Residue extraction procedures

Commercial Immunoassay Kits. Most commercially available immunoassay kits have been developed for determination of pesticides in water. At a minimum, appropriate residue extraction procedures must be developed before these kits can be applied to pesticide residue analysis of foods. Encouraging results have been obtained in preliminary FDA evaluations of several kits (12). For example, in studies of a kit for detection of triazine herbicides, a typical residue extraction solvent (acetonitrile) was used and then diluted with water to levels tolerated by the immunoassay. Visual comparison of color developed for extract, standard, and reagent blank was made for qualitative analysis. Spectrophotometric readings of the color were made for quantitative analysis. 

Solvent extraction in batch or continuous systems is used to recover most of the residual oil from the presscake. Heptane, hexane, or a mixture of these solvents is used to recover the oil. The solvent-extracted presscake is steam stripped to recover solvent and a residual meal known as castor pomace, containing 1% residual oil. The solvent extracted oil is also processed for solvent recovery (qv). The oil from the extraction procedure is darker than the mechanically pressed oil and has a higher free fatty acid content. It is sometimes referred to as a No. 3 castor oil and is used for blending with higher quaUty oils that are well above No. 1 specifications. 

The ability to identify and quantify cyanobacterial toxins in animal and human clinical material following (suspected) intoxications or illnesses associated with contact with toxic cyanobacteria is an increasing requirement. The recoveries of anatoxin-a from animal stomach material and of microcystins from sheep rumen contents are relatively straightforward. However, the recovery of microcystin from liver and tissue samples cannot be expected to be complete without the application of proteolytic digestion and extraction procedures. This is likely because microcystins bind covalently to a cysteine residue in protein phosphatase. Unless an effective procedure is applied for the extraction of covalently bound microcystins (and nodiilarins), then a negative result in analysis cannot be taken to indicate the absence of toxins in clinical specimens. Furthermore, any positive result may be an underestimate of the true amount of microcystin in the material and would only represent free toxin, not bound to the protein phosphatases. Optimized procedures for the extraction of bound microcystins and nodiilarins from organ and tissue samples are needed. 

Pyronin Y [3,6-bis(dimethylamino)xanthylium chloride] [92-32-0] M 302.8, m 250-260, Cl 45005, Xmax 522nm, pKesi Commercial material contained a large quantity of zinc. Purified by dissolving Ig in 50mL of hot water containing 5g NaEDTA. Cooled to 0, filtered, evapd to dryness and the residue extracted with EtOH. The soln was evaporated to 5-lOmL, filtered, and the dye pptd by addition of excess dry diethyl ether. It was centrifuged and the crystals were washed with dry ether. The procedure was repeated, then the product was dissolved in CHCI3, filtered and evapd. The dye was stored in a vacuum. 

The usual extraction procedure is to roast the crushed ore, or vanadium residue, with NaCl or Na2C03 at 850°C. This produces sodium vanadate, NaV03, which is leached out with water. Acidification with sulfuric acid to pH 2-3 precipitates red cake , a polyvanadate which, on fusing at 700°C, gives a black, technical grade vanadium pentoxide. Reduction is then necessary to obtain the metal, but, since about 80% of vanadium produced is used as an additive to steel, it is usual to effect the reduction in an electric furnace in the presence of iron or iron ore to produce ferrovanadium, which can then be used without further refinement. Carbon was formerly used as the reductant, but it is difficult to avoid the formation of an intractable carbide, and so it has been superseded by aluminium or, more commonly, ferrosilicon (p. 330) in which case lime is also added to remove the silica as a slag of calcium silicate. If pure vanadium metal is required it can... 

Complex ( + )-(FeS)-3 is obtained by extraction of the diastereomeric mixture with 50 mL of pentane in portions at r.t. The specific rotation of the solid product obtained from the first fraction is [a]jJ6 +50 (o = 0.001. benzene). The rotation of the remaining mixture becomes more negative as the (+)-isomer is removed. After concentration of the combined fractions, the resulting material, enriched in the ( + )-isomcr, is further purified by extraction with 10 mL of pentane in portions. Repetition of the pentane extraction procedure affords ( + )-(FeS)-3 yield-. 750 mg (30%) mp 120"C (dee) [a] +72 (c = 0.001, benzene). The residue, enriched in (—)-(FeR)-3 is extracted for 1 h with boiling pentane. After separation from the extraction liquor, a yellow-orange solid is obtained yield I -35 g (50%) mp 120°C (dec) — 120... 

For pesticide extraction procedures pertaining to food samples, refer to U.S. government manuals on pesticide residue analysis. 

