples were first reduced, alkylated, tryptically digested and then diluted 10 times before the subsequent extraction using an n-hexane solution. Resultant pesticide residues were determined by capillary gas chromatography coupled with a mass spectrometer. Mean recoveries of the 5 organophosphorus pesticides pre-spiked in fish blood including diazinon, parathion-methyl, malathion, parathion-ethyl and ethion were 85%, 95%, 84%, 103%, and 43% respectively using eDOSE strategy but only 24%, 45%, 40%, 27%, and 29% respectively using direct solvent extraction approach. The eDOSE approach was effective for demonstrating the critical role of folded native structure of serum albumin in adsorption of exogenous chemicals. It provides an alterative means for denaturation of proteins when the target analytes are not stable in acidic solution or entrapped within the protein aggregates caused by organic solvents such as acetone that have been applied for protein denaturation. The eDOSE approach should be able to combine with other advanced techniques of enrichment and separation for more efficient and accurate measurement of target compounds present in the context of complex biological systems. This approach can provide wide applications to the analysis of a variety of small molecules including environmental pesticide residues and metabolites as well as other toxins present in cells, tissues and biofluids.

A Rapid Direct Solvent Extraction Method for the Extraction of 2‐Dodecylcyclobutanone from Irradiated Ground Beef Patties Using Acetonitrile

The amount of irradiated beef in the U.S. market is growing, and a reliable, rapid method is needed to detect irradiated beef and quantify the irradiation dose. The official analytical method (BS EN 1785 2003) that has been adopted by the European Union is time consuming. The objective of this study was to develop a rapid method for the analysis of 2-dodecylcyclobutanone (2-DCB) in irradiated beef. A 5 g sample of commercially irradiated ground beef patty (90/10) was extracted with n-hexane using a Soxhlet apparatus or with acetonitrile via direct solvent extraction. The Soxhlet hexane extract was evaporated to dryness, and the sample was dissolved in a mixture of ethyl acetate and acetonitrile (1:1). The defatted extract was purified with a 1 g silica cartridge. Another 5 g aliquot of the same patty was mixed with 50 mL acetonitrile and either blended for 1 min with a hand blender or crushed for 10 min with a glass rod. The extraction procedure was repeated 3 times, and the acetonitrile was collected and evaporated to dryness. Eluants from both methods were concentrated under nitrogen and injected into a gas chromatography-mass spectrometry. The 2-DCB concentration in the commercial samples was 0.031 +/- 0.0026 ppm (n = 5) for the Soxhlet method and 0.031 +/- 0.0025 ppm (n = 10) for direct solvent extraction. Recovery of 2-DCB from spiked beef samples in the direct solvent extraction method was 93.2 +/- 9.0% (n = 7). This study showed that the direct solvent extraction method is simple and as efficient and reproducible as the Soxhlet method.

Detecting gravesoil with headspace analysis with adsorption on short porous layer open tubular (PLOT) columns

Victims of crimes are often buried in clandestine graves. There are several techniques for finding buried bodies or the scattered remains of a victim; however, none of these methods are very reliable or work in all scenarios. One way to detect gravesoil is to detect the biochemical changes of the surrounding soil due to cadaver decomposition, for example, the release of nitrogenous compounds. A simple and low-cost way to detect these compounds is based on the reaction of alpha amino groups with ninhydrin to form Ruhemann's purple. This test for ninhydrin-reactive nitrogen (NRN) has, to date, only been performed by direct solvent extraction of soil samples. Here, we present a method that detects trace quantities of NRN in the headspace air above gravesoil. Our method is based on an improved purge and trap method developed in our lab for sampling low volatility compounds, as well as volatile compounds at trace quantities, by applying low temperature collection on short alumina-coated porous layer open tubular (PLOT) columns. We modified this method to sample the headspace air above gravesoil with a motorized pipetter and a PLOT column at ambient temperatures. We generated gravesoil using rat cadavers and local soil. Trace quantities of NRN were successfully detected in the headspace air above gravesoil. We report the quantities of NRN recovered for buried rats, rats laid on top of soil, and blank graves (no rats) as a function of time (weeks to months). This work is the first (and thus far, only) example of a method for detecting NRN in the vapor phase, providing another tool for forensic investigators to aid in locating elusive clandestine graves.

