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Lim and Cha: Proposal of Modified HIL-indices for Determining Hemolysis, Icterus and Lipemia Interference on the Beckman Coulter AU5800 Automated Platform
Original Article

Lab Med Online 2017;7(2):66-72.

Published online: 17 April 2017

DOI: https://doi.org/10.3343/lmo.2017.7.2.66

Proposal of Modified HIL-indices for Determining Hemolysis, Icterus and Lipemia Interference on the Beckman Coulter AU5800 Automated Platform

Yong Kwan Lim, M.D., Young Joo Cha, M.D.

Department of Laboratory Medicine, Chung-Ang University College of Medicine, Seoul, Korea

Corresponding author: Young Joo Cha Department of Laboratory Medicine, Chung-Ang University College of Medicine, 102 Heukseok-ro, Dongjak-gu, Seoul 06973, Korea Tel: +82-2-6299-2720, Fax: +82-2-6298-8630, E-mail: chayoung@cau.ac.kr

Received 4 July 201617 August 2016       Accepted 30 August 2016

Copyright © 2017 The Korean Society for Laboratory Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

The amount of interference due to hemolysis, bilirubin, and lipemia can be measured on the AU5800 autoanalyzer (Beckman Coulter, USA) by spectrophotometry. This is reported as semi-quantitative indices, specifically H-index, I-index, and L-index, respectively. In this study, we evaluated the impact of interference using chemistry assays and established the concentration of interfering substances and HIL-index above which analytically significant interference exists, according to CLSI guidelines C56-A and EP7-A2.

Methods

Pooled sera including different concentrations of analytes were prepared and mixed with hemoglobin, bilirubin, or Intralipid. These samples were then tested for 35 clinical chemistry analytes by AU5800 and the bias based on interferent concentrations was computed. The interferent concentration above which significant interference exists was calculated from the 50% within-subject biological variation (desirable analytic goal), and the corresponding index was assigned.

Results

Among 35 items evaluated, interference was detected for 12 analytes by hemoglobin, 7 analytes by bilirubin, and 12 analytes by Intralipid. We proposed HIL-index1 and HIL-index2 for each analyte according to 2 different medical decision levels. HIL-index1 and HIL-index2 were considered more reasonable criteria than the HIL-index from the manufacturer's technical document (HIL-indexTD). This is because HIL-indexTD was empirically set to 5% or 10%, and had a wide tolerance range, which was not sufficient to reflect the presence of interference, compared to HIL-index 1 and HIL-index2.

Conclusions

We have demonstrated hemoglobin, bilirubin, and Intralipid interferences according to CLSI guidelines using the desirable analytic goal. Our results provide applicable information for Beckman Coulter automated chemistry analyzers.

Keywords: Interference, Hemolysis, Icterus, Lipemia, Chemistry, AU5800

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Fig. 1.
Bilirubin interference in the measurement of total protein. Analyte bias was plotted by linear regression analysis, and the interferent concentrations at which interference began were calculated with the desirable analytic goal (Bcutoff).
lmo-7-66f1.tif
Table 1.
The relationship between the concentration of the interfering substance and the corresponding HIL-index, according to manufacturer's specification
HIL-index Hemoglobin (mg/dL) Bilirubin (mg/dL) Intralipid (mg/dL)
0 <50 <2.5 <40
1 50-99 2.5-4.9 40-99
2 100-199 5.0-9.9 100-199
3 200-299 10-19.9 200-299
4 300-500 20-40 300-500
5 >500 >40 >500
Table 2.
Average concentration of tested analytes and desirable bias cutoff criteria for interference
Analyte Unit Average concentrations Bcut- off(%) Analyte Unit Average concentrations Bcut- off(%)
Level 1 Level 2 Level 1 Level 2
Total protein g/dL 7 8.6 1.4 LDH IU/L 146.8 332.3 4.3
Albumin g/dL 4.1 4.7 1.6 GGT IU/L 33.2 201.5 6.7
Total bilirubin mg/dL 0.6 12.7 10.9 Total calcium mg/dL 9 9.6 1.1
Direct bilirubin mg/dL 0.2 8.2 18.4 Amylase IU/L 65 363.3 4.4
BUN mg/dL 18 45 6.1 CK IU/L 62.2 1197.5 11.4
Uric acid mg/dL 4.9 7.5 4.3 Sodium mEq/L 143.8 148.8 0.3
ALP IU/L 55.9 300.7 3.2 Potassium mEq/L 4.5 4.9 2.3
IP mg/dL 3.8 3.9 4.1 Chloride mEq/L 106.2 115.7 0.6
Magnesium mg/dL 2.1 2.7 1.8 Iron μg/dL 90.5 147.2 13.3
Triglyceride mg/dL 108.7 547.8 10.0 UIBC μg/dL 209.5 384 10∗
HDL mg/dL 42.7 67.5 3.7 Total cholesterol mg/dL 176 279.7 3.0
LDL mg/dL 105.2 180.3 3.9 Bicarbonate mmol/L 16.1 27.6 2.0
Lipase IU/L 24.5 427 16.1 CRP mg/L 9.8 154.5 21.1
Phospholipid mg/dL 188 262.5 3.3 hs-CRP mg/L 10.6 30.4 24.9
Glucose mg/dL 128 204.7 2.8 Ethanol mg/dL 7.6 87.4 10∗
Creatinine mg/dL 1.2 2.9 3.0 Ammonia μg/dL 200 394 10∗
AST IU/L 22.7 269.5 6.2 KB μmol/L 85.5 479 10∗
ALT IU/L 9.5 83.2 9.7          

