Synthesis and Analysis of Cholesterol by Bromination and Debromination of Commercial Cholesterol with Physical, Chemical, and Chromatographic Techniques

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Synthesis and Analysis of Cholesterol by Bromination and Debromination of Commercial Cholesterol with Physical, Chemical, and Chromatographic Techniques The purpose of this experiment was to purify commercial grade cholesterol from impurities using organic reactions. Bromination of cholesterol yielded a product with a significantly different structure that was more easily separated from unwanted cholesterol derivatives. This reaction was reversed by debromination which yielded cholesterol now largely free of impurities. Commercial cholesterol was dissolved in t-butylmethyl ether, a bromine mixture was added, and an electrophilic addition reaction proceeded. This product, dibromocholesterol, was washed and debrominated by an elimination reaction using zinc dust. Percent yield and recovery evaluated the efficiency of this synthesis. Melting point and thin layer chromatography (TLC) analysis assessed the purity of commercial cholesterol, dibromocholesterol, and synthesized cholesterol by physical analysis. Chemical tests evaluated the purity of these samples by assessing the presence of specific functional groups. Silver nitrate tested for 2o and 3o alkyl halides expected in dibromocholesterol and sulfuric acid tested for alkenes expected in cholesterol. Bromination had 68.8% yield, debromination 73.7% yield, and overall 53.3% of commercial cholesterol was recovered as synthesized cholesterol. For commercial cholesterol, dibromocholesterol, and synthesized cholesterol, melting points were 145.2-148.1oC, 109.0-110.1oC, 146.3-148.2oC and retention factor (Rf) values were 0.69, 0.80 respectively. Chemical tests confirmed expectations, though some alkenes were present in dibromocholesterol. The similar melting points of the synthesized product as compared to the literature value; and similar Rf values for commercial and synthesized cholesterol showed the purity of

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