1. A
PROJECT
ON
“Study of organic constituents present of the urine in
different age group”
Submitted To
TEERTHANKER MAHAVEER UNIVERSITY
COLLEGE OF PARAMEDICAL SCIENCES
MORADABAD
In Partial Fulfillment of the Requirement of the Degree of
Bachelor of Science in Medical Lab Technology
SESSION: 2015-16
GUIDE BY: -
MR. VIKASH GAUR
CO-ORDINATOR
Dept. of Medical Lab Technology
TMU College of Paramedical Sciences
Teerthanker Mahaveer University
SUBMITTED BY:-
DEEPANSHU GUPTA (TPS1301011)
2. ii
TEERTHANKER MAHAVEER UNIVERSITY
COLLEGE OF PARAMEDICAL SCIENCES
CERTIFICATE
This is to certify that DEEPANSHU GUPTA has carried out the Project work for the award of
Bachelor Degree in Medical Lab Technology from Teerthanker Mahaveer University,
Moradabad as regular student, in session (2015-16).
In compliance with the provision/guidelines of Teerthanker Mahaveer University, Moradabad, He
has been assigned a project entitled “STUDY OF ORGANIC CONSTITUENTS IN URINE IN
DIFFERENT AGE GROUP” under the guidance of Mr. VIKASH GAUR. The project work
has been genuinely carried out by the student for the duration specified by the university.
He has made sincere efforts in the completion of the project work.
HOD/ Coordinator
Dept. of Medical Lab Technology
TMU College of Paramedical Sciences
Teerthanker Mahaveer University
3. iii
TEERTHANKER MAHAVEER UNIVERSITY
COLLEGE OF PARAMEDICAL SCIENCES
CERTIFICATE
This is to certify that DEEPANSHU GUPTA has carried out the Project work for the award of
Bachelor Degree in Medical Lab Technology from Teerthanker Mahaveer University,
Moradabad as regular student, in session (2015-16).
In compliance with the provision/guidelines of Teerthanker Mahaveer University, Moradabad, He
has been assigned a project entitled “STUDY OF ORGANIC CONSTITUENTS IN URINE IN
DIFFERENT AGE GROUP” under the guidance of Mr. VIKASH GAUR. The project work
has been genuinely carried out by the student for the duration specified by the university.
He has made sincere efforts in the completion of the project work.
GUIDE: - MR. VIKASH GAUR
CO-ORDINATOR
Dept. of Medical Lab Technology
TMU College of Paramedical Sciences
Teerthanker Mahaveer University
4. iv
ACKNOWLEDGEMENT
If words are consider as symbol of approval and tokens of acknowledgement, then let words play
the role of expression of my gratitude.
First the foremost I would like to thank the Almighty, who showered his blessing in all walks of
my life.
I am highly indebted to my guide Mr. Vikash Gaur (Course -Coordinator) T.M.U College of
Paramedical Sciences, who took his real personal interest in providing me proper guidance,
encouragement and support at all levels.
I am very thankful My Class Coordinator Mr. Devendra Singh and other faculty members for
giving their excellent guidance and constant encouragement in every step of this study.
My entire efforts stand credited at this movement only because of my family who whole heartedly
stood beside me always in each step of my career.
Last but not the least, I would like to thank my friend for their valuable suggestions and support in
the completion of my project work.
5. SR.NO CONTAIN PAGE NO
1 INTRODUCTION 1 - 2
2 AIM OF THE STUDY 3
3 LITERATURE REVIEW
FORMATION OF URINE
NORMAL PROPERTIES
PATHOLOGICAL DISORDER
4 -20
4 METHODOLOGY
EXAMINATION OF URINE
PHYSICAL EXAMINATION
CHEMICAL EXAMINATION
MICROSCOPIC EXAMINATION
21 - 42
5 DATA COLLECTION 43 - 55
6 DATA ANALYSIS 56 - 62
7 CONCLUSION 63 - 65
8 REFERENCE 66 - 68
INDEX
7. 2
INTRODUCTION
Urine is the fluid containing water soluble waste products excreted from the blood through the
urinary system The urinary system consists of the kidneys, ureters, urinary bladder, and urethra.
The system produces urine by a process of filtration, reabsorption, and tubular secretion. Urine is
a liquid by-product of the body secreted by the kidneys through a process called urination (or
micturition) and excreted through the urethra. Urine is mainly composed of 95% water and the rest
being made up 5% of organic (urea, protein, sugar, uric acid) and inorganic (sodium, potassium,
calcium, magnesium) substance.
Kidney is most important excretory organ and most of the metabolic wastes are excreted in urine.
