PHYSICAL PROPERTIES

 Color: Typically yellow-amber, but varies according to recent diet and the concentration of
the urine. Drinking more water generally tends to reduce the concentration of urine, and
therefore causes it to have a lighter color. Dark urine may indicate dehydration. Red urine
indicates red blood cells within the urine, a sign of kidney damage and disease.

 Smell: The smell of urine may provide health information. For example, urine of diabetics
may have a sweet or fruity odor due to the presence of ketones (organic molecules of a
particular structure) or glucose. Generally fresh urine has a mild smell but aged urine has
a stronger odor similar to that of ammonia.

 The pH of normal urine is generally in the range 4.6 – 8, with a typical average being around
6.0. Much of the variation occurs due to diet. For example, high protein diets result in more
acidic urine, but vegetarian diets generally result in more alkaline urine (both within the
typical range of 4.6 – 8).

 Density: Density is also known as “specific gravity.” This is the ratio of the weight of a
volume of a substance compared with the weight of the same volume of distilled water. The
density of normal urine ranges from 0.001 to 0.035.

 Turbidity: The turbidity of the urine sample is gauged subjectively and reported as clear,
slightly cloudy, cloudy, opaque or flocculent. Normally, fresh urine is either clear or very
slightly cloudy. Excess turbidity results from the presence of suspended particles in the
urine, the cause of which can usually be determined by the results of the microscopic urine
sediment examination. Common causes of abnormal turbidity include: increased cells,
urinary tract infections or obstructions.

BIURET TEST

The biuret test uses an alkaline mixture, or reagent, composed of potassium hydroxide and
copper sulfate. The normal color of biuret reagent is blue. The reagent turns violet in the presence
of peptide bonds -- the chemical bonds that hold amino acids together. The proteins detected
must have at least three amino acids, which means that the protein must have at least two peptide
bonds. The reagent’s copper ions, with a charge of +2, are reduced to a charge of +1 in the
presence of peptide bonds, causing the color change. The techniques of absorption spectroscopy,
which identify the electromagnetic frequencies a sample will absorb, allow testers to quantify the
concentration of protein in a sample.

We describe a method for measuring urinary protein with a centrifugal analyzer. Biuret reagent is
used, and blanking with an ultrafiltrate of urine eliminates interferences from the nonprotein,
biuret-positive chromogens in urine. We compare results by this new method with those by a
manual method in which trichloroacetic acid precipitation and biuret reagent are used. The new
method shows good precision and excellent correlation (r = 0.997) with the manual method. The
ease and convenience of this assay should make this a useful method for the routine clinical
laboratory.

BENEDICT’S TEST

Benedict’s test is used as a simple test for reducing sugars. A reducing sugar is a carbohydrate
possessing either a free aldehyde or free ketone functional group as part of its molecular structure.
This includes all monosaccharides (eg. glucose, fructose, galactose) and many disaccharides,
including lactose and maltose.
Benedict’s test is most commonly used to test for the presence of glucose in urine. Glucose
found to be present in urine is an indication of Diabetes mellitus.

Principle of Benedict’s Test

Reducing sugars under alkaline condition tautomerise and form enediols. Enediols are powerful
reducing agents. They can reduce cupric ions (Cu2+) to cuprous form (Cu+), which is responsible
for the change in color of the reaction mixture. This is the basis of Benedict’s test. When the
conditions are carefully controlled, the colouration developed and the amount of precipitate
formed (Cuprous oxide) depends upon the amount of reducing sugars present.

Composition and Preparation of Benedict’s reagent

One litre of Benedict’s Solution can be prepared from 100 g of anhydrous sodium carbonate, 173
g of sodium citrate and 17.3 g of copper(II) sulfate pentahydrate.

Constitutent Amount Functions

Copper 17.3 gm Furnishes cupric ions (Cu++)

sulphate
 Sodium 100 gm Makes medium alkaline
carbonate

Sodium citrate 173 gm Complexes with the copper (II) ions so that they
do not deteriorate to copper(I) ions during
storage

Distilled water Upto 1000 Solvent

ml

Quality Checking : Benedict’s solution is blue in color. In order to check purity of Benedict’s
solution take 5 ml of Benedict’s solution in test tube and heat it. If is does not change color, it
means it is pure.

