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Chapter 2

Soxtec: Its Principles and Applications
Shirley Anderson
Foss North America, Eden Prairie, MN 55344

The classical Soxhlet method provides the fundamental basis for a modern-day solvent extraction system, the SoxtecTM. Using the Randall modification, sometimes
called the submersion method, the Soxtec provides a faster approach to solvent
extraction for the gravimetric quantitation of fat and oil. Typically, the Soxtec methods require only 20–25% of the time required for traditional Soxhlet extraction.
Sample preparation, general extraction procedures, method considerations, and optimization are addressed. By definition, the procedure to determine “crude fat” is an
empirical method in which the result is determined by the conditions of the procedure.
Several aspects of the extraction process, such as solvent type, time, and temperature,
are explored. Several standardized Soxtec methods are discussed, including the
recently approved AOAC method for determining crude fat in feeds, cereal grains,
and forages. Many Soxtec applications are routinely used in food, feed, industrial, and
environmental laboratories for the measurement of fats, oils, semivolatiles, and other
solvent “extractables.” For the determination of crude fat, descriptions are given for
various sample pretreatment and extraction procedures. Practical guidelines for handling challenging samples as well as general suggestions are presented.

The foundation of today’s automated solvent extraction systems can be traced to
1879 to a German chemist, Franz Von Soxhlet. He devised a liquid/solid extraction apparatus in which a sample is placed in a cellulose thimble and stationed
over boiling solvent. Condensed solvent would then drip into the sample, solubilizing extractable material and then siphon back into the boiling solvent, where this
cycle would then repeat. After several cycles over many hours, the apparatus is
disassembled and the solvent, now containing extract (fat), is evaporated off, leaving the residue for further analysis. The Soxhlet procedure remains the most
exhaustive extraction technique, and today it is still widely used.
Over the years, there had been some improvements to the basic technique but
the procedure remained long, tedious, and prone to variability. In the early 1970s,
Edward Randall (1) developed an accelerated extraction technique that cut the
extraction time to as little as 30 min. In the Randall method, the sample is lowered

Copyright © 2004 AOCS Press

condensed solvent is flowing continuously through the sample and thimble back into the boiling solvent. The simple principle is that the material to be extracted. In 1975.and totally immersed in the boiling solvent. then raising it out of the solvent for the rinsing step. (a) Condenser (b) sample thimble (c) solvent flask (d) siphon tube (e) solvent vapor tube (f) thimble positioning mechanism (g) heater (not shown on the Soxhlet). In the original Randall method. This was first commercialized as the RaFaTec and later became the Soxtec systems of today. Automated Soxhlet for Crude Fat Automated extraction methods using the Soxtec have gained widespread acceptance and have a number of regulatory agency approvals worldwide. is more soluble in hot solvent than in cold or room temperature solvent. in this case. from Foss-Tecator. Sample Preparation. Crude fat by solvent extraction is classified as an empirical method (2). It therefore becomes critical that all aspects of the procedure be followed strictly. Copyright © 2004 AOCS Press . Lowering the thimble (b) into the boiling solvent for the boiling step. Proper handling of the sample and attention to detail are extremely important parts of the analytical process. 2. Tecator AB of Höganäs Sweden acquired the rights to what had become known as the “Randall modification” of the Soxhlet method. An improperly or sloppily preFIG. Demonstrating excellent agreement with Soxhlet and improved precision. Figure 2. fats and waxes. Procedural Overview of Soxtec. The Soxtec method has been used on literally hundreds of different sample types for many extracts. This means that the final result can be arrived at only according to the terms or variables of the method. Original Soxhlet (left) and Randall Extraction Apparatus. 2.1). the thimble is positioned by use of the slide rod (f).2 is a photograph of the SoxtecTM Avanti.1. this discussion will be limited mainly to crude fat extraction. In both stages. For our purposes. an automated Soxhlet extraction system using the Randall submersion technique. the Randall method has become the basis of many automated extraction systems. His procedure included this new boiling step followed by a rinsing step to flush residual extract from the sample (Fig.