Urine is formed in kidney by ultrafiltration of blood plasma. Nephron is a basic filtration
(functional) and structure unite of kidney that take part in urine formation. Each kidney has about
one million nephrons. Each nephron consist of glomerulus, surrounded by a double walled
epithelium sac called Bowman’s capsule and a long renal tubule divided into proximal convoluted
tubule (PCT), loop of Henle and distil convoluted tubule (DCT), that opens finally into the
collecting duct. The loop of Henle consist of three tubular segments i.e. descending limb thin
segment and ascending limb.
8. 3
AIM OF STUDY
To estimate the amount of organic constituents in urine of patients of different age group to
determine the prevalence of pathological disorder in the Moradabad district.
10. 5
FORMATION OF URINE
A large number of metabolic waste are produced in various metabolic processes that take place in
our body.
Metabolic activities
In
Hepatic & extra – hepatic tissues
Metabolic wastes
WATER &CO2
INORGANIC: PHOSPHATE, SULPHATE,
AMMONIA, CHOLORIDES ETC.
INORGANIC: UREA, URIC ACID,
CREATININE, HIPPURIC ACID,
OXALATES, UROBILINOGEN, ETC.
CHOLESTEROL, BILE ACIDS,
BILE PIGMENTS, ETC.
Bile
Gastric intestine tract
Excreted outside the
body through faeces
Lung
Co2 is exhaled in
gaseous from
Urine formation
Glomerular Filtration Tubular
Reabsorption Tubular Secretion
Urine containing
water and water –
soluble metabolic
wastes
BLOOD
KIDNEY
11. 6
The various metabolic wastes normally formed in different metabolic activities, are water, co2
organic substance and inorganic substance. Because of their harmful effect, e.g. retention of co2
in lungs causes respiratory acidosis, retention of ammonia and urea in blood leads to
hyperammonemia and uremia, respiratory, and metabolic wastes cannot be retained in body for
longer time and, therefore, they are regularly excreted outside the body. Although there are many
routes for the disposal of metabolic wastes, but lungs and kidneys are the two important orange
that are actively involved in excretion of wastes. Co2 is largely eliminated from the body through
lungs and most of the water – soluble metabolic wastes are excreted through kidneys, in urine.
Some of the metabolic wastes are also excreted through faeces, sweat and saliva
Nephron
Kidney is the most important excretory organ and most of the metabolic wastes excreted in urine.
Urine is formed in kidney by ultrafiltration of blood plasma. Nephron is basic filtration functional
and structure unit of the kidney that takes part in urine formation. Each kidney have about one
million nephron. Each nephron consist of glomerulus surrounded by a double walled epithelial sac
called bowman’s capsule and long renal tubule divided into proximal convoluted tubule, loop of
Henle and distal convoluted tubule that open finally into the collecting duct that loop of Henle
consist of three tubular segments-a descending limb a thin segment and an ascending limb.
12. 7
About 1200ml blood (i.e. about 650ml plasma) flow per min. through both kidney and filtrate to
form urine (1-2ml/min).The urine formation occurs in three secretion
1. Glomerular filtration
2. Selective tubular reabsorption
3. Tubular secretion
13. 8
Glomerular Filtration
Renal artery supply about 1200ml blood to both kidney per minute i.e. 1700litre/24 hrs. In kidney
blood filtrate passively through glomeruli are nephron as blood passes through glomeruli the water
substance of low molecular weight (<69kD) are filtrated out from the blood with the retention of
plasma protein. Each glomerulus consist of a network of fine blood capillary lined by a single layer
of a simple cuboidal epithelium. The lining epithelial cell of glomerulus and bowman capsule serve
a filtrate. The fluid which is filtrate by glomeruli is called glomerulus filtrate are primary urine.
14. 9
The volume of glomerulus filtrate form per minute by both kidney is known as glomerular filtrate
rate (GFR). In the adult normal GFR 120ml/minute under normal condition about 170litre
glomerular filtrate is formed per 24 hrs. Out of which only 1.5-1.8 liter is excreted as urine. The
glomerular filtrate the normally contain water Na+
, K+
, Ca++
, Mg++
, Phosphate sulphate amino
acid, glucose, urea, uric acid, creatinine etc. the glomerular filtrate also contain very small quantity
of low molecular weight protein. Since the plasma constituents of high molecular weight (>60kD)
cannot be filtrated through glomeruli high molecular protein in urine .example albumin Gama
globulin etc. are normally found absent in glomerular filtrate the presence of protein in urine
(protein, urea) indicate renal damage or dysfunction in renal damage lining epithelial cell of
glomeruli permeable to high molecular weight protein which pass in urine.
