Ice-water baths

Add 20 g. of /)-bromoaniline to 20 ml. of water in a 250 ml. beaker, and warm the mixture until the amine melts. Now add 23 ml. of concentrated hydrochloric acid and without delay stir the mixture mechanically in an ice-water bath, so that a paste of fine /> bromo-aniline hydrochloride crystals separates. Maintain the temperature of the stirred mixture at about 5° whilst slowly adding from a dropping-funnel a solution of 8 5 g. of sodium nitrite in 20 ml. of water con tinue the stirring for 20 minutes after the complete addition of the nitrite. 

Transfer the diazotised solution to a 600 mi. beaker, add 150 ml. of benzene, and stir the mixture vigorously to obtain an intimate mixture of the two liquids. Then again maintain the temperature at about 5° (by ice-salt cooling because of the heat evolved) whilst 27 ml. of 20% aqueous sodium hydroxide solution are added from a dropping-fiinnel during 40-50 minutes. When the addition of the alkali is complete, remove the ice-water bath, and allow the stirred mixture to reach room temperature. 

A 1500 ml. flask is fitted (preferably by means of a three-necked adaptor) with a rubber-sleeved or mercury-sealed stirrer (Fig. 20, p. 39), a reflux water-condenser, and a dropping-funnel cf. Fig. 23(c), p. 45, in which only a two-necked adaptor is shown or Fig. 23(G)). The dried zinc powder (20 g.) is placed in the flask, and a solution of 28 ml. of ethyl bromoacetate and 32 ml. of benzaldehyde in 40 ml. of dry benzene containing 5 ml. of dry ether is placed in the dropping-funnel. Approximately 10 ml. of this solution is run on to the zinc powder, and the mixture allowed to remain unstirred until (usually within a few minutes) a vigorous reaction occurs. (If no reaction occurs, warm the mixture on the water-bath until the reaction starts.) The stirrer is now started, and the rest of the solution allowed to run in drop-wise over a period of about 30 minutes so that the initial reaction is steadily maintained. The flask is then heated on a water-bath for 30 minutes with continuous stirring, and is then cooled in an ice-water bath. The well-stirred product is then hydrolysed by the addition of 120 ml. of 10% sulphuric acid. The mixture is transferred to a separating-funnel, the lower aqueous layer discarded, and the upper benzene layer then... 

The suspension of phenylacetamide may be further hydrolysed to phenylacetic acid by refluxing with stirring until the solid dissolves. The mixture becomes turbid after 30 minutes and the product begins to separate as an oil refluxing is continued for 6 hours, the mixture is cooled first with tap water and then by an ice-water bath for about 4 hours. The crude phenylacetic acid is filtered at the pump, washed with two 50 ml. portions of cold water, and dried in a desiccator. The resulting crude acid melts at 69- 70° it may be purified by recrystallisation from light p>etroleum (b.p. 40-60°) or, better, by vacuum distillation. 

To a solution of 0.5D mol of KO-tert.-C Hg (uncomplexed base, commercially available) and 180 ml of dry THF was added at room temperature 0.25 mol of the Mannich product. A weakly exothermic reaction took place. The mixture was heated in a bath at 50°C and the THF started to reflux (occasional cooling in an ice-water bath may be necessary). When the refluxing had ceased, the mixture was heated for 30 min in a bath at 70°C, then cooled to room temperature and 300 ml of redistilled, dry pentane were added. The precipitate of potassium raethoxide and... 

To a solution of 8 g of lithiim alanate in 250 ml of diethyl ether was added in 15 min 24 g (0.3 mol) of 2-penten-4-yn-l-ol (III, Exp. 57). The diethyl ether began to reflux and a rubber-like greyish precipitate was formed. After heating for 1 h under reflux the flask was placed in an ice + ice-water bath and water (150 ml) was added dropwise with vigorous stirring. After this hydrolysis procedure the ethereal solution was decanted and the aqueous jelly layer was extracted ten times with diethyl ether. The ethereal extracts were dried (without washing) over magnesium sulfate and subsequently concentrated in a water-pump vacuum. 

After the flask has been cooled by immersion in ice water, 44 g- (o-5 mole) of dry ethyl acetate (Note i) in an equal volume of ether is added over a period of twelve minutes. The cooling bath is removed and stirring is continued for ten minutes. When the reaction flask has again been cooled by the ice-water bath, a previously prepared ammonium chloride solution (Note 2) is added, very slowly at first, over a period of ten minutes, during which a pasty solid separates. 

D-(-H)-Biotin 4-nitrophenyl ester [33755-53-2] M 365.4, m 160-163 , 163-165 , [a]n -n47 (c 2, MeaNCHO containing 1% AcOH). It has been recrystd by dissolving 2g in 95% EtOH (30mL), heated to dissolve, then cooled in an ice-water bath. The crystals are collected, washed with ice-cold 95% EtOH (5mL) and dried over P2O5. The Rp on silica plates (CHCl3 MeOH-19 l) is 0.19 [Bodanszky and Fagan J Am Chem Soc 99 235 1977]. 

In a 3-I. round-bottomed, three-necked flask fltted with a liquid-sealed mechanical stirrer, a thermometer, and a 500-cc. separatory funnel are placed 1700 cc. of dry ben2ene and 160 g. (1.2 moles) of powdered, anhydrous aluminum chloride (Note i). The mixture is cooled to 10° by means of an ice-water bath and maintained at 10-20 during the addition of a solution of 120 g. (0.58 mole) of benzalacetophenone (Note 2) (Org. Syn. Coll. Vol. I, 71) in 300 cc. of dry benzene. This addition requires about thirty minutes. The cooling bath is then removed and stirring continued at room temperature until all the dense, yellow precipitate formed at first has gone into solution (Note 3). The reaction is complete after stirring for an additional hour. 