The mill or milling process should not contribute to any loss of moisture or fat from the sample. pared sample can invalidate even the most carefully performed extraction procedure. The SoxtecTM Avanti 2050 automated extraction system. and glycerol can be extracted with fat yielding falsely high recoveries. Samples are weighed into the extraction thimbles and are then typically dried at 102 ± 2°C for 1–2 h (3–5. The weight of the sample is dependent on its approximate fat content. resulting in low fat recoveries. salts.1 can be used a guideline. Samples should be weighed using a calibrated 4-place analytical balance and in most cases. 2. Water in the sample can decrease the efficiency of the solvent extraction. the sample preparation may incorporate different procedures. Particular attention should be paid to the type of grinding mill used. carbohydrates. Pretreatment Drying. Depending on the type and nature of the sample. Conversely. Because the samples are weighed before drying. For grinding and weighing. the sample should be homogenous and finely ground.2. Most samples should be predried to optimize the fat extraction. water-soluble components in the samples such as urea. Copyright © 2004 AOCS Press . usually to pass through a 1-mm sieve (~18 mesh). it is critical that this small sample be representative of the larger sample lot. Table 2. Because only a few grams of sample are normally used for the analysis. can be weighed directly into the cellulose thimble.FIG.7).

5–1 the results are on an as-is basis.3). breaks these bonds. The boiling step is the key to accelerating the extraction process compared with the Soxhlet method. Copyright © 2004 AOCS Press . followed by a thorough drying step can be used to obtain an acceptable recovery by removing these water-soluble components. allowing the fat to be solvent extracted (see below: Total fat).1 Expected Fat Content and Sample Weights Fat content (%) 0–10 10–25 >25 Sample weight (g) 2–3 1–2 0. (3. in which a sample is boiled with hydrochloric acid. The solvent simply solubilizes the extract faster in hot solvent. Results expressed on a dry matter basis must be calculated from a separate moisture determination. The samples and extraction cups are then positioned in the extractor. 2. an alarm signals completion. Samples that have been processed. The weights of clean and dry extraction cups must also be obtained for later use in the final calculation.TABLE 2. sand may be mixed with the sample before drying. carbohydrates. The solvent is added through a closed-loop addition process and the extraction begins. In this step. A preextraction with water. and/or minerals. The sample is dried and the extraction procedure is carried out as usual. The solvent vapor refluxes against a water-jacketed cooling column and the condensed solvent flows back continuously through the sample returning to the boiling solvent (Fig.4) Solvent Extraction Once the samples have been properly prepared and pretreated. For very moist samples. rinsing. or extruded often have fat that is bound to proteins. cooked. Hydrolysis. Boiling. making it unavailable for solubilization. thus decreasing the time required for extraction. they can now be placed in the Soxtec for fat extraction. The procedure specifies washing the weighed sample with 5 aliquots of 20 mL of deionized water. Samples that contain a large amount of water-soluble components may exhibit poor solvent extraction efficiency. Water Rinse. This prevents the sample from becoming caked during drying and improves the solvent flow for optimal extraction (see below: Crude fat in meat and meat products). At the end of the cycle. the sample and thimble are lowered and totally immersed in the boiling solvent contained in the extraction cup. Acid hydrolysis. The steps of boiling. and evaporation/solvent recovery then proceed in an automated manner.

. The evaporation step is complete when all solvent is driven from the cup.3.FIG. During rinsing. Immediately after the boiling step. Copyright © 2004 AOCS Press . This step is used in applications in which the extract is extremely heat labile. In the boiling and rinsing steps. Once the extraction process is completed. Three-step extraction procedure. Extended drying. Excessive drying may oxidize the extract. The Soxtec Avanti stores the evaporated solvent in a common collection tank for reuse. causing weight changes and erroneous readings. should be avoided because it can cause oxidation of the fat extract and falsely high results. the condensate is redirected out of the condenser. solvent flow is blocked from returning to the extraction cup and flows out tube (a) into a collection tank (not shown). 70–90 mL of solvent is used. This step is usually 10–20 min longer than the boiling step to ensure complete extraction. The Foss-Tecator Soxtec Avanti automated extraction system is based on the Randall modification of the Soxhlet technique. using an internal valve. solvent is refluxed within the condenser. Typical extraction times range from 20 to 40 min depending on the solvent and sample characteristics. The condensed solvent continues to boil and evaporate and. cup predrying step. The Soxtec Avanti offers an optional fourth.e. A plug of defatted cotton is frequently placed on top of the sample to keep it in the thimble during extraction. Postextraction. concentrating the extract. 2. The last step in the crude fat extraction process is evaporation/solvent recovery. the rinsing step begins. especially at higher temperatures. i. residual traces of the extractable material are flushed out of the sample and are retained in the extraction cup. the extraction cups are raised a few millimeters off the heating surface allowing radiant heat to complete the drying cycle. During evaporation. Extraction cups are cooled completely to room temperature in a desiccator before final weights are taken. This usually requires 7–10 min depending on the solvent. To ensure optimal extraction. the level of boiling solvent must be higher than the sample in the thimble. With the Soxtec Avanti system. the cups are taken off the Soxtec and placed in a drying oven at 103°C for 30 min to drive off any moisture or solvent residuals. The sample is raised and suspended over the boiling solvent.