15. 10
Tubular Reabsorption
The glomerular filtrate pass through renal tubule. It is concentrate due to reabsorption of water.
About 99% of the total water glomerular filtrate is reabsorbed during its passage through different
segment of the renal tubule. This is based on the fact the under normal condition glomerular filtrate
formed in adult is about 170litre/24 hrs. Of which only 1.5-1.8 liters is excreted as urine after its
passes through the renal tubules. A long with water high renal threshold substance such as sugar
(glucose, fructose, and galactose) and amino acid are selectively reabsorbed by the renal tubule to
the maximum extent. The term tubular maximum is used to indicate the maximum capacity of
kidney reabsorbed a particular substance e.g. maximal rate of glucose reabsorption by renal tubules
is expressed as TMG (tubular maximum for glucose). The normal TMG is about 350mg/minute.
In renal function impairment, Tm of a substance decrease due to its diminished tubular
reabsorption .e.g. in renal glycosuria, glucose begins to appear in urine as it cannot be completely
16. 11
reabsorbed due to the renal tubular transport defect. Small quantity of low threshold substance
such as urea, uric acid creatinine etc. are also reabsorbed from renal tubule. Renal tubule also
reabsorbed phosphate, sulphate, sodium, Cl-
, HCO3, Ca etc. some extent.
Tubular Secretion
Although renal tubule are mainly involved in reabsorption of water and other substance as
described above but certain substance such as K+
, Creatinine, uric acid ,and hydrogen ion are
secreted by the tubular epithelium in urine ammonia formed in distal tubule is also secreted in
urine
17. 12
NORMAL PROPERTIES OF URINE
Constituent grams/liter
Dry weight 55-70 g/24 hrs.
pH 4.6-8.0
Specific gravity 1.005-1.030
Volume per day 600-2500 ml/24 hrs.
INORGANIC SUBSTANCE
Chloride 9.0
Phosphorus 2.0
Total sulfur 1.5
Sodium 4.0
Potassium 2.0
Calcium 0.2
18. 13
Magnesium 0.2
Iron 0.003
ORGANIC SUBSTANCE
Urea 25.0
Uric acid 0.6
Creatinine 1.5
Ammonia 0.6
Undetermined N 0.6
Sugar < 0.5
Protein 0-0.1
ketone bodies -
Carbonates, bicarbonates and free carbonic
acid.
-
Pigment -
Mucin and mucin like substance -
Diastase -
19. 14
PATHOLOGICAL DISORDER IN THE URINE
Proteinuria: Presence of protein in abnormal concentration in urine.
Some common causes of proteinuria
A. Pre-renal causes : Due to impairment of renal circulation:
I. Vomiting in pyloric obstruction.
II. Intestinal obstruction
III. Severe diarrhea
IV. Addison’s disease
V. Heart disease with passive venous congestion of internal organs including kidney.
VI. Ascites and intraabdominal tumour.
VII. Fever
VIII. Malignant hypertension.
B. Renal causes: All causes of glomerulonephritis, nephrotic syndrome and other glomerular
and tubular disease, pyogenic or tubular pyelonephritis.
C. Post renal causes: Lesions of renal pelvis, urinary bladder, prostate, urethra and severe
urinary tract infection., severe lower urinary tract infection, Urinary schistosomiasis
20. 15
Bacterium Endocarditis
It simply refers to the inflammation of endocardium, the innermost layer of the heart. It is more
prominent in patients having congenital heart diseases and acquired valvular diseases. This
infection is usually occurred in damaged part of endothelium as it attracts the floating platelets
and fibrin. The deposition of platelets and fibrin along with microbial growth on damaged part of
endothelium is referred as vegetation. The vegetation formation blocks the passage and damages
the adjacent tissues.it is mainly two types –
1, acute endocarditis
2. Subacute endocarditis
Clinical feature
Symptoms
a. Fever with chills
b. Loss of appetite
c. Unexplained weight loss
d. Discomfort
e. Blurring of vision
Signs
a. Clubbing of fingers
b. Roth’s spots
c. Splinter hemorrhages
21. 16
d. Pleural rub
e. Splenomegaly
f. Poor dentition
g. Osler’s nodules
Treatment
The benzyl Penicillin and is the choice of drug as it inhibit the cell wall synthesis of the
microorganism in combination with the gentamicin in case of streptococcus bacteria .in case of
staphylococcus aureus or If the patient is allergic or resistant to penicillin then vancomycin and
gentamicin are used.
