Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
-
Nick Neuwinger
Abstract
Background
Lactate dehydrogenase (LD) activity is routinely monitored for therapeutic risk stratification of malignant diseases, but is also prone to preanalytical influences.
Methods
We systematically analyzed the impact of defined preanalytical conditions on the hemolysis-susceptible parameters LD, potassium (K) and hemolysis index in vacuum blood collection tubes (serum [SE], heparin plasma [HP]). Blood was collected by venipuncture from healthy volunteers. Tubes were either filled or underfilled to approximately 50%, then processed directly or stored at room temperature for 4 h. Potassium (K), sodium (Na), chloride (Cl), LD, creatine kinase (CK), total cholesterol, and indices for hemolysis, icterus, and lipemia were analyzed. Filling velocity was determined in a subset of tubes. Findings in healthy volunteers were reconfirmed in an in-patient cohort (n = 74,751) that was analyzed for plasma yield and LD data distribution.
Results
LD activity was higher in HP compared to SE. Underfilling led to higher LD values (SE: +21.6%; HP: +28.3%), K (SE: +4.2%; HP: +5.3%), and hemolysis index (SE: +260.8%; HP: +210.0%), while other analytes remained largely unchanged. Filling velocity of tubes was approximately 3-fold higher in the first half compared to the second half in both HP and SE collection tubes. Importantly, plasma yield also inversely correlated with LD in routine patients. By calculating reference limits, the lowest plasma yield quartile of the patient cohort displayed LD values clearly exceeding current reference recommendations.
Conclusions
Underfilling of tubes leads to a higher proportion of blood aspirated with high velocity and relevant elevations in LD. This finding should be considered in cases of clinically implausible elevated LD activities.
Acknowledgments
We thank the technical staff of Labor Berlin – Charité Vivantes GmbH for expertise measurement of clinical chemistry analytes. We thank Dr. Karin Hensel-Wiegel for critical intellectual input.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) 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.
Authors’ disclosures or potential conflicts of interest: Upon manuscript submission, all authors completed the author disclosure form. The authors declare: nothing to disclose.
References
1. Burgner 2nd JW, Ray Jr WJ. On the origin of the lactate dehydrogenase induced rate effect. Biochemistry 1984;23:3636–48.10.1021/bi00311a010Search in Google Scholar PubMed
2. Carobene A, Roraas T, Solvik UO, Sylte MS, Sandberg S, Guerra E, et al. Biological variation estimates obtained from91 healthy study participants for 9 enzymes in serum. Clin Chem 2017;63:1141–50.10.1373/clinchem.2016.269811Search in Google Scholar PubMed
3. Zierk J, Arzideh F, Rechenauer T, Haeckel R, Rascher W, Metzler M, et al. Age- and sex-specific dynamics in 22 hematologic and biochemical analytes from birth to adolescence. Clin Chem 2015;61:964–73.10.1373/clinchem.2015.239731Search in Google Scholar PubMed
4. Petrelli F, Cabiddu M, Coinu A, Borgonovo K, Ghilardi M, Lonati V, et al. Prognostic role of lactate dehydrogenase in solid tumors: a systematic review and meta-analysis of 76 studies. Acta Oncol 2015;54:961–70.10.3109/0284186X.2015.1043026Search in Google Scholar PubMed
5. Nieder C, Marienhagen K, Dalhaug A, Norum J. Towards improved prognostic scores predicting survival in patients with brain metastases: a pilot study of serum lactate dehydrogenase levels. ScientificWorldJ 2012;2012:609323.10.1100/2012/609323Search in Google Scholar PubMed PubMed Central
6. Lu R, Jiang M, Chen Z, Xu X, Hu H, Zhao X, et al. Lactate dehydrogenase 5 expression in non-Hodgkin lymphoma is associated with the induced hypoxia regulated protein and poor prognosis. PLoS One 2013;8:e74853.10.1371/journal.pone.0074853Search in Google Scholar PubMed PubMed Central
