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Protein Lab PDF

The document summarizes several common protein tests, including Xanthoproteic, Millon's, Hopkin's Cole, Lead Acetate, Ninhydrin, Biuret, and precipitation tests. It describes the principle, reagents, and results for each test. For example, the Xanthoproteic test detects benzene rings and gives a positive result for all proteins tested except gelatin. The Biuret test detects peptide bonds and is only positive for albumin. Precipitation can be caused by heavy metal salts and alkaloidal reagents.

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90 views19 pages

Protein Lab PDF

The document summarizes several common protein tests, including Xanthoproteic, Millon's, Hopkin's Cole, Lead Acetate, Ninhydrin, Biuret, and precipitation tests. It describes the principle, reagents, and results for each test. For example, the Xanthoproteic test detects benzene rings and gives a positive result for all proteins tested except gelatin. The Biuret test detects peptide bonds and is only positive for albumin. Precipitation can be caused by heavy metal salts and alkaloidal reagents.

Uploaded by

Hopeu Store
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Proteins

Xanthoproteic Test

 Principle:  Reagents:
 Detects benzene ring  Sample + HNO3 + NH4OH
Xanthoproteic Test

 Positive result:
 White ppt. turns yellow
EXCEPT for gelatin
Millon’s Test

 Principle:
 Detects phenolic –OH
group

 Reagents:
 Sample + Millon’s rgt.
(mercurous nitrate in
nitric acid) + 0.1% NaNO2
 Phenol: positive control
Millon’s Test

 Positive result:
 White ppt. turns red
EXCEPT for
phenylalanine
Hopkin’s Cole Test

 Principle:  Reagents:
 Detects indole ring  Sample + Hopkin’s Cole
rgt. (Glyoxylic acid) +
H2SO4
Hopkin’s Cole Test

 Positive result:
 Blue violet ring ONLY for
tryptophan
Lead Acetate (a.k.a. Sulfur) Test

 Principle:
 Detects sulfur-containing
amino acids

 Reagents:
 Sample + 10% NaOH +
lead acetate

 Positive result:
 Black ppt. (lead
sulfide/PbS) ONLY for
albumin
Ninhydrin Test

 Principle:  Reagents:
 Detects amine group  Sample + Ninhydrin
reagent
Ninhydrin Test

 Positive result:
 Purple color
 Albumin more positive
than urea
Biuret

 Principle:  Reagents:
 Detects at least two  Sample + NaOH + CuSO4
peptide bonds
Biuret

 Positive result:
 Purple color ONLY for
albumin
Denaturation

 Destruction of peptide
bonds, breaking down
proteins into amino acids
(negative Biuret test)

 Caused by: Heat, acid or


alcohol
Denaturation

 Results to:
 Coagulation and
appearance of white ppt.
 Inactivation of enzymes
and proteins in cells and
pathogens
Precipitation w/ Heavy Metal Salts

 Free carboxylate anions


of acidic amino acids
precipitate heavy metals
(cations).

 Milk and egg albumin


serve as antidotes in
heavy metal posioning.
Precipitation w/ Alkaloidal Rgts.

 Alkaloidal reagents:
tannic and picric acid

 The anion of the acid


reacts with the amino
groups of the protein
resulting to precipitation.
 Tannic acid – brown ppt.
 Picric acid – yellow ppt.
Precipitation w/ Alkaloidal Rgts.

 Used to treat burns as


these reagents form a
protective covering w/
skin proteins over the
wound.
Results Summary

 Xanthoproteic – All samples tested positive EXCEPT


gelatin

 Millon’s – All samples tested positive EXCEPT


phenylalanine

 Hopkin’s Cole – Only tryptophan was positive.

 Lead acetate – Only albumin was positive.

 Ninhydrin – albumin > urea; ammonia water (-)


Results Summary

 Biuret – Only albumin was positive.

 Heat, HCl, H2SO4 and ethanol causes protein


denaturation/coagulation.

 Heavy metal salts (lead acetate, AgNO3 and HgCl2)


and alkaloidal rgts (tannic and picric acids) causes
protein to precipitate.

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