Using established extraction and cleanup methods, followed by GC/FPD and GC/thermionic detection, Carey et al. (1979) obtained detection limits in the ppb range and recoveries of 80-110% in soil and 70-100% in plant tissue. Good sensitivity and recovery were maintained in a simplified extraction procedure of sediments followed by GC/FPD analysis (Belisle and Swineford 1988). Bound methyl parathion residues that were not extracted with the usual methods were extracted using supercritical methanol by Capriel et al. (1986). They were able to remove 38% of the methyl parathion residues bound to soil, but 34% remained unextractable, and 28% could not be accounted for. 

Extraction procedures must be adjusted when separated anthocyanins will be tested in biological studies. We have found that the types of acids used for anthocyanin extraction as well as their residual concentrations in the final extract may affect the results obtained from biological tests. The growth inhibitory effect of anthocyanins on HT29 (human colonic cancer) cells may be overestimated if the residual acid in the extract exerts a toxic effect on the cells. Acetic acid residues in anthocyanin extracts showed less toxicity to HT29 cells than hydrochloric acid when samples were prepared under the same extraction procedure and subjected to the same tests on HT29 cells. In addition, the procedure to remove acids affected the acid residual concentration as well in final anthocyanin extracts, with lyophilization being more successful than rotary evaporation. 

Once several target methods employing, e.g., LC/MS/MS techniques have been combined, a multi-residue method will evolve which includes the DEC S19 extraction procedures in combination with the generally applicable GPC cleanup and requires automatic multiple injections to circumvent the limitations of the limited HPLC peak capacity and the target-specific MS/MS methods. 

The definitions of method detection and quantification limits should be reliable and applicable to a variety of extraction procedures and analytical methods. The issue is of particular importance to the US Environmental Protection Agency (EPA) and also pesticide regulatory and health agencies around the world in risk assessment. The critical question central to risk assessment is assessing the risk posed to a human being from the consumption of foods treated with pesticides, when the amount of the residue present in the food product is reported nondetect (ND) or no detectable residues . 

Validation of true extraction efficiency normally requires the identification and quantitation of field-applied radiolabeled analyte(s), including resulting metabolites and all other degradation products. The manufacturer of a new pesticide has to perform such experiments and is able to determine the extraction efficiency of aged residues. Without any identification of residue components the calculation of the ratio between extracted radioactivity and total radioactivity inside the sample before extraction gives a first impression of the extraction efficiency of solvents. At best, this ratio is nearly 1 (i.e., a traceability of about 100%) and no further information is required. Such an efficient extraction solvent may serve as a reference solvent for any comparison with other extraction procedures. 

Both multi-residue methods are presented in several parts, which separate general considerations from procedures for extraction, cleanup and determination/ confirmation. Whereas in EN 12393 several extraction and cleanup steps cannot be combined arbitrarily, the modular concept is utilized to a greater extent in EN 1528. In the latter standard, there is no limitation to the combination of several extraction procedures, mostly designed for different commodities, e.g., milk, butter, cheese, meat or fish, with different cleanup steps. Both standards, EN 1528 and EN 12393, do not specify fixed GC conditions for the determination and confirmation. All types of GC instruments and columns, temperature programs and detectors can be used, if suitable. 

Specifically for triazines in water, multi-residue methods incorporating SPE and LC/MS/MS will soon be available that are capable of measuring numerous parent compounds and all their relevant degradates (including the hydroxytriazines) in one analysis. Continued increases in liquid chromatography/atmospheric pressure ionization tandem mass spectrometry (LC/API-MS/MS) sensitivity will lead to methods requiring no aqueous sample preparation at all, and portions of water samples will be injected directly into the LC column. The use of SPE and GC or LC coupled with MS and MS/MS systems will also be applied routinely to the analysis of more complex sample matrices such as soil and crop and animal tissues. However, the analyte(s) must first be removed from the sample matrix, and additional research is needed to develop more efficient extraction procedures. Increased selectivity during extraction also simplifies the sample purification requirements prior to injection. Certainly, miniaturization of all aspects of the analysis (sample extraction, purification, and instrumentation) will continue, and some of this may involve SEE, subcritical and microwave extraction, sonication, others or even combinations of these techniques for the initial isolation of the analyte(s) from the bulk of the sample matrix. 