7. Jin Y, Ye X, Shao L, Lin BC, He CX, Zhang BB, et al. Serum lactic dehydrogenase strongly predicts survival in metastatic nasopharyngeal carcinoma treated with palliative chemotherapy. Eur J Cancer 2013;49:1619–26.10.1016/j.ejca.2012.11.032Search in Google Scholar PubMed
8. Hauschild A, Larkin J, Ribas A, Dreno B, Flaherty KT, Ascierto PA, et al. Modeled prognostic subgroups for survival and treatment outcomes in BRAF V600-mutated metastatic melanoma: pooled analysis of 4 randomized clinical trials. JAMA Oncol 2018;4:1382–8.10.1001/jamaoncol.2018.2668Search in Google Scholar PubMed PubMed Central
9. Kim HS, Lee HE, Yang HK, Kim WH. High lactate dehydrogenase 5 expression correlates with high tumoral and stromal vascular endothelial growth factor expression in gastric cancer. Pathobiology 2014;81:78–85.10.1159/000357017Search in Google Scholar PubMed
10. Xu HN, Kadlececk S, Profka H, Glickson JD, Rizi R, Li LZ. Is higher lactate an indicator of tumor metastatic risk? A pilot MRS study using hyperpolarized (13)C-pyruvate. Acad Radiol 2014;21:223–31.10.1016/j.acra.2013.11.014Search in Google Scholar PubMed PubMed Central
11. Pantel K, Hille C, Scher HI. Circulating tumor cells in prostate cancer: from discovery to clinical utility. Clin Chem 2019;65:87–99.10.1373/clinchem.2018.287102Search in Google Scholar PubMed
12. Solal-Celigny P, Roy P, Colombat P, White J, Armitage JO, Arranz-Saez R, et al. Follicular lymphoma international prognostic index. Blood 2004;104:1258–65.10.1182/blood-2003-12-4434Search in Google Scholar PubMed
13. Drent M, Cobben NA, Henderson RF, Wouters EF, van Dieijen-Visser M. Usefulness of lactate dehydrogenase and its isoenzymes as indicators of lung damage or inflammation. Eur Respir J 1996;9:1736–42.10.1183/09031936.96.09081736Search in Google Scholar PubMed
14. Haselmann V, Gebhardt C, Brechtel I, Duda A, Czerwinski C, Sucker A, et al. Liquid profiling of circulating tumor DNA in plasma of melanoma patients for companion diagnostics and monitoring of BRAF inhibitor therapy. Clin Chem 2018;64:830–42.10.1373/clinchem.2017.281543Search in Google Scholar PubMed
15. Farnsworth CW, Webber DM, Krekeler JA, Budelier MM, Bartlett NL, Gronowski AM. Parameters for validating a hospital pneumatic tube system. Clin Chem 2019;65:694–702.10.1373/clinchem.2018.301408Search in Google Scholar PubMed
16. Carraro P, Plebani M. Errors in a stat laboratory: types and frequencies 10 years later. Clin Chem 2007;53:1338–42.10.1373/clinchem.2007.088344Search in Google Scholar PubMed
17. Boyanton Jr BL, Blick KE. Stability studies of twenty-four analytes in human plasma and serum. Clin Chem 2002;48:2242–7.10.1093/clinchem/48.12.2242Search in Google Scholar
18. Thomas L, Müller M, Schumann G, Weidemann G, Klein G, Lunau S, et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. LaboratoriumsMedizin 2005;29:301.10.1515/JLM.2005.041Search in Google Scholar
19. Schumann G, Bonora R, Ceriotti F, Clerc-Renaud P, Ferrero CA, Ferard G, et al. IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37 degrees C. Part 3. Reference procedure for the measurement of catalytic concentration of lactate dehydrogenase. Clin Chem Lab Med 2002;40:643–8.10.1515/CCLM.2002.111Search in Google Scholar PubMed
20. Lorentz K, Röhle G. Einführung der neuen standardmethoden 1994 zur bestimmung der katalytischen enzymkonzentration bei 37 °C. [Introduction of the new standard methods 1994 for the determination of the catalytic enzyme concentration at 37 °C.] Klin Chem Mitt 1995;26:290–3.Search in Google Scholar
21. Arzideh F, Wosniok W, Gurr E, Hinsch W, Schumann G, Weinstock N, et al. A plea for intra-laboratory reference limits. Part 2. A bimodal retrospective concept for determining reference limits from intra-laboratory databases demonstrated by catalytic activity concentrations of enzymes. Clin Chem Lab Med 2007;45:1043–57.10.1515/CCLM.2007.250Search in Google Scholar PubMed
