Abstract
Background
The accurate and precise measurement of triglycerides is important due to the adverse effects associated with hypertriglyceridaemia. Most laboratory methods are based on enzymatic hydrolysis of triglycerides with measurement of the total glycerol. An elevated free glycerol concentration may result in overestimation of triglyceride concentrations. The removal of free glycerol by blanking may therefore be of clinical importance. The aim of this study was to compare the glycerol blanking and non-glycerol blanking triglyceride methods.
Methods
This was a method comparison study of 1518 samples from both in-patients and out-patients at Charlotte Maxeke Johannesburg Academic Hospital. Triglycerides were measured in each sample using both the blanking and the non-blanking methods. Analytical performance was assessed based on the National Cholesterol Education Program (NCEP) goals. Clinical impact was assessed according to the NCEP Adult Treatment Program III (ATP III) risk classification.
Results
The method median was significantly higher in the non-blanking compared to the blanking method (1.33 vs. 1.12 mmol/L, p < 0.0001) in all patients. The average bias was above the total allowable error of 15% across all groups. There was a significant change in NCEP ATP III risk classification, with fewer patients classified as normal (67.6% vs. 74.6%, p < 0.0001) with the non-blanking method compared to the blanking method.
Conclusions
There was a significant error when glycerol blanking for triglyceride determination was not performed. The non-blanking triglyceride method overestimates triglyceride concentrations. This does not only exceed analytical performance goals, but also impacts on patient categorisation and clinical decision making in all patients.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: The reagents for this research study were sponsored by Roche Diagnostics South Africa.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organisation played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
References
1. Berglund L, Brunzell JD, Goldberg AC, Goldberg IJ, Sacks F, Murad MH, et al. Evaluation and treatment of hypertriglyceridemia: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 2012;97:2969–89.10.1210/jc.2011-3213Search in Google Scholar
2. Miller M, Stone NJ, Ballantyne C, Bittner V, Criqui MH, Ginsberg HN, et al. Triglycerides and cardiovascular disease: a scientific statement from the American Heart Association. Circulation 2011;123:2292–333.10.1161/CIR.0b013e3182160726Search in Google Scholar
3. Friedewald WT, Levy RI, Fredrickson DS. Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 1972;18:499–502.10.1093/clinchem/18.6.499Search in Google Scholar
4. Freiberg JJ, Tybjærg-Hansen A, Jensen JS, Nordestgaard BG. Nonfasting triglycerides and risk of ischemic stroke in the general population. J Am Med Assoc 2008;300:2142–52.10.1001/jama.2008.621Search in Google Scholar
5. Bansal S, Buring JE, Rifai N, Mora S, Sacks FM, Ridker PM. Fasting compared with nonfasting triglycerides and risk of cardiovascular events in women. J Am Med Assoc 2007;298:309–16.10.1001/jama.298.3.309Search in Google Scholar
6. Nordestgaard BG, Benn M, Schnohr P, Tybjaerg-Hansen A. Nonfasting triglycerides and risk of myocardial infarction, ischemic heart disease, and death in men and women. J Am Med Assoc 2007;298:299–308.10.1001/jama.298.3.299Search in Google Scholar
7. Alberti KG, Eckel RH, Grundy SM, Zimmet PZ, Cleeman JI, Donato KA, et al. Harmonizing the Metabolic Syndrome: A Joint Interim Statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and BloodInstitute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity. Circulation 2009;120:1640–5.10.1161/CIRCULATIONAHA.109.192644Search in Google Scholar
8. Hegele RA, Ginsberg HN, Chapman MJ, Nordestgaard BG, Kuivenhoven JA, Averna M, et al. The polygenic nature of hypertriglyceridaemia: implications for definition, diagnosis, and management. Lancet Diabetes Endocrinol 2014;2:655–66.10.1016/S2213-8587(13)70191-8Search in Google Scholar
9. Expert Panel on Detection Evaluation and Treatment of High Blood Cholesterol in Adults. Executive summary of the third report (NCEP) – adult treatment panel III. J Am Med Assoc 2001;285:2486–97.10.1001/jama.285.19.2486Search in Google Scholar
