Analytical performance of Abbott C-16000 analyser haemolysis index and its potential use in measuring plasma cell-free haemoglobin.

Abstract

Measurement of the haemolysis index (HI) is usually performed in clinical chemistry laboratories in order to inform about whether biological analyses are influenced by in vivo or in vitro haemolysis of the specimen. Our aim was to evaluate the analytical performance of Abbott C-16000 analyser HI measurement in order to determine whether this could be used to reliably measure cell-free haemoglobin (fHB) in plasma samples. The repeatability, reproducibility, lower limit of detection (LLOD) and lower limit of quantification (LLOQ) of C-16000 HI measurement were determined as well as the potential interference of bilirubin, triglycerides and myoglobin. C-16000 HI values of biological samples with various ranges of fHB were compared to those measured using the established reference method, second-derivate spectroscopy. Results: C-16000 HI determination showed excellent linear correlation with the reference method (y = 1.0043x – 1.248, R² = 0.998), a broad analytical measurement range (400-20,000 mg/L; y = 0.9904x + 72.972, R² = 0.999), clinically relevant LLOD (56 mg/L) and LLOQ (84 mg/L), good repeatability (coefficient of variation (CV) = 1-15%) and good reproducibility (CV = 5-7%). No interference was observed with myoglobin at concentrations as high as 35,447 mg/L, unconjugated and conjugated bilirubin (at concentrations up to 500 mg/L and 375 mg/L, respectively) or triglycerides up to 6.8 mmol/L. However, a significant underestimation of fHB concentrations was observed at higher triglyceride levels. This study demonstrates that Abbott C-16000 analyser HI is reliable and accurately measures plasma fHB concentrations under pathophysiological conditions except when there are high blood concentrations of triglycerides.