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Sample text
This is conveniently done in a glass-stoppered graduate or bottle, which is chilled in an ice b a t h . With the graduate still in the ice b a t h and containing the required amount of acetic anhydride, 1 ml. of concentrated sulfuric acid is added for each 20 ml. of acetic anhydride and mixed during the addition. T h e stopper is then inserted, and the graduate is removed from the b a t h , shaken vigorously for a few moments, and returned to the bath. About 10 minutes later, when the reagent is thoroughly chilled, the first tube is removed from the 25° C.
Extraction of blood serum: A glass-stoppered 25-ml. volumetric flask is filled about half-way with acetone-ethanol, and 1 ml. of serum is carefully measured and added slowly with rotation of the TOTAL AND F R E E CHOLESTEROL 45 flask. A finely divided precipitate should result. The flask is immersed in water, which has been heated to boiling, and agitated to prevent bumping until the solvent is brought just to a boil. Caution: There should be no open flames! After the flask has cooled, acetone-ethanol is added to the mark, the glass stopper is inserted, and the contents are throughly mixed and filtered through a rapid filter paper into a test tube.
Biol Chem. 61: 523 (1924). To clean the manometer, open the cock at the top, draw the mercury as completely as possible into the leveling bulb, and disconnect the rubber tubing. Attach the tube leading to the suction flask where the leveling bulb was attached. Draw water through the manometer at the open top and, if the tube is greasy, alcoholic K O H , then acetone. Instead of K O H , cleaning solution, water, 34 STANDARD METHODS O F CLINICAL CHEMISTRY and acetone, or nitric acid, water, and acetone m a y be used.