Partition Coefficients of Organic Contaminants with Carbohydrates

In view of the current lack of reliable partition coefficients for organic compounds with carbohydrates (K(ch)), carefully measured values with cellulose and starch, the two major forms of carbohydrates, are provided for a wide range of compounds: short-chain chlorinated hydrocarbons, halogenated benzenes, alkyl benzenes, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls, and organochlorine pesticides. To ensure the accuracy of the K(ch) data, solute concentrations in both water and carbohydrate phases are measured by direct solvent extraction of the samples. For a given compound, the observed partition coefficient with cellulose (K(cl)) is virtually the same as that with starch (K(st)). This finding expedites the evaluation of organic contamination with different forms of carbohydrates. The presently determined K(ch) values of 13 PAHs are substantially lower (by 3-66 times) than the literature data; the latter are suspect as they were obtained with (i) presumably impure carbohydrate samples or (ii) indirectly measured equilibrium solute concentrations in carbohydrate and water phases. Although the K(ch) values are generally considerably lower than the respective K(ow) (octanol-water) or K(lipid) (lipid-water), accurate K(ch) data are duly required to accurately estimate the contamination of carbohydrates by organic compounds because of the abundance of carbohydrates over lipids in crops and plants. To overcome the current lack of reliable K(ch) data for organic compounds, a close correlation of log K(ch) with log K(ow) has been established for predicting the unavailable K(ch) data for low-polarity compounds.

Recovery of nickel and cobalt from laterite leach solutions using direct solvent extraction: Part 2: Semi- and fully-continuous tests

In Part 1 of this paper, two synergistic solvent extraction systems consisting of Versatic 10/LIX63/TBP and Versatic 10/4PC were assessed in batch tests for the separation and purification of nickel and cobalt from synthetic laterite leach solution after iron removal. In Part 2, semi- and fully-continuous tests are reported for the Versatic 10/LIX63/TBP system, with conditions optimised for separating nickel and cobalt from manganese, magnesium and calcium. Semi-continuous extraction tests were conducted using the synergistic organic system consisting of 0.50 M Versatic 10, 0.45 M LIX63 and 1.0 M TBP in Shellsol D70. With a pH profile of 5.5/6.1/6.5 for the three stages EX1/EX2/EX3 at 40 °C, the nickel and cobalt extractions were 99.9% with only 5 mg/L nickel and < 1 mg/L cobalt left in the raffinate. With two stages of scrubbing and a pH profile of 5.4/5.0 at 40 °C, about 2 mg/L manganese and less than 1 mg/L magnesium and calcium were left in the scrubbed organic solution. With two stripping stages and an O/A ratio of 10 at 40 °C using 50 g/L H 2SO 4 as strip solution, the stripping efficiencies of nickel and cobalt were over 95%. A fully-continuous pilot plant was operated for 280 h. With an O/A ratio of about 2 and a pH profile of 5.5/5.8/6.0/6.3 for the four stages EX1/EX2/EX3/EX4 at 40 °C, both nickel and cobalt were almost completely extracted. The nickel and cobalt concentration in the raffinate was lower than detection limit of 0.2 mg/L. The manganese, magnesium and calcium concentrations in the loaded organic solution were 34, 8 and 1 mg/L, respectively. Using a pH profile of 5.4/5.0 for SC1/SC2 at an O/A ratio of 10 and 40 °C, the manganese scrubbing efficiency was over 96% and the concentrations of manganese and magnesium in the scrubbed organic solution were < 5 mg/L and that of calcium 1 mg/L. Using three strip stages and a strip solution containing 50 g/L H 2SO 4 and 55 g/L Ni at an O/A ratio of 10 and 40 °C, over 98% Ni and 99% Co were stripped with only 64 mg/L Ni in the stripped organic solution. The nickel concentration in the loaded strip liquor was 86 g/L, giving a ΔNi of 31 g/L. The loaded strip liquor contained less than 1 g/L acid.

Recovery of nickel and cobalt from laterite leach solutions using direct solvent extraction: Part 1 — selection of a synergistic SX system