the most significant of these is the specific solvent that is being used. aromatic. and general Soxtec operating conditions is important. For crude fat extractions. a sample extraction will have somewhat different fat yields depending on the solvent. The versatility of the Soxhlet/Soxtec extraction method allows for the use of various classes of organic solvents.1 mg (0. i. As mentioned earlier. In these instances. it can act as a physical barrier preventing dissolution of the fat into the solvent. % Fat = (W2 – W1)/W3 × 100 where W1 = weight of the extraction cup W2 = weight of the extraction cup + extract W3 = weight of the sample All weights should be recorded to 0. Keeping in mind the empirical nature of the analysis. water in the sample can contribute to error in two ways. are examples in which failing to predry the sample can have a marked effect on recovery. many laboratories do not predry samples. There are several factors that influence the extraction. Solvent Choices. in the interest of time and productivity. extraction timing. and the feedlot concentrate.0001g).e. The final result is calculated from the original sample weight and the weights of the extraction cup before and after the extraction.. These include ethers. it can also contribute to falsely high apparent fat recoveries by allowing water-soluble components such as urea or carbohydrates to be co-extracted with the fat. Due to the different solubility characteristics of various solvents. thus generating low fat recoveries. aliphatic. [%Recovery is defined as (%crude fat from the undried sample/% crude fat from the dried sample) × 100]. Note that this is also dependent on the solvent used. the influence of sample preparation. Nevertheless. as well as alcohols. and chlorinated hydrocarbons. Moisture in the samples ranges from 5 to 25%. Optimizing the Extraction Process The Soxtec/Soxhlet extraction method for crude fat relies on separating sample components on the basis of physical and chemical (solubility) properties.Calculation of Results. undried samples. consistency with all aspects of the procedure is strongly recommended. diethyl ether and petroleum ether are most commonly used. An often overlooked aspect of a fat extraction method is the predrying of the sample. which contains urea. Figure 2. which contain molasses. Samples such as the texturized feeds. Some samples show a “water effect” more than others. Unfortunately.4 illustrates apparent fat recovery on dried vs. The peroxide-forming Copyright © 2004 AOCS Press . Crude fat by solvent extraction is a gravimetric method. the error potential for each type of sample should be investigated fully by carrying out extractions both with and without predrying.

to that of diethyl ether in terms of crude fat recovery. This is consistent with the use of petroleum ether in the AOAC method 991. Extraction Times. In Soxtec extraction procedures.9925. nature of diethyl ether causes it to be a less than desirable choice for routine use in the laboratory. Commonly. This can cause a change in the properties of petroleum ether because its more volatile components may be driven off. 2. petroleum ether is directly substituted. From these results. The results are shown in Table 2. undried samples with moisture content ranging from 5 to 25%.2. An experiment was undertaken (3. The objective was to find a solvent that is safer and has recovery statistically equivalent to diethyl ether. the extraction will likely not adequately recover the fat in the sample. are not true ethers but mixtures of aliphatic hydrocarbons and can be purchased in various formulations and boiling point ranges. hexanes.36 (7) for crude fat in meat and meat products. petroleum ether. However.4. Most method development protocols will define the extraction times at which the results closely match Copyright © 2004 AOCS Press . Considering the innate variability of petroleum ether and the relative lower recovery of plant-based lipids. M edica ted Med icate d Go at Fe ed Fat S upple men t Textu rized F (Mola eed sses ) Mixe d Bir d Se ed Corn Silag e Dehy drate d Alfa lfa % Recovery Hexanes FIG. the timing for the boiling and the rinsing steps is important. Further complicating the “pet ether” issue is that solvents are often recycled and reused in the Soxtec. It should also be noted that for meat and bone meal. This can cause a change or drift in the fat results. If the boiling or rinsing step is too short. Apparent fat recovery from dried vs.Undried vs. and pentane. it is not a suitable substitution for diethyl ether. Med icate d Meat Meal/ Hulls Mixtu re Feed lot C onc. This has caused many laboratories to look for an alternative solvent.4) to compare the recovery of three common solvents. petroleum ethers or ligroin. Dried Diethyl ether Corn High Oil Broil er Sta rter Swin e Fe ed Calf Feed . petroleum ether also yields a good recovery. (Urea ) Calf Start er. it can be seen that hexanes yield a recovery equivalent to that of diethyl ether with an R2 of 0.

