Water-autoclave extraction

Water-autoclave extractions were also much more efficient than neutral EDTA extractions in recovering sorbed/fixed phenolic acids from plant tissues/residues (Blum et al. 1992). For example HPLC analysis of EDTA extracts for wheat stubble collected after a wheat harvest contained no detectable phenolic acid peaks. Water-autoclave extracts of this wheat stubble had 11 distinct peaks. Concentrations for ferulic acid, vanillic acid, p-coumaric acid, and p-hydroxybenzoic acid were 33, 22, 1034, and 47 p,g/g dry weight, respectively (Fig. 3.8). On the other hand water and... 

One hundred days after the addition of 1,000 p.g/g (approximately 5 timol/g soil) ferulic acid to sterile Cecil soil samples, water, neutral EDTA, and water-autoclave extractions recovered 28% (277 tig/g), 37% (373 tig/g), and 32% (322 p.g/g) of the ferulic acid added, respectively, from Cecil A soil samples (3.7% organic matter) and 17% (167 tig/g), 52% (524 tig/g), and 30% (304 p.g/g) of the ferulic acid added, respectively, from Cecil B soil samples (0.2% organic matter Blum et al. 1992). The recovery of femlic acid from soil by neutral EDTA extraction was thus more effective than the water-autoclave extraction. 

We suspect that both neutral EDTA and water-autoclave extraction procedures readily recovered the free phenolic acid fraction. So how effective were they in extracting reversibly sorbed phenolic acids (i.e., sorbed recovered by neutral EDTA or water-autoclave extraction minus free recovered by water extraction) For the 1,000 p.g/g added 723 p.g/g was sorbed by Cecil A and 833 p,g/g by Cecil B. For Cecil A samples, 13% (96 of 723 tig/g) and 6% (45 of 723 p-g/g) of the sorbed ferulic acid was recovered by the neutral EDTA and the water-autoclave procedures, respectively. Of this 96% (92 pg/g) and 55% (25 pg/g), respectively, was utilized by microorganisms over the 30 days (Blum et al. 1992). For Cecil B samples, 43% (357 of 833 pg/g) and 16% (137 of 833 pg/g) of the sorbed ferulic acid was recovered by the neutral EDTA and the water-autoclave procedures, respectively. Of this 100% (357 pg/g) and 97% (133 pg/g), respectively, was utilized by microorganisms over the 30 days. 

In conclusion, the water-autoclave extraction procedure when compared to the EDTA extraction procedure underestimated the total available ferulic acid in the soil by roughly 5% for Cecil A and 22% for Cecil B. In addition to the quantitative difference there also appeared to be a difference in the types of the sorbed ferulic acid recovered. The water-autoclave-procedure recovered some irreversibly sorbed phenolic acids from Cecil A soil since only 55% of the sorbed phenolic acid recovered was utilized by microbes. This difference should not be surprising since the physical and chemical processes of the two extraction procedures, i.e., chelation vs. 

All total phenolic acid values in ferulic acid equivalence were determined by water-autoclave extraction and the Folin Cicoltaeu s phenol reagent approximately biweekly during the 1992 and 1993 experimental periods. Mean total phenolic acid for the upper 2.5 cm of the soil in 1992 were as follows plots without cover crops (reference plots) 41 1 rg/g soil (mean standard error), crimson clover plots 55 1 JLg/g soil, subterranean clover plots 58 1 p,g/g soil, rye plots 50 1 irg/g soil, and wheat plots 45 1 rg/g soil (Fig. 3.14 Blum et al. 1997). In 1993 soil total phenolic acid values for the upper 2.5 cm of the soil were higher than in 1992 (Fig. 3.14 Blum et al. 1997). The values for the April glyphosate desiccated plots were plots without cover crops (reference plots) 62 2 t,g/g soil, crimson clover plots 83 4 jtg/g soil, subterranean clover plots 83 3 t,g/g soil. 

Pro 4 Total phenolic acid concentrations estimated by water-autoclave extraction and Folin Ciocalteu s phenol reagent are not absolute values and thus should not be viewed as such, they are relative values. The following suggest that these relative values for total phenolic acid content can actually be useful and meaningful ... 

A) Preparation of 1-Methyl-2-Picolinium Chloride 98 ml of cx-picoline is dissolved in 200 ml of methanol, cooled and 85 ml (at -68°C) of methyl chloride is added. The solution is charged to an autoclave, sealed and the nitrogen pressure of 300 psig is established. The mixture is heated at 120° to 130°C for 2 hours, cooled and opened. The resulting solution is then evaporated to dryness in vacuo, yielding a residue of 110 g. This residue is then dissolved in 50 ml of water and extracted with two 50 ml portions of ether. The aqueous phase is then diluted to 150 ml with water and an assay for ionic chloride is performed which indicates the presence of chloride ion equivalent to 721 mg/ml of 1-methyl-2-picolinium chloride. 