The separation of nickel and cobalt from impurities such as manganese, magnesium and calcium using solvent extraction with Versatic 10 was largely improved by the addition of a synergistic reagent LIX63 (an α-hydroxyoxime) or 4PC (a pyridine carboxylate ester). With the organic systems containing Versatic 10 alone, the separation factors of nickel and cobalt over manganese were 6 and 15 respectively. When 4PC was added to the system, these increased to 147 and 1870 respectively, and with LIX63, they were even higher at 534 and 7720 respectively. This indicates that the synergistic solvent extraction (SSX) system with Versatic 10 and LIX63 performed very well and better than that with Versatic 10 and 4PC. The SSX system consisting of 0.5 M Versatic 10, 0.45 M LIX63 and 1.0 M TBP in Shellsol D70 performed the best among the systems tested containing LIX63. After a single contact, the extraction of Ni and Co was 99.6% and 96.9%, respectively. Only 6 mg/L Mn, 8 mg/L Mg and 1 mg/L Ca were found in the loaded organic solution. The manganese scrub efficiency was 97.7% at pH 5.3, resulting in a scrubbed organic solution containing only 0.8 mg/L Mn. Over 99% nickel, cobalt and manganese were stripped at pH 2.0, indicating easy stripping of these metals. The SSX system consisting of 0.5 M Versatic 10 and 1.0 M 4PC in Shellsol D70 performed the best among the systems tested containing 4PC. After a single contact, the extraction of Ni and Co was 99.4% and 89.4%, respectively. Some 200 mg/L Mn, 10 mg/L Mg and 48 mg/L Ca were found in the loaded organic solution. The manganese could not be scrubbed at the tested pH range of 5.4–6.0. Very fast Ni and fast Co stripping kinetics were observed, however, the Mn stripping kinetics were very slow. After 2 min of stripping, only 1.22% Mn was stripped. It is concluded that the SSX system containing 0.5 M Versatic 10, 0.45 M LIX63 and 1.0 M TBP performed much better than the SSX system containing 0.5 M Versatic 10 and 1.0 M 4PC in terms of both manganese and calcium behaviour in extraction, scrubbing and stripping.

Mate Recognition and Sex Differences in Cuticular Hydrocarbons of the Diurnal Firefly Ellychnia corrusca (Coleoptera: Lampyridae)

Abstract There are several genera of diurnally active fireflies (Coleoptera: Lampyridae), and these adults generally do not produce bioluminescent signals. We investigated whether contact sex pheromones play a role in mate recognition in the diurnal firefly Ellychnia corrusca (L.). In laboratory behavioral assays, after antennal contact &amp;gt;70% of males attempted copulation with freeze-killed females, whereas no copulation attempts occurred when freeze-killed females had been washed with hexane. This shows that E. corrusca mate recognition relies on contact chemoreception and suggests that male mating responses are mediated by contact sex pheromones on the female cuticle. Using direct contact solid phase microextraction (SPME) and solvent extraction, we sampled cuticular hydrocarbons (CHCs) from both sexes of E. corrusca adults. Gas chromatography-mass spectrometry showed that SPME was more effective than solvent extraction in detecting CHCs (a two-fold increase in the number of compounds detected). Although E. corrusca males and females showed similar CHCs profiles, we documented several quantitative and qualitative differences between the sexes that may play a role in mate recognition. This report provides the first behavioral evidence for the existence of contact sex pheromones in any diurnal firefly.

Commercial Runner Peanut Cultivars in the United States: Tocopherol Composition

Tocopherols in commercially grown normal, mid- and high-oleic Runner peanuts from 2005 and 2006 were quantified to give accurate vitamin E contents. Tocopherols were extracted from raw peanuts by a direct solvent extraction procedure using 10% ethyl acetate in hexanes that provided percent recoveries of 105.4, 101.2, 103.9, and 102.8 for alpha-tocopherol (T), beta-T, gamma-T, and delta-T, respectively. No significant (P > 0.05) differences were noted in total tocopherol levels in normal- (22.4 mg/100 g), mid- (23.9 mg/100 g), and high-oleic (22.4 mg/100 g) Runner peanuts. alpha-T levels did vary significantly among the Runner cultivars classified by their oleic acid content (mid, 11.7 mg/100 g; normal, 10.9 mg/100 g; high, 9.8 mg/100 g). Cultivar effects were highly significant (P < 0.001) for alpha-, beta-, gamma-, and delta-T and total tocopherol contents, whereas production year effects were highly significant for alpha- and beta-T levels. Year x cultivar interactions were not significant (P > 0.05). Cluster analysis segregated the cultivars into two major groups represented by lower alpha-T and higher gamma-T levels (cluster A) and high alpha-T and low gamma-T levels (cluster B) (P < 0.05). The mean alpha-T level in Runner peanuts (151 samples) was 10.5 +/- 1.5 mg/100 g, which is 26.7% greater than the imputed value for peanuts, all types (NBD 16087) provided by the USDA National Nutrient Database for Standard Reference.

