Hematocrit (HCT)

Mondal H, Budh DP.

Publication Details

Introduction
The term "hematocrit (HCT)" originated from English “hemato-“ and Greek
“krites.” HCT measures the volume of packed red blood cells (RBC) relative to
whole blood. Hence, it is also known and reported as a packed cell volume (PCV).
It is a simple test to identify conditions like anemia or polycythemia and also to
monitor response to the treatment. A glass tube and a centrifuge machine are
sufficient to measure HCT. After centrifugation, the component of blood separates
into three distinct parts. From below upwards, the layers are - a layer of red blood
cells (RBC), a layer of white blood cells(WBC) and platelets, and a layer of plasma
at the top. This method of determining HCT by Wintrobe hematocrit tube is known
as the “macro-hematocrit” method.[1] A Wintrobe tube is a narrow glass tube
measuring 110-mm-long, with graduation from 0 to 100 mm in both ascending and
descending order. This method has been succeeded by the “micro-hematocrit”
method which uses a small capillary tube instead of a Wintrobe hematocrit tube. It
requires less quantity of blood as well as less time requirement for the testing
procedure. It is beneficial for patients from whom blood collection is difficult (e.g.,
pediatric patients/hypovolemia). However, the principle of the test remains the
same as the “macro-hematocrit” method. HCT calculation is by dividing the
lengths of the packed RBC layer by the length of total cells and plasma. As it is a
ratio, it doesn’t have any unit. Multiplying the ratio by 100 gives the accurate
value, which is the accepted reporting style for HCT. A normal adult male shows
an HCT of 40% to 54% and female shows 36% to 48%.[2] Though these two
methods are still in use in some settings of primary care and medical teachings,
they are widely replaced in the majority of settings by an automated analyzer,
where HCT reports get generated along with the complete blood count.

Specimen Requirements and Procedure

For the macro-hematocrit method, venous blood is taken as a random sample (i.e.,
no special precautions needed) maintaining proper aseptic precautions. The blood
is either collected in a commercially available vacutainer containing
ethylenediaminetetraacetic acid (EDTA) or in a vial/test tube with EDTA where
vacutainer is not available[3]. For the microhematocrit method, the blood
requirement is less, and single finger-prick blood is sufficient. Heparin filled
capillary tube may is used to collect blood. However, if there is available blood
with anticoagulant for other hematologic tests, a capillary tube without heparin can
be used. For measurement of HCT in automated hematologic cell counter, blood
with anticoagulant used for CBC is necessary.

Testing Procedures
The macro-hematocrit method uses a Wintrobe hematocrit tube, a centrifuge
machine, and a Pasteur pipette. Blood is filled in Wintrobe hematocrit tube up to
100 mm mark by the help of Pasteur pipette. Care is taken not to leave any bubble
in the blood column. For this, the tube is filled slowly with the tip of the pipette
being always below the highest position of the blood column. Then, the tube gets
placed in the centrifuge machine. When testing a single specimen, another blood-
filled Wintrobe hematocrit tube is kept on the opposite holder to counterbalance.
The spin setting is 3000 rpm for 30 min. After completion of the centrifugation, the
tube is taken out, and RBC column height is reported as HCT. During the
reporting, special precaution is necessary to omit the buffy coat, which is a
combination of WBC and platelets. This layer should not be included in the HCT,
as it may lead to false positive results.
For the microhematocrit method, after filling the blood in a capillary tube, the two
ends of the tube (commonly 75 mm long, 1 mm diameter) are sealed with clay
sealant or heat. Then, it is centrifuged at a rate of 11000 to 12000 rpm for 4 to 5
min. Reading is with the help of the scale on a tube holder or microhematocrit card
reader.
The automated analyzer measures the average RBC size and number by the
“Coulter principle”[4]. In this method, the size and number of the RBCs is
measurable by detecting impedance while the blood passes through a passage
between two electrodes.

Interfering Factors
There are several physiological and pathological conditions where the HCT may
deviate from its normal range. New-born babies show a high HCT, and it gradually
decreases during the neonatal period[5]. Adult male shows higher HCT than an
adult female.[6] Pregnant women show lower HCT due to hemodilution. In high
altitude, the number of RBC becomes high due to persistent hypoxia; hence, the
 inhabitant of high altitude shows higher HCT. Methodological variation may
provide a minor deviation of HCT tested for the same sample. In the macro-
hematocrit method, there is an increased amount of trapped plasma (approximately
2%) in the packed RBC, which may give a higher HCT. This factor becomes
minimized in the microhematocrit method, where the amount of trapped plasma is
less as the diameter of the capillary tube is less than that of the Wintrobe
hematocrit tube. Blood collected from different sources may also show variation.
Venous blood shows higher HCT than arterial blood. However, there is no
difference in HCT between venous blood and finger prick blood.[7]

Results, Reporting, Critical Findings

Wintrobe hematocrit tube has graduation 0 to 100 from below upwards. Hence, the
highest level of packed RBC is the HCT in percentage. For the microhematocrit
method, the reading is from the HCT card or scale. However, for these two
methods, even without any graduation or scale, the HCT value can be calculated
with a simple scale as we compare the length of the RBC column with the total
length of the fluid column. The final report is a percentage. HCT, along with RBC
count and hemoglobin (Hb) concentration, is used to report other blood indices
manually as follows:
The mean corpuscular volume (MCV) calculation uses HCT and RBC count.

