BLOOD SAMPLES

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 Blood
• Blood is a constantly circulating fluid.
• Functions:
1. Supply of oxygen to tissues.
2. Supply of nutrients such as glucose and amino acids.
3. Removal of waste such as carbon dioxide, urea, and lactic
acid.
4. Immunological functions, including circulation of white
blood cells, and detection of foreign material by
antibodies.
5. Coagulation, which is one part of the body's self-repair
mechanism.
6. Messenger functions, including the transport of
hormones.
7. Regulation of body pH.
8. Regulation of body temperature.
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 Composition
5L
54.3%

0.7% 45%

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 Blood cells
• Blood cells are formed in the bone marrow.
• All blood cells arise from the same bone
marrow stem cells (hematopoietic stem cell).

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 Blood Conditions
• Hemorrhage (bleeding): Blood leaking out of blood vessels
and may be obvious (external) or internal.

• Hematoma: Collection of blood inside the body tissues.

• Leukemia: Form of blood cancer, in which white blood cells

multiply abnormally and circulating in the blood.

• Lymphoma: Form of blood cancer, in which white blood

cells multiply abnormally inside lymph nodes and other
tissues.

• Multiple myeloma: Form of blood cancer of plasma cells.

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 Blood Conditions
• Anemia: An abnormally low number of red blood cells in the blood.

• Hemolytic anemia: Anemia caused by rapid bursting of large

numbers of red blood cells (hemolysis).

• Sickle cell disease: A genetic condition in which red blood cells

periodically lose their proper shape (appearing like sickles, rather
than discs).

• Polycythemia: Abnormally high numbers of red blood cells in the

blood and can result from low blood oxygen levels.

• Hemochromatosis: A disorder causing excessive levels of iron in the

blood. The iron deposits in the liver, pancreas and other organs,
causing liver problems and diabetes.

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 Blood Conditions
• Leukopenia: Abnormally low numbers of white blood cells in the
blood.

• Leukocytosis: Abnormally high numbers of white blood cells in the

blood.

• Thrombocytopenia: Abnormally low numbers of platelets in the

blood.

• Thrombocytosis: Abnormally high numbers of platelets in the

blood.

• Disseminated intravascular coagulation (DIC): An uncontrolled

process of simultaneous bleeding and clotting in very small blood
vessels. DIC usually results from severe infections or cancer.

• Hemophilia: Genetic deficiency of certain blood clotting proteins.

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 Blood Conditions
• Hypercoaguable state: Abnormally increased tendency toward
blood clotting (coagulation). A heart attack, stroke, or blood clots in
the legs or lungs can result.

• Deep venous thrombosis (DVT): A blood clot in a deep vein, usually

in the leg.

• Myocardial infarction (MI): Commonly called a heart attack, a

myocardial infarction occurs when a sudden blood clot develops in
one of the coronary arteries, which supply blood to the heart.

• Bacteremia: Bacterial infection of the blood.

• Malaria: Infection of red blood cells by Plasmodium; parasite

transmitted by mosquitos.

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 Whole
blood

Blood
samples

Plasm
a
Serum
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 Plasma Serum
Definition and how it’s Liquid part obtained Liquid part obtained
obtained from centrifugation of from centrifugation of
anti-coagulated blood coagulated blood
processing Can be obtained Time consuming
immediately (completion of blood
coagulation for about
30-60 min)
Fibrinogen Fibrinogen is present!! Absent

Yield Higher yield from blood less

Applications Blood transfusion • Blood typing

• Diagnostic testing
• Supplementation of
culture media
• Testing for antibiotics
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 Blood samples
• The difference for calcium, glucose, inorganic
phosphorus, potassium, and total protein
assessments in whole blood, plasma, or serum
is sufficient to alter clinical interpretation.

• Glucose concentrations were noted to be 5%

higher in serum than plasma as a result of
fluid shift from erythrocytes during the
clotting process.

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 Blood samples
• A slight increase in total protein may be seen in plasma as a
result of fibrinogen.

• High levels of lithium and sodium may be observed in

plasma samples due to contamination with cations from
the anticoagulants.

• Use of plasma samples for blood tests in the dialysis

population eliminates delays in sample processing while
waiting for clotting to complete.

• ESR, blood gas, and ammonia determination must done in

whole blood sample.
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Anti-coagulants

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 Heparine EDTA Sodium citrate Oxalate

MOA Increase activity of Chelating agent: Chelating agent : Forming insoluble

antithrombin III binding of Ca+2 binding of Ca+2 complex with
calcium

Advantages Causes least Preserves the cellular Its effect is reversible

interference with tests component of blood by addition of Ca+2

Disadvntages - High cost - Can’t used for - Can't be used for - It inhibits acid
Ca+2 and Fe+2 Ca+2 analysis. & alkaline
- Temporary action analysis. phosphatase,
- Citrates complex amylase and
- Inhibit acid - Inhibit alkaline with molybdate, lactate.
phosphatase, phosphatase, so produce low dehydrogenase.
lactate & creatine kinase, results.
hydroxybutyrate leucine - Can't be used
dehdrogenses aminopeptidase - Inhibit alkaline for C+2 analysis.
phosphatase and
- Affect binding of aminotransferase
triiodothyroxin to
carrier proteins
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 Other preservatives
Fluoride:
• Best preservative for glucose (antiglycolytic).
• Without antiglycolytic agent, blood glucose conc.
decreases 10 mg/dL/hour at 25ºC
• The rate of decrease is faster in newborns because
of the increased metabolic activity of their RBCs
and in leukemic patients because of high metabolic
activity of white blood cells.