Comparison of the Volatile Components in Two Chinese Wines, Moutai and Wuliangye

Volatile and odorous components in two Chinese wines, Moutai and Wuliangye, were partially investigated by direct solvent extraction. Sample extracts were analyzed by gas chromatography/ mass spectrometry and gas chromatography-olfactometry. Fifty-six and thirty-eight compounds were found in Moutai and Wuliangye, respectively, including esters (19, 20), alcohols (8, 7), acids (6, 3), furans (5, 2), acetals (5, 2), phenols (2, 2), ketones (7, 0) and miscellaneous compounds (4, 2). Based on olfactometry, ethyl acetate, furfural and 2-furanmethanol were perceived only in the Moutai whereas diethyl succinate was recognized only in the Wuliangye. However, 1,1-Diethyoxyehtane, ethyl hexanoate, ethyl benzoate and pentanoic acid were found in both wines.

Determination of eighty-one volatile organic compounds in dietary Rumexinduratus leaves by GC/IT-MS, using different extractive techniques

The volatile compounds of Rumex induratus leaves were studied for the first time. In order to achieve the most complete volatile profile four different extractive techniques (hydrodistillation, solid-phase microextraction, Soxhlet system and direct solvent extraction) were applied. The different extracts were analysed by gas chromatography/ion trap-mass spectrometry (GC/IT-MS) which allowed the identification of 81 compounds, distributed by several chemical classes: esters, terpenes, aldehydes, acids, norisoprenoids, ketones, naphthalene derivatives, steroids derivatives and alcohols, among others. The different techniques lead to different compounds' extraction. Using hexane and dichloromethane, 9 and 4 compounds were extracted, respectively. Hydrodistillation, solid-phase microextraction and Soxhlet extraction allowed the extraction of 43, 24 and 29 compounds, respectively. Hydrodistillation was revealed to be the most effective technique, allowing the extraction of a higher number of compounds, both in the hydrolat (20 compounds) and essential oil (34 compounds), having 11 compounds in common.

Manuka Oil and Phoebe Oil are Attractive Baits for &lt;I&gt;Xyleborus glabratus&lt;/I&gt; (Coleoptera: Scolytinae), the Vector of Laurel Wilt

Redbay ambrosia beetle, Xyleborus glabratus Eichhoff, is a native of Southeast Asia recently established in coastal forests of Georgia, SC and Florida. It vectors a wilt fungus, Raffaelea sp., lethal to redbay trees, Persea borbonia L. Spreng, and certain other Lauraceae. No practical monitoring system exists for this beetle so we conducted studies to identify host attractants and develop lures. Volatiles were collected from redbay wood and bark by steam distillation, direct solvent extraction, and dynamic headspace sampling with a Poropak Q cartridge. Steam, methanol, and pentane extracts were tested as baits in trapping trials but were not attractive to X. glabratus. Major constituents in Poropak aerations identified by gas chromatography-mass spectrometry included alpha-pinene, beta-pinene, delta-3-carene, eucalyptol, p-cymene, alpha-copaene, terpinene-4-ol, linalool, calamenene, and nonanoic acid. We assayed several of these compounds (including eucalyptol, p-cymene, terpinene-4-ol, linalool, nonanoic acid, and caryophyllene oxide) both individually and in combination, but none were attractive at tested doses. Two other redbay odor components, alpha-copaene and calamenene, were unavailable in sufficient quantities commercially so we substituted manuka oil, the essential oil extracted from Leptospermum scoparium Forst. and Forst., which contains high proportions of both compounds. Manuka oil was equally attractive as redbay wood to X. glabratus, but increasing release rates >10-fold did not enhance its activity. Phoebe oil, an extract of Brazilian walnut (Phoebe porosa Mez.), which contains significant quantities of alpha-copaene and calamenene, was also attractive. Fractions of manuka oil were not more attractive than the whole oil. Manuka and phoebe oil are readily available and are good alternatives to redbay wood as a trap bait for monitoring X. glabratus distribution and population trends.

Improved method for fatty acid analysis in herbage based on direct transesterification followed by solid-phase extraction

Direct transesterification (DT) and solvent extraction with acid or basic derivatization procedure for fatty acid (FA) analysis in herbage were compared. The highest total FA, α-linolenic and linoleic acid contents were obtained with DT. However, DT also produced the highest amount of interfering compounds, identified as phytadienes and sugar derivative products, which may co-elute with FA. An additional step based on solid-phase extraction to produce clean samples was proposed. This procedure is simple and gives good recoveries for the FA fortified samples. Additionally, structural characterization of 16:1 trans-3 was conducted by covalent adduct chemical ionization tandem mass spectrometry.

