Methanol silicates

To a mixture of ethyl 5a-cholestan-3-one 2a-xanthate (2 g, 3.95 mmol) and 100 ml methanol is added sufficient ether to completely dissolve the solids. Sodium borohydride (90 mg, 2.36 mmol) is added directly to the reaction flask and the solution is stirred at room temperature for 4 hr. (The use of an excess of sodium borohydride and an extended reaction time produces 5oc-cholestan-2a,3a-thiirane.) The reaction is diluted with 200 ml ether and washed several times with ca. 100 ml water, dried (MgS04) and the solvent is removed under vacuum. The crude sticky gum is chromatographed on a column of 85 g silicic acid. The hexane eluates contain 5a-cholest-2-ene. Ethyl 5a-cholestan-3a-ol 2a-xanthate is obtained in ca. 30% yield by subsequent elution with benzene hexane (1 7) and the desired ethyl 5a-cholestan-3 -ol 2a-xanthate is eluted with ether hexane (1 3) in ca. 30% yield. 

Twenty grams of the resulting oily substances were mixed with 20 grams of silicic acid (Mal-linckrodt Chemical Co.),applied to a column 40 cm in length and 4.5 cm in diameter filled with silicic acid,and eluted with a be nzene-ecetone-methanol mixture. The initial eluate which eluted with a 1 1 0 mixture was discarded and the active fractions eluted with 1 3 0 and 1 3 0.3 mixtures were collected and concentrated to dryness in vacuo. 11 g of this crude substance was then dissolved in a small amount of ethyl acetate and applied to the same silicic acid column as above. After discarding the initial eluates by the 1 1 and 2 1 benzene-acetone mixtures, aclacinomycin 8 fractions were first eluted with the above mixtures of 1 3 and 1 5 ratio, and aclacinomycin A fractions were then eluted with the 1 5 0.5 and 1 5 1 benzene-acetone-methanol mixtures. The eluates were dried over anhydrous sodium sulfate and concentrated to dryness in vacuo. 4,8 g of crude aclacinomycin A and 3.5 g of aclacinomycin 8 were obtained as yellow powder. 

This reaction mixture is kept between 0°C and -i-5°C for six hours, with agitation and under an inert atmosphere, then 5 cc of a 0.2N solution of acetic acid in toluene are added. The mixture is extracted with toluene, and the extracts are washed with water and evaporated to dryness. The residue is taken up in ethyl acetate, and then the solution Is evaporated to dryness in vacuo, yielding a resin which is dissolved in methylene chloride, and the solution passed through a column of 40 g of magnesium silicate. Elution is carried out first with methylene chloride, then with methylene chloride containing 0.5% of acetone, and 0.361 g Is thus recovered of a crude product, which is dissolved in 1.5 cc of isopropyl ether then hot methanol Is added and the mixture left at 0°C for one night. 

A solution of sodium methoxide (80 mmol) in methanol (40 mL) is added to silica gel (2.7 g, 45 mmol) followed by a solution of catechol (13.2 g, 120 mmol) in methanol (40 mL). The resulting mixture is stirred and heated under reflux for 18 h. The methanol is then evaporated and the solid residue washed with ether. The black sohd is dissolved in THF (400 mL) and the resulting solution is heated for 1 h in the presence of charcoal. After filtration and evaporation of the solvent the sodium tris(benzene-l,2-diolato)silicate 80 is isolated as a white powder (12.53 g, 70% Scheme 2.20) [93]. 

The darkness associated with dense interstellar clouds is caused by dust particles of size =0.1 microns, which are a common ingredient in interstellar and circum-stellar space, taking up perhaps 1% of the mass of interstellar clouds with a fractional number density of 10-12. These particles both scatter and absorb external visible and ultraviolet radiation from stars, protecting molecules in dense clouds from direct photodissociation via external starlight. They are rather less protective in the infrared, and are quite transparent in the microwave.6 The chemical nature of the dust particles is not easy to ascertain compared with the chemical nature of the interstellar gas broad spectral features in the infrared have been interpreted in terms of core-mantle particles, with the cores consisting of two populations, one of silicates and one of carbonaceous, possibly graphitic material. The mantles, which appear to be restricted to dense clouds, are probably a mixture of ices such as water, carbon monoxide, and methanol.7... 

