urine

BIOLOGICAL EVIDENCE
 INTRODUCTION

• Urine can be defined as the liquid waste product secreted by kidneys by a

process known as urination and is excreted through the urethra.

• Cellular metabolism produces various waste products that are needed to be

cleared from the bloodstream and are also rich in nitrogen.

• Healthy urine is not toxic and only undesirable substances are eliminated by the
body.

• The urine is transparent pale yellow solution but its color also ranges from
colorless to pale yellow over-hydration is indicated by the presence of colorless
urine.
 CONSTITUENTS OF
URINE
Urine is a liquid byproduct of the body secreted by the kidneys through a
process called urination and excreted through the urethra. The normal
chemical composition of urine is mainly water content, but it also includes
nitrogenous molecules, such as urea, as well as creatinine and other metabolic
waste components.

Normal constituent of Urine

• Urine is an aqueous solution of greater than 95% water, with a
minimum of
these remaining constituents, in order of decreasing
concentration:
• Urea 9.3 g/L.
• Chloride 1.87 g/L.
• Sodium 1.17 g/L.
• Potassium 0.750 g/L.
• Creatinine 0.670 g/L .
• Other dissolved ions, inorganic and organic compounds
(proteins ,metabolites & hormones)
 GENERAL URINE TEST

Colour
Normal: Pale to dark yellow in colour
Abnormal: Various foods and medicines and drugs may affect the urine
color. Long-term kidney disease or uncontrolled diabetes may also result in
color change of urine. Dark yellow urine can be caused by the dehydration.
Blood in urine results in its red color.

Odour
Normal: bad odour
Abnormal: Consumption of some food, vitamins, and antibiotics can cause
urine to have a different odour. A sweet, fruity odour may be caused by
uncontrolled diabetes
pH
Normal: Ph between 4.6–8.0
Abnormal: Some foods and medicines can affect urine pH; it can be caused
by severe vomiting, a kidney disease, some urinary tract infections, and
asthma. A low pH may be caused by severe lung diseases.

Protein
Normal: None
Abnormal: Protein in the urine indicates kidney damage, an infection,
cancer, high blood pressure, diabetes.Protein in the urine may also indicate
that the person is suffering from leukemia.

Ketones
Normal: None
Abnormal: Ketones in the urine shows uncontrolled diabetes, a very lowcarbohydrate
diet, starvation or eating disorders.
 Test of Degraded Protein in Urine
Bilirubin
Bilirubin is a bile salt formed by the breakdown of red blood cells and is passed on from
the body in feces. Generally, urine does not consist of bilirubin. If it is present, it means
that the liver is damaged or that the flow of bile from the gallbladder is blocked and
passes in blood streams. During the process of normal heme catabolism, the final
breakdown product results in the yellow colored bilirubin

The circulation of bilirubin in the bloodstream is carried out in two forms:

 Indirect Bilirubin. The indirect form of bilirubin is not soluble in water but once
it travels through the bloodstream into the liver, it is changed into the soluble
form.
 Direct Bilirubin. It is formed from the indirect bilirubin in liver and readily
dissolves in water
Urobilinogen
Urobilinogen is formed by the breakdown of bilirubin. It is also passed from the body in
stool but only small amount of it is present in urine. High amount of Urobilinogen may be
a sign of disease caused in liver or it may also be that the flow of bile has been blocked
from the gallbladder.

Bence Jones protein

In about 50% individuals suffering with a rare type of cancer, this abnormal protein
is
found in their urine. A urine test is often done when multiple myeloma is suspected.
 PRELIMINARY TESTS FOR URINE
Detection of Urea by Gee’s Method
The suspected stain is extracted with acetone and then the acetone extract is concentrated
by evaporation. The concentrated extract is then filtered and then it is evaporated to
dryness.

The residue is dissolved in acetone mixed with the help of glass rod. A drop of the
solution is then taken on a microscopic slide and then again it allowed to evaporate, after
this the separation of the urea crystals may be observed. The crystals are long colourless,
4 or 6 sided rhombic crystals. The nitric acid solution is added by the help of glass rod,
the urea nitrate crystals may be observed.
Detection of Creatinine
Concentrated stain is taken on a chromatographic paper and it is treated with 2N NaOH
solution and then 1 drop of picric acid. The red color crystal indicates the presence of
creatinine.

Detection of Indican
1 ml of resorcinol reagent is added to the small quantity of the extracted stain then
the
1ml of cupric bromide solution is added mixed and whole mixture is extracted with
amylacetate.The red colour of the crystal indicates the presence of Indican.
 CONFIRMATORY TEST FOR URINE

Urea nitrate crystal test:

 Aquous extract of stain on slide

 Add one drop of conc. Nitric acid
 Urea nitate crystal are observed
 Hexagonal stacked crystals formed
 ANALYSIS OF DRUGS IN URINE

• Urine tests use a sample of urine to test for the

presence of alcohol, certain illegal drugs and
prescriptions medications. Urine tests are
painless and only requires a urine sample of the
person getting tested. The urine drug test
usually screens for alcohol, amphetamines,

benzodiazepines, marijuana, cocaine, PCP, and

opioids (narcotics).

• Urine analysis can test for alcohol use by

detecting the presence of Ethyl Glucuronide
(EtG), a metabolite found in alcoholic beverages.
The test can show ethanol ingestion within the
previous 3-4 days.
DETERMINATION OF TIME FOR DIFFERENT DRUGS IN URINE

TYPES OF DRUG Detection of drug in urine

o Marijuana – single use • 1-7 DAYS

o Marijuana- regular use • 7-100 DAYS
o Amphetamines • 1-3 DAYS
o Cocaine • 1-3 DAYS
o Heroine • 1-4 DAYS
o PCP • 3-7DAYS
(Phenylcyclohexyl piperidine)
Thank you