Hemoglobin Determination

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Hemoglobin Determination
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Hemoglobin

• Hemoglobin
• Found in the RBC.
• Responsible for carrying oxygen to all cells in
  the body.
• Also binds to carbon dioxide and carries it to
  the lungs from the cells to
• be released.
• An accurate measurement of hemoglobin in the body
  gives the health care worker an idea of the
  patients oxygen carrying capacity.
• Hemoglobin detection can also give the health
  care worker of current blood loss and recovery
  from blood loss.
• Monitoring hemoglobin can also aid in the
  treatment of RBC disorders, like anemia.

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Hemoglobin

• Hemoglobin
• Makes up 98 of the protein found in the RBC.
• Gives blood its red color.
• Composed of two parts.
• Heme ? 4 iron atoms in the ferrous state (Fe2)
• Globin ? 4 protein chains (genetically
  determined)
• The most common globin forms are alpha and beta
  chains.
• Hemoglobin's are named based on the structure of
  the
• protein chain that is present.
• Hemoglobin A ? 95-98 of adult hemoglobin
• Hemoglobin A2 ? 3-5 of adult hemoglobin
• Hemoglobin F ? normal fetal hemoglobin, lt1 of
  adult hemoglobin
• Hemoglobin S ? abnormal hemoglobin found in
  sickle cell anemia

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Hemoglobin

• Each hemoglobin molecule contains four heme
  groups (iron molecules), each associated with its
  own protein chain.
• Iron is a must for hemoglobin synthesis.
• Iron is reused by the body as needed.
• When old RBCs die the iron is recycled to make
  new hemoglobin molecules.
• If the body doesnt have enough iron, hemoglobin
  production decreases and the RBCs will not have
  enough hemoglobin.
• This can result in anemia causing patients to
  experience fatigue and paleness of skin.
• Excessive blood loss can also lead to low iron
  levels and poor hemoglobin production.

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Reference Values for Hemoglobin

• Reference values for hemoglobin depend on gender
  and age.
• Hemoglobin is higher at birth and lowers with
  childhood and increases again as you get older.
• At birth hemoglobin levels range from 16-23 g/dL
• During childhood hemoglobin levels range from
  10-14 g/dL
• Adult males have hemoglobin levels ranging from
  13-17 g/dL
• Adult females have hemoglobin levels ranging from
  12-16 g/dL
• Your hemoglobin level should be 1/3 of your
  hematocrit level.
• Exercise as well as diet affect the level of
  hemoglobin you have in your system.
• If you exercise a lot your hemoglobin will be
  higher because you need more oxygen carrying
  capacity.
• If you have a diet low in iron your hemoglobin
  level may be lower than normal.

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Reference Values for Hemoglobin

• Determining hemoglobin value
• There are several ways to determine a persons
  hemoglobin
• concentration.
• Methods include..
• Specific Gravity Technique
• Cyanmethemoglobin
• Azidemethemoglobin
• Some methods can be performed manually while
  others are performed on a hematology or POCT
  analyzer.

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Reference Values for Hemoglobin

• Determining hemoglobin value
• Specific Gravity Technique
• Estimates the hemoglobin concentration.
• Drop of blood is placed into a cupper sulfate
  solution.
• The specific gravity of the solution must be
  from 1.052 to 1.054.
• This is the same specific gravity of blood with
  a hemoglobin
• concentration at the low end of the
  reference range.
• If the drop falls to the bottom quickly the
  specific gravity is
• higher than the solution and the hemoglobin
  is in the
• normal range.
• If the drop floats or falls slowly the specific
  gravity is lower
• than the solution and the hemoglobin is
  below the normal range.

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Reference Values for Hemoglobin

• Determining hemoglobin value
• Cyanmethemoglobin
• Blood is mixed with a reagent (Drabkins
• reagent) containing iron, potassium, cyanide,
  
• and sodium bicarbonate.
• The Drabkins reagent and the hemoglobin in
• the blood combin to form a colored end-
• product known as cyanmethemoglobin.
• A hematology analyzer or hemoglobinometer
• measures the end product photometrically.

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Reference Values for Hemoglobin

• Determining hemoglobin value
• Azidemethemoglobin
• The azidemethemolgobin reagent contains a
• lysing chemical, an oxidizing chemical and
  azide.
• The oxyhemoglobin which is in the form of
• ferrous iron is oxidized to ferric iron to
  form
• methemoglobin.
• The methemoglobin combines with azide to form
• azidemethemoglobin, that can also be measured
  
• photometrically by a hematology analyzer.

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Reference Values for Hemoglobin

• Determining hemoglobin value
• POCT analyzers or hematology analyzers use the
• before mentioned methods to determine ones
• hemoglobin.
• POCT analyzers require only a small drop or two
  of
• blood to determine the samples hemoglobin
• concentration.
• Hematology analyzers require more blood and test
  
• for more parameters than the hemoglobin alone
• and often times run an entire CBC.