NONPROTEIN NITROGEN (NPN) EXPERIMENTAL RESULTS:

1. Urea: Generation of Ammonia

- Contains Nitrogen but they are not
proteins
- Used to monitor renal function
- Increased NPNs may cause renal
failure of associated diseases.
1. AMMONIA
- colorless gas with a pungent odor
- water soluble
- produced from CHON deamination
- liver converts Ammonia to Urea
Diseases:
- Liver failure
- Reye’ syndrome
o Affects children
o Swelling of brain & liver
o ↑ Ammonia in blood
2. UREA
- highest conc. of NPN in blood
- excretory product (urine)
- water soluble
- odorless and colorless
- salty taste
Diseases:
- Azotemia
- Uremia
- Urea Cycle disorders
- NaOH hydrolyzes Urea to produce
Ammonia.
- Ammonia has pungent odor
- Red litmus paper turned to blue
(basic).
2. Hypobromite test
- used to detect Urea
- Urea decomposes to form Nitrogen
- Effervescence: Presence of
Nitrogen gas
3.Urease test
- Urease: an enzyme that catalyzes
break down of Urea into Ammonia
and CO2.
- pHpH: Turned pink due to basicity
4. Biuret formation
- Biuret is a compound formed when
Urea is heated at high temperature.
- Biuret when treated with Copper
Sulfate and NaOH, purple-solution is
formed.

3. URIC ACID
- purine nucleotide breakdown
- excretory product
- can be found in blood and urine
Diseases:
- Gout (“King’s/ Richman’s disease”)
- Kidney stones

4. CREATININE 5. Uric Acid Phosphotungstic Acid

- degradation product of Creatine Reduction Test
- majority produced in muscles - detection of Uric acid
- NPN waste product - Uric acid is used as a reducing
Diseases: agent in strong alkaline condition.
- Kidney diseases - It reduces colorless
- Muscle weakness Phosphotungstic acid into Tungsten
blue.
- (+) result: Blue solution
6. Schiff’s test • Monocytes – Immune
- Uric acid reduces Ammoniacal defenses
Silver Nitrate into metallic silver. • Basophils – Hematologic
- (+) result: Black solution diseases
• Lymphocytes – viral infection
7. Jaffe’s test o T cells – HIV
- detection of Creatinine o B cells – Antibodies
- Solution: Picric acid in NaOH
- (+) result: Orange solution 3. Platelets
- Color intensity is directly - measures from 2 to 4µm
proportional to the amount - round, oval & anucleate
Creatinine present. - Hemostasis
• Series of process for wound
BLOOD healing, vessel wall repairs
- Transports O2 from lungs to tissues, BLOOD COLLECTION:
clears CO2, transports biomolecules a. Anticoagulants
and moves waste to the kidneys and b. Blood tests
liver. c. Collection:
- Humans possess 5L of blood • Venipucture
- 3 families: o Preferred site is
o RBC (Erythrocytes) median cubital vein
o WBC (Leukocytes) • Skin puncture
o Platelets (Thrombocytes) o Plantar surface
EXPERIMENTAL RESULTS:
1. Red Blood Cells 1. Benzidine test
- anucleated cells with hemoglobin - detection of blood
- Life span: 120 days/ 4 months - Principle involved: Peroxidase
- appears pink to red activity of hemoglobin decomoposes
- measures 6 to 8µm H2O2 releasing O2 thus oxidizing
- Hematocrit Benzidine.
• Packed cell volume - (+) result: Blue solution
- Hemoglobin
• A protein responsible for O2 2. Hemin test
transport - detection of blood traces
• Contains Iron bound to the - Hemin is a part of heme present in
heme group hemoglobin
• A conjugated enzyme: - (+) result: yellowish crystals
o Heme: Prosthetic
group
o Globin: Protein part

2. White Blood Cells

- Protects host from infection and
injury.
- Types of WBCs:
• Neutrophils – bacterial
infection
• Eosinophils – allergic/
parasitic infection
3. Guaiac test B.2. Pathologic Urine turbidity
- test for occult blood RBCs
- Principle: Heme portion of the WBCs
hemoglobin reacts with Guaiac, Bacteria
oxidizing it when H2O2 is added. Yeast
- (+) result: Blue solution Nonsquamous epithelial cells
Abnormal crystals
4. Confimatory test Lymph fluid
- Ether Lipids
• highly flammable & explosive
• used to confirm presence of C. Specific Gravity
blood - Density of solution compared
• (+) result: blue solution to the density of water.
- Urinometer
URINE o Classical method
- 95% water and 5% solutes o Instrument used to
- Odor: Aromatic determine specific
- Normal urine excretion: 1200 to gravity.
1500mL
- Ultrafiltrate of plasma D. pH
- Biohazardous substance - Normal pH ranges: 6.5 to 8.0
- Morning urine: 5.0 to 6.0
EXPERIMENTAL RESULT: - After meals: Alkaline pH
1. Physical properties: - Random samples: 4.5 to 8.0
A. Color
Colorless Recent fluid consumption 2. Pathologic urine
Pale yellow Dilute urine A. Sugar
Dark yellow Conc. urine - Fehling’s A & B
Orange Bilirubin o Test for reducing
Green Bacterial infection sugars
Pink RBCs - Fehling’s A
Red Hemoglobin o Copper (II) Sulfate
Black Melanin/ Melanogen - Fehling’s B
o Potassium Sodium
B. Clarity Tartrate (Rochelle
Clear No particulates salt) and NaOH
Hazy Few particulates - Glucose threshold: 120 to
Cloudy Phosphate & Carbonates 180 mg/ dL
Turbid Many particulates - Result:
Milky Clotted or precipitated Color Percent sugar:
Blue Absent
B.1. Nonpathologic Urine Turbidity Green 0.5 to 1%
Squamous epithelial cells Yellow 1 to 2%
Mucus Brick red 2% above
Phosphates, Carbonates & Urates
Semen, spermatozoa
Fecal contamination
Radiographic contrast media
Talcum powder
Vaginal creams
B. Blood
- Benzidine test
o Used to detect blood
- Result:
o (+): Blue solution
o (-): No color change

C. Proteins
I. Heat and Acetic acid test
o Turbidity – presence of
Albumin, Phosphates and
Carbonates.
o Addition of Acetic acid:
▪ Turbid: Albumin
▪ Clear: Phosphates
and Carbonates
II. Heller’s Nitric acid test
o Nitric acid causes the
precipitation of Proteins
o (+) result: White ring at a
junction of 2 layers.

D. Ketone bodies
- Ketones react with Sodium
Nitroprusside in an alkaline
condition.
- (+) result: Purple ring at a junction of
2 layers.
- (+) Ketones: Acetoacetate and
Acetone (w Glycine)
- (-) Ketone: β-Hydroxybutyrate