Malignant hypertension
Malignant hypertension is extremely high blood pressure that’s develops rapidly and causes some
type of organ damage. Normal blood pressure id below 140/90. A person with malignant
hypertension has a blood pressure that’s typically above 180/120. Malignant hypertension should
be treated as a medical emergency.
What Causes Malignant Hypertension?
In many people, high blood pressure is the main cause of malignant hypertension. Missing doses
of blood pressure medication can also cause it. In addition there are a certain medical conditions
that can cause it. They include.
22. 17
Collagen vascular disease, such as scleroderma
Kidney disease
Spinal cord injuries
Tumor of the adrenal gland
Use of certain medication including birth control pills and MAOIs
Use illegal drugs, such as cocaine hypertension high blood pressure health care.
What are the symptoms of malignant hypertension?
The main symptoms of malignant hypertension are a rapidly increasing blood pressure of 180/120
or higher and signs of organ damage. Usually, the damage happens to the kidneys or the eyes.
Other symptoms depend on how the rise in blood pressure affects organs. A common symptoms
is bleeding and swelling in the tiny blood vessels in the retina. The retina is the layer of nerves that
line the back of the eye. It senses light and sends signals to the brain through the optic nerve, which
can also be affected by malignant hypertension. When the eye is involved, malignant hypertension
can cause changes in vision.
Other symptoms of malignant hypertension include:
Blurred vision and severe headache.
Chest pain (angina)
Difficulty breathing
Dizziness and shortness of breath
Numbness in the arms, legs and face
23. 18
Glycosuria: Presence of glucose in abnormal concentration in urine.
Conditions in which glycosuria is detected.
A. Diabetes mellitus
B. Certain endocrine disorders like thyrotoxicosis, Cushing’s syndrome
C. Iv infusion of glucose
D. Increased intracranial pressure
E. Phloridzin poisoning
F. Alimentary glycosuria
G. Fanconi syndrome
Diabetes mellitus
Diabetes mellitus is a group of metabolic disorder that occurs due to the hyperglycemia which is
marked by deficiency of insulin secretion and defects in insulin action. Diabetes is a chronic,
lifelong condition that affects your body’s ability to use the energy found in food. There are three
major types of diabetes: types of diabetes,
Type 1 diabetes,
Type 2 diabetes,
Gestational diabetes.
24. 19
All types of diabetes mellitus have something in common. Normally, your body breaks down the
sugars and carbohydrates you eat into a special sugar called glucose. Glucose fuels the cells in
your body. But the cells need insulin, a hormone, in your bloodstream in order to take in the
glucose and use it for energy. With diabetes mellitus, either your body does not make enough
insulin, it cannot use the insulin it does produce or a combination of both.
Since the cells cannot take in the glucose, it builds up in your blood. High levels of the blood
glucose can damage the tiny blood vessels your kidneys, heart, eye, it nervous system.
That’s why diabetes- especially if left untreated-can eventually cause heart disease, stroke, kidney
disease, blindness and nerve damage to nerves in the feet.
Clinical feature
a. Polyuria
b. Polyphagia
c. Polydipsia
d. Blurring of vision
e. Loss of weight
Treatment
Diet is of utmost importance with regard to regularity of meals and total caloric intake in IDDM.
Oral hypoglycemic agents are used to lower the blood sugar level in case of NIDDM. Metformin
often the first choice as it lowers the liver glucose production and increased glucose peripheral
utilization. Sulfonylurea acts to stimulate insulin secretion from the pancreas. The patient is taught
the technique of insulin administration beneath the skin.
25. 20
Cushing syndrome
Cushing’s syndrome, also known as hypercortisolism, itsenko-cushing syndrome, and
hyperadrenocortcism, is a collection of signs and symptoms due to prolonged exposure to cortisol.
Signs and symptoms may include
High blood pressure, abdominal obesity but with thin arms and legs, reddish stretch, marks, a
round red face, a fat lump between the shoulders, week muscles, week bones, acne, and fragile
skin that heals poorly. Women may have more hair and irregular menstruation. Occasionally there
may be changes in mood, headache and a chronic feeling of tiredness.
Cushing’s syndrome is caused by either excessive cortisol-like medications such as prednisone or
a tumor that either produce or result in the production of excessive cortisol by the adrenal glands.
Cause due to a pituitary adenoma are known as Cushing’s disease. It is the second most common
cause of causing syndrome after medication. A number of other tumors may also cause Cushing’s.
Some of they are associated with inherited disorders such as multiple endocrine neooplasia type 1
and carney complex. Diagnosis requires a number of steps. The first step is to check the medication
a person takes. The second step is major level of cortisol in the urine, saliva or in the blood after
taking dexamethasone. If this test id abnormal, the cortisol may be measured late at night. If the
cortisol remains high, a blood test for ATCH may done to determine if the pituitary I involved.
