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Received April 2, 2019, accepted April 14, 2020, date of publication May 2, 2020

Proposed Calibration of Apheresis Equipment 

By A. A. dos Santos1, M. A. Marciano2 and R. L. Rezer1

1 Hospital Moinhos de Vento/Clinical Engineering, Porto Alegre, Brazil
2 Hospital Moinhos de Vento/ Clinical and Hospital Engineering, Porto Alegre, Brazil

ABSTRACT
The health establishment is currently developing quality control through the calibration of biomedical equipment, sys-
tematically and comprehensively throughout the wide range of available hospital technology. Thus, this work aims to 
propose and demonstrate a method of qualifying apheresis equipment through equipment calibration before releasing it 
for first-time use. Results show the values obtained in calibration of apheresis equipment, relating to the MNC protocol 
(removal of mononuclear cells), the pressure of access, and return pressure.

Keywords – Qualification of Incorporation, Biomedical Equipment, Calibration of Apheresis Machines. 

Copyright © 2021. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY): Creative Commons - Attribution 4.0 International 
- CC BY 4.0. The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original 
publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

INTRODUCTION
Among the human pathologies are hematological 

diseases, immunodeficiency, genetic diseases and some 
tumors such as from breast cancer. One of the available 
treatments is donor (autologous) and donor (allogeneic) 
donor transplantation of peripheral blood progenitor cells. 
These cells, classified as CD34 +, have a high capacity for 
self-renewal and proliferative potential, which makes it 
possible to differentiate them into progenitor cells from 
all blood strains and to reconstitute the population from 
a single cell hematopoietic. Also called a blood stem 
cell. They comprise 0.05% to 0.1% of circulating hu-
man bone marrow hematopoietic cells.1,2 For collection 
and treatment in this invasive and safe procedure, a cell 
separator, called the apheresis machine, is used, which 
in Greek means separation. Apheresis therapy is the 
removal of one or more components of a person's whole 
blood. One of the most important steps in the process is 
the introduction of a sufficient amount of anticoagulant 

sufficient to prevent blood components from clotting or 
clumping together when they are processed through the 
apheresis device. The anticoagulant flow rate cannot be 
high, to avoid adverse reactions in the donor/patient. 
Depending on the amount of anticoagulant returned, 
you can generate physical symptoms such as tingling in 
the extremities until the potential damage to the donor/
patient.3 The apheresis equipment can be used to remove 
plasma (plasmapheresis), leukocytes (leukapheresis/
lymphocyteapheresis), platelets (thrombocyte-apheresis 
centre) or the red cells (erythrocyte apheresis).4

The apheresis machine performs the separation of 
peripheral blood and blood products through a technol-
ogy that utilizes centrifugal effect forces. Cells with higher 
densities are targeted at specific layers, where they can 
be identified and collected by a cardiopulmonary by-
pass induced by the equipment itself. The basic steps of 
apheresis, are the removal of whole blood from a donor 

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5 J Global Clinical Engineering Vol.2 Issue3: 2020

Santos, Marciano and Rezer: Proposed Calibration of Apheresis Equipment 

or patient, separating blood components, plasma reten-
tion, or one of its cellular components and returning the 
remaining elements to the donor or patient.5 The principle 
of operation of this equipment is shown in Figures 1A and 
1B.6 These centrifugal forces act on only a small portion 
of the blood, causing the separation of blood elements, 
called apheresis.

After the end of the cycle, which takes around three 
to five hours, the desired component is directed into a 
collection bag. The apheresis machine procedure is initi-
ated by a physician who performs the programming of 
the parameters based on the mononuclear cells removal-
MNC protocol. (The protocol's calculation is based on 
the amount of hematocrit collected from autologous or 
allogeneic donator, on the day of the procedure.7) The 
physician keeps up with the process and monitors the 
physical state of the patient. The physician inputs the 
age, sex, and weight of the patient or donor and also as-
sesses the need to change the length time or flow rate. At 
the end of processing, it is possible to collect the plasma 
cells, platelets, lymphocytes, granulocyte, and erythrocyte 
as needed. Figure 2 demonstrates the specific blood cell 
densities.8

Considering the given relevance, criticality, and ca-
re necessary in the method of separation of cells by 
apheresis, we need to understand how testing with this 
equipment is performed. When evaluating the methods 
of certification effectiveness of operating and the results 
of the procedure performed by the apheresis machine, it 
was verified that an item of Brazilian Resolution number 
57,9 called calibration is not necessarily being performed 
in a systematized way. This evaluation was done with the 
manufacturer of the equipment, specialists, and general 
research. It is critical to confirm the proper functioning of 
apheresis equipment using calibration, before releasing 
use throughout the course of its lifecycle. This paper aims 
to propose and demonstrate a method of accomplishing 
the qualification of apheresis equipment through calibra-
tion, in addition to the usual quantitative testing, before 
it is released for use.

