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General Proteins Color Tests - KSU...

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Done by :Sahar AL-subaie General Proteins Color Tests
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Page 1: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Done by :Sahar AL-subaie

General Proteins Color Tests

Page 2: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Proteins are polymers of amino acids

covalently linked through peptide

bonds into a chain.

protein structure

Page 3: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 4: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Amino acid structure

Page 5: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Peptide bound

Page 6: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Biuret Test

Ninhydrin Test

Xanthoproteic Test

General Proteins Color Tests

Page 7: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

It is a general test used for detecting the

presence of proteins and peptides.

Biuret Test

Page 8: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

(CU++) + peptide bonds (CO-NH)

pink-violet colored complex

Principle:

alkaline solution

Page 9: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 10: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

at least two peptide bonds are required

for a positive test.

Page 11: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 12: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

1)Do amino acids answer the test?

2)What it is the smallest size of peptides

that can answer the test?

(some students can act as amino acids and make peptide bonds by holding

there hands like this

to answer the 2nd question)

Page 13: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

1)The free amino acids lack the peptide bonds ,so

they do not answer this test.

2)Tripeptid =3 amino acids or 2 peptide bonds

Answers:

Page 14: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 15: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Procedure:

Page 16: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

1%casein 1%glucose 1%sucrose 1%alanine 1%egg albumin H2O

1 ml 1 ml 1 ml 1 ml 1 ml 1 ml

10% NaOH

1 ml 1 ml 1 ml 1 ml 1 ml 1 ml

0.1% CUSO4

5 drops 5 drops 5 drops 5 drops 5 drops 5 drops

Page 17: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

It is a general test used for detecting the presence of proteins and peptides and amino acids.

Ninhydrin Test

Page 18: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

α-amino group (in amino acids)+ninhydrin

blue colored complex (librate NH3 with ninhydrin).

principle

Page 19: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Ninhydrin is used to locate the α-amino

acid in paper chromatography as a blue to

purple spots.

Page 20: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 21: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 22: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

It also permits the quantitative estimation

of α-amino acid and peptides in

column chromatography

Page 23: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Proline give yellow color due to lack of α-

amino group

Page 24: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

It is a general test for proteins depending

on the presence of 2 amino acids (tyrosine

& tryptophan)

Xanthoproteic Test

Page 25: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Tyrosine or Tryptophan + con.HNO3 heat Yellow color

Principle:

Page 26: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Tyrosine structure:

Page 27: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Tryptophan structure:

Page 28: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

Phenylalanine does not produce the color

because the benzene ring is not activated

for nitration.

Page 29: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color
Page 30: General Proteins Color Tests - KSU Facultyfac.ksu.edu.sa/sites/default/files/general_proteins_color_tests_1.pdf · Biuret Test Ninhydrin Test Xanthoproteic Test General Proteins Color

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