Procedure of Benedict’s Test

1. Pipette 5 ml of Benedict’s reagent in a test tube (20x150mm).

2. Add 8 drops of urine to the Benedict’s reagent.

3. Heat carefully on a flame of a gas burner or place in a boiling water for 5-10 minutes.

4. Cool under tap water or by placing in a beaker containing tap water.

5. Observe the color change and precipitate formation and analyse the test result.

Result Interpretation of Benedict’s Test

The colour of the mixture serves as a guide to the amount of sugar in the urine. Remove the tubes
and examine the solution in each tube for precipitate and change of colour. Report the sugar
concentration as follows:
 Color Approximate glucose mg/dl Indication

Blue solution Nil

Green solution <500 mg/dl Trace

Green ppt 500-1000 mg/dl +

Yellow ppt 1000-1500 mg/dl ++

Orange ppt 1500-2000 mg/dl +++

Red to Brick red ppt >2000 mg/dl ++++

Factors affecting Benedict’s Test

False positive reactions may also be obtained if certain drugs are present, e.g. salicylates,
penicillin, streptomycin, isoniazid, and p-aminosalicyclic acid.

Chemicals present in a concentrated urine which may reduce Benedict’s reaction include
creatinine, urate, and ascorbic acid (reduction is slight).

TEST FOR KETONE BODIES

The test measures ketone levels in your urine. Normally, your body burns glucose (sugar) for
energy. If your cells don't get enough glucose, your body burns fat for energy instead. This
produces a substance called ketones, which can show up in your blood and urine. High ketone
levels in urine may indicate diabetic ketoacidosis (DKA), a complication of diabetes that can lead
to a coma or even death. A ketones in urine test can prompt you to get treatment before a medical
emergency occurs.
A urine test will show that you have:

 No ketones

 Trace amounts of ketones

 Moderate levels of ketones

 Large amounts of ketones

TEST FOR BICARBONATE

Bicarbonate is an electrolyte, a negatively charged ion that is used by the body to help maintain
the body's acid-base (pH) balance. It also works with the other electrolytes (sodium, potassium,
and chloride) to maintain electrical neutrality at the cellular level. This test measures the total
amount of carbon dioxide (CO2) in the blood, which occurs mostly in the form of bicarbonate
(HCO3-). The CO2 is mainly a by-product of various metabolic processes.

Measuring bicarbonate as part of an electrolyte or metabolic panel may help diagnose an

electrolyte imbalance or acidosis or alkalosis. Acidosis and alkalosis describe the abnormal
conditions that result from an imbalance in the pH of the blood caused by an excess of acid or
alkali (base). This imbalance is typically caused by some underlying condition or disease.

The lungs and kidneys are the major organs involved in regulating blood pH through the removal
of excess bicarbonate.

 The lungs flush acid out of the body by exhaling CO2. Raising and lowering the respiratory
rate alters the amount of CO2 that is breathed out, and this can affect blood pH within
minutes.

 The kidneys eliminate acids in the urine and they regulate the concentration of bicarbonate
(HCO3-, a base) in blood. Acid-base changes due to increases or decreases in
HCO3- concentration occur more slowly than changes in CO2, taking hours or days.

Any disease or condition that affects the lungs, kidneys, metabolism, or breathing has the
potential to cause acidosis or alkalosis.

The bicarbonate test gives a healthcare practitioner a rough estimate of a patient's acid-base
balance. This is usually sufficient, but measurements of gases dissolved in the blood (blood
gases) may be done if more information is needed. Bicarbonate is typically measured along
with sodium, potassium, and possibly chloride in an electrolyte panel as it is the balance of these
molecules that gives the healthcare practitioner the most information.
 TEST FOR CHLORIDE

Urine chloride test measures the amount of chloride in your urine.

Your body tries to keep its acid-base (pH) level in balance. But certain conditions can cause an
imbalance. If your body tissues become too alkaline, you may get metabolic alkalosis. Alkaline is
the opposite of acidic.

You can get metabolic alkalosis from taking medicines that make you urinate more (diuretics). It
can also happen after you have been vomiting. Or it can happen if you've had the contents of
your stomach suctioned.

It can also be caused by a rare medical condition, such as Bartter syndrome or Gitelman
syndrome. It can also be caused by having low levels of potassium.

If you have metabolic alkalosis, seeing how much chloride is in your urine gives your healthcare
provider more information about your condition.

Test results may vary depending on your age, gender, health history, the method used for the
test, and other things. Your test results may not mean you have a problem. Ask your healthcare
provider what your test results mean for you.

Results are given in milliequivalents per liter (mEq/L). Normal results range from 25 to 40 mEq/L.

If your levels are below 25 mEq/L, you may have a lack of chloride in your diet. It may also mean
that you have been vomiting or had medical suctioning of your stomach contents. It may also
mean that you have used diuretics in the past.

If your levels are higher than 40 mEq/L, you may have:

 Bartter syndrome or Gitelman syndrome

 Severely low potassium levels

Higher levels of chloride may also be caused by recently taking medicines used to increase
urination (diuretics).