METHOD

Search
During this process, we checked for quality control 

regulations for this equipment and standards relevant to 
its use. It was requested for the equipment manufacturer 
to provide the operating limits and tolerances of pressure 
and volume measured parameters as well as procedures 
to confirm conformity of post-factory output specifica-
tion. It was verified in article 11 of the Brazilian Resolu-
tion 57/2010 that "The Hematology services must have 
compatible equipment activities and establish a program 
that includes initial validation, qualification, calibration, 
preventive and corrective maintenance of equipment 
and instruments, keeping their schedules and records."

When evaluating the methods of certification effec-
tiveness for operation and results of the machine, it was 
found that the method used is often a six-month check 
to compare technical specifications to actual physical pa-
rameters. The parameters evaluated are those relating to 
access pressure, back pressure, pressure leaks, pressure 
sensors, rotation of centrifugal, circulation pumps, red 
sensor, and the digital conversion of these parameters.

According to information from the medical care spe-
cialists in the subject, normally the form of verification 
of the performance of the machine is the analysis of 

FIGURE 1. Apheresis circuit.

FIGURE 2. Demonstration of specific cell densities.



Santos, Marciano and Rezer: Proposed Calibration of Apheresis Equipment 

J Global Clinical Engineering Vol.2 Issue 3: 2020  6

the count of the blood products and blood products of 
patients, post-procedure. In this case, when the quality 
of processing is not adequate, there is a need to inform 
the patient of the need to perform a new procedure and 
consequently expose themselves again. Figure 3 has the 
flow method, with job steps.

Figure 3 shows the proposed inclusion of equipment 
calibration step as a way to raise the level of confidence 
of apheresis.

RESULTS

Calibration Planning
A qualified company was hired to perform the cali-

bration which evaluated the uncertainties involved and 
the standard deviations. This was done in conjunction 
with hospital technicians who assisted in this activity by 
providing access to the service equipment.

The following parameters were evaluated: pressure 
(mmHg), back pressure (mmHg), and Protocol MNC in 
volume (mL). The calibration method for measuring the 

volume was to measure the volume by weighing the liquid 
by time. For pressure measurement was performed com-
pared to the default. The materials used for calibration 
were: digital scale, digital Timer and Pressure Analyzer.

The calibration results were presented according to 
the following Table 1.

TABLE 1. Presentation of the Measured Data for the Equip-
ment: a) MNC protocol collection bag, b) access pressure and 
c) back pressure

a) Apheresis - MNC Protocol - Collection Bag

Set. 
Volume 
(mL)

Value 
Measured

Average
Error

Uncertainty
Total 
Error

Tolerance
Limits

175 179,10 4,10 ±0,01 | 4,11| 6%

b) Apheresis - MNC Protocol - Access Pressure

Set. 
Pressure 
(mmHg)

Value 
Measured

Average
Error

Uncertainty
Total 
Error

Tolerance
Limits

− 265 −270,00 −5,00 ±0,01  | 5,01| 12%

− 150 −147,10 3,00 ±0,01 | 3,01| 12%

− 50 −54,00 −4,00 ±0,01 | 4,01| 12%

c) Apheresis - MNC Protocol - Return Pressure

Set. 
Pressure 
(mmHg)

Value 
Measured

Average
Error

Uncertainty
Total 
Error

Tolerance
Limits

52 50,00 −2,00 ±0,01 | 2,01| 12%

254 256,00 2,00 ±0,01 | 2,01| 12%

403 408,00 5,00 ±0,01 | 5,01| 12%

In this way, the calibration procedure was added to 
the quantitative tests that were conducted for this type 
of equipment and complemented the proposed qualifying 
procedure apheresis equipment before release technique 
to use.

Performing Verification Technician
Among the quantitative tests also performed are: check-

ing the battery voltage of master boards; endurance tests 

FIGURE 3. The flow method, with job steps.



7 J Global Clinical Engineering Vol.2 Issue3: 2020

Santos, Marciano and Rezer: Proposed Calibration of Apheresis Equipment 

of the protection earth leakage current; verification of AC 
and DC voltages; verification and calibration of pressure 
sensors; functional check of the RBC detector; verification 
of pump rotation; and simulation with saline.

CONCLUSIONS
Evidence shows that the parameters compared were 

calibrated within the tolerances stated by the manufacturer. 
It was noted also that all measurements fell between the 
variations set and measured (even if within the range of 
tolerance). In this way, there may be situations where 
the parameters are very close to the tolerance allowed 
or even outside of it. Also, we can apply techniques of 
probability (considering the uncertainties of measure-
ment) to evaluate whether the value measured is within 
the maximum allowable limit.

Another point to assess is the type of protocols and 
tests that must be performed in this equipment so that 
they can reflect, in the most appropriate way, the effec-
tiveness of the process of blood cell separation carried 
out by the machine.

Considering the details above, there is a need to con-
solidate criteria and do a greater scope of tests and calibra-
tions on apheresis equipment to avoid initial or routine 
use of equipment that has not been calibrated.  This may 
help ensure that the equipment performs properly and 
thus avoid the risk of errors.

CONFLICT OF INTEREST
The authors declare that they have no conflict of interest.

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