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February 8, 2026The Journal of Applied Laboratory Medicine0 citationsOpen Access

Hemolysis, Icterus, and Lipemia Interference Limits for Serum β-hydroxybutyrate Testing

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MCMark A CervinskiDartmouth CollegeMSMacey J SmithDartmouth–Hitchcock Medical CenterGFGracie ForguesDartmouth–Hitchcock Medical Center

Key Points

  • This research aims to evaluate the interference limits for hemolysis, icterus, and lipemia in β-hydroxybutyrate testing.
  • Combined patient samples to create pools with specific BOHB concentrations
  • Added varying amounts of hemolysate, bilirubin, or Intralipid® to test interference
  • Measured interference and BOHB concentration on Roche cobas c502, c501, and c311 analyzers
  • Hemolysis tolerated up to a hemolysis index of 400 for BOHB ≤1.00 mmol/L
  • The assay remained unaffected by bilirubin up to an icterus index of 25 for all BOHB concentrations
  • Lipemia interference limit set at a lipemia index of 800

Abstract

Abstract Background Measurement of β-hydroxybutyrate (BOHB) is a valuable tool for the assessment of metabolic acidoses. In this study we evaluated hemolysis, icterus, and lipemia interference thresholds for a commonly used BOHB assay, the Stanbio β-Hydroxybutyrate LiquiColor reagent, on 3 different Roche cobas analyzers. Methods Remnant patient samples were combined to generate pools with target BOHB concentrations of approximately 0.20, 3.00, and 7.00 mmol/L into which increasing amounts of hemolysate, bilirubin, or Intralipid® were added. Samples were measured in triplicate for both interferences and BOHB concentration on Roche cobas c502, c501, and c311 analyzers. Results We determined that the Stanbio β-Hydroxybutyrate LiquiColor reagent will tolerate hemolysis to a hemolysis index of 400 (approximately 400 mg/dL hemoglobin) for BOHB concentrations ≤1.00 mmol/L, or 1010 for BOHB concentrations of 1.00 mmol/L. For icterus, the assay was unaffected by bilirubin to an icterus index of 25 approximately 25 mg/dL bilirubin (427 μmol/L) for all concentrations tested. Using the most conservative data from our study, the lipemia index limit was set at 800. Conclusions Our studies across 3 laboratories running 3 different cobas modules allowed our health system to better define permissible limits of sample quality and these updated values will reduce the number of unnecessary cancelations as compared to the limits defined by the manufacturer. Our data also demonstrated heterogeneity of interference between analyzer models, which is an important consideration for those seeking to adopt the limits determined on analyzers other than those in their own laboratories.

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Cite This Study

Cervinski et al. (2026) studied this question.

synapsesocial.com/papers/6988291e0fc35cd7a8849345https://doi.org/10.1093/jalm/jfag001
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