10. National Cholesterol Education Program. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) Final Report [Internet]. Bethesda, MD: National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH). NIH Publication No. 02-5215, 2002. Available from: https://www.nhlbi.nih.gov/health-topics/all-publications-and-resources/third-report-expert-panel-detection-evaluation-and-0.Search in Google Scholar
11. Cole TG, Klotzsch SG, McNamara JR. Measurement of Triglyceride Concentration. In: Rifai N, Warnick GR, Dominiczak MH, editors. Handbook of Lipoprotein Testing, 2nd ed. Washington, DC: AACC Press, 2000:207–19.Search in Google Scholar
12. Warnick GR, Kimberly MM, Waymack PP, Leary ET, Myers GL. Standardization of measurements for cholesterol, triglycerides, and major lipoproteins. Lab Med 2008;39:481–90.10.1309/6UL9RHJH1JFFU4PYSearch in Google Scholar
13. Walmsley TA, Potter HC, George PM, Florkowski CM. Pseudo-hypertriglyceridaemia: A measurement artefact due to glycerol kinase deficiency. Postgrad Med J 2008;84:552–4.10.1136/pgmj.2008.068577Search in Google Scholar PubMed
14. Naito HK. Reliability of lipid and lipoprotein testing (I). Am J Cardiol 1985;56:6J–9J.10.1016/0002-9149(85)90470-9Search in Google Scholar
15. Speeckaert MM, Segers H, Van Biesen W, Verstraete A, Langlois MR, Delanghe JR. An unusual case of (pseudo)hypertriglyceridaemia. NDT Plus 2010;3:570–2.10.1093/ndtplus/sfq148Search in Google Scholar PubMed PubMed Central
16. Jessen RH, Dass CJ, Eckfeldt JH. Do enzymatic analyses of serum triglycerides really need blanking for free glycerol? Clin Chem 1990;36:1372–5.10.1093/clinchem/36.7.1372Search in Google Scholar
17. Backes JM, Dayspring TD, Hoefner DM, Contois JH, Mcconnell JP, Moriarty PM. Identifying pseudohypertriglyceridemia in clinical practice. Clin Lipidol 2014;9:625–41.10.2217/clp.14.52Search in Google Scholar
18. Nauck M, Winkler K, Siekmeier R, Marangos N, Richter B, Marz W, et al. Pseudo-pseudohypertriglyceridemia: A case of increased free glycerol without evidence for glycerol kinase deficiency. Clin Chem 1995;41:619–20.10.1093/clinchem/41.4.619Search in Google Scholar
19. Cole TG. Glycerol blanking in triglyceride assays: Is it necessary? Clin Chem 1990;36:1267–8.10.1093/clinchem/36.7.1267Search in Google Scholar
20. Nakamura M, Iso H, Kitamura A, Imano H, Noda H, Kiyama M, et al. Comparison between the triglycerides standardization of routine methods used in Japan and the chromotropic acid reference measurement procedure used by the CDC Lipid Standardization Programme. Ann Clin Biochem 2016;53:632–9.10.1177/0004563215624461Search in Google Scholar PubMed PubMed Central
21. Jeon CH, Kim SK, Lee SC. Glycerol interference on the measurement of triglyceride concentration. Korean J Clin Pathol 1997;17:703–10.Search in Google Scholar
22. Roche Cobas® c501/502. TRIGB Triglycerides/Glycerol Blanked. Order info (2016-06 V6.0):Ref 11877771 216.Search in Google Scholar
23. Roche Cobas® c701/702. TRIGL Triglycerides. Order info(2017-11 V12.0):Ref 05171407 190.Search in Google Scholar
24. Stein EA, Myers GL. National cholesterol education program recommendations for triglyceride measurement: executive summary. The National Cholesterol Education Program Working Group on Lipoprotein Measurement. Clin Chem 1995;41:1421–6.10.1093/clinchem/41.10.1421Search in Google Scholar
25. Jensen AL, Kjelgaard-Hansen M. Method comparison in the clinical laboratory. Vet Clin Pathol 2006;35:276–86.10.1111/j.1939-165X.2006.tb00131.xSearch in Google Scholar PubMed
26. Simundic A. Statistical analysis in method comparison studies – Part one. AcutecaretestingOrg [Internet], 2016:1–7. Available from: https://acutecaretesting.org/en/articles/statistical-analysis-in-method-comparison-studies-part-one.Search in Google Scholar
27. Choi JS, Son BR, Kim YK. A clinical study on the need for glycerol-blanking in triglyceride measurement. Korean J Clin Pathol 1995;15:204–11.Search in Google Scholar
28. Backes JM, Dayspring T, Mieras T, Moriarty PM. Pseudohypertriglyceridemia: two cases of probable glycerol kinase deficiency. J Clin Lipidol 2012;6:469–73.10.1016/j.jacl.2012.02.001Search in Google Scholar PubMed