Fig. 3.12 Model of an agglomerate consisting of many small interstellar dust particles. Each of the rod-shaped particles consists of a silicate nucleus surrounded by yellowish organic material. A further coating consists of ice formed from condensed gases, such as water, ammonia, methanol, carbon dioxide and carbon monoxide. Photograph Gisela Kruger, University of Bremen...

The polarity values of binary acetonitrile/water and methanol/water mobile phases used in RPLC were measured and compared with methylene selectivity (acH2) for both traditional siliceous bonded phases and for a polystyrene-divinylbenzene resin reversed-phase material [82], The variation in methylene selectivity for both was found to correlate best with percent organic solvent in methanol/water mixtures, whereas the polarity value provided the best correlation in acetonitrile/water mixtures. The polymeric resin column was found to provide higher methylene selectivity than the siliceous-bonded phase at all concentrations of organic solvent. 

Procedures for determining fatty acids in sediments involved liquid-liquid extraction, liquid-solid adsorption chromatography followed by gas liquid chromatographic analysis [10-12], Liquid extractions have been performed with methanol-chloroform [13], methylene chloride [14] and benzene-methanol [15, 16]. Typical liquid-solid adsorbents are silicic acid. Standard gas chromatographic separations for complex mixtures employ non-polar columns packed with OV-1, OV-17, OV-101, SE-30, or glass capillary columns containing similar phases. 

Mono-, di- and triacylglycerols may all be measured by determination of the amount of glycerol released by hydrolysis. The lipid is first extracted into chloroform-methanol (2 1) and saponification is performed under conditions that will not affect any phosphate ester bonds, otherwise glycerol originating from phosphoglycerides would also be measured. Heating at 70°C for 30 min with alcoholic potassium hydroxide (0.5 mol l-1) has been shown to be satisfactory. However, the phospholipids may be removed prior to saponification either by extraction or by adsorption on activated silicic acid. 

Isolation of toxins. The digestive glands of shellfish were extracted with acetone at room temperature. After removal of the acetone by evaporation, the aqueous suspension was extracted with diethyl ether. The ether soluble residue was successively chromatographed twice over silicic acid columns with following solvents benzene to benzene-methanol (9 1), and diethyl ether to diethyl ether-methanol (1 1). To avoid degradation of dinophysistoxin-3 by contaminant acid, the silicic acid was washed with dilute sodium hydroxide solution and then with water prior to activation at 110 C. Toxic residue obtained in the second eluates was separated into two fractions... 

GT-1, the smallest of the three toxic fractions, was separated on silicic acid with chloroform methanol (9 1). At a concentration of 40 ng/ml it was found to be a reversible inhibitor of the guinea pig ileum. At concentrations less than 8 ng/ml the ileum response to histamine was augmented. 

GT-2, separated on silicic acid with chloroform methanol (1 1), constituted the largest fraction. At a concentration of 4 ng/ml, GT-2 was found to be a reversible inhibitor of the guinea pig ileum response to histamine. The separation of all three fractions up to the last step (see d. Table 1) was exactly the same procedure previously utilized by Tachibana (22) Scheuer ( ) and others ( 3, ). In the last step, when we attempted to utilize a Sephadex column and water for further purification of GT-2, we lost a considerable amount of toxin and also appeared to lose toxicity. 

GT-3 was separated from GT-1 and GT-2 on silicic acid using 100% methanol. The separation of this fraction is the same as that used by Tachibana (22) and Yasumoto (13) for maitotoxin. Unlike GT-1 and GT-2, this fraction was found to be an irreversible inhibitor of the guinea pig ileum preparation. The time studies of GT-3 are important in that they characterize GT-3 as a very toxic fraction but one which is very slow in its action. 

Fig. 13. Effect oftheciiainleiKthofn-alkyllisatet in siliceous bond phases on the retention of (A) shoit-chain aliphatic alcohols and (B) lona-chain alkanes. Tne number on the curve indicates the carbon number of the solute. The stationary phases w e prepared from silica gel having 10 nm average pore size with n-alkyltrichlorosilanes. Mobile phase (A) methanol (B) water. Replotted thorn data by Kerch el al, (73).

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