different blood samples

1. BLOOD SAMPLES
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Dr. Mohamed Badr
Lecturer of Biochemistry
Faculty of pharmacy Menoufia university

2. 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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3. Composition
5 L
45%
54.3%
0.7%
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4. 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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5. 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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6. 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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7. 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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8. 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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9. Blood
samples
Whole
blood
SerumPlasma
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10. 10

11. Plasma Serum
Definition and how it’s
obtained
Liquid part obtained
from centrifugation of
anti-coagulated blood
Liquid part obtained
from centrifugation of
coagulated blood
processing Can be obtained
immediately
Time consuming
(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 antibiotics11

12. 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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13. 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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14. Anti-coagulants
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15. Heparine EDTA Sodium citrate Oxalate
MOA Increase activity of
antithrombin III
Chelating agent:
binding of Ca+2
Chelating agent :
binding of Ca+2
Forming insoluble
complex with
calcium
Advantages Causes least
interference with tests
Preserves the cellular
component of blood
Its effect is reversible
by addition of Ca+2
Disadvntages - High cost
- Temporary action
- Inhibit acid
phosphatase,
lactate &
hydroxybutyrate
dehdrogenses
- Affect binding of
triiodothyroxin to
carrier proteins
- Can’t used for
Ca+2 and Fe+2
analysis.
- Inhibit alkaline
phosphatase,
creatine kinase,
leucine
aminopeptidase
- Can't be used for
Ca+2 analysis.
- Citrates complex
with molybdate,
so produce low
results.
- Inhibit alkaline
phosphatase and
aminotransferase
- It inhibits acid
& alkaline
phosphatase,
amylase and
lactate.
dehydrogenase.
- Can't be used
for C+2 analysis.
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16. 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.
Other preservatives
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17. 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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18. Blood tubes’ color system
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19. Blood tubes’ color system
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20. 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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21. Common causes of unacceptable serum or plasma
specimens and inaccurate test results
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22. 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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23. 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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24. 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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25. Determination of serum albumin
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26. 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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27. Pre-albumin (transthyretin)
• Prealbumin is so named because it migrates
before albumin in electrophoresis.
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28. Biological functions of Pre-albumin
Transport protein
for thyroid
hormones
Binds with retinol-
binding protein to
form a complex
that transports
retinol (vitamin A)
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29. Clinical significance of Pre-albumin
↓↓↓
• Hepatic damage.
• Tissue necrosis.
• A low prealbumin level is
a sensitive marker of poor
nutritional status.

- Patients receiving
steroids.
- Alcoholism.
- Chronic renal failure
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30. 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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31. Biological functions of albumin
Responsible for
nearly 80% of the
colloid osmotic
pressure of the
intravascular fluid.
Bind various
substances
(hormones and
medications) in
the blood.
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32. Clinical significance of albumin
↓↓↓
• Liver disease
• malnutrition and
malabsorption
• Protein-losing
enteropathy
• renal disease
• Hypothyroidism
↓↓↓
• Skin loss in the
absence of the
skin barrier such
as in burns or
exfoliative
dermatitis
• Dilution by excess:
polydipsia or
excess
administration of
intravenous fluids.
↓↓↓
• Redistribution by
hemodilution,
increased capillary
permeability
(increased
interstitial
albumin).
• Rarely, Mutation
→ analbuminemia
or bisalbuminemia

-Dehydration.
-Excessive
albumin
infusion
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33. 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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34. 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:
conc. of test (g/dL) = 𝐴 𝑡𝑒𝑠𝑡
𝐴 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑
𝑋𝑐𝑜𝑛𝑐. 𝑂𝑓 𝑠𝑡𝑎𝑛𝑑𝑎𝑟𝑑
 What to do when concentration exceed readout of the
spectrophotometer or kit sensitivity?!!
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