29. Hellerud C, Burlina A, Gabelli C, Ellis JR, Nyholm PG, Lindstedt S. Glycerol metabolism and the determination of triglycerides – clinical, biochemical and molecular findings in six subjects. Clin Chem Lab Med 2003;41:46–55.10.1515/CCLM.2003.009Search in Google Scholar PubMed
30. Klug EQ, Raal FJ, Marais AD, Smuts CM, Schamroth C, Jankelow D, et al. South African dyslipidaemia guideline consensus statement: 2018 update a joint statement from the South African Heart Association (SA Heart) and the Lipid and Atherosclerosis Society of Southern Africa (LASSA). S Afr Med J 2018;108:973–1000.10.7196/SAMJ.2018.v108i11.13383Search in Google Scholar
31. Catapano AL, Graham I, De Backer G, Wiklund O, Chapman MJ, Drexel H, et al. 2016 ESC/EAS guidelines for the management of dyslipidaemias. Eur Heart J 2016;37:2999–3058.10.1093/eurheartj/ehw272Search in Google Scholar PubMed
32. Ford ES, Li C, Zhao G, Pearson WS, Mokdad AH. Hypertriglyceridemia and its pharmacologic treatment among US adults. Arch Intern Med 2009;169:572–8.10.1001/archinternmed.2008.599Search in Google Scholar PubMed
©2020 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
- Frontmatter
- Editorial
- Quality controls for serology: an unfinished agenda
- A modern and pragmatic definition of Laboratory Medicine
- Reviews
- Blood biochemical characteristics of patients with coronavirus disease 2019 (COVID-19): a systemic review and meta-analysis
- ISO/TS 20914:2019 – a critical commentary
- Mini Review
- Reporting of D-dimer data in COVID-19: some confusion and potential for misinformation
- Opinion Paper
- Implementation of metrological traceability in laboratory medicine: where we are and what is missing
- IFCC Recommendation
- Recommendation for performance verification of patient-based real-time quality control
- Genetics and Molecular Diagnostics
- Comparison of BCR-ABL1 quantification in peripheral blood and bone marrow using an International Scale-standardized assay for assessment of deep molecular response in chronic myeloid leukemia
- General Clinical Chemistry and Laboratory Medicine
- Risk assessment of the total testing process based on quality indicators with the Sigma metrics
- Determination of hemolysis cut-offs for biochemical and immunochemical analytes according to their value
- A computer model for professional competence assessment according to ISO 15189
- Traceability validation of six enzyme measurements on the Abbott Alinity c analytical system
- Evaluating the need for free glycerol blanking for serum triglyceride measurements at Charlotte Maxeke Johannesburg Academic Hospital
- Challenges of LC-MS/MS ethyl glucuronide analysis in abstinence monitoring of liver transplant candidates
- Changes in the result of antinuclear antibody immunofluorescence assay on HEp-2 cells reflect disease activity status in systemic lupus erythematosus
- Reference Values and Biological Variations
- Long-term biological variation estimates of 13 hematological parameters in healthy Chinese subjects
- Age-specific reference values improve the diagnostic performance of AMH in polycystic ovary syndrome
- Establishment of reference intervals for immunoassay analytes of adult population in Saudi Arabia
- Hematology and Coagulation
- Total haemoglobin – a reference measuring system for improvement of standardisation
- Laboratory testing for activated protein C resistance: rivaroxaban induced interference and a comparative evaluation of andexanet alfa and DOAC Stop to neutralise interference
- Cancer Diagnostics
- Identification of a four-gene methylation biomarker panel in high-grade serous ovarian carcinoma
- Performance comparison of two next-generation sequencing panels to detect actionable mutations in cell-free DNA in cancer patients
- Diabetes
- Availability and analytical quality of hemoglobin A1c point-of-care testing in general practitioners’ offices are associated with better glycemic control in type 2 diabetes
- Infectious Diseases
- Validation of a chemiluminescent assay for specific SARS-CoV-2 antibody
- Dynamic profile and clinical implications of hematological parameters in hospitalized patients with coronavirus disease 2019
- Does a change in quality control results influence the sensitivity of an anti-HCV test?