Transfer the residue prepared as in Section 6.1.1 into a 300-nL separatory funnel with 25 mL of phosphate buffer solution (0.1 M, pH 7.4). Add 10 mL of saturated aqueous sodium chloride and 50 mL of 0.5 M sodium hydrogen carbonate to the funnel and shake the funnel vigorously for 1 min. Add 70 mL of ethyl acetate to wash the aqueous layer to the funnel, shake, separate, and discard the ethyl acetate layer. Repeat this extraction procedure three times. Add 2 mL of phosphoric acid and 20 mL of an acetate buffer solution (0.1 M, pH 4) to the aqueous layer and extract the mixmre with 50 mL of ethyl acetate three times. Combine the extracts and filter into a 500-mL round-bottom flask through 60 g of anhydrous sodium sulfate supported by a plug of cotton wool in a funnel. Concentrate the filtrate to dryness under reduced pressure. 

The extraction efficiencies using a blender and a shaker were compared and both methods gave similar results. A corn sample treated with radiolabeled carfentrazone-ethyl and collected from a metabolism study was used for comparison. Multiple samples can be extracted simultaneously if extraction is performed by shaking. In addition, since the extraction procedures in the residue study closely followed the extraction scheme in the metabolism study, the resulting extraction efficiencies from both studies were almost identical. 

Transfer the filtrate from Section 6.1.1 or 6.1.2 to a 500-mL separatory funnel and add 150 mL of 5% aqueous sodium chloride solution. Rinse the filter flask from the extraction procedure with two 40-mL portions of dichloromethane. Add both 40-mL rinses to the separatory funnel. Partition the residue into the dichloromethane. Filter the dichloromethane extract through a 10-cm filter funnel containing ca 50 g of anhydrous sodium sulfate supported on a plug of glass wool. Collect the dichloromethane in a 500-mL round-bottom flask. Repeat the partition and filtration steps with an additional 60 mL of dichloromethane. Rinse the sodium sulfate filter cake with 20 mL of dichloromethane and combine the partition and rinse solvents. Concentrate the combined dichloromethane solvents to dryness in a rotary evaporator under reduced pressure at <40 °C. 

Two different extraction procedures were developed, a manual and an automated method. A work Aow diagram of this residue analytical method is presented in... 

The sample extraction procedures are similar to those for the Multi-residue analysis method (see Section 3.3.1). 

The fundamental issue is to describe how much of the residue can be characterized accurately and whether an accounting of the applied mass of pesticide can be maintained throughout the course of the experiment. A series of environmental fate studies is required for pesticide registration in order to characterize the degradation pathways and formation and decline patterns of each major degradate. These studies are typically conducted in the laboratory under controlled conditions, applying radiolabeled pesticides to evaluate the extraction efficiency of various procedures. When standard extraction methods fail to release a significant amount of the applied radioactivity, more efficient and exhaustive extraction procedures are tried in a stepwise fashion... 

Some agrochemicals bind strongly to the soil component as bound residues, which cannot be extracted without vigorous extraction procedures. In this case, an acidic (e.g., hydrochloric acid, sulfuric acid) or alkaline solution (e.g., sodium hydroxide, potassium hydroxide) can be used as an extraction solvent, and also heating may be effective in improving the extraction of the residues. Analytical procedures after the extraction are the same as above, but a filtration procedure may be troublesome in some of these situations. However, these procedures are rare exceptions or are needed for specific chemicals that are stable under such harsh extraction conditions. 

The residue analysis of CPMA and CPMF in vegetables and fruits is carried out as follows. A 20-g amount of the sample is homogenized for 3 min with 100 mL of acetone and filtered. The extraction procedure is repeated once with 50 mL of acetone and filtered. The combined filtrate is concentrated to 10 mL at 50 °C and, after addition of 0.2 mL of triethylamine to the concentrate, the reaction mixture is allowed to stand for 30 min at 50 °C. The mixture is applied to an Extrelut column (Extrelut 14-g). CPF from CPMA via CPMF is eluted with 50 ml of diethyl ether after washing the... 

Extraction Procedure. A flow chart of the isolation and identification procedure is presented in Figure 1. Field-grown rye ( Abruzzi, harvested at early flowering stage on March 24, 1983, from the Central Crops Research Station, Clayton, NC) was air-dried for 7 days. The tissue (150 g) was extracted with 3 L of distilled water for 10 hr with agitation. The extract was filtered through cheesecloth and then centrifuged at 28,000 x g for 20 min. The supernatant was reduced in volume to 300 ml jim vacuo at 50°C. Sixty ml of the concentrated aqueous extract was dried in vacuo, the residue extracted with 20 ml of methanol and filtered. The metha-nolic extract was stored at 0°C until use. 

The extraction procedure described by Aepli, Munter, and Gall (1) may be used, if preferred, as an alternate extraction procedure for the residue after ether extraction. 

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