M. A. Ahmad Presentation FMC

THE INTERFERANCE EFFECTS OF HAEMOLYSIS,
ICTERUS AND LIPEMIA ON ROUTINE CLINICAL

CHEMISTRY PARAMETERS
BY
MUSTAPHA ABUBAKAR AHMAD
Supervised by;
MLS. ABUBAKAR MUSA
Chemical Pathology Unit, Pathology Department,

Federal Medical Center Nguru 1
OUTLINE
 Introduction
 Haemolysis
 Causes of haemolysis
 Haemolysis Interference
 Icterus
 Causes of Icterus
 Icterus Interference
 Lipemia
 Causes of Lipemia
 Lipemia Interference
 Precautions and Control of Interference
 Conclusion
 Recommendation
 References 2
Introduction
 Interference in clinical laboratory occur when a substance
present in a sample leads to a change in the measured value of
another parameter.
 Interference can be from endogenous source (substances
originate from the patient’s sample) or exogenous source
(substances introduce into the specimen).
(Mainali et al., 2021)
3
Introduction Cont.
 Interference from endogenous substances is one of the
leading source of errors that clinical laboratories frequently
encounter at the pre-analytical phase of testing.
 Pre-analytical phase encompasses all the procedures before
carrying out laboratory testing.
 This phase of the testing process is responsible for majority
of laboratory errors. (Asmelash et al., 2020)
4
Haemolysis
 Haemolysis is defined as the destruction of red blood cells
(RBC) which leads to the release of haemoglobin and other
intracellular components of erythrocytes into the extracellular
space of blood.
 Haemolysis may occur in vivo as a results of a number of
circumstances and haemolytic diseases, while in vitro as a
results of preanalytical errors.
(Thomas, 2010) 5
Causes of Haemolysis
Preanalytical causes may be associated with;

 Improper sampling,
 Improper tube mixing
 Sample transport method
 Delayed processing
 Prolonged storage (Lippi et al., 2011)

6
(Lippi et al., 2011) 6
Haemolysis Interference
 Increased absorbance
 Release of intracellular constituents that are directly

measured
 Release of membrane constituents that participate in
reactions
 Water release
(Lippi et al., 2018) 8

Icterus (Jaundice)
 Icterus is one of the most frequent endogenous interferences in
clinical laboratories
 Icterus is a yellowish pigmentation of the skin and mucous
membrane, mainly caused by an increased bilirubin level due to
overproduction in the liver.
 The presence of elevated levels of bilirubin (icterus) in serum or
plasma specimens has the potential to interfere with clinical chemistry
and other laboratory assays. (Alvarez et al., 2022)
9
Cause of Icetrus
There are two main ways that bilirubin can increase in the blood;
 Intrinsic Icterus; caused by liver disease, blood disorders, or
other conditions that lead to increased Rbc destruction.
 Extrinsic Icterus; mostly caused by UDP glucuronyl transferase
deficiency and Dubin-Johnson syndrome.
(Farrell and Carter, 2016) 10

(Farrell and Carter, 2016)
11
Icterus Interference
There are two common mechanisms by which icterus can cause
assay interference;
 Chemical reaction of bilirubin with the reagents used in some
enzymatic assays (e.g. glucose, cholesterol, uric acid,
triglyceride, albumin e.t.c)
 Spectral interference due to absorption at wavelengths used in
assays by bilirubin and/or bilirubin breakdown products.
(Nicolay et al., 2018) 12
Lipemia
 Lipemia is a term used to describe an increase in the amount
of lipids in the blood.
 Lipemia plays a significant role in causing pre-analytical
errors by increasing the turbidity of the sample due to the
presence of high concentrations of lipoprotein particles.
(Nikolac, 2014)
13
Causes of Lipemia
Lipemia is caused by increased triglycerides, and the common
cause is inadequate fasting and patient preparation before blood
sampling.
Other common causes include;
 Genetic background
 Diabetes mellitus
 Acute pancreatitis
 Renal failure
 Alcoholism
 Hypothyroidism (Nelson, 2013)
 Some drugs 14
(Nelson, 2013) 15
Lipemia Interference
Lipemia can cause interference in biochemistry results through a
variety of mechanisms such as;
 Interference with spectrophotometric methods
 interference due to non-homogeneity of the sample
 Volume Displacement Effect (VDE).
(Ji & Meng, 2011; Sen et al., 2013) 16