Aroma‐Active Components ofLycii fructus(kukija)

The fruit of Lycii fructus has been used as a tonic medicine and a long-term healthy food without side effect in Asia. An increase in the demand for natural healthy food, L. fructus has been thought as a source of healthy foods. For its value adding, its character impact aromas were isolated by using direct solvent extraction with vacuum transfer and identified by using gas chromatography olfactometry (GC-O) and GC-mass spectrometry (GC-MS). Thirty-three odor compounds were sniffed at GC-O, each trial equipped with DB-5MS and HP-WAX capillary column. The most contributing odor compounds in L. fructus were (E)-2-heptenal (green, mushroomy), 1-heptanol (planty, oily), hexanal (planty), 3-octanol (mushroomy, planty), 1-octen-3-ol (mushroomy), and 2-methyl-2-butenoic acid (pungent, planty), which might be produced by enzymatic oxidation and/or oxidation of lipids and carotenes, resulting in undesirable aromas.

Inter-laboratory Trial to Validate the Direct Solvent Extraction Method for the Identification of 2-dodecylcyclobutanone in Irradiated Chicken and Whole Liquid Egg

The main objective of this interlaboratory trial was to assess the qualitative performance of new rapid direct solvent extraction (DSE) method as potential detection method to differentiate between irradiated and unirradiated food samples. The trial was designed and carried out between four European laboratories that agreed to use 2-dodecylcyclobutanone (DCB) as radiolytic marker to detect irradiated minced chicken and liquid whole egg. Chicken samples were irradiated at 3 and 7 kGys while egg samples were irradiated at 3 and 6 kGys. Every laboratory had received 12-blind coded `unknown' and 4-known coded samples to be analyzed. All four laboratories were able to correctly identify all 12-blind coded samples as either irradiated (at medium and/or high doses) or unirradiated samples. The tested DSE method is regarded as fast, cheap, suitable and recommended for use in laboratories which are involved in screening large number of food samples for irradiation.

Extraction and identification of cyclobutanones from irradiated cheese employing a rapid direct solvent extraction method

2-Alkylcyclobutanones (cyclobutanones) are accepted as chemical markers for irradiated foods containing lipid. However, current extraction procedures (Soxhlet–florisil chromatography) for the isolation of these markers involve a long and tedious clean-up regime prior to gas chromatography–mass spectrophotometry identification. This paper outlines an alternative isolation and clean-up method for the extraction of cyclobutanones in irradiated Camembert cheese. The newly developed direct solvent extraction method enables the efficient screening of large numbers of food samples and is not as resource intensive as the BS EN 1785:1997 method. Direct solvent extraction appears to be a simple, robust method and has the added advantage of a considerably shorter extraction time for the analysis of foods containing lipid.

A rapid direct solvent extraction method for the extraction of cyclobutanones from irradiated chicken and liquid whole egg

Summary2‐Alkylcyclobutanones (cyclobutanones) are accepted as radiolytic markers in lipid‐containing foods (LCF). To date, cyclobutanones have not been detected in non‐irradiated foods. Their identification is the basis of a standardised detection test for irradiated LCF (BS EN 1785, 2003). This paper reports work to develop and refine a new rapid method for the analysis of cyclobutanones in irradiated LCF (e.g. chicken meat and liquid whole egg). Direct solvent extraction (DSE) enables the efficient screening of large numbers of food samples and is not as resource intensive as the BS EN 1785 (2003) method. The new DSE appears to be a promising, rapid, simple and robust method for the analysis of irradiated lipid‐containing foods.

Comparison of extraction methods for determining tocopherols in soybeans

AbstractWe evaluated the usefulness of saponification, direct solvent extraction, and Soxhlet extraction as extraction methods to determine the amounts of tocopherols in soybeans. Soxhlet extraction yielded the highest analytical values for each tocopherol homolog and was the best method for quantifying tocopherols in soybeans. Coupling of simple Soxhlet extraction with HPLC provided a highly reproducible procedure to quantify tocopherols in soybeans. The percent mean recovery ± standard deviation (n = 5) was 103.2 ± 1.21, 109.8 ± 4.18, 93.8 ± 1.12, and 106.9 ± 1.54% for α‐, β‐, γ‐, and δ‐tocopherol, respectively. The linearity test for quantification was carried out over the ranges of 0.4–10.0, 0.2–4.0, 2.0–16.0, and 0.4–10.0 µg/mL for α‐, β‐, γ‐, and δ‐tocopherol, respectively. Regression analysis showed an excellent linear relationship (R2 &gt;0.998), and the results of the validation parameters were generally reliable and satisfactory.