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Preparation of In-House Quality Control Human Serum for Urea and its Use in
Clinical Chemistry

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DOI: 10.7754/Clin.Lab.2019.190704)

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Clin. Lab. 2020;66:365-368
©Copyright
ORIGINAL ARTICLE

Preparation of In-House Quality Control Human Serum for Urea and

its Use in Clinical Chemistry
Berhanu Haile 1, Demiraw Bikila 1, 2, Haymanot Tewabe 1, Mistire Wolde 1
1
Addis Ababa University, Department of Medical Laboratory Sciences, Addis Ababa, Ethiopia
2
Ethiopian Public Health Institute, Addis Ababa, Ethiopia

SUMMARY

Background: Quality control materials play a vital role in the laboratory internal and external quality assessment
program. However, Ethiopia and other developing countries are challenged by the unavailability and high cost of
commercial control material. Therefore, preparing in-house quality control human serum will be a cost-effective
way to obtain QC material for use in poor settings in a country like Ethiopia. To prepare urea in-house control
human serum and scientifically evaluate it with commercially available sera already in use in the clinical chemis-
try laboratory of the Ethiopian Public Health Institute.
Methods: The urea in-house quality control human serum was prepared as per ISO Guide 80, a guideline docu-
ment protocol from 57 participants’ normal serum specimens at the Ethiopian Public Health Institute clinical
chemistry laboratory. The quality control material was analyzed on a fully automated chemistry analyzer (Cobas
6000). The initial 20 values were used for calculation of means, standard deviation (SD) and coefficient of varia-
tion (CV). Results were compared with those of commercially available lyophilized human sera.
Results: The average concentrations of urea were found to be near the middle of the physiological range of healthy
subjects and the in-house serum could be a good substitute for the commercial serum of normal range. The pre-
pared in-house quality control human serum is stable for about three months without any alterations in the con-
centration of urea.
Conclusions: Well prepared in-house quality control human serum is a good substitute for commercially available
control serum of normal range, especially in a developing country like Ethiopia.
(Clin. Lab. 2020;66:365-368. DOI: 10.7754/Clin.Lab.2019.190704)

Correspondence: KEY WORDS

Berhanu Haile
Addis Ababa University urea nitrogen, quality control, in-house control
Addis Ababa
Ethiopia
Mobile Phone: +251 916350525
Email: birex06@gmail.com INTRODUCTION

Urea is synthesized in the liver from ammonia produced

as a result of deamination of amino acids. This biosyn-
thetic pathway is the human body's chief means of ex-
creting surplus nitrogen. Urea measurements are used in
the diagnosis of certain renal and metabolic diseases.
The determination of serum urea is the most widely
used test for the evaluation of kidney function. The test
is frequently requested in conjunction with the serum
____________________________________________ creatinine test for the differential diagnosis of pre-renal,
Manuscript accepted August 25, 2019 renal, and post-renal uremia. High urea levels are asso-