Most cause can be treated and cured. If due to medications, these can often be slowly stopped. If
caused by a tumor, it may be treated by a combination of surgery, chemotherapy, and/or radiation.
If the pituitary was affected, other medications may be required to replace its lost functions. With
treatment, life expectancy is usually normal. Some in women surgery id unable to remove the
entire tumor have an increased risk of death.
27. 22
EXAMINATION OF URINE
Urine examination is indicated in various kidney disease {e.g. glomerulonephritis, nephritic
syndrome, renal failure pyelonephritis etc.}.It is also indicated for detection of urinary tract
infection, for diagnosis of pregnancy, for various metabolic derangements in diabetes mellitus, for
differential diagnosis of jaundice etc.
Routine examination of urine is divided into three parts:
1. Physical examination
2. Chemical examination
3. Microscopic examination
Types of urine sample
First voided morning urine: This specimen is preferred for routine examination including
glucose, protein, microscopy and bacteriological analysis.
24 hrs. Urine specimen: discard first morning urine and subsequently collect urine in a large
container during rest of the day and night and then collect first morning urine on the next day.
Preserve urine at 4-60
c after collection. This method is used for quantitative estimation of
protein and hormones.
Random midstream urine: For routine examination.
Post-Prandial urine specimen: For estimation of glucose.
28. 23
Notes
For routine urine examination, collect 1-25ml of urine in wide mouthed, clean, dry, leak proof
glass bottle with tight fitted stopper.
For bacterial culture, urine should be collected sterile container without any preservative.
PRESERVATION OF URINE
Urine sample should ideally be examine within 1-2 hrs. voiding
Alternatively keep the specimen for maximum of 8 hrs. In refrigerator at 4-60
c.
Chemically preservative (e.g. hydrochloric acid, toluene boric acid, thymol and formalin can
also be used for 24 hrs. urine sample.
COLLECTION OF URINE
Mid-stream urine specimen:
This method is useful in most types of examination. After voiding initial part of the urine in toilet;
a part of urine is collected in a container. First half of stream server to flush out contaminating
cells and microbes form urethra and perineum. Subsequent stream is used for various test.
Urine container
29. 24
PHYSICAL EXAMINATION
Parameter to be examine for physical examination of urine include
1. Volume
2. Color
3. Odour
4. Specific gravity
5. pH.
Volume
Average 24 hrs. Urinary volume in adult is 600-2400ml.
Urine volume >2000ml/hrs. Is polyuria.
Volume <400ml is oliguria
<100/24hrs complete cessation of urine output is anuria
POLYURIA
Diabetes mellitus
Diabetes insipidus
Chronic renal failure
Diuretic therapy
Primary aldosteronism
30. 25
OLIGURIA
Febrile states
Acute glomerulonephritis
Congestive heart failure
Burns
Crush syndrome.
ANURIA
Shock
Complete urinary tract obstruction
Hemolytic transfusion reaction
Acute glomerulonephritis
Color
Normal fresh urine color is amber to pale yellow. Normal color of urine is due to presence of
various pigment known as urochrome. Urine can be different color in various pathological
condition.
Note:
Many drugs may also alter the color of urine and therefore drug history is very important for urine
examination.
31. 26
DIFFERENT COLOR OF URINE
Odour
Fresh urine has typical aromatic odour due to volatile organic acids. After standing, ammoniacal
odor may develop due to
Decomposition odor urea into ammonia by bacteria. Abnormal odor may be associated various
conditions like:
COLOR CONDITION
Colorless Dilute Urine (Diabetes Mellitus, Diabetes Insipidus)
Red Hematuria, Hemoglobinuria, Myoglobinuria, Porphyria
Blackish Alkaptonuria
Yellow Concentrated Urine
Deep Yellow Bilirubin
Orange Urobilinogen
Milky White Chyluia
Cola Colored Hemoglobinuria
32. 27
Fruity: ketoacidosis, starvation
Fishy : urinary tract infections (UTI ) by proteus
Ammoniacal: UTI by E.coli, old standing urine
Putrid: UTI.
Specific gravity
This is also knows as relative mass density and depends on the amount of solutes in urine. Normal
specific gravity of urine is 1.003 to 1.030. Specific gravity of urine is the measure of concentrating
ability of kidney. Since concentration of urine is the major function of tubules, therefore specific
gravity is the measure of tubular function of kidney
Methods of measurement of Specific gravity
(a) Urinometer method.
(b) Refractometer method.
(c) Reagent strip method.