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 Other preservatives
Iodoacetate:
• Antiglycolytic agent.
• It doesn't affect urease; so it can be used for glucose
and urea in single specimen.
• It inhibit creatine kinase.

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Blood tubes’ color system

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 Blood tubes’ color system

For more info, visit the following link:

file:///C:/Users/pc/Downloads/User%20Guide_English%20Leaflet.pdf 19
 Preservation of specimen
• Specimen must be kept at 4OC from the time
of collection until specimens are analyzed or
until serum or plasma is separated.
• For thermolabile constituents tests, serum or
plasma should be separated in cooling
centrifuge.
• To avoid photo-degradation of light sensitive
constituents e.g. bilirubin or carotene, keep
sample in brown containers.

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Common causes of unacceptable serum or plasma
specimens and inaccurate test results

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 Common causes of unacceptable serum or plasma
specimens and inaccurate test results
❖ Hemolysis
• Serum shows visual evidence of hemolysis when
hemoglobin concentration >20 mg/dL.
• Hemolysis occurs when the membrane surrounding red
blood cells is disrupted and hemoglobin and other
intracellular components escape into the serum or plasma.
• Hemolyzed specimens may be rejected and even slight
hemolysis may alter certain test results
❑ Examples for affected biochemical tests:
✓ K+, LDH, AST and ALT (+)
✓ Troponin T and Amylase (-)

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 Common causes of unacceptable serum or plasma
specimens and inaccurate test results

❖ Turbidity (lipemia)
• Turbid, cloudy or milky serum (lipemic serum) may be
produced by the presence of fatty substances (lipids) in
the blood.
• Bacterial contamination may also cause cloudy serum.
• Moderately or grossly lipemic specimens may alter
certain test results.
❑ Examples for affected biochemical tests:
✓ Mg++, direct bilirubin (+)
✓ K+ and Na+ (-)

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 Common causes of unacceptable serum or plasma
specimens and inaccurate test results

❖Hyperbilirubinemia
• Icteric serum or plasma varies in color from
dark to bright yellow, rather than the normal
straw color.
• Icterus may affect certain determinations.
❑Examples for affected biochemical tests:
✓Mg++ (+)
✓Cholesterol and TG (-)
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Determination of serum albumin

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 Introduction
• The major measured plasma proteins are:
albumin and globulins.
• A typical blood panel will provide four
different measurements; total protein,
albumin, globulins, and the albumin/globulin
ratio.

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 Pre-albumin (transthyretin)
• Prealbumin is so named because it migrates
before albumin in electrophoresis.

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Biological functions of Pre-albumin

Binds with retinol-

Transport protein binding protein to
for thyroid form a complex
hormones that transports
retinol (vitamin A)

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 Clinical significance of Pre-albumin

↓↓↓

• Hepatic damage.
• Tissue necrosis. - Patients receiving
• A low prealbumin level is steroids.
a sensitive marker of poor
nutritional status. - Alcoholism.
- Chronic renal failure

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 Albumin
• Albumin is synthesized in the liver.
• It is the protein present in highest concentration
in the plasma.
• Albumin also exists in the extravascular
(interstitial) space.
• The total extravascular albumin exceeds the total
intravascular amount by 30%, but the
concentration of albumin in the blood is much
greater than its concentration is in the interstitial
space.
• Normal values (adult): 3.5-5.5 g/dL.
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 Biological functions of albumin

Responsible for Bind various

nearly 80% of the substances
colloid osmotic (hormones and
pressure of the medications) in
intravascular fluid. the blood.

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 Clinical significance of albumin

↓↓↓ ↓↓↓ ↓↓↓

• Liver disease • Skin loss in the • Redistribution by
• malnutrition and absence of the hemodilution,
malabsorption skin barrier such increased capillary 
as in burns or permeability
• Protein-losing -Dehydration.
exfoliative (increased
enteropathy
dermatitis interstitial
• renal disease
• Dilution by excess: albumin). -Excessive
• Hypothyroidism polydipsia or • Rarely, Mutation albumin
excess → analbuminemia infusion
administration of or bisalbuminemia
intravenous fluids.

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 Chemical determination of albumin
by Biuret method
• Principle
• Cupric ion in alkaline medium forms a violet
colored complex with nitrogens of the peptide
bonds.

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 Procedures
Blank Standard Test
DW 2 mL ----- -----
Standard ----- 2 mL -----
Sample ----- ----- 2 mL
Biuret reagent 2 mL 2 mL 2 mL

✓ Mix and leave for 5 minutes at room temperature.

✓ Read absorbance of test and standard against blank at 540 nm.
✓ Calculation:
𝐴 𝑡𝑒𝑠𝑡
conc. of test (g/dL) = 𝐴 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑 𝑋𝑐𝑜𝑛𝑐. 𝑂𝑓 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑
✓ What to do when concentration exceed readout of the
spectrophotometer or kit sensitivity?!!
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