The automation of the Soxtec offers consistent extraction timing for each batch of samples. <20°C at 2 L/min. weight. The purpose of this exercise was to determine whether the method can tolerate minor variations in the procedure that might be encountered in the laboratory. For the extraction of crude fat. sample weight of 1 g is described as variable “E” and 3 g as “e. several variables or factors are identified and are selectively used in a defined protocol or matrix. sample handling. Robustness of a Method.9880 those obtained by classical Soxhlet methods using suitable reference materials.54 3. it will usually cause slow reflux rates and can result in low fat recovery.3. In areas in which cold tap water is seasonal or not obtainable or in which water conservation is an issue.2 Crude Fat Recovery of Four Common Solvents Diethyl ether Sample Alfalfa hay Beet pulp Meat/bone meal Cattle protein supplement Corn Average R2 Petroleum ether Hexanes Pentane (% Crude fat) 1.06 3. In this model. This is described in Table 2.65 3.54 2. refrigerated circulating water baths are a convenient way to regulate the temperature and flow. Ideally.47 0. should be used so that the condensers feel cool to the touch.76 — 1.00 3. In some cases. For example.59 3. Extraction Temperature.10 3.30 10.TABLE 2. Condenser Temperature.” Using the schedule in Table 2. If the water temperature is too warm.67 0. warm condensers can cause the loss of solvent during the boiling and rinsing stages.16 3.36 0.24 10.88 3. This helps ensure optimal condensation or reflex rates.9925 0. Using the statistical model described by Youden (8) the “ruggedness” of the extraction method was evaluated. and solvent types were defined as variables.3.52 3.19 10.25 10.9878 1.69 2. this is typically 3–5 drops/s coming off the condenser. The temperature of the extraction system should be set to the recommendations provided by the manufacturer. and implementing the variables in this manner allows the evaluation of the ruggedness of the method while minimizing the number of determinations.97 0.29 0. The temperature of the condenser cooling water plays an important role in establishing the condensation or reflux rate of the solvent.40 2. extraction parameters.45 0. Cold tap water. Copyright © 2004 AOCS Press .00 0.

103°C. 2 h B Boil time. Cereal Grains. 2/s 1 2 A b c d e F G V a B C d e F g W a B c d E f G X a b C D e f G Y a b c D E F g Z a b c d e f g Sample predry 103°C. or >10% of other water-soluble Copyright © 2004 AOCS Press . Values for the individual factors are averages from all results in which this variable was used. Crude Fat in Feed. 6/s The results from three different samples. 3 g Cup dry. 40 min Petroleum ether Rinse time. lactic acid. A glass stir rod is used to thoroughly mix the sample and sand together. cattle. The glass rod is left in the sample during drying and subsequently used to break up any sample/sand clumps before extraction. 60 min Sample weight. 20 min C Diethyl ether D Rinse time. A 1.4. 1 g F Cup dry. A 2-g sample of homogenized meat is mixed with acid-washed sand and dried at 125°C for 1 h. swine. The sand is used to maintain porosity of the sample after drying for optimal solvent penetration. The sand is added directly to the thimble at approximately double the sample weight.05 and 2003. 4 h Solvent drip rate. 103°C. 2 h G Solvent drip rate. 30 min E Sample weight. >15% glycerol. Common Applications Crude Fat in Meat and Meat Products: AOAC Method 991. the most significant variable is the choice of solvent. or amino salts. and mixed feeds. Considering the averages of the differences.36.06. A petroleum ether Soxtec extraction is then performed with 25-min boil and 35-min rinse periods.TABLE 2. 4 h Boil time. thus making the method robust. from three different laboratories are seen in Table 2. and Forages: AOAC Methods 5-g ground sample is weighed into an extraction thimble.3 Variables Used to Determine Method Robustness Combination or determination number Factor value 3 4 5 6 7 8 A or a A A A B or b B B b C or c C c C D or d D D d E or e E e E F or f F f f G or g G g g Observed result S T U The chosen variables (factors): A Sample predry 103°C. If the sample contains quantities of water-soluble components such as >5% carbohydrates. Other variables in the method do not contribute to a significant variation. Table 2.5 compares recovery and repeatability of classical Soxhlet to Soxtec (6) showing that the Soxtec offers better precision with the same results as the classical Soxhlet method.