- Letters to the Editor
- Variability between testing methods for SARS-CoV-2 nucleic acid detection 16 days post-discharge: a case report
- L-index, more than a screening tool for hypertriglyceridemia
- Neutralization of biotin interference: preliminary evaluation of the VeraTest Biotin™, VeraPrep Biotin™ and BioT-Filter®
- Counting and reporting band count is unreliable practice due to the high inter-observer variability
- Cigarette smoking prior to blood sampling acutely affects serum levels of the chronic obstructive pulmonary disease biomarker surfactant protein D
- How reliable is the detection of anti-mitochondrial antibodies on murine triple-tissue?
- Further advices on measuring lipoprotein(a) for reducing the residual cardiovascular risk on statin therapy
Articles in the same Issue
- Frontmatter
- Editorial
- Quality controls for serology: an unfinished agenda
- A modern and pragmatic definition of Laboratory Medicine
- Reviews
- Blood biochemical characteristics of patients with coronavirus disease 2019 (COVID-19): a systemic review and meta-analysis
- ISO/TS 20914:2019 – a critical commentary
- Mini Review
- Reporting of D-dimer data in COVID-19: some confusion and potential for misinformation
- Opinion Paper
- Implementation of metrological traceability in laboratory medicine: where we are and what is missing
- IFCC Recommendation
- Recommendation for performance verification of patient-based real-time quality control
- Genetics and Molecular Diagnostics
- Comparison of BCR-ABL1 quantification in peripheral blood and bone marrow using an International Scale-standardized assay for assessment of deep molecular response in chronic myeloid leukemia
- General Clinical Chemistry and Laboratory Medicine
- Risk assessment of the total testing process based on quality indicators with the Sigma metrics
- Determination of hemolysis cut-offs for biochemical and immunochemical analytes according to their value
- A computer model for professional competence assessment according to ISO 15189
- Traceability validation of six enzyme measurements on the Abbott Alinity c analytical system
- Evaluating the need for free glycerol blanking for serum triglyceride measurements at Charlotte Maxeke Johannesburg Academic Hospital
- Challenges of LC-MS/MS ethyl glucuronide analysis in abstinence monitoring of liver transplant candidates
- Changes in the result of antinuclear antibody immunofluorescence assay on HEp-2 cells reflect disease activity status in systemic lupus erythematosus
- Reference Values and Biological Variations
- Long-term biological variation estimates of 13 hematological parameters in healthy Chinese subjects
- Age-specific reference values improve the diagnostic performance of AMH in polycystic ovary syndrome
- Establishment of reference intervals for immunoassay analytes of adult population in Saudi Arabia
- Hematology and Coagulation
- Total haemoglobin – a reference measuring system for improvement of standardisation
- Laboratory testing for activated protein C resistance: rivaroxaban induced interference and a comparative evaluation of andexanet alfa and DOAC Stop to neutralise interference
- Cancer Diagnostics
- Identification of a four-gene methylation biomarker panel in high-grade serous ovarian carcinoma
- Performance comparison of two next-generation sequencing panels to detect actionable mutations in cell-free DNA in cancer patients
- Diabetes
- Availability and analytical quality of hemoglobin A1c point-of-care testing in general practitioners’ offices are associated with better glycemic control in type 2 diabetes
- Infectious Diseases
- Validation of a chemiluminescent assay for specific SARS-CoV-2 antibody
- Dynamic profile and clinical implications of hematological parameters in hospitalized patients with coronavirus disease 2019
- Does a change in quality control results influence the sensitivity of an anti-HCV test?
- Letters to the Editor
- Variability between testing methods for SARS-CoV-2 nucleic acid detection 16 days post-discharge: a case report
- L-index, more than a screening tool for hypertriglyceridemia
- Neutralization of biotin interference: preliminary evaluation of the VeraTest Biotin™, VeraPrep Biotin™ and BioT-Filter®
- Counting and reporting band count is unreliable practice due to the high inter-observer variability
- Cigarette smoking prior to blood sampling acutely affects serum levels of the chronic obstructive pulmonary disease biomarker surfactant protein D
- How reliable is the detection of anti-mitochondrial antibodies on murine triple-tissue?
- Further advices on measuring lipoprotein(a) for reducing the residual cardiovascular risk on statin therapy