Reaction ofvinylacetylene with Te-KOH-DMSO triad A mixtnre of Te (25.9.), KOH (20 g), H2O (10.9 mL) and DMSO (100 mL) was heated (110°C) with vinylacetylene (31.7 g) in 1 L steel-rotating autoclave for 3 h. The reaction mixtnre was ponred into water, and extracted with Et20. The extracts were washed with H2O. The solvent was stripped off and the residne was distilled in vacuum to collect the rnn with b.p. 80-86°C (2.2 g) consisting of bis(l,3-butadienyl) telluride (1.1 g). 

First, the effects of aerobic and anaerobic culture conditions on toxaphene degradation were studied with washed P. putida cells grown on camphor and incubated with no readily usable carbon source. The radioactivities remaining in water after extraction with n-hexane were used as an indicator of metabolic activity. This was further extracted with ethyl acetate after acidification to divide this "total polar metabolites" fraction into aqueous buffer phase and ethyl acetate phase, i.e., the total polar metabolites reported refer to summation of the aqueous buffer and ethyl acetate soluble phases (Table 4). All radioactivities have been corrected by zero time controls and autoclaved 8 hr controls are included in each experiment. 

The product from Step 6 (0.315 mol), l-methyl-5-hydroxypyrazole (0.315 mol), potassium carbonate (0.63 mol), triethylamine (0.63 mol) and bis(triphenylphosphine)palladium dichloride (0.016 mol) were dissolved in 2.2 L of 1,4-dioxane and added to a 3.51 autoclave. Carbon monoxide at a pressure of 10-20 kg/cm was introduced and the mixture heated to 130°C 24 hours. The mixture was concentrated, the residue dissolved in water and extracted with CH2CI2. The aqueous pH phase was lowered to 4 with 18% HCl, the precipitate filtered, washed 3 times with water, and dried. The filtrate was extracted with CH2CI2, dried, concentrated, and the product isolated in 85.6% yield, mp = 215-219°C. H-NMR data supplied. 

Dibenzyl (1.82 g, 10 mmol) was added to a solution of Lewis acids with HF or CF3 SO3 H in a Hastelloy autoclave (100 mL) equipped with a Hastelloy magnetic stir bar under temperature contfol. The autoclave was sealed, and CO was introduced with vigorous stirring until reaching the pressure of 20 atm. After the reaction was over, the autoclave was depressurized and opened. The reaction was quenched with ice water and extracted with benzene. After removal of the solvent, 95% aromatic aldehyde was obtained, where 92% of the aldehyde was p,p -bisformyl dibenzyl. 

To extract phenolic acids in plant tissues/residues ground plant tissues/residues were extracted with water, EDTA, citrate plus or minus imidazole, KCl, or dibasic sodium phosphate and the water-autoclave procedure (Blum et al. 1992 Blum 1997). For procedures used for " C-labeled phenolic acids see Section 2.2.10... 

So what were the concentrations of phenolic acids in the Cecil A soil wheat stubble (Triticum aestivum L. Coker 916 )/soybean (Glycine maxL. Deltapine417 ) systems Subsamples taken from wheat stubble/soybean (no-till), wheat stubble tilled under/soybean (conventional-till), and fallow/soybean soil (conventional-till) cores were extracted by the water-autoclave procedure and analyzed for 7 common phenolic acids (ferulic, caffeic, p-coumaric, p-hydroxybenzoic, sinapic, syringinc, and vanillic) and total phenolic acid (Blum et al. 1991). With minor exception, individual phenolic acids were correlated with each other, with the sum of the 7 phenolic acids identified by HPLC analysis, and total phenolic acid as determined by the Folin Ciocalteu s phenol reagent method. 

Yeast-rich medium, YPD Add 5 g of Bacto yeast extract and 10 g of Bacto peptone to 450 mL of deionized water, autoclave, and then add a sterilized 50 mL glucose solution containing 10 g of glucose in deionized water. 

A hst of polyol producers is shown in Table 6. Each producer has a varied line of PPO and EOPO copolymers for polyurethane use. Polyols are usually produced in a semibatch mode in stainless steel autoclaves using basic catalysis. Autoclaves in use range from one gallon (3.785 L) size in research faciUties to 20,000 gallon (75.7 m ) commercial vessels. In semibatch operation, starter and catalyst are charged to the reactor and the water formed is removed under vacuum. Sometimes an intermediate is made and stored because a 30—100 dilution of starter with PO would require an extraordinary reactor to provide adequate stirring. PO and/or EO are added continuously until the desired OH No. is reached the reaction is stopped and the catalyst is removed. A uniform addition rate and temperature profile is required to keep unsaturation the same from batch to batch. The KOH catalyst can be removed by absorbent treatment (140), extraction into water (141), neutralization and/or crystallization of the salt (142—147), and ion exchange (148—150). 

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