Bcutoff: Cut-off bias calculated based on the desirable analytic goal.

∗ Arbitrarily set to 10% owing to a lack of within-subject biological variation data.

Table 3.
Averages of reported H-, I-, and L-index according to interferent concentrations
Tested hemoglobin concentration (mg/dL) Averages of H-index Tested Bilirubin concentration (mg/dL) Averages of I-index Tested Intralipid concentration (mg/dL) Averages of L-index
0 0 0 0 0 0
30 0.04 2 1.13 20 0.27
60 0.88 4 2.04 70 1.08
120 1.52 8 2.92 150 2.00
240 2.02 16 3.65 250 2.58
480 4.04 32 4.10 400 3.96
720 4.96 48 5 600 4.13
960 5 64 5 1,000 5
Table 4.
Items affected by hemoglobin; comparing the H-indexTD obtained with AU5800 and the H-index calculated in this study for each analyte
Analyte H-indexTD H-index1 H-index2
Increasing effect      
   Total protein 4 1 2
   Albumin 4 2 3
   IP 4 5 4
   Magnesium 2 0 0
   AST NS∗ 1 4
   ALT 5 2 5
   LDH NS∗ 2 2
   Iron 2 0 1
   Total cholesterol 5 3 5
Decreasing effect ALP 5 3 5
   GGT 3 2 4
   UIBC 3 1 2

H-indexTD: H-index from the technical document provided by the manufacturer.

H-index1: H-index calculated from the level 1 sample in this study.

H-index2: H-index calculated from the level 2 sample in this study.

∗ Not specified in the technical document.

Table 5.
Items affected by bilirubin; comparing the I-indexTD obtained with AU5800 and the I-index calculated in this study for each analyte
Analyte I-indexTD I-index1 I-index2
Increasing effect      
   Total bilirubin NS∗ 0 0
   Direct bilirubin NS∗ 0 0
   Magnesium 4 2 3
   Ammonia NS∗ 5 5
Decreasing effect      
   Total protein 4 1 2
   Creatinine 4 4 4
   Total cholesterol 2 1 1

I-indexTD: I-index from the technical document provided by the manufacturer.

I-index11: I-index calculated from the level 1 sample.

I-index2: I-index calculated from the level 2 sample.

∗ Not specified in the technical document.

Table 6.
Items affected by Intralipid; comparing the L-indexTD obtained with AU5800 and the L-index calculated in this study for each analyte
Analyte L-indexTD L-index1 L-index2
Increasing effect      
   Albumin 5 1 3
   IP 5 3 5
   Magnesium 5 1 1
   Triglyceride NS∗ 0 1
   Phospholipid NS∗ 1 3
   Iron 2 2 5
   Ammonia NS∗ 3 5
Decreasing effect      
   Total protein 5 2 4
   Uric acid 5 4 5
   HDL 5 2 4
   LDL NS∗ 1 2
   Creatinine 5 0 5

L-indexTD: L-index from the technical document provided by the manufacturer.

L-index1: L-index calculated from the level 1 sample.

L-index2: L-index calculated from the level 2 sample.

∗ Not specified in the technical document.

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