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Hematocrit
Authors

Himel Mondal1; Saran Lotfollahzadeh2.

Affiliations
1 Bhima Bhoi Medical College and Hospital, Balangir
2 Renal and Vascular Section, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA

Last Update: January 2, 2023.

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 a 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, and 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.

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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: PMC1995040] [PubMed: 17948084]
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: 21250102]
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: 18678135]
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: 17824917]
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: 19171584]
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: 11324238]
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: 11553055]
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: PMC5511531] [PubMed: 28790546]

Disclosure: Himel Mondal declares no relevant financial relationships with ineligible companies.

Disclosure: Saran Lotfollahzadeh declares no relevant financial relationships with ineligible companies.
Figures

Wintrobe hematocrit tube containing components of blood after centrifugation. Contributed by Shaikat Mondal,
MD

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