In our biochemistry practical class on osazones, we had seen osazones under the microscope. Osazones are produced when reducing sugars are treated with phenyl hydrazine hydrochloride in acidic medium at 100°C. We actually mix some sugar solution with some osazone buffer (containing phenyl hydrazine, sodium acetate and acetic acid). We then keep the test tube in a boiling water bath for 20 minutes and then cool the test tubes to room temperature. Osazones are crystals that have a particular shape. Osazones of monosaccharides are formed immediately in the boiling water bath while those of disaccharides are formed after gradual cooling. We then observe the osazones microscopically to tell if it is lactosazone, maltosazone or glucosazone.
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