Urinometer method
This method is based on the principle of buoyancy (i.e.
ability of the fluid to exert an upward thrust on a body
placed over it).
This method is as follows:
Urinometer method for measuring specific gravity of urine.
33. 28
Method
Fill measuring cylinder with 50ml urine. Lower Urinometer gently into urine and let it float
freely.
Allow urinometor to settle. Urinometor should not touch the slide or bottom of cylinder.
Take S.G reading on the scale which is the lowest point of meniscus on the surface of urine.
Take out urinometor and measure the temperature of urine immediately by thermometer.
i. CORRECTION OF SPECIFIC GRAVITY FOR TEMPERATURE
Add 0.001 to the reading for every 30
c (if urine temperature is above the temperature. Of
calibration). Similarly subtract 0.001 from the reading for every 30
c (if urine temperature is below
the calibration temperature).
The above corrections are made because density of urine increases at low temperature and
decreases at higher temperature.
ii. CORRECTION FOR SPECIFIC GRAVITY FOR SOLUTION
Proteinuria and glycosuria may freshly raise specific gravity of urine. Therefore correction should
be made. Subtract 0.003 fore very. 1 gm. protein/dl urine and subtract 0.004 for every 1 gm.
glucose/dl urine.
34. 29
Case of increased & decreased specific gravity
Increase specific gravity
Dehydration
Decreased fluid in take
Diarrhea
Vomiting
Fever
Diabetes mellitus
Albuminuria
Acute glomerulonephritis
Decrease specific gravity
Excessive mellitus
Diabetes insipidus
End stage kidney disease (like chronic glomerulonephritis, bilateral polycystic kidneys,
chronic pyelonephritis and hypertensive glomerulopathy).
Low and fixed specific gravity (also known as isosthenurea) of 1.008-1.010 result when
concentration power of kidney is severely affected and is a sign of end stage kidney disease.
35. 30
pH.
Normally ph. Of urine is 4.6- 8.0 (average 6.0 or slightly acidic). pH. Of urine depends on
diet, acid base balance. Water balance and renal tubular function.
For correct estimation of pH. Fresh urine should be examined because on standing urine
turns alkaline due to production of ammonia from urea. Method for determination of pH.
Are:-
a. Litmus paper test
Dip a small strip of litmus paper in the urine & note any color change. If blue litmus
turns red, it indicate acidic urine and if red litmus paper turns blue, it indicate alkaline
urine.
b. Reagent strip test
The test area contain polyionic polymer bond to H+ on reaction with cations in urine,
H+ is released causing change in color of pH. Sensitive dye.
Cause of acidic urine: ketoacidosis, starvation, fever, urinary tract infection by E.coli,
high protein diet.
Cause of alkaline urine: urinary tract infection by urea splitting organisms (proteus,
pseudomonas etc.) several vomiting, chronic renal failure.
36. 31
CHEMICAL EXAMINATION OF URINE
Protein
Glucose
Ketone
Bilirubin
Bile salts
Urobilinogen
Blood
Hemoglobin
Myoglobin
Nitrite or leukocyte esterase
PROTEIN
Normally very less amount (up to 150mg/24hr.) of proteins are excreted from the kidney. These
include plasma proteins derived from urine tract, Tamm-Horshfall protein, secretory IgA and
protein from desquamated tubular epithelium cell. Protein normally excreted from the kidney
cannot be detect by routine test. Presence protein in urine indicate injury to glomerulus resulting
in crossing of albumin through the basement membrane.
CAUSES OF PROTEINURIA
37. 32
Massive proteinuria
Nephrotic syndrome
Renal vein thrombosis
SLE
Diabetes mellitus
Moderate proteinuria
Chronic glomerulonephritis
Nephrosclerosis
Multiple myeloma
Pyelonephritis
Mild proteinuria
Hypertension
Polycystic kidney
Urinary Tract Infection
Fever
Chronic Pyelonephritis
Types of protein urea
Glomerular protein urea: This occurs due to increased permeability of glomerular capillary
wall in glomerular disease.
38. 33
Tubular proteinuria: This type of proteinuria occurs in acute and chronic pyelonephritis,
heavy metal poisoning, interstitial nephritis, and transplant rejection tuberculosis of kidney.
Overflow proteinuria: when concentration of proteins in plasma rises it overflow into the
urine known as overflow proteinuria. These protein can be Bence Jones proteins (multiple
myeloma). Hemoglobin (intravascular hemolysis) and myoglobin (skeletal muscle injury).
Hemodynamic proteinuria: when there is alteration of blood flow through the glomeruli, it
results in transient proteinuria e.g. in high fever, heavy exercise, exposure to cold etc.
