Experiment 6: Proteins Flashcards

1
Q

The source of protein used for Experiment 6

A

Egg albumin

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2
Q

6.A1. Biuret Test

A
  1. Add 1 ml of albumin, then 1/2 mL of 10% NaOH. Then shake.
  2. Add CuSO4 drop wise.
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3
Q

6.A2. What was the physical result of the Biuret test?

A

Solution turned a violet color

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4
Q

6.A3. What group is responsible for Biuret test’s results?

A

The presence of peptide linkage in amide group (CONH)

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5
Q

6.A4. What is Biuret Test a good index for determining?

A

Biuret test is a good index for determining the extent of protein hydrolysis.

Hydrolytic products give colors ranging from violet to pink depending on extent of hydrolysis progression.

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6
Q

6.B1. XANTHROPROTEIC TEST

A

Egg albumin + HNO3 (nitric acid) + conc. NH4OH (ammonium hydroxide)

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7
Q

6.B2. Results of Xanthroproteic Test

A

Layers separated; yellow-orange solution forms

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8
Q

6.B3. What is the role of NH4OH in Xanthroproteic test?

A

It is the alkali used to neutralize the solution

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9
Q

6.B4. What was responsible for the formation of yellow-orange color in Xanthroproteic test?

A

This is due to the nitration of the benzene ring / C6H5 in amino acids

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10
Q

6.C1. Millon’s Test

A

2 ml of albumin + Hg (NO3)2 (Millon’s reagent)

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11
Q

6.C2. Results of Millon’s Test

A

Produces flesh-colored —> red precipitate

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12
Q

6.C3. How is Millon’s reagent made?

A

By dissolving Hg / mercury in HNO3 / nitric acid = Hg (NO3)2

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13
Q

6.C4. What causes the reaction in Millon’s test?

A

The phenol group (C6H5-OH) in tyrosine

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14
Q

6.D1. Molisch’s Test

A

Albumin + Molisch reagent + H2SO4

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15
Q

6.D2. Results of Molisch’s Test

A

Violet ring develops at junction of liquid layers

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16
Q

6.D3. What causes the reaction in Molisch’s test?

A

Presence of glyco-protein

17
Q

6.E1. Sulfur Test

A

albumin + NaOH + CH3COOH

Cover above solution with filter paper covered in Pb (C2H3O2)2. Expose to heat.

18
Q

6.E2. What happened to the solution? The paper dipped in Pb (C2H3O2)2 when exposed to the solution?

A

Black precipitates formed

The bottom of the paper turned brown

19
Q

6.E3. Albumin when heated with NaOH split up the sulfur to form _____

A

Na2S / Sodium sulfide

20
Q

6.E4. What happens when Na2S is acidified with CH3COOH and treated with Pb (C2H3O2)2?

A

Black precipitate or PbS / lead sulfide forms

21
Q

6.E5. What makes the reaction in Sulfur Test possible?

A

Presence of cystine, cysteine, or methionine

22
Q

6.F1. EFFECT OF HEAT IN PROTEINS

A

Albumin + heat in water bath

23
Q

6.F2. What happens to albumin after being heated?

A

Albumin coagulates and the solution becomes cloudy

24
Q

6.G1. HELLER’S RING TEST

A

Albumin + conc. HNO3

25
Q

6.G2. What is the reaction after albumin is added with HNO3 in Heller’s ring test?

A

White precipitate ring forms, and liquid at the bottom turns yellow

26
Q

6.G3. What type of acid is HNO3 used in the test?

A

Mineral Acid

27
Q

6.G4. What does presence of white precipitate ring indicate?

A

It indicates the presence of coagulated protein

28
Q

6.H1. HEAVY-METAL SALTS

A

Albumin + (1) CuSO4 / copper sulfate, (2) AgNO3 / silver nitrate, (3) Pb (C2H3O2) / lead acetate

CSL

29
Q

6.H2. What is the pH level and the % of the three heavy metal salts?

A
  • pH level above 6
  • all 1% solution
30
Q

6.H3. What do heavy metal salts and protein form?

A

Insoluble metallic proteinates

31
Q

6.H4. Reaction of heavy metal salts

A

Albumin + CuSO4: light blue color

Albumin + AgNO3: cloudy dirty white solution w/ ppt

Albumin + Pb (C2H3O2)2: Very White ppt is obtained

32
Q

6.I1. ALKALOIDAL REAGENTS

A

Albumin + (1) C76H52O46 / tannic acid, (2) C6FeK4N6 / potassium ferrocyanide, and (3) saturated C6H3N3O7 / picric acid

TPP

33
Q

6.I2. What is the pH level and percentage of all alkaloidal reagents?

A
  • pH level at below 4
  • Percentage at 5%
34
Q

6.I3. Results of alkaloidal reagents:

A

Albumin + C76H52O45: Greyish Brown ppt forms

Albumin + C6FeK4N6: Metallic silver color forms

Albumin + C6H3N3O7: Bright yellow ppt forms