Extraction protocol

One can find a number of methods of quinolizidine alkaloid extraction within existing literature. The most frequently applied methods are extraction by alcohols (50% methanol or 96% ethanol, or these alkaloids with a 1% addition of glacial acetic acid). 

Wysocka and Przybyl presented an efficient extraction method of quinolizidine alkaloids. The authors themselves developed and investigated this method, in which includes the following steps of extraction  

This method has since been modified for efficiency, resulting in the following steps  

Degreasing of the ground seeds with petroleum ether 

Mixing of the macerated meal with diatomaceous earth 

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Hey That really wasn t a lot of work. Just a lot of talk on Strike s part. All one did was mix an oil with some acid, added water and isolated. One gets some pure propenylbenzene without distillation. Done on a massive scale, this is a cheap method for getting lots of small concentration allylbenzene compounds out of complex oil mixes. And since Strike blew so much dough on this glorified extraction protocol, someone better damn well use it (In an academic lab of course). 

Nucleic acid extraction protocols using guanidine hydrochloride, sodium sarco-syl, and ethanol have been developed to quantify viral RNA by bDNA in lymph node tissue, liver tissue, and peripheral blood monocytes (Wilber and Urdea, 1995). 

The protocol involving NaOAc-HOAc at pH 5 was first proposed and used by Jackson (1958) to remove carbonates from calcareous soils to analyze soil cation exchange characteristics (Grossman and Millet, 1961). Other researchers used HOAc for the extraction of metals from sediments and soils (Nissenbaum, 1972 Mclaren and Crawford, 1973). Tessier et al. (1979) first used the NaOAc-HOAc solution at pH 5 to dissolve the carbonate fraction from sediments. Since then, the NaOAc-HOAc buffer has been widely used as a specific extractant for the carbonate phase in various media (Tessier et al., 1979 Hickey and Kittrick, 1984 Rapin et al., 1986 Mahan et al., 1987 Han et al., 1992 Clevenger, 1990 Banin et al., 1990). Despite its widespread use, this step is not free from difficulties, and further optimization is required in its application. Questions arise with regard to this step in the elemental extraction from noncalcareous soils, the dissolution capacity and dissolution rates imposed by the buffer at various pHs, and the possibility that different carbonate minerals may require different extraction protocols (Grossman and Millet, 1961 Tessier et al., 1979). 

It is difficult, at present, to get a clear idea of the efficiency of the various plant PHA extraction protocols. In a patent application by Metabolix [74], it was reported that the efficiency of purifying poly(3HB) which was mixed with ground rapeseed was approximately 24% when using an oil-extraction step with hexane followed by refluxing in chloroform for 22 h and evaporating the chloroform to recover poly(3HB). The co-extraction of oil with a PHA contain-... 

We performed a study to apply the a-CGH technique to test the quality of DNA extracted from FFPE tissues by different methods, using a nonheating protocol, a heat-induced extraction protocol, based on AR as applied to IHC, and comparing the findings to extracts from paired fresh frozen tissue samples (unpublished data). The study was conducted in two stages. 

In conclusion, the data described demonstrate the reproducible quality of DNA samples extracted by using a heat-induced retrieval protocol from FFPE tissue sections, based on careful comparison of a-CGH analysis data. This simple and effective DNA extraction protocol may provide an alternative technique for DNA analysis for CGH as well as for other methods of DNA... 

TABLE 19.1 Protein Extraction Protocols from FFPE Tissues Documented in the Literature... 

Phillips N, Smith CM, Morden CW (2001) An effective DNA extraction protocol for brown algae. Phycol Res 49 97-102... 

The interlaboratory results obtained from the analysis of defined standard solutions, but also from the analysis of sediment extracts prepared either by the coordinator of the study or by the participants themselves, also provide a measure of the variation between laboratories. The results show that the interlaboratory reproducibility ranges from 6.5% for the defined dioxin sample to 27.9% for the sediment sample extracted by the participants themselves. As was mentioned before, the reproducibility for this last sample is relatively high and most presumably due to the introduction of extra handlings (extraction and cleanup) to the total procedure. In addition, the fact that not all the participants had prior experience with the extraction protocol to be used could have added to the increase in variability of the process. Furthermore, the dilution factor was not dictated. This also introduces a certain degree of variation. For the reproducibility of the DR CALUX bioassay itself and not caused by differences in operating extraction conditions, the maximum variation between laboratories was observed to be 18.0%. The results for the sediment extract samples can also be used to estimate the method variability for extracts, that is, based on samples of unknown composition. Again, given the intra-as well as the interlaboratory variations observed in this study, it appears justified to conclude that the standard deviation of the means provides a reasonable estimate of the method variability, based on the overall aver-... 

Chlorophyll sample (1 to 100 mg/liter 2 to 200 (lM see UNITF4.2 for extraction protocols) dissolved in volatile oiganic solvent such as acetone, diethyl ether, or ethyl acetate (store in airtight glass vial)... 

Unlike many of the common extraction protocols used in organic chemistry the extraction of odorants from complex foods will result in seemingly unbreakable emulsions because of the fats and proteins they often contain therefore, shaking the phases is seldom recommended. 

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