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II. Tests for Lipids A. Bromine Test Table 3. Results for Bromine test. SAMPLE Ranking of the number of drops of Bromine* Degree of unsaturation* Oleic oil Coconut oil Soybean oil Rank (1", 2nd, 31d) – 1ª as the highest number of drops; 3rd as the lowest number of drops ** minimal, moderate, highest B. Acrolein Test Table 4. Results for the Acrolein test. OBSERVATION INFERENCE SAMPLE Oleic oil Coconut oil Beef fat Egg yolk * Pungent odor or no observable result * positive or negative for glycerol c. Zak's Test Table 5. Results for Zak's test. SAMPLE Oleic oil Coconut oil Egg yolk Purple product or no observable result * positive or negative for cholesterol OBSERVATION INFERENCE**

II. Tests for Lipids A. Bromine Test Table 3. Results for Bromine test. SAMPLE Ranking of the number of drops of Bromine* Degree of unsaturation* Oleic oil Coconut oil Soybean oil Rank (1", 2nd, 31d) – 1ª as the highest number of drops; 3rd as the lowest number of drops ** minimal, moderate, highest B. Acrolein Test Table 4. Results for the Acrolein test. OBSERVATION INFERENCE SAMPLE Oleic oil Coconut oil Beef fat Egg yolk * Pungent odor or no observable result * positive or negative for glycerol c. Zak's Test Table 5. Results for Zak's test. SAMPLE Oleic oil Coconut oil Egg yolk Purple product or no observable result * positive or negative for cholesterol OBSERVATION INFERENCE**

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
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II. Tests for Lipids A. Bromine Test Table 3. Results for Bromine test. SAMPLE Ranking of the number of drops of Bromine Degree of unsaturation Oleic oil Coconut oil Soybean oil * Rank (1", 2nd, 3rd) – 1ª as the highest number of drops; 3d as the lowest number of drops ** minimal, moderate, highest B. Acrolein Test Table 4. Results for the Acrolein test. INFERENCE** SAMPLE Oleic oil Coconut oil Beef fat Egg yolk Pungent odor or no observable result * positive or negative for glycerol OBSERVATION c. Zak's Test Table 5. Results for Zak's test. SAMPLE OBSERVATION INFERENCE*. Oleic oil Coconut oil Egg yolk * Purple product or no observable result * positive or negative for cholesterol D. Saponification Table 6. Results for saponification. SAMPLE REMARKS* Soap suspension (Step 2) Purified soap (Step 3) Acid hydrolysis product of soap (Step 4) Rank (14, 2nd, 3rd) as to the estimated pH value -1 as the highest pH value; 3rd as the lowest pH value
Transcribed Image Text:II. Tests for Lipids A. Bromine Test Table 3. Results for Bromine test. SAMPLE Ranking of the number of drops of Bromine Degree of unsaturation Oleic oil Coconut oil Soybean oil * Rank (1", 2nd, 3rd) – 1ª as the highest number of drops; 3d as the lowest number of drops ** minimal, moderate, highest B. Acrolein Test Table 4. Results for the Acrolein test. INFERENCE** SAMPLE Oleic oil Coconut oil Beef fat Egg yolk Pungent odor or no observable result * positive or negative for glycerol OBSERVATION c. Zak's Test Table 5. Results for Zak's test. SAMPLE OBSERVATION INFERENCE*. Oleic oil Coconut oil Egg yolk * Purple product or no observable result * positive or negative for cholesterol D. Saponification Table 6. Results for saponification. SAMPLE REMARKS* Soap suspension (Step 2) Purified soap (Step 3) Acid hydrolysis product of soap (Step 4) Rank (14, 2nd, 3rd) as to the estimated pH value -1 as the highest pH value; 3rd as the lowest pH value
B. Acrolein Test for Glycerol 1. Get 4 test tubes and label them as A (oleic acid), B (coconut oil), C (melted beef fat), and D (egg yolk). 2. Place 5 drops of each sample (coconut oil, egg yolk, oleic acid, and beef fat/tallow) into the designated labelled test tubes. 3. Add 1-2 crystals of potassium hydrogen sulfate. 4. Gently heat each test tube, one at a time, over a flame, while vigorously shaking it until the evolution of fumes. 5. Carefully smell the test tubes. A characteristic pungent odor of acrolein, resembling burnt hamburgers, indicates the presence of glycerol. (NOTE: Do not overheat the samples to avoid the adherence of a black residue in the test tubes which is difficult to remove.) C. Zak's Test for Cholesterol 1. Get 3 test tubes and label them as A (oleic acid), B (coconut oil), and C (egg yolk). 