4 Evaluating the Robustness of the Crude Fat Method Using Cattle.61 11.35 2.02 2.23 10.29 0.60 2.64 11.35 10.70 2.96 –0.47 11.98 11.06 2.22 10.50 10.03 10.17 2.32 10.35 –0.78 9.13 11.00 11.29 –0.66 11.22 11.23 0.05 11.28 10.74 2.54 –0.60 0.38 2.34 Swine Mixed Average 10.86 10.81 2.60 11.68 0.60 9.41 2.43 2.81 10.49 10.28 10.00 components.02 10.20 2.62 10.45 10.60 –0.73 11.00 2.24 10.53 10. The Soxtec method was compared with the AOAC Soxhlet method on data from 90 AAFCO check samples.84 –0.35 2. Regression analysis generated an R2 correlation coefficient of 0.47 0.29 2. Swine.76 11.79 11.82 11.46 2.79 10.38 3.41 2.59 11.02 2.36 –0.25 2.58 0.61 11.11 3.31 11.73 –0.45 2.25 10.72 2.60 10.82 2.77 –0.01 2.66 –0.53 –0.46 2.27 11.28 0.29 10.43 2.44 0.99 0.06 10.57 2.73 2.02 10.22 10.53 11.70 10.52 2.11 2.59 11.98 –0. Extraction is performed with either diethyl ether or hexanes using a 20-min boil and 40-min rinse cycle.57 11.45 0.28 11.45 11.22 11.39 0.75 10.30 10.04 0.27 0.53 0.05 –0.49 11.56 –0.99 11.03 0.27 10.13 2.05 –0.16 10.59 9.35 10.40 2.75 –0.50 10.74 11.94 11.38 2.11 11.09 11.24 0.47 10.70 2.39 10.70 2.19 2.63 11.64 2.80 10.17 2.28 0.02 –0.34 0.17 0.22 2.01 10.61 –0. slope Copyright © 2004 AOCS Press .55 1.27 10.34 11.43 11.46 4.04 0.14 0.01 11.9946.02 11.43 10.24 10.87 11.43 11.03 11.52 2.91 11.23 10.33 0.79 10.12 2.16 11.26 2.73 0.15 10.12 11.60 2.16 10.17 2.20 10.87 11.27 0.33 –0.35 –0.02 10.60 9.57 10.17 0.64 0.06 2.24 11.49 2. and Mixed Feeds from Three Different Laboratories Laboratory 1 Laboratory 2 Laboratory 3 (% crude fat) Feed type Cattle Swine Mixed Cattle Swine Mixed Cattle S T U V W X Y Z Predry 2h 4h Difference Boil time 20 min 40 min Difference Ether Diethyl Petroleum Difference Rinse time 30 min 60 min Difference Sample wt 1g 3g Difference Cup dry 30 m 2 hr Difference Drop rate 2/s 6/s Difference 11.31 0.98 –0.06 2.76 0.00 2.08 –0.70 0. the sample is washed with 5 aliquots of 20 mL deionized water.80 11.91 10.28 2.31 –0.97 10.08 0.05 10.29 2.02 11.10 2.91 11.20 0.67 10.59 11.24 10.70 11.66 0.26 0.82 11.29 2.15 11.37 10.80 –0.32 11.94 10.39 0.20 0.30 11.62 –0.10 2.66 3.63 0.73 2.02 10.48 –0.48 –0.00 2.55 –0.84 10.15 3.04 0.97 10.76 10.00 10.02 11.88 10.36 0.05 11.81 11.14 11.13 2.93 2.06 11.57 11.75 0.61 11.46 –0.00 11. The sample is dried at 102°C for 2 h.69 11.55 11.51 10.10 11.84 11.04 2.76 10.03 2.99 2.36 2.46 0.69 10.75 0.79 11.TABLE 2.04 10.

