Qualitative Tests for Proteins
Dr. Abbasi
Hamadan Medical University
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Proteins
Proteins are large, complex molecules that play many critical roles
in the body.
They do most of the work in cells and are required for the
structure, function, and regulation of the body’s tissues and
organs. Some function:
• Antibody
• Enzyme
• Messenger
• Structural component
• Transport/storage
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Importance of this test?????? Detection of proteinuria
PROTEINURIA
Presence of greater than normal amounts of protein in the urine
The condition is often a sign of kidney disease.
Healthy kidneys do not allow a significant amount of protein to pass through their filters.
But filters damaged by kidney disease may let proteins such as albumin leak from the blood
into the urine.
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Urine Albumin - A urine test checks for albumin in urine.
Albumin is a small protein that can pass into the urine
when the filters in the kidneys are damaged.
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What are the symptoms of kidney disease?
Most people with kidney disease do not have symptoms unless the
disease is very advanced. However, patients may feel:
• Foamy, frothy or bubbly-looking urine
• Swelling in hands, feet, abdomen or face
• Shortness of breath
• Needing to urinate more often
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• Hiccups
• Fatigue (feeling tired)
• Sleep problem
• Nausea and vomiting
• Dry, itchy skin.
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What are the stages of Chronic Kidney Disease (CKD)?
CKD is divided into 5 stages. Each stage tells you how well the kidneys are working.
Doctors often estimate kidney function based on the glomerular filtration rate (GFR), a test
based on levels of creatinine (a waste product that is normally removed by the kidneys) in
the blood. As kidney disease gets worse, GFR decreases.
when the kidneys are working normally, they can be described as working at 100%.
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Cause of Proteinuria
• Damage to the kidneys
• Diabetes
• High blood pressure (hypertension)
• Infection - Inflammation in the kidneys (Nephritis)
• Toxins
• Immune system disorders
• Other risk factors include:
• Age over 65
• Family history of kidney disease
• Preeclampsia (high blood pressure and proteinuria in pregnancy)
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If doctor suspects that you have kidney disease, the urine test
for albumin will be repeated.
Three positive results over three months or more is a sign of
kidney disease.
Subjects may also be given:
• Imaging tests. (An ultrasound or CT scan). This produces a
picture of kidneys and urinary tract. It can show whether
kidneys have kidney stones or other problems.
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Treatment
Because proteinuria is a symptom and not a disease itself,
medical care focuses on treating the underlying condition,
such as normalizing blood pressure in people with
hypertension or controlling blood sugar levels in those with
diabetes.
Treatment may include:
• Medications (blood pressure reducing)
• Changes in diet
• Lifestyle changes such as losing extra weight, exercising, and
stopping smoking.
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Diet For Proteinuria
Limit Carbohydrates - Carbohydrates have the greatest impact on blood sugar
levels, which is important if you have kidney problems due to your diabetes. In
general, your diet should include around 50% carbohydrates.
Eat less red meat; instead, have more lean poultry and seafood.
Choose grilled or baked protein instead of fried.
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Renal Diet - If you are diagnosed with Proteinuria, you may be
advised to follow a "renal diet". This includes foods that are low
in sodium, potassium, magnesium and protein.
Some specific that have high amount of potassium include:
• Oranges and orange juice
• Leafy green vegetables, such as spinach and
greens (collard and kale)Potatoes
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Avoid excess magnesium in your diet, often found in laxatives
(such as milk of magnesia), or antacids( neutralizes stomach
acidity) unless specified by your healthcare provider.
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Increase fiber intake - Up to 55 grams of fiber per day is
recommended.
There are many types of "good and bad" fats. The easiest thing to
remember is to limit your intake of saturated fats and oils.
Limit high-fat dairy, including milk, yogurt, frozen desserts, cream
cheese and sour cream.
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Denaturation of Proteins
• Denaturation is the disruption of secondary, tertiary and
quaternary structure of proteins leading to loss of their biological
activity. precipitation
Acids, pH, Heat
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Causes of denaturation
Physical factors Chemical factors such
such as heating as strong acid or base
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Biuret Test
To detect the presence of a protein or peptides.
Alkaline Copper sulfate reacts with all compounds containing peptide bonds to
give a purple color in a strongly alkaline solution.
When proteins and peptides (i.e peptide bonds) treated with an alkaline solution
of dilute copper sulfate a violet color is formed .
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Method
Test tube 1: add 1ml of protein Albumin +1ml of NaOH
mix well and then add 500ul(0.5ml) of Copper sulfate (CuSO4)
>>>> purple color (positive).
Test tube 2: add 1ml of water +1ml of NaOH
mix well and then add 500ul(0.5ml) of CuSO4 >>>> blue color
(negative).
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Heller's test
• Heller's test is commonly used to test for the presence of proteins
in urine. This test was invented by the Austrian Chemist,
Johann Florian Heller (1813-1871).
• Heller's test is a chemical test that shows that strong acids cause
the denaturation of proteins.
• Concentrated nitric acid is added to a protein solution from the
side of the test tube to form two layers.
• A white ring appears between the two layers if the test is positive.
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Method
Test tube 1: add 1ml of protein Albumin +1ml of nitric acid along
the inner sides of the tube >>>> white ring appears (positive).
• Test tube 2: add 1ml of water +1ml of nitric acid along the
inner sides of the tube >>>> without change (negative).
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Sulfosalicylic acid test
Sulfosalicylic acid is used in urine to determine urine
protein content.
The chemical causes the precipitation of dissolved proteins,
which is measured from the degree of turbidity.
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Method
Test tube 1: add 1ml of protein Albumin +1ml of Sulfosalicylic acid
>>>> precipitation (positive).
Test tube 2: add 1ml of water+1ml of Sulfosalicylic acid >>>>
without precipitation (negative).
Results:
Neg. No turbidity
(1+) Turbidly, No granules.
(2+) Turbidity, granulation, flocculation.
(3+) Turbidity, granulation
(4+) Clumps of proteins.
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