A 1-liter flask is equipped with a magnetic stirrer, a thermometer immersed in the reaction mixture, and a drying tube. In the flask is placed 100 ml of anhydrous pyridine, and the flask is cooled in an ice-water bath to 15-20° (lower temperatures impede the complex formation). Chromium trioxide (80 g) is added in small portions to the stirred solvent at a rate so as to keep the temperature below 30°. After about one-third of the chromium trioxide has been added, the yellow complex begins to precipitate. At the end of the addition (about 1 hour), a slurry of the yellow complex in pyridine remains. (This form of the complex is apparently a microcrystalline form and is very difficult to handle.)... 

A 500-ml flask is equipped with a thermometer, a magnetic stirrer, and a dropping funnel, and all openings are protected by drying tubes. The system is flushed with nitrogen and a solution of 2.84 g (0.075 mole) of sodium borohydride in 150 ml of diglyme is introduced followed by 28.3 g (0.30 mole) of norbornene. The flask is immersed in an ice-water bath and the hydroboration is achieved by the dropwise addition of 27.4 ml (0.10 mole) of boron trifluoride diglymate. The solution is stirred... 

The residual gum was dissolved in 360 ml of ethanol and cooled to 15°C in an ice/water bath, 8 g of sodium borohydride was then added in portions over 30 minutes while maintaining the temperature at 15°-20°C. After a further 30 minutes at 20°C the solution was stirred at room temperature for 2 hours. The solution was again cooled In ice and 250 ml of 2 N sulfuric acid were slowly added, then the solution was evaporated In vacuo until the ethanol had been removed. The clear aqueous solution was then treated with 250 ml of 10% sodium carbonate solution and the oil which precipitated was extracted into ethyl acetate. The ethyl acetate layer was washed with sodium carbonatesolution, then with water, and was dried over anhydrous sodium sulfate and evaporated in vacuo, to a small volume. Petroleum ether (BP 40°- 0°C) was added, and after standing overnight a white solid was obtained. This was filtered off to give 23 g of the product, MP 110°-114°C. 

The resulting suspension is placed in a round flask equipped with a scraper-agitator device, and agitation is effected for 48 hours in an ice water bath. The antibiotic is isolated from the mother liquor by filtration through a Buchner filter. The filter cake is washed with 5 liters of a methyl alcohol and water mixture (1/2.5 by volume) cooled to 4°C. After drying in an oven at reduced pressure, 2.805 kilograms of a greyish-yellow crude product is obtained. 

The temperature 0°C is readily obtained and maintained with an ice water bath. The temperature is one at which thermometers are calibrated this aids measurement. A temperature that is easy to maintain and easy to measure makes a good standard temperature. 

Fig. 4.3.1 Effect of pH on the total light emission of phialidin (A), and the temperature stability profiles of phialidin (minute open circles) and aequorin (solid line) (B). In A, each buffer contained 0.1 M CaCl2 plus 0.1 M Tris, glycine or sodium acetate, the pH being adjusted with NaOH or HC1. In B, the photoprotein samples in 10 mM Tris-EDTA buffer solution, pH 8.0, were maintained at a test temperature for 10 min, and immediately cooled in an ice water bath. Then total luminescence activity was measured by injecting 1ml of 0.1 M CaCl2/Tris-HCl, pH 7.0, to 10 pd of the test solution. From Levine and Ward (1982), with permission from Elsevier.

A solution of 1.0 g (33 mmol) of the mixture of heptofuranoside diastereomers (14) in 5 ntL of THE is stirred in an ice-water bath. 3.4 mL of 1 M soln ofTBAF (3.4 mmol) in THF are added, and Ihe reaction is monitored by TLC (CHCI,/CH, OH, 9 1). After ca. 15 min, the reaction mixture is carefully neutralized with dil sulfuric acid, then diluted with water and extracted with CHC13. The extract is dried over Na,S04 and concentrated to give an off-white solid which is a single stereoisomer (by NMR spectroscopy). Column chromatography gives the inositol as a white solid yield 0.61 g (70%) mp 186-187 C (benzene/CH,OH) [a]D + 118 (c = 03, CH, OH). 

B. l0-Bromo-ll-hydroxy-10,ll-dihydrofarnesyl Acetate [2,6-Dodeca-diene-1,11-diol, 10-bromo-3,7, -trimethyl-, 1-acetate, (E,E)-]. Farnesyl acetate (29 g., 0.11 mole) is dissolved in 1 1. of /erf-butyl alcohol (Note 4) contained in a 3-1. Erlenmeyer flask. Water is added (500 ml.), and the solution is cooled to about 12° using an external ice water bath. Maintaining this temperature, rapid magnetic stirring is begun, and more water is added until a saturated solution is obtained. The second addition of water may be rapid initially, but the saturation point must be approached carefully, like the end point of a titration. A total of about 1200 ml. of water is required for the above amounts of farnesyl acetate and ferf-butyl alcohol. The solution must remain clear and homogeneous at about 12°, and if the saturation point is accidentally passed by adding too much water, ferf-butyl alcohol should be added to remove the turbidity. 

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