Clin. Lab. 3/2020 365

 B. Haile et al.
ciated with impaired renal function, increased protein
catabolism, nephritis, intestinal obstruction, urinary ob-
struction, metallic poisoning, cardiac failure, peritonitis,
dehydration, malignancy, pneumonia, surgical shock,
Addison's disease, and uremia. Low urea levels are as-
sociated with amyloidosis, acute liver disease, pregnan-
cy, and nephrosis. Normal variations are observed ac-
cording to a person's age and gender, the time of day,
and diet, particularly protein intake [1].
The urea in-house control human serum preparation for
control of the quality measurements is not products (not
for sale). It is an important activity which provides ma-
terials suitable for the day-to-day demonstration that a
particular (part of a) measurement system is under sta-
tistical control and can be prepared "in-house", i.e., by
laboratory staff familiar with their behavior, to fulfill
specific quality control requirements [2].
ISO Guide 80 outlines the essential characteristics of
quality control materials and describes the processes by
which they can be prepared by competent staff within
the facility in which they will be used (i.e., where insta-
bility due to transportation conditions is avoided). The
content of the Guide also applies to inherently stable
materials, which can be transported to other locations
without the risk of any significant change in the proper-
ty values of interest. The requirements for ‘‘in-house''
QCMs intended for in-house use only are less demand-
ing than those for a CRM, for example, with respect to
transport issues. Before use, the quality assessment of
QCMs should involve homogeneity and stability assess-
ments and a limited characterization of the material to
indicate its relevant property values and their variation
[3]. In a developing country like Ethiopia, preparation
of in-house quality control materials was less practiced.
Therefore the study that had been undertaken would be
able to contribute highly in such circumstances. The
study aim is to prepare urea in-house control human se-
rum and scientifically evaluate it with commercially
available sera already used in the clinical chemistry lab-
oratory of the Ethiopian Public Health Institute.
MATERIALS AND METHODS
Study Area: Ethiopian Public Health Institute Clinical
Chemistry Laboratory, Addis Ababa, Ethiopia.
Study Period: The study period was from March 10 to
June 11, 2019.
Sample size: Fifty-Seven study participants’ concentra-
tions were the specimens were used.
The in-house liquid human quality control serum was
prepared with a modification of the ISO Guide 80, a
guiding document or protocol [3]. All the 57 normal se-
ra were pooled together in a graduated test tube and
their total volume was measured (5,700 µL). Once the
pooled sera have been sourced, there was a processing
stage (e.g., mixing and blending) to ensure the material
had the appropriate homogeneity and stability for its in-
tended purpose.
366
Once the pooled serum had been processed, the pre-
pared quality control serum was subdivided into differ-
ent Nunc tubes and the pooled sera were stored in dif-
ferent refrigerators (2 to 8°C and -20 to - 30°C). The
stability of pooled sera of urea was seven days at 15 -
25°C and one year at -20°C or -25°C.
To store the quality control serum, 1.8 mL polystyrene
capped tubes (Nunc tubes) were labeled as Normal con-
trol with the date. Then, 200 µL quality control serum
was pipetted into each tube making a total of 5,700 ali-
quots. Half of the aliquots were stored in a deep freezer
(-20 to -30°C) and the other half was stored at 2 - 8°C
until analysis. The prepared liquid quality control hu-
man serum along with the commercial lyophilized con-
trol sera was analyzed on a fully automated chemistry
analyzer (Cobas 6000) for comparison of the result of
urea.
Per day, four quality control serum samples were ana-
lyzed, two quality control serum samples from 2 - 8°C
and two quality control serum samples from -20 to -
30°C were run for five consecutive days; the five days
initial 20 results were obtained and this prospective
study continued for three months. The mean (first target
average), standard deviation, and coefficient of varia-
tion (CV) for urea analyte were then calculated for each
lot of the control sera and the results are presented in
the table.
Ethical considerations
The study was approved by Addis Ababa University
College of Health Sciences, Department of Medical
Laboratory Sciences, Ethical review committee. Permis-
sion letter was written from Addis Ababa University to
Ethiopian Public Health Institute Clinical Chemistry
Laboratory. For privacy reasons, all data were kept con-
fidential. The anonymity of result records was maintain-
ed by using client registration number and unique code
numbers used by service providers at the Ethiopian
Public Health Institute Clinical Chemistry Laboratory.
RESULTS
As shown in Table 1, the coefficient of variation (CV)
of the new tube/day (1.71) was lower than in-out tube/
day stored at similar temperatures (2 - 8°C) which de-
picted that the variability of the in-out tube/day was
higher than the new tube/day under similar storage tem-
peratures. For quality controls stored at -20 to-30°C, the
CV of the new tube/day was higher than that of the in-
out tube/per day under similar storage temperatures
which depicted less variability in the in-out tube/per
day. Overall the coefficient of variation of the in house
quality control stored at -20 to-30°C has good stability
as evidenced by less variability observed among mea-
surements.
As shown in Table 2, the prepared urea in house control
human serum was more stable at -20 to -30°C for about
three months. The mean urea in house quality control
Clin. Lab. 3/2020
 Preparation of In-House Quality Control Human Serum for Urea and Its Use in Clinical Chemistry

Table 1. Mean, standard deviation (SD), and coefficient of variation (CV) of routinely measured results of urea in-house con-
trol human serum stored at different temperatures.