4).34 27. Application Sub Notes (ASN) are available from Foss Tecator.09 2. sludge.25 1. and minerals. which takes 8–24 h.51 33. thus eliminating any sample transfer errors.10). 2-h drying time.TABLE 2. SW 846.10 26.00062 and a y-intercept of 0.6 28. carbohydrates. 30-min drying time. and hazardous waste samples (9.137. The extract/solvent mixture is further concentrated and the final analysis performed by GC or GC/MS techniques. the methods appear to be comparable (3.98 34.64 2. dried and then extracted.55 1.52 2.32 2. An acid hydrolysis before the solvent extraction step is needed to “free” the fat in the sample.83 4.44 1.57 26.21 1.6. The sample is then rinsed.35 28.74 3. Environmental EPA 3541.5 Average Recoveries and Relative Standard Deviations (RSD) for Meat Samples with Petroleum Ether Extraction Average recovery RSDr Average recovery Soxhleta Meat sample 1 2 3 4 5 6 Average RSDr Soxtecb (%) 4. Soxtec extraction methods were found to be suitable for use in industrial applications.20 25. In samples that are baked. A summary of some of these applications appears in Table 2. 2. the Soxtec extraction procedure is used widely in environmental laboratories to extract organic compounds such as pesticides or PCB from soils.01 1. Industrial Applications. extruded. On the basis of these data. The Soxtec method with a 2-h extraction replaces the traditional Soxhlet. the Soxtec is used as a sample preparation device. Total Fat.5) for acid hydrolysis from Foss Tecator enables the acid hydrolysis step and extraction step to occur in the same sample vessel.95 2.18 2. The process is stopped during the evaporation/solvent recovery step while there is still 15–20 mL of solvent (containing the semivolatiles) left in the cup.63 8.29 27. or with some commercial processing.75 5. of 1. Typically 1–2 g of sample is boiled with strong hydrochloric acid solutions.21 34. 55-min extraction. Copyright © 2004 AOCS Press .81 26. The sample is weighed directly into a cellulose or fritted glass thimble and placed in the Soxtec where the extraction of semivolatile organics is done using a mixture of hexane and acetone (1:1). The SoxCapTM (Fig.91 aSoxhlet: bSoxtec: 4-h extraction. sediments. In these applications. Of note. making it available for solvent extraction. the fat becomes bound to other components in the sample such as proteins.95 34.

6 Summary of Soxtec Extraction Methods Used in Industrial Applicationsa ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN ASN aApplication 3516 3602 3603 3604 3605 3606 3607 3608 3611 3612 3613 3614 3615 3616 3617 3618 3619 3622 3700 Extraction of aromatic hydrocarbons in soil Extraction of resins from paper pulp Extraction of finish from textiles Extraction of starch containing finish from textiles Extraction of surfactant from detergents Extraction of paraffin from detergent Extractable matter in leather Extraction of petroleum source rock Extraction of explosives and propellants Extraction of plastics and polymers Extraction of rubber and rubber compounds Extraction of finish oils from textiles and synthetic fibers Extraction of migration components in plastic packaging Extraction of organic dyestuffs Extraction of leather Extraction of core material in petroleum exploration Extraction of tobacco Extraction of solubles in paper pulp Extraction of fecal fat sub notes (ASN) are available from Foss Tecator.FIG. 2.5. TABLE 2. Copyright © 2004 AOCS Press . The 2047 SoxCapTM hydrolysis system.

