Of 2- acetic acid

Cyanoandrost-5-ene-3jS,17-diol 3-Acetate Acetic acid (385 ml) is added during a 40 min period at 10° to a stirred mixture of 60 g of 3) -liydroxy-androst-5-en-17-one acetate and 360 g of potassium cyanide in 2 liters of alcohol. The mixture is stirred for 1 hr at 10° and then for 2 hr at room temperature. After dilution with water the precipitated product is collected by filtration, washed with 3 liters of 2 % acetic acid and dried at room temperature. The yield of epimeric cyanohydrins (mp 124° dec [ajj, —93°) is essentially quantitative. 

Pour exactly lOmL of 1% ammonia in dimethylsulfoxide (DMSO) into the ground powder and sonicate it in an ultrasonic bath for two minutes. Add exactly 90mL of 2% acetic acid in water to the mixture. Stir it with a magnetic stir bar for 1 min. 

Rinsing is first done with an aqueous solution of 2 % acetic acid and finally with an acetate buffer at pH 3.6, since the influence of pH on this dye is considerable (D15). 

The reaction contained 0.1 mM sodium laurate, 0.7 mM NADPH, 50 mAf Tris-HCl (pH 7.5) and microsomes (0.5 mg from liver, or 0.3 mg from kidney) in a final volume of 1.0 mL. The reaction was initiated by adding NADPH and terminated after 10 minutes at 37°C by adding 0.25 mL of 4 M HQ. The internal standard, 0.25 mL of 2.5 /ug/mL 10-hydroxycapric acid, was then added. The mixture was extracted with 4.5 mL of diethyl ether. The organic phase was washed once with 4 mL of water and evaporated to dryness under reduced pressure. The residue was dissolved in 0.1 mL of acetonitrile containing 10 mAf 3-bromomethyl-7-methoxy-l,4-benzoxazin-2-one and 0.1 mL of 15 mg/mL 18-crown-6 in acetonitrile saturated with K2C03. The resulting solution was allowed to stand at 40°C for 30 minutes before 0.05 mL of 2% acetic acid was added. Two-microliter aliquots were used for HPLC analysis. 

Isolation. The fresh crushed potato sprouts are allowed to remain for 4S hours with twice their weight of 2% acetic acid. The acid solution is separated with the aid of a centrifuge, filtered with a little celite, and rendered distinctly alkaline by the addition of ammonia. After 24 hours the precipitate is separated by filtration, washed, and dried. The admixed solanidine is separated by exhaustive ether extraction and may be obtained in impure form by evaporation of the ether. The ether-insoluble portion is extracted with hot 80% ethanol and, upon concentrating and cooling the extract, the crude solanine separates. It may be recrystallized from 80% ethanol or from dioxane (39, 56). As thus purified it melts at 285° and has [a]D — 42 (dilute hydrochloric acid), —56.5° (pyridine), —60° (pyridine) (31, 62). 

D) Spraying solution. A solution of 0-2 g. of ninhydrin in a mixture of 99 ml. of redistilled n-butanol and i ml. of glacial acetic acid. 

Alternatively the semi micro apparatus shown in Fig. 38 (p. 63) may be used. Heat the anilide and sulphuric acid under reflux for 15 minutes in such a manner that the vapour does not rise higher than half-way up the vertical condenser through which water is passed. Then dilute the solution in the flask with 5 ml. of water. Empty the vertical condenser and run cold water through the inclined condenser. Now increase the rate of heating and distil off 1-2 ml. of aqueous acetic acid. 

Dissolve I g. of glucose (or fructose) in 5 ml. of water in a boiling-tube. In another tube dissolve 2 ml. of glacial acetic acid in 5 ml. of water, add 2 ml. of phenylhydrazine, and shake until... 

Dissolve 0 3 ml. of glacial acetic acid in 2 ml. of water in a 25 ml. conical flask, and add 0 4 ml. (0 44 g.) of phenylhydrazine. Mix thoroughly to obtain a clear solution of phenylhydrazine acetate and then add 0 2 ml. (0 21 g.) of benzaldehyde. Cork the flask securely and shake the contents vigorously. A yellow crystalline mass of the hydrazone soon begins to separate. Allow to stand for 15 minutes, with occasional shaking, and then filter the solid product at the pump, wash first with very dilute acetic acid and then with water, and finally drain thoroughly. Recrystallise the material from rectified or methylated spirit, the benzaldehyde phenylhydrazone being thus obtained in fine colourless needles, m.p. 157 yield, 0 4 g. 