Postural proteinuria: it is seen only on standing but absent on recumbent position.
Post renal proteinuria: it is caused by inflammatory or neoplastic conditions of renal pelvis,
ureter, bladder, prostate or urethra.
TEST FOR DETECTION OF PROTEINURIA
(a) Heat-acetic acid test.
(b) Sulpho-salicylic acid test.
(c) Heller’s method.
(d) Reagent strip test.
39. 34
HEAT-ACETIC ACID TEST
Principal: Protein get precipitated on boiling in an acidic solution.
Method:
2/3rd
of a test tube is filled with urine.
Incline the test tube and boil the upper portion over the flame.
Compare the heated part with lower part.
If cloudiness or turbidity appears, it indicates either phosphates or protein in urine.
Add few drop of 10% acetic acid and boil again.
If turbidity disappears, it is due to phosphates and if it remains it is due to protein.
Heat test for protein
40. 35
SULPHOSALICYLIC ACID TEST
Principle: addition of Sulphosalicylic acid to urine causes formation of white precipitate if protein
are present.
Method
Take 2 ml of clean urine in a test tube.
Add 2-3 drop of Sulphosalicylic acid (3-5%) & look for turbidity against a dark back
ground.
INTERPRETATION OF PROTEIN IN URINE
NEGATIVE
TRACES
WHITE CLOUDINESS + (<0.10%)
GRANULAR WHITE PPT ++ (0.1-0.25%)
FLOCCULAR PRECIPITATE +++ (0.25-0.50%)
THICK OPAQUE PRECIPITE ++++ (>0.5%)
41. 36
REAGENT STRIP TEST
Stripe is dipped in urine: Change in the color is compared with the color chart for proteins for
proteins, provide on the container.
Principle: reagent strip is coated with citrate buffer at ph. 3.0 and bromothymol blue as indicator.
If the dye gets absorbed to protein there is change in ionization (and hence ph.) of the indicator.
The intensity of the color is proportional to the concentration of protein.
42. 37
GLUCOSE
Normally very less amount of glucose is excreted in urine (<15mg/dl) which cannot be detected
by routine test.
Case of glycosuria
Diabetes
Renal glycosuria
Pregnancy
Alimentary glycosuria
Increased intracranial tension
Test for detection of glucose in urine
(a) Benedict’s test
(b) Multistix reagent strips test
BENEDICT’S TEST
Principle: urine when boiled with Benedict’s reagent, reduce the alkaline cupric hydroxide of the
reagent to cuperous oxide. The color of Benedict’s reagent turn from blue to red brown, if reducing
sugar is present.
43. 38
Cu (OH) 2 + Reducing substance Cu2O (colored ppt.)
This test is not specific for glucose. Other carbohydrates (e.g. glucose, fructose, lactose, pentose,
etc.) and certain drugs (salicylates, cephalosporin, penicillin etc.) can also reduce alkaline cupric
hydroxide.
Method
Take 5ml of Benedict’s reagent in a test tube.
Add 0.5ml (or 8 drop) of urine and mix thoroughly.
Boil over the flame.
Allow to cool at room temperature.
Note any change in color.
Benedict’s test for glucose
Comparison B/w normal benedict reagent (blue) $ glucose positive test (brick red)
44. 39
Glucose oxidase test & Multistix reagent strips test
This test is specific for glucose and is therefore preferred over Benedict’s test.
Principle: Diastix/ Multistix /Dipstix contains:
i. Glucose oxidase
ii. peroxidise
iii. Chromogen –o-tolidine and potassium iodide.
If glucose is present in urine, glucose oxidase acts on it and the following reaction take place:
NIL NO COLOR CHANGE.
TRACE GREEN WITHOUT PRECIPITATE
+ GREEN WITH PRECIPITATE (0.5 mg /dl)
++ BROWN PRECIPITATE (1 mg /dl)
+++ YELLOW-ORANGE PRECIPITATE (1.5 mg /dl)
++++ BRICK RED PRECIPITATE (2 mg /dl)
INTERPRETATION OF GLUCOSE IN URINE
45. 40
Glucose +O2
glucose oxidase
gluconic acid + H2O2
H2O2 + chromogen peroxidase
oxidized chromogen + H2O
Multistix use for glucose presence in urine.
Note: This test is used for estimation of glucose in a short period of time.
46. 41
MICROSCOPIC EXAMINATION
Microscopic examination of urine includes examination for red bloods cells, white blood cells,
epithelium cells, casts, bacteria, parasites, crystals and amorphous material.
Method
Urine sample is centrifuged for 5 min. for 1500 rpm & supernatant is poured off. Place a drop of
this sediment
On a glass slide and cover it with a cover slip. Examine the slid under microscope immediately,
using low & high power objectives.