 MCV (fL) = HCT (%) × 10 / RBC count (10^12/L)

Mean corpuscular hemoglobin concentration (MCHC) is calculated with Hb

concentration and HCT.

 MCHC (%) = Hb concentration (g/dL) / HCT (%) × 100

Clinical Significance
In primary health care settings, especially in resource-limited settings, macro-
hematocrit and micro-hematocrit methods are two low-cost and simple tests for
determining RBC in blood. Clinically, HCT is used to identify anemia and
polycythemia along with other parameters (e.g., RBC count, Hb concentration). In
anemia, where there are fewer RBCs in the circulating blood relative to the total
volume of the blood, the HCT decreases.[8] In polycythemia, there is a higher
number of RBCs in the blood; HCT increases. Smokers and chronic obstructive
pulmonary disease (COPD) patients also have high HCT due to chronic
hypoxia. The increase in HCT increases the viscosity of the blood, so does the
peripheral resistance. Hence, patients with higher HCT may have higher blood
pressure.

Quality control and Lab Safety

Handling of blood should take place with the maintenance of proper aseptic
precautions. The collected blood should be tested as soon as possible after
collection. Prolonged storage at the room temperature would result in a change in
the shape of the RBCs due to metabolism. After about 6 hours, the chances of
hemolysis increase, which would give an erroneous result. In the macro-hematocrit
method, the filing of the Wintrobe hematocrit tube requires proper care. In the
microhematocrit method, the sealing of the capillary tube should be secure to
prevent any leakage. The centrifuge machine should not be opened during the test
to avoid erroneous results. The chances of error in the result will increase if the
centrifugation is interrupted. Immediately after completion of the rotation, the
operator should not open the lid should until after the complete stoppage of the
rotation. For reuse of the Wintrobe hematocrit tube, proper cleaning is necessary as
any foreign particle inside the tube would be counted either in the RBC column or
plasma column.

Enhancing Healthcare Team Outcomes

After the introduction of the automated hematology cell counters, the usage of
macro-hematocrit and microhematocrit method is less used nowadays. However, in
many resource-poor settings, it is still used for the diagnosis of anemia and
polycythemia and monitoring response to treatment. In a rural healthcare facility, it
is a more accurate method for identifying anemia than the total RBC count, as
chances of error in manual RBC count is very high. The micro-hematocrit method
is more acceptable as it requires less amount of blood, and the testing time is very
less in comparison to the macro-hematocrit method. Due to the requirement of less
amount of blood and less time for the test, it may be useful in a mass survey.

Questions
To access free multiple choice questions on this topic, click here.
Figure
Wintrobe hematocrit tube containing components of blood after centrifugation. Contributed
by Shaikat Mondal, MD

References
1.
Fred HL. Maxwell Myer Wintrobe: new history and a new appreciation. Tex
Heart Inst J. 2007;34(3):328-35. [PMC free article] [PubMed]
2.
Billett HH. Hemoglobin and Hematocrit. In: Walker HK, Hall WD, Hurst
JW, editors. Clinical Methods: The History, Physical, and Laboratory
Examinations. 3rd ed. Butterworths; Boston: 1990. [PubMed]
3.
Bamberg R, Gwyn T, Miller J, Thompson M, Transou P. The effects of
over-anticoagulated blood on hematocrit values by the microcentrifuge
method. Clin Lab Sci. 2008 Summer;21(3):146-50. [PubMed]
4.
Peng L, Wang W, Bai L. Performance evaluation of the Z2 coulter counter
for WBC and RBC counting. Int J Lab Hematol. 2007 Oct;29(5):361-
8.[PubMed]
5.
Jopling J, Henry E, Wiedmeier SE, Christensen RD. Reference ranges for
hematocrit and blood hemoglobin concentration during the neonatal period:
data from a multihospital health care system. Pediatrics. 2009
Feb;123(2):e333-7. [PubMed]
6.
Zeng SM, Yankowitz J, Widness JA, Strauss RG. Etiology of differences in
hematocrit between males and females: sequence-based polymorphisms in
erythropoietin and its receptor. J Gend Specif Med. 2001;4(1):35-
40.[PubMed]
7.
 Yang ZW, Yang SH, Chen L, Qu J, Zhu J, Tang Z. Comparison of blood
counts in venous, fingertip and arterial blood and their measurement
variation. Clin Lab Haematol. 2001 Jun;23(3):155-9. [PubMed]
8.
Malenica M, Prnjavorac B, Bego T, Dujic T, Semiz S, Skrbo S, Gusic A,
Hadzic A, Causevic A. Effect of Cigarette Smoking on Haematological
Parameters in Healthy Population. Med Arch. 2017 Apr;71(2):132-
136.[PMC free article] [PubMed]

Publication Details
Author Information

Authors
Himel Mondal1; Deepa P. Budh2.

Affiliations
1 Bhima Bhoi Medical College and Hospital, Balangir
2 Sardar Patel University

Publication History
Last Update: February 20, 2020.

Copyright
Copyright © 2020, StatPearls Publishing LLC.

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Publisher
StatPearls Publishing, Treasure Island (FL)

NLM Citation
Mondal H, Budh DP. Hematocrit (HCT) [Updated 2020 Feb 20]. In: StatPearls [Internet]. Treasure Island (FL):
StatPearls Publishing; 2020 Jan-.