Precautions and Control of Interferences
Interference can be avoided at the pre-analytical phase through
the following ways;
 Promoting optimal phlebotomy
 Promoting specimen transport/processing
 Adequate fasting and patient preparation before blood
sampling
(Khasawneh and Bani, 2018)
17
Cont’d
Interference can also be controlled in the clinical laboratory by
the following methods;
 Ultracentrifugation
 Sample dillution
(Dias et al., 2017)
18
Conclusion
Analytical interferences caused by pre-analytical factors, such as
icterus, are significant source of errors in clinical laboratory
measurements.
Hemolysis, Icterus and Lipemia can cause falsely high/low results for
electrolytes, enzymes, glucose, cholesterol, triglycerides, bilirubin e.t.c.
Results for other analytes can also be affected, depending on the
specific test being performed.
19
Recommendation
It is recommended that phlebotomist and laboratory staffs should be
aware of the potential of these interferences so that they can be
detected and corrected. It is also crucial to focus on standardization of
blood collection practices, staff training, and quick transportation of
blood specimens to reduce hemolysis rates in the pre-analytical
process.
Additionally, it’s important to have good quality control procedures in

place to detect and correct any errors that may occur. 20
Alvarez, Y. F., Kelliher, M. T., Hubbard, J. A., Nerenz, R. D., & Cervinski, M. A. (2022). Reevaluating the Icterus Index Cutoff of a Jaffé
Creatinine Method. The Journal of Applied Laboratory Medicine, 7(5), 1151-1157.
Asmelash, D., Worede, A., & Teshome, M. (2020). Extra-analytical clinical laboratory errors in Africa: a systematic review and meta-
analysis. Ejifcc, 31(3), 208.
Dias, C. B., Moughan, P. J., Wood, L. G., Singh, H., & Garg, M. L. (2017). Postprandial lipemia: factoring in lipemic response for ranking
foods for their healthiness. Lipids in health and disease, 16(1), 1-11.
Farrell, C. J. L., & Carter, A. C. (2016). Serum indices: managing assay interference. Annals of clinical biochemistry, 53(5), 527-538.
Ji, J. Z., & Meng, Q. H. (2011). Evaluation of the interference of hemoglobin, bilirubin, and lipids on Roche Cobas 6000 assays. Clinica
chimica acta, 412(17-18), 1550-1553.
Khasawneh, W., & Bani Hani, S. (2018). Intravenous lipid emulsion overdose in infancy: a case report and overview of opportunities,
challenges and prevention. Drug safety-case reports, 5, 1-4.
Lippi, G., Avanzini, P., Dipalo, M., Aloe, R., & Cervellin, G. (2011). Influence of hemolysis on troponin testing: studies on Beckman Coulter
UniCel Dxl 800 Accu-TnI and overview of the literature. Clinical Chemistry and Laboratory Medicine (CCLM), 49(12),
2097-2100.
Lippi, G., Favaloro, E. J., & Franchini, M. (2018). Haemolysis index for the screening of intravascular haemolysis: a novel diagnostic
opportunity?. Blood Transfusion, 16(5), 433.
Mainali, S., Merrill, A. E., & Krasowski, M. D. (2021). Frequency of icteric interference in clinical chemistry laboratory tests and causes of
severe icterus. Practical laboratory medicine, 27, e00259.
Nelson, R. K. (2013). Influence of Habitual Physical Activity and Acute Exercise on Lipid Metabolism and Insulin Action (Doctoral
dissertation).
Nicolay, A., Lorec, A. M., Gomez, G., & Portugal, H. (2018). Icteric human samples: icterus index and method of estimating an
interference‐free value for 16 biochemical analyses. Journal of Clinical Laboratory Analysis, 32(2), e22229.
Nikolac, N. (2014). Lipemia: causes, interference mechanisms, detection and management. Biochemia medica, 24(1), 57-67.
Thomas, L. (2010). Haemolysis as influence & interference factor. Ejifcc, 13(4), 95.
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THE INTERFERANCE EFFECTS OF HAEMOLYSIS,

ICTERUS AND LIPEMIA ON ROUTINE CLINICAL

Chemical Pathology Unit, Pathology Department,

Preanalytical causes may be associated with;

 Improper tube mixing

 Sample transport method

 Prolonged storage (Lippi et al., 2011)

 Release of intracellular constituents that are directly

(Lippi et al., 2018) 8

 Intrinsic Icterus; caused by liver disease, blood disorders, or

other conditions that lead to increased Rbc destruction.

 Extrinsic Icterus; mostly caused by UDP glucuronyl transferase

deficiency and Dubin-Johnson syndrome.

(Farrell and Carter, 2016) 10

 Interference with spectrophotometric methods

 interference due to non-homogeneity of the sample

 Volume Displacement Effect (VDE).

(Ji & Meng, 2011; Sen et al., 2013) 16

(Dias et al., 2017)

Additionally, it’s important to have good quality control procedures in

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