Dissolve 1 g. of anthracene in 10 ml. of glacial acetic acid and place in 50 ml. bolt head flask fitted with a reflux water-condenser. Dissolve 2 g. of chromium trioxide in 2 ml. of water and add 5 ml. of glacial acetic acid. Pour this solution down the condenser, shake the contents of the flask and boil gently for 10 minutes. Cool and pour the contents of the flask into about 20 ml. of cold water. Filter off the crude anthraquinone at the pump, wash with water, drain well and dry. Yield, 1 g. Purify by re crystallisation from glacial acetic acid or by sublimation using the semi-micro sublimation apparatus (Fig. 35, p. 62, or Fig. 50, p. 70). 

Formation of a Quinoxaline. Heat together for 5 minutes under reflux 0 2 g. of phenanthraquinone dissolved in i ml. of glacial acetic acid and 0-2 g. of O -phenylene diamine also dissolved in i ml, of glacial acetic acid. The yellow substituted quinoxaline (p. 305) separates rapidly. Cool, filter and recrystallise from benzene m.p. 225 . 

Ester formation. Heat under very efficient reflux 1 ml. of diethyl ether, 4 ml. of glacial acetic acid and i ml. of cone. H2SO4 for ro minutes. Distil off 2 ml. of liquid. Use a few drops of this liquid for the hydroxamic add test for esters (p. 334). Use the remainder for other tests for esters (p. 354). 

Fit two similar 250 ml. conical flasks, A and B, with reflux water-condensers (using ground-glass joints or rubber stoppers) and connect the condensers in series as before over two water-baths. Prepare a mixture of 2 volumes of acetic anhydride and i volume of glacial acetic acid,... 

At the end of about an hour,f filter the solution and tfansfer 20 ml. of the clear filtrate to a boiling-tube. Dissolve 2 5 ml. of phenylhydrazine in 3 ml. of glacial acetic acid diluted with 3 ml. of water, and add this solution to that in the boiling-tube. Mix well and place the tube in a boiling water-bath. After about 15 minutes, crystals of glucosazone begin to separate. At the end of about i hour, filter off the precipitate and identify (p. 319). 

Treat the distillate with 2 drops of glacial acetic acid (to destroy the phosphorus esters present) and redistil using the same apparatus as before except that the separatory funnel is replaced by a thermometer. Collect the liquid which passes over at 50-56°. Transfer the acetyl chloride to a weighed glass-stoppered bottle (since cork and rubber stoppers are attacked) and determine the weight. The yield is 22 g. 

Furfuryl acetate. Reflux a mixture of 39 2 g. (34-8 ml.) of redistilled furfuryl alcohol, 48 g. of glacial acetic acid, 150 ml. of benzene and 20 g. of Zeo-Karb 225/H in a 500 ml. bolt-head flask, using the apparatus described under iaoPropyl Lactate. After 3 hours, when the rate of collection of water in the water separator is extremely slow, allow to cool, separate the resin by suction filtration, and wash it with three 15 ml. portions of benzene. Remove the benzene, etc., from the combined filtrate and washings under reduced pressure (water pump) and then collect the crude ester at 74-90°/10 mm. a small sohd residue remains in the flask. Redistil the crude ester from a Claisen flask with fractionating side arm pure furfuryl acetate passes over at 79-80°/17 mm. The yield is 14 -5 g. 

Some amides do not dissolve in glacial acetic acid in such cases a mixture of 2 ml. of glacial acetic acid and 3 ml. of water may be used as a solvent for the reaction. 

Diphenyl. Reflux a mixture of 1 g, of diphenyl, 2 ml. of glacial acetic acid and 0 -5 ml. of fuming nitric acid for 10 minutes. Pour into 20 ml. of cold water, filter oflF the precipitate, wash it with cold water imtil free from acid, and recrystallise from alcohol. The product is 4-nitrodiphenyl, m.p. 114°,... 

Reflux 1 g. of the sulphonamide with 2-5 ml. of acetyl chloride for 30 minutes if solution is not complete within 5 minutes, add up to 2-5 ml. of glacial acetic acid. Remove the excess of acetyl chloride by distillation on a water bath, and pour the cold reaction mixture into water. Collect the product, wash with water and dissolve it in warm sodium bicarbonate solution. Acidify the Altered solution with glacial acetic acid Alter oflF the precipitated sulphonacetamide and recrystaUise it from aqueous alcohol. 

The best results are obtained with freshly prepared xanthhydrol (reduction of xanthone with sodium amalgam. Section VII,16). Dissolve 0 -25 g. of xanthhydrol and 0 -25g. of the primary sulphonamide in 10 ml. of glacial acetic acid. Shake for 2-3 minutes at the laboratory temperature and allow to stand for 60-90 minutes. Filter oflf the derivative, recrystallise it from dioxan-water (3 1), and dry at room temperature under water pump suction for 30 minutes. 

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