62. 57
Around 100 male patients were analyzed.
54 male patients were found to be falling in the age group of 20 - 40 years.
35 male patients were found to be falling in the age group of 40 - 60 years.
9 male patients were found to be falling in the age group of 60 -80 years.
2 male patients were found to be falling in the age group of 80 - 100 years.
54%35%
9% 2%
EVALUATION OF URINE SAMPLE IN DIFFERENT
AGE GROUPS OF MALE PATIENTS.
20-40 YEARS 40-60 YEARS 60-80 YEARS 80-100 YEARS
63. 58
13 male patients out of 100, were found to be suffering from pathological disorder in the
age group of 20 – 40 years.
10 male patients out of 100, were found to be suffering from pathological disorder in the
age group of 40 – 60 years.
6 male patients out of 100, were found to be suffering from pathological disorder in the
age group of 60 –80 years.
1 male patient out of 100, were found to be suffering from pathological disorder in the age
group of 80 – 100years.
54%
13%
35%
10%9%
6%
2% 1%
0%
10%
20%
30%
40%
50%
60%
PATIENT DISEASE
Prevalence of pathological disorder in
male patients in different age groups
20-40 YEARS 40-60 YEARS 60-80 YEARS 80-100 YEARS
64. 59
Out of 100 male patients, 7% patients were found to be having Glycosuria while 23%
patients were found to be having proteinuria.
70% male patients were found to be dealing with any other pathological disorder.
70%
23%
7%
Prevalence of proteinuria and
glycosuria in males
Patients with other
pathological disorder
Patients
withProteinuria
Patients withglycosuria
65. 60
69%
33%
8% 0%
Around 100 female patients were analyzed.
69 female patients were found to be falling in the age group of 20 - 40 years.
33 female patients were found to be falling in the age group of 40 - 60 years.
8 female patients were found to be falling in the age group of 60 -80 years.
None of the female patient were found to be falling in the age group of 80 - 100 years.
69%
33%
8% 0%
20 - 40 YEARS 40 - 60 YEARS 60 - 80 YEARS 80 - 100 YEARS
66. 61
26 female patients out of 100, were found to be suffering from pathological disorder in
the age group of 20 – 40 years.
6 female patients out of 100, were found to be suffering from pathological disorder in
the age group of 40 – 60 years.
4 female patients out of 100, were found to be suffering from pathological disorder in
the age group of 60 –80 years.
None of the female patient out of 100, were found to be suffering from pathological
disorder in the age group of 80 – 100 years
0%
10%
20%
30%
40%
50%
60%
70%
PATIENT DISEASE
69%
26%
33%
16%
8%
4%
0% 0%
Prevalence of pathological disorder in female patients in different
age groups
20 - 40 YEARS 40 - 60 YEARS 60 - 80 YEARS 80 - 100 YEARS
67. 62
Out of 100 female patients, 7% patients were found to be having Glycosuria while 35%
patients were found to be having proteinuria.
58% female patients were found to be dealing with any other pathological disorder
58%
35%
7%
Prevalence of proteinuria and
glycosuria in females
Patients with other
pathological disorder
patients with protienuria
Patients with glycosuria
69. 64
CONCLUSION
IN CASE OF MALE PATIENTS:
OUT OF 100 MALE PATIENTS, 30 WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL DISORDER.
13 MALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 20 – 40 YEARS.
10 MALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 40 – 60 YEARS.
6 MALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 60 – 40 YEARS.
1 MALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 80 – 100 YEARS.
AMONG ALL THE MALE PATIENTS, 7% PATIENTS WERE FOUND TO HAVE GLYCOSURIA WHILE 23%
WERE FOUND TO HAVE PROTEINURIA.
70. 65
IN CASE OF FEMALE PATIENTS:
OUT OF 100 FEMALE PATIENTS, 36 WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL DISORDER.
26 FEMALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 20 – 40 YEARS.
6 FEMALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 40 – 60 YEARS.
4 FEMALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM PATHOLOGICAL
DISORDER IN THE AGE GROUP 60 – 40 YEARS.
NONE OF THE FEMALE PATIENT OUT OF 100, WERE FOUND TO BE SUFFERING FROM
PATHOLOGICAL DISORDER IN THE AGE GROUP 80 – 100 YEARS.
AMONG ALL THE FEMALE PATIENTS, 7% PATIENTS WERE FOUND TO HAVE GLYCOSURIA WHILE 35%
WERE FOUND TO HAVE PROTEINURIA.
71. 66
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