high-fat seeds • A two-phase extraction protocol is used: After the first extraction boil and rinse. • Do replicate analysis to generate reportable results. • Diethyl ether can be purchased with stabilizers to minimize the formation of peroxides. Moist samples • Mix sample with sand and predry. • Wear gloves during handling of thimbles and cups to avoid errors. Solvent overboiling • Use 3–5 boiling beads in the extraction cups. Incomplete fat recovery. The results are added together. • Decrease the temperature setting on the extractor. Such ether should be used with the label guidelines. • Test strips are available to check for peroxide formation in diethyl ether.7 summarizes some common approaches to aid the analyst in extracting crude fat from these types of samples.TABLE 2. • Weigh cups at room temperature. Table 2. Difficult Samples. Nonhomogeneous samples • Optimize sample preparation step. Weighing errors will result from warm cups. Evaluating the performance of the extraction process is normally achieved by running a reference or check sample. General practice • Place a plug of defatted cotton on top of the sample to ensure that the sample is retained in the thimble.7 Common Approaches to Help Extract Crude Fat from Difficult Samples High-fat samples that melt during drying • Place the thimble containing the sample into a preweighed extraction cup. Samples that represent special challenges and handling are frequently encountered. Semisolid • Depending on the nature of the sample. Quality Control. Many commercial check sam- Copyright © 2004 AOCS Press . • When using recovered petroleum ether. ground with a mortar and pestle and returned to the thimble for a secondary extraction. • Use larger sample size to obtain a representative sample. mix with either sand and dry or mix with sodium sulfate. the samples are removed from the thimbles. • Mix sample with equal weight of sodium sulfate to bind water. Any sample that melts out of the thimble will be retained in the cup. This allows for better solvent flow through the sample. supplement with fresh ether to help maintain desired boiling point range. Sample becomes impacted during extraction • Mix equal volumes of acid-washed sand or Celite and sample in the extraction thimble. Low-fat samples • Use larger sample weights.

Kansas State Board of Agriculture. DC. U.01. (1992) Soxtec Fat Analyzer for Determination of Total Fat in Meat: Collaborative Study. 1997. and Gildemeister. Washington.36.L. Official Methods of Analysis of AOAC International..S. Physical/Chemical Methods (1996) SW-846.5. (2003) Crude Fat. Hexanes Extraction. 4. JAOAC Int. Thiex. AOAC International. MD. in Feed. 16th ed. Diethyl Ether Extraction. Las Vegas. 7. & Forage (Randall/ Soxtec/Submersion Method): A Collaborative Study. Anderson.L. Italy. Thiex. Foster. V. Cereal Grain.. (1975) Statistical Manual of the AOAC. section 39. 86: 888–898. AOAC International. Chapter 39. (Beckert. & Forage (Randall/Soxtec/Submersion Method): A Collaborative Study. and Steiner. 139. 6th edn. Copyright © 2004 AOCS Press .. section 4. E. 6. MD. S. B.08. U. N. Rome. M. EPA. 9. W. 75: 288–292. References 1. Gaithersburg. 25. E.. Environmental Protection Agency. such as the SoxtecTM Avanti. Lopez-Avila. N. 10.1. They offer convenient and useful tools with which to improve productivity in the laboratory. Results should be compared only to those from similar instruments running the same extraction procedure and sample protocol. Test Methods for Evaluation of Solid Waste. Codex Alimentarius Commission (1986) Procedural Manual. Official Methods of Analysis of AOAC International.. JAOAC Int. S. Fat (Crude) in Meats and Meat Products. 3. Food and Agricultural Organization.H. Office of Solid Waste. JAOAC 57: 1165–1168. (1974) Improved Method for Fat and Oil Analysis by a New Process of Extraction. Project Officer) (1991) Development of a Soxtec Extraction Procedure for Extracting Organic Compounds from Soils and Sediments. 86: 899–908. 2.. in Feed.. S.. Fat (Crude) or Ether Extract in Animal Feed. Method 3541. AOAC Official Method 920. Chapter 4. 5. 16th ed. Environmental Monitoring Systems Laboratory.. 8. Conclusion The long history of solvent extraction has led to automated Soxhlet extraction systems.J. 3. B. Cereal Grain. and Gildemeister. p. (2003) Crude Fat. Randall. W. Youden. Modern instrumentation provides application flexibility and improved economy as well as enhancement of the precision and recovery of the extraction.. EPA600/X91/140. Association of the Official Analytical Chemists. AOAC Official Method 991. Gaithersburg.S.E. and Gonzales.39. JAOAC Int. p.ple services are available for different sample types. Anderson. VA. Arlington. p. 1997.