22. Wollowitz A, Bijur PE, Esses D, John Gallagher E. Use of butterfly needles to draw blood is independently associated with marked reduction in hemolysis compared to intravenous catheter. Acad Emerg Med 2013;20:1151–5.10.1111/acem.12245Search in Google Scholar
23. Huyghe T, Buntinx F, Bruyninckx R, Besard V, Vunckx J, Church S, et al. Studies on the use of BD Vacutainer® SST II and RST in general practice: investigation of artefactual hyperkalaemia. Ann Clin Biochem 2014;51:30–7.10.1177/0004563213488758Search in Google Scholar
24. Hamilton PK. Tube under-filling and artefactual hyperkalaemia. Ann Clin Biochem 2014;51:717.10.1177/0004563214532239Search in Google Scholar
25. Golemanov K, Tcholakova S, Denkov ND, Ananthapadmanabhan KP, Lips A. Breakup of bubbles and drops in steadily sheared foams and concentrated emulsions. Phys Rev E Stat Nonlin Soft Matter Phys 2008;78:051405.10.1103/PhysRevE.78.051405Search in Google Scholar
26. Landi EP, Roveri EG, Ozelo MC, Annichino-Bizzacchi JM, Origa AF, de Carvalho Reis AR, et al. Effects of high platelet concentration in collecting and freezing dry platelets concentrates. Transfus Apher Sci 2004;30:205–12.10.1016/j.transci.2004.03.003Search in Google Scholar
27. Ryan JB, Stuart LA, Southby SJ, Than MP, Mackay R, Florkowski CM, et al. Comparison of BD Vacutainer® rapid serum tube and plasma for haemolysis markers in the emergency department. Ann Clin Biochem 2015;52:293–6.10.1177/0004563214533317Search in Google Scholar
28. Dimeski G, Masci PP, Trabi M, Lavin MF, de Jersey J. Evaluation of the Becton-Dickinson rapid serum tube: does it provide a suitable alternative to lithium heparin plasma tubes? Clin Chem Lab Med 2010;48:651–7.10.1515/CCLM.2010.141Search in Google Scholar
29. Hafkenscheid JC, Hectors MP. Serum versus heparinized plasma for alanine aminotransferase and aspartate aminotransferase of normal individuals. Clin Chim Acta 1977;78:23–7.10.1016/0009-8981(77)90334-5Search in Google Scholar
30. Dupuy AM, Cristol JP, Vincent B, Bargnoux AS, Mendes M, Philibert P, et al. Stability of routine biochemical analytes in whole blood and plasma/serum: focus on potassium stability from lithium heparin. Clin Chem Lab Med 2018;56:413–21.10.1515/cclm-2017-0292Search in Google Scholar PubMed
31. Wörmann D, Driessen C, Einsele H, Goldschmidt H, Gunsilius E, Kortüm M, et al. Leitlinie: Multiples Myelom ICD10: C90.0. onkopediacom 2018.Search in Google Scholar
32. Palumbo A, Avet-Loiseau H, Oliva S, Lokhorst HM, Goldschmidt H, Rosinol L, et al. Revised international staging system for multiple myeloma: a report from international myeloma working group. J Clin Oncol 2015;33:2863–9.10.1200/JCO.2015.61.2267Search in Google Scholar PubMed PubMed Central
33. Huijgen HJ, Sanders GT, Koster RW, Vreeken J, Bossuyt PM. The clinical value of lactate dehydrogenase in serum: a quantitative review. Eur J Clin Chem Clin Biochem 1997;35:569–79.Search in Google Scholar
34. Levato F, Martinello R, Campobasso C, Porto S. LDH and LDH isoenzymes in ovarian dysgerminoma. Eur J Gynaecol Oncol 1995;16:212–5.Search in Google Scholar
©2019 Walter de Gruyter GmbH, Berlin/Boston
Articles in the same Issue
- Frontmatter
- Editorial
- Towards a personalized assessment of vitamin D status
- Reviews
- Circulating tumor DNA and their added value in molecular oncology
- Telomere length determinants in childhood
- Opinion Papers
- Serum or plasma? An old question looking for new answers
- Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
- General Clinical Chemistry and Laboratory Medicine
- Simultaneous measurement of 25(OH)-vitamin D and 24,25(OH)2-vitamin D to define cut-offs for CYP24A1 mutation and vitamin D deficiency in a population of 1200 young subjects
- How well do Croatian laboratories adhere to national recommendations for laboratory diagnostics of chronic kidney disease (CKD)?
- Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
- Calcium state estimation by total calcium: the evidence to end the never-ending story
- The use of faecal immunochemical testing in the decision-making process for the endoscopic investigation of iron deficiency anaemia
- Measurement uncertainty of β-lactam antibiotics results: estimation and clinical impact on therapeutic drug monitoring
- Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
- Plasma neurofilament light chain is associated with mortality after spontaneous intracerebral hemorrhage
- Comparison of the diagnostic performance of two automated urine sediment analyzers with manual phase-contrast microscopy
- Development and validation of LC-MS/MS methods to measure tobramycin and lincomycin in plasma, microdialysis fluid and urine: application to a pilot pharmacokinetic research study
- Reference Values and Biological Variations
- Cord blood S100B: reference ranges and interest for early identification of newborns with brain injury
- Hematology and Coagulation
- Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need
- Cancer Diagnostics
- Next-generation sequencing for tumor mutation quantification using liquid biopsies
- Cardiovascular Diseases
- Evolution of the slopes of ST2 and galectin-3 during marathon and ultratrail running compared to a control group
- Letters to the Editor
- Biological variation of two serum markers for preeclampsia prediction
- Daily monitoring of a control material with a concentration near the limit of detection improves the measurement accuracy of highly sensitive troponin assays
- Are patients adequately informed about procedures for 24-h urine collection?
- Interferences in free thyroxine concentration using the Roche analytical platform: improvement of the third generation?
- Procedures for the diagnosis of macro-follicle stimulating hormone (FSH) in a patient with high serum FSH concentrations
- False-positive result of immunochromatographic (IC) strip test for the diagnosis of α-thalassemia in samples with autoantibodies
- Sample dilution for soluble interleukin-2 receptor α measurement: comparison of two different matrices
- The growing problem of predatory publishing: a case report
- Cerebrospinal fluid lactate levels according to the site of puncture
Articles in the same Issue
- Frontmatter
- Editorial
- Towards a personalized assessment of vitamin D status
- Reviews
- Circulating tumor DNA and their added value in molecular oncology
- Telomere length determinants in childhood
- Opinion Papers
- Serum or plasma? An old question looking for new answers
- Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
- General Clinical Chemistry and Laboratory Medicine
- Simultaneous measurement of 25(OH)-vitamin D and 24,25(OH)2-vitamin D to define cut-offs for CYP24A1 mutation and vitamin D deficiency in a population of 1200 young subjects
- How well do Croatian laboratories adhere to national recommendations for laboratory diagnostics of chronic kidney disease (CKD)?
- Underfilling of vacuum blood collection tubes leads to increased lactate dehydrogenase activity in serum and heparin plasma samples
- Calcium state estimation by total calcium: the evidence to end the never-ending story
- The use of faecal immunochemical testing in the decision-making process for the endoscopic investigation of iron deficiency anaemia
- Measurement uncertainty of β-lactam antibiotics results: estimation and clinical impact on therapeutic drug monitoring
- Practical approach to method verification in plasma and validation in cerebrospinal fluid under accreditation using a flexible scope in molecular virology: setting up the HIV, HBV and HCV Aptima™ Quant Dx assays
- Plasma neurofilament light chain is associated with mortality after spontaneous intracerebral hemorrhage
- Comparison of the diagnostic performance of two automated urine sediment analyzers with manual phase-contrast microscopy
- Development and validation of LC-MS/MS methods to measure tobramycin and lincomycin in plasma, microdialysis fluid and urine: application to a pilot pharmacokinetic research study
- Reference Values and Biological Variations
- Cord blood S100B: reference ranges and interest for early identification of newborns with brain injury
- Hematology and Coagulation
- Validation and standardization of the ETP-based activated protein C resistance test for the clinical investigation of steroid contraceptives in women: an unmet clinical and regulatory need
- Cancer Diagnostics
- Next-generation sequencing for tumor mutