Hematocrit

- Packed cell volume

- Proportion of the volume of a blood that is occupied by RBCs
- Reflects the concentration of red cells
- Relative volume percentage of RBC in whole blood
- Millimeters of PCV/100ml of blood indicating the volume (%) of PCV/100ml of blood
- Rule of three
o 3 x RBC= hemoglobin (g%)
o 3 x Hemoglobin = hematocrit (%)
- Normal value
o At birth: 45-60%
Two types of Hematocrit
- Body hematocrit
o Ratio of total red cell volume to plasma in the circulating blood
o 9% higher than venous blood
o Body hct= Red cell volume/(RCV + plasma volume)
- Venous hematocrit
o Ratio of red cells to plasma in venous blood
o Measures the proportion of RBC to plasma in the peripheral blood
o Layers formed when anticoagulated blood is centrifuged
 From top to bottom
 Fatty layer
o barely visible
o Presence of lipidemia, layer is several mm thick
 Plasma layer
o Pale yellow and fairly clear (normal)
o Cherry red color (excessive hemolysis)
o Deep yellow color (jaundice)
 Buffy coat layer
o Thin, yellowish white layer
o Consist of platelets, WBCs, young RBC (nucleated RBC)
o 1mm buffy coat layer= 10,000 WBC/cu.mm.
 Red cell layer
o Retics can be found in the top most portion
Methods of Hematocrit determination
- Macromethod
Macromethod Anticoagulant used
Wintrobe method Double oxalate
Haden’s modification 1.1% Sodium oxalate
Van Allen’s 1.6% Sodium oxalate
Sanford & Magath 1.3% Sodium oxalte
Bray’s Method Heparin

o Wintrobe method
 Principle
 Anticoagulated whole blood is centrifuged in a wintrobe tube to
completely pack the red cells
  Procedure:
 The tube is filled with blood up to the 10th mark of the right white side of the
tube using capillary pipet
 Centrifuge the tube at 3500 rpm for 30 minutes
 Read the volume of the packed RBC at the lower right white side calibration of
the tube
 Volume % Hct= (height of packed RBC/height of whole blood used) x 100
 Normal values
 Adult male
o 42 to 52%, CU
o 0.42 to 0.52L/L, SI
 Adult female
o 37 to 47 %, CU
o 0.37 to 0.47L/L, SI
- Micromethod
o Adam’s microhematocrit method
 Procedure:
 Fill two heparinized capillary tubes about 2/3 full with blood from the finger
 Mix tubes several times and seal with sealing clay, 3-6 mm depth
 Centrifuge for 5 minutes at 10,000 rpm
*Should be read within 10 minutes to avoid merging of the layers
 Place the lower end of the blood column on the 0 line or red line and upper end
of the plasma column on the 100% (1.0 line) from a microhematocrit reading
device
 Read the upper end of the RBC column and record the results
 1% hematocrit=0.34 g% (Hemoglobin), 107, 000 RBC/mm3
 Normal values
 Male
o 40-54% (CU)
o 0.40-0.54L/L (SI)
 Female
o 35-49% (CU)
o 0.35-0.49L/L (SI)
o Capillary tube
 Diameter
 1.2 – 2.4mm
 Bore
 1mm
 Length
 75mm
Sources of Error
- Insufficient centrifugation
- Insufficient mixing of the sample
- Inappropriate concentration of the anticoagulant
- Specimen collection error
Physiologic factors that can affect hematocrit
- Abnormal erythrocyte shapes
 - Trapped plasma (causes to be 1-3% or 0.01 to 0.03 L/L higher hematocrit)
o Amount of plasma that still remains in RBC portion after the microhematocrit has been
spun
o Affected by centrifugation time, centrifugal force, smoothness of centrifugal unning,
temperature, sample of erythrocyte column studied, plasma label used
o Increased in macrocytic anemias, spherocytosis, thalassemia, hypochromic anemia,
sickle cell anemia
- Immediately after blood loss (low hematocrit)
o Plasma is replaced faster than erythrocytes
- Dehydration (high hematocrit)
o As the volume of fluid in the blood drops, the RBCs per volume of fluid artificially rises
Poikilocytes that cause trapped plasma
- Spherocytes
o Hereditary spherocytosis
o Immune hemolytic anemias
o Severe burns
o RBCs lost their biconcavity due to decrease surface to volume ratio
o Hyperchromic (dense hemoglobin)
- Knizocyte
o Spherocytosis
o With more than two concavities
o Associated with spherocyte
- Drepanocyte
o Sickle cell anemia
o Thin elongated cells with pointed ends, may be curved, straight, or S, V, L shaped
o Polymerized Hemoglobin S showing various shapes, crescent or boat shaped
- Elliptocyte
o Thalassemia
o Elongated cells with central area of palor and hemoglobin at both ends
- Leptocyte
o Thalassemia
o Thin flat cell with hemoglobin at the periphery
Pathologic conditions
- Increase hematocrit
o Polycythemia vera
 Increased RBC production
o Hypoxia
 increase erythropoietin production in response to low blood O2 level
o Congenital heart disease
- Decrease hematocrit
o RBC hemolysis (hemolytic anemia)
 Increased RBC destruction=decreased RBC in whole blood
o Overhydration
 Too much volume of fluid (water) causes decreased RBC concentration

How hematocrit is derived using automated hematology analyzers

- Hematocrit is calculated from the determination of MCV and RBCs
- The computation makes no allowance for trapped plasma