Utility of HIL in Clinical Chemistry

Yachana Kataria, PhD, DABCC

Clinical Chemistry Fellow

Boston Children’s Hospital

DOI: 10.15428/CCTC.2016.264754

© Clinical Chemistry
What are serum indices?

• Hemolysis, icterus and lipemia (HIL) are the most

common specimen integrity issues
• Objective way to detect interferences compared
to visual inspection
• Standardized and reproducible tool

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 Limitations of HIL on automated analyzers

Method Limitations

Hemolysis • Spectral Interference • Different manufacturers have

different cut-off values
• More than one HIL interferent
Icterus • Spectral Interference may be present simultaneously in
a patient sample
• Other interferents may still be
present
Lipemia • Light Scattering
• Does not replace assays of
hemoglobin, bilirubin, or
triglycerides

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 Determination of HIL Cut-Off Limits

Bilirubin Interference on ABC Bilirubin Interference on DEF

9 20
18
ABC Response (mg/dL)

DEF Response (mg/dL)

7 16
6 14
High Level 12 High Level
5
10
4 Low Level Low Level
8
3 6
2 4
1 2
0 0
0 5 10 15 20 25 0 5 10 15 20 25
Bilirubin (mg/dL) added Bilirubin (mg/dL) added

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Hemolysis index (H) is assessed by the
amount of red pigmentation associated
with free hemoglobin

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CLSI 2012
Analytes affected by hemolysis

Positive Interference Negative Interference

• Elevated intracellular • Haptoglobin

concentration
• Potassium,
magnesium and
phosphate
• Lactate Positive or Negative
dehydrogenase (LDH) Interference
• Aspartate • Troponin
aminotransferase
(AST)

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 How to deal with hemolyzed specimens
Over the cut-off limit?

No Yes

Proceed to testing Dilution as possibility?

Yes No

Proceed to testing In-vitro or in-vivo?

In-vitro In-vivo

Determine 7
Reject
Alternatives
Icteric index (I) is assessed by yellow
pigmentation due to increased
bilirubin concentration

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CLSI 2012
Analytes affected by icterus

• Peroxidase catalyzed reactions

• Examples: cholesterol, glucose and triglycerides

• Creatinine – Jaffe Method

Creatinine + Picric Acid → Janovsky Complex (orange-red color)

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 How to deal with icteric specimens
Over the cut-off limit?

No Yes

Proceed to testing Dilution as possibility?

Yes No

Determine available
Proceed to testing methods (i.e. Creatinine -
enzymatic method)
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Lipemic index (L) is assessed by turbidity
due to elevated lipoproteins

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CLSI 2012
Lipemia causes volume displacement

‘Normal’ Lipemic
Plasma Sample

Water Content 93% 84%

Lipids 7% 16%

Na [mmol/L] 140 126

Indirect ISE

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 How to deal with lipemia

Dilution

Endogenous (i.e.
Determine Cause

Physiologically Remove Lipids* Lipid Clearing Agent

elevated)

Ultracentrifugation
Exogenous (i.e. TPN
Reject specimen
administration)

*For indirect ISEs, use direct ISEs for

comparison
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Automated assessment of hemolysis,
icterus, & lipemia (HIL) provides the
laboratory a standardized, reproducible
and efficient tool to detect possible
interference related to sample integrity

© Can Stock Photo / aurielaki

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References
1. Clinical and Laboratory Standards Institute (CLSI) ” Hemolysis,
icterus, and lipemia/turbidity indices as indicators of interference in
clinical laboratory analysis; Approved guideline." CLSI document
C56-A, Wayne, Clinical and Laboratory Standards Institute (2012).
2. Dimeski, Goce. "Interference testing." The Clinical Biochemist
Reviews 29.Suppl 1 (2008): S43.
3. Nikolac, Nora. "Lipemia: causes, interference mechanisms, detection
and management." Biochemia medica 24.1 (2014): 57-67.
4. Bowen, Raffick AR, et al. "Impact of blood collection devices on
clinical chemistry assays." Clinical biochemistry 43.1 (2010): 4-25.

http://www.clinchem.org/site/info_ar/info_authors.xhtml#References

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Disclosures/Potential Conflicts of Interest
Upon Pearl submission, the presenter completed the Clinical Chemistry
disclosure form. Disclosures and/or potential conflicts of interest:
▪ Employment or Leadership: No disclosures
▪ Consultant or Advisory Role: No disclosures
▪ Stock Ownership: No disclosures
▪ Honoraria: No disclosures
▪ Research Funding: No disclosures
▪ Expert Testimony: No disclosures
▪ Patents: No disclosures

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