Refrigerator 1 (2 - 8°C) Refrigerator 2 (-20 to -30°C)

NC-01 NC-02 NC-03 NC-04
(new tube/day) (a freez-thaw tube/day) (new tube/day) (a freez-thaw tube/day)
Mean 27.1 27.9 27.38 27.8
SD 0.46 0.95 0.73 0.43
% CV 1.71 3.43 2.68 1.56

Table 2. Results of mean concentrations of the in-house quality control human serum and Cobas® c 501 commercial sera for
the sample stored at 2 - 8°C and -20 to -30°C over a period of three months.

Cobas® c 501 In-house control at 2 - 8°C In-house control at -20 to -30°C Reference
Test commercial
April May June April May June range
sera
Urea
36.5 27.50 29.6 30.9 27.59 28.5 27.7 16.6 - 48.5
(mg/dL)

was 27.5, 29.6, 30.9 and 27.59, 28.5, 27.7 for the sam-
ples stored at 2 - 8°C and -20 to -30°C during the three
months study period, respectively. The reference range
used in the Ethiopian Public Health Institute Clinical
Chemistry laboratory was from 16.6 - 48.5 which en-
abled us to understand the level of control material
compared to physiological human concentrations. The
results showed that the urea in-house quality control
was near the middle of the physiological ranges com-
pared to the commercial sera.
DISCUSSION
Commercial control sera are prepared at two or three
levels of concentration. These can be used for all rou-
tine analyses. The disadvantages of commercial materi-
al are vial to vial variation in the concentration of their
constituents, no matter how carefully the vials are filled.
Reconstitution of material can introduce additional er-
ror. They are also very expensive [4].
The prepared urea in-house quality control sera showed
a narrower coefficient of variation contributing to better
error detection in the normal ranges. Therefore, well
prepared in house quality control human serum is a
good substitute for the normal commercial serum being
used in the laboratory. Furthermore, errors that occur as
a result of reconstitution of lyophilized sera and poten-
tial for introduction of an additional pipetting error dur-
ing reconstitution process are abolished as the in house
quality control serum was appropriately apportioned
during the initial preparation into vials adequate for urea
one-day usage in the daily analytical runs.
Additional advantages of the in-house control human
serum include easy preparation using normal laboratory
expertise. It is inexpensive and very cost-effective re-
sulting in cost-savings compared to the import of com-
mercial serum [4].
CONCLUSION
Well-prepared in-house quality control human serum is
a good substitute for the commercial control serum of
normal range especially in a developing country like
Ethiopia. Thus it is important to encourage the prepara-
tion of in-house quality control material as a substitute
for the commercially prepared quality control serum.
Further studies need to be conducted on in-house quali-
ty control human serum preparation that includes differ-
ent study sites and for a longer duration of time.
Acknowledgment:
We would like to thank the clinical chemistry laboratory
staff of the Ethiopian Public Health Institute for their
cooperation and willingness in analyzing the prepared
in house quality control serum.
Funding:
The research received no specific grant from any fund-
ing agency in the public, commercial, or not-for-profit
sectors.

Clin. Lab. 3/2020 367

 B. Haile et al.

Author Contributions:
Manuscript Preparation: Berhanu Haile, Mistire Wolde,
Haymanot Tewabe, Demiraw Bikila, Laboratory Analy-
sis: Berhanu Haile, Demiraw Bikila, Data analysis: Ber-
hanu Haile, Demiraw Bikila, Funding acquisition: No
funding source found, Investigation: Berhanu Haile, De-
miraw Bikila, Methodology: Berhanu Haile, Haymanot
Tewabe, Manuscript administration: Mistire Wolde,
Writing - review & editing: Berhanu Haile, Demiraw
Bikila, Haymanot Tewabe, Mistire Wolde.

Declaration of Interest:
We, the undersigned, declare no conflict of interest.

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368 Clin. Lab. 3/2020

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