1.

MAJOR EXPERIMENT (12 marks)

A 45 year old male has come to the Out-patient Department with complaints of weight loss, polyuria,
polydipsia and polyphagia for the past one month.
a. What is your probable diagnosis? (1 mark )
b. Write relevant investigations for the above mentioned case and the reference ranges for the relevant
parameters. (2 marks )
c. Write the principle and procedure for the estimation of GLUCOSE. (3 marks )
d. Estimate the concentration of GLUCOSE present in the given sample. (3 marks)
e. Discussion. ( 3 marks )

MINOR EXPERIMENT (12 marks )

Find out TWO abnormal constituents present in the given sample of urine.
1.(a) PROBABLE DIAGNOSIS - DIABETES MELLITUS

(b)RELEVANT INVESTIGATIONS REFERENCE RANGES:

Fasting Blood Glucose 70-110 mg/dl
Post Prandial Blood Glucose <140 mg/dl
HbA1C 4- 5.6 % of total Hb
Urine Examination Sugar – Nil Ketones – Nil

(c) PRINCIPLE & PROCEDURE

Glucose is oxidized by glucose oxidase to give gluconic acid and hydrogen peroxide. The H2O2 that is
formed is broken down by peroxidase to water and oxygen. O2 oxidizes phenol which combines with 4 amino-
antipyrine to give a pink colored complex. The intensity of the colored complex is measured colorimetrically at
515nm which is directly proportional to concentration of glucose.

Glucose oxidase
Glucose + 2H2O + O2 Gluconic acid + H2O2 + CO2
Peroxidase
H2O2 + 4amino-antipyrine Pink colored compound

REAGENT BLANK (REAGENT STANDARD TEST

+DISTILLED WATER)
Working reagent 1ml 1ml 1ml
Serum -------- ------ 10µl
Standard -------- 10 µl ------
Distilled water 10µl ------ -------

Mix the content of the test tube thoroughly and keep them for 15 minutes. Measure the optical density using
calorimeter at 515 nm.

(d) CALCULATION:
Glucose present in 100 ml of plasma / serum = OD (T) – OD (B) X Concentration of Standard
OD (S) - OD (B)

OD (T) – OD (B) X 100 mg/dl

OD (S) - OD (B)

RESULT :
The concentration of glucose present in the given sample is ________ mg/dl
 2. MAJOR EXPERIMENT (12 marks)

A 60 year old male has been treated for hypertension and diabetes for 10 years and
was admitted in the hospital with the complaints of oliguria and puffiness of face for the past 10 days.

a. What is your probable diagnosis? (1 mark )

b. Write relevant investigations for the above mentioned case and the reference ranges for the
relevant parameters. (2 marks )
c. Write the principle and procedure for the estimation of UREA. (3 marks )
d. Estimate the concentration of UREA present in the given sample.
(3 marks )
e. Discussion. (3 marks )

MINOR EXPERIMENT (12 marks)

Find out TWO abnormal constituents present in the given sample of urine.
2. (a) PROBABLE DIAGNOSIS - CHRONIC KIDNEY DISEASE

(b) RELEVANT INVESTIGATIONS REFERENCE RANGES

Blood Urea 15 - 40 mg/dl

Serum Creatinine Male 0.9 - 1.3 mg/dl
Female 0.6 -1.1 mg/dl

Creatinine Clearance Male 95 -140 ml/min

Female 85 - 120 ml/min
24 hrs urine Protein <150mg/day

(c) PRINCIPLE :
Urea reacts with DAM in hot acidic medium in the presence of catalyst namely FeCl3 and
thiosemicarbazide (TSC) to give pink colored substance. The intensity of the color is directly proportional to the
concentration of urea present in the sample which is measured at 540 nm.

PROCEDURE

ANALYSIS
Reagent Blank Standard Test

Protein Free Filtrate 0.5 ml 0.5ml 0.5ml

DAM/ TSC reagent 3ml 3ml 3ml

Acid mixture 3ml 3ml 3ml

Mix and keep it in boiling water bath for 15 minutes and read at 540 nm

(d) CALCULATION
Amount of urea present in 100 ml of plasma or serum

OD (T) – OD (B)
= X Concentration of Standard
OD (S) - OD (B)

OD (T) – OD (B)
= X 50 mg/dl
OD (S) - OD (B)

RESULT
The concentration of urea present in the given sample is ________ mg/dl
3. MAJOR EXPERIMENT (12 marks)

A 60 year old male has been treated for hypertension and diabetes for 10 years and
was admitted in the hospital with the complaints of oliguria and puffiness of face for the past 10 days.

a. What is your probable diagnosis? (1 mark )

b. Write the relevant investigations for the above mentioned case and the reference ranges for the
relevant parameters. (2 marks )
c. Write the principle and procedure for the estimation of CREATININE.
(3 marks )
d. Estimate the concentration of CREATININE present in the given sample.
(3 marks )
e. Discussion. (3 marks )

MINOR EXPERIMENT (12 marks)

Find out TWO abnormal constituents present in the given sample of urine
3. (a) PROBABLE DIAGNOSIS - CHRONIC KIDNEY DISEASE

(b) RELEVANT INVESTIGATIONS REFERENCE RANGES

Blood Urea 15 - 40 mg/dl

Serum Creatinine Male 0.9 - 1.3 mg/dl
Female 0.6 -1.1 mg/dl

Creatinine Clearance Male 95 -140 ml/min

Female 85 - 120 ml/min
24 hrs urine Protein <150mg/day

(c) PRINCIPLE:

Creatine is estimated by Jaffe’s reaction. Creatinine with picric acid in presence of alkaline medium form
orange red creatinine picrate complex.