2. Place 1 ml or 100 mg of the lipid samples into separate labeled test tubes. 3. Add 1 ml of Zak's reagent. 4. Tilt the test tubes to 45° and add 2 drops of concentrated sulfuric acid. 5. Observe the test tubes. The appearance of a deep red color indicates the presence of cholesterol. D. Saponification Test 1. Place about 1 ml of oil into a test tube. Add 2% w/v sodium hydroxide dropwise with continued shaking until it traces. Trace is the point at which the soap has thickened and blended enough to have formed a stable emulsion. When the mixture traces, it has formed soap. 2. Pipet out the soap obtained from Step 1 and mix with distilled water until the entire solid has dissolved. This should make a little above 5 ml of soap solution. Test the pH of soap using pH paper. 3. Salting out of soap. Add solid Nacl to the solution obtained in Step 2 above until no more Nacl dissolves. Soaps can be forced out of the solution by dissolving NaCl in the soap solution. This is a way of purifying soap from extra NaOH left from the saponification process. Pipet out the soap again and mix with distilled water until the entire solid has dissolved. Test the pH using pH paper. 4. Acid hydrolysis. Get about 1 ml of soap solution obtained in Step 2 and add 6 M HCI drop by drop until no more changes occur. Record your observations. Pipet out a small sample of the product and dissolve in distilled water. Test the pH using рH раper. * Rank (1", 2nd, 3rd) as to the estimated pH value of the soap solution (Step 2), the product of the salting-out (Step 3), and the product of hydrolysis (Step 4): 1ª – the highest pH value and 3rd – the lowest pH value
Transcribed Image Text:B. Acrolein Test for Glycerol 1. Get 4 test tubes and label them as A (oleic acid), B (coconut oil), C (melted beef fat), and D (egg yolk). 2. Place 5 drops of each sample (coconut oil, egg yolk, oleic acid, and beef fat/tallow) into the designated labelled test tubes. 3. Add 1-2 crystals of potassium hydrogen sulfate. 4. Gently heat each test tube, one at a time, over a flame, while vigorously shaking it until the evolution of fumes. 5. Carefully smell the test tubes. A characteristic pungent odor of acrolein, resembling burnt hamburgers, indicates the presence of glycerol. (NOTE: Do not overheat the samples to avoid the adherence of a black residue in the test tubes which is difficult to remove.) C. Zak's Test for Cholesterol 1. Get 3 test tubes and label them as A (oleic acid), B (coconut oil), and C (egg yolk). 2. Place 1 ml or 100 mg of the lipid samples into separate labeled test tubes. 3. Add 1 ml of Zak's reagent. 4. Tilt the test tubes to 45° and add 2 drops of concentrated sulfuric acid. 5. Observe the test tubes. The appearance of a deep red color indicates the presence of cholesterol. D. Saponification Test 1. Place about 1 ml of oil into a test tube. Add 2% w/v sodium hydroxide dropwise with continued shaking until it traces. Trace is the point at which the soap has thickened and blended enough to have formed a stable emulsion. When the mixture traces, it has formed soap. 2. Pipet out the soap obtained from Step 1 and mix with distilled water until the entire solid has dissolved. This should make a little above 5 ml of soap solution. Test the pH of soap using pH paper. 3. Salting out of soap. Add solid Nacl to the solution obtained in Step 2 above until no more Nacl dissolves. Soaps can be forced out of the solution by dissolving NaCl in the soap solution. This is a way of purifying soap from extra NaOH left from the saponification process. Pipet out the soap again and mix with distilled water until the entire solid has dissolved. Test the pH using pH paper. 4. Acid hydrolysis. Get about 1 ml of soap solution obtained in Step 2 and add 6 M HCI drop by drop until no more changes occur. Record your observations. Pipet out a small sample of the product and dissolve in distilled water. Test the pH using рH раper. * Rank (1", 2nd, 3rd) as to the estimated pH value of the soap solution (Step 2), the product of the salting-out (Step 3), and the product of hydrolysis (Step 4): 1ª – the highest pH value and 3rd – the lowest pH value
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