Alkali
Creatinine + picric acid Creatinine picrate

The intensity of the colour is directly proportional to the concentration of creatinine present in the given sample,
which is measured at 515 nm.

Analysis
Reagent Blank Standard Test

Protein Free Filtrate 3ml 3ml 3ml

0.04M picric acid 1ml 1ml 1ml

0.75 N NaOH 1ml 1ml 1ml

Wait for 20 minutes and read at 515 nm.

(d) CALCULATION
Amount of creatinine present in 100 ml of sample is

OD (T) – OD (B)
= X Concentration of Standard
OD (S) - OD (B)

OD (T) – OD (B)
= X 2 mg/dl
OD (S) - OD (B)

RESULT :
The concentration of creatinine present in the given sample is ________ mg/dl
 4. MAJOR EXPERIMENT (12 marks)

A middle aged female suddenly developed acute pain and swelling in the second metatarso-
phalangeal joint and pain in other joints
a. What is your probable diagnosis? (1 mark )
b. Write relevant investigations for the above mentioned case and the reference ranges for the
relevant parameters. (2 marks )
c. Write the principle and procedure for the estimation of URIC ACID.
(3 marks )
d. Estimate the concentration of URIC ACID present in the given sample.
(3 marks )
e. Discussion. (3 marks )

MINOR EXPERIMENT (12 marks)

Find out TWO abnormal constituents present in the given sample of urine.
4.(a)PROBABLE DIAGNOSIS - GOUTY ARTHRITIS

RELEVANT INVESTIGATIONS REFERENCE RANGES

Serum Uric acid Male - 4.4 – 7.6 mg/dl
Female- 2.3-6.6 mg/dl
Urinary Uric acid – 250-750 mg/day

(c) PRINCIPLE:

Uric acid is acted on by uricase forming allantoin and hydrogen peroxide. The hydrogen peroxide reacts with
phenolic chromogens in presence of peroxidase forming a red colored compound.

Uricase
Uric acid + 2H2O + O2 Allantoin + H2O2 + CO2
Peroxidase
H2O2 + Phenolic Chromogen Red colored compound

This compound has maximum absorption at 520 (500 – 530) nm.Absorption is directly proportional to
concentration of uric acid

PROCEDURE
Reagent Sample Standard Blank
Working reagent 1ml 1ml 1ml
Sample 25 µl -- -- ---
Standard --- 25 µl ---

Incubate for 10 minutes at room temperature. After completion of incubation measure absorbance of acyl
mixture against blank at 520nm

CALCULATION
Amount of uric acid present in 100 ml of plasma or serum

OD (T) – OD (B)
= X Concentration of Standard
OD (S) - OD (B)

OD (T) – OD (B)
= X 6 mg/dl
OD (S) - OD (B)

RESULT :

The concentration of uric acid present in the given sample is ________ mg/dl
 5. MAJOR EXPERIMENT (12 marks)
A 5 year old male child was brought to the Out-patient department with difficulty in
passing urine, puffiness of face and generalized edema for the past 10 days.
a. What is your probable diagnosis? (1 mark )
b. Write relevant investigations for the above mentioned case and the reference ranges for the
relevant parameters.(2 marks )
c. Write the principle and procedure for the estimation of PROTEIN.
(3 marks )
d. Estimate the concentration of PROTEIN present in the given sample.
(3 marks )
e. Discussion. (3 marks )

MINOR EXPERIMENT (12 marks)

Find out TWO abnormal constituents present in the given sample of urine.
5.(a) PROBABLE DIAGNOSIS - NEPHROTIC SYNDROME

RELEVANT INVESTIGATIONS REFERENCE RANGES

Serum Total Protein 6.4-8.3 g/dl
Serum Albumin 3.5-5.2 g/dl
Serum Globulin 2.5 – 3.5 g/dl
A:G ratio 1.2 - 1.5 : 1
Serum Cholesterol 150-200mg/dl
Urine Protein < 150 mg/day

(c) PRINCIPLE
Substances which contain two or more peptide bonds (CO-NH) form a bluish violet color compound
with cupric ion in alkaline medium. Intensity of the colour is directly propotional to the concentration of the
total protein present in the given sample which is measured at 540 nm
Alkaline
Protein + CuSO4 Violet colored compound

PROCEDURE
Reagent Blank Standard Test

Biuret 1ml 1ml 1ml

Distilled water 10µl
Protein standard -- 10µl --
Serum -- -- 10µl
Incubate for 20 minutes at 370C . Take reading at 540 nm

CALCULATION
Amount of protein present in 100 ml of plasma or serum

= OD( T) – OD (B) X Concentration of Standard

OD (S) - OD (B)

= OD( T) – OD (B) X 6 g/dl

OD (S) - OD (B)

RESULT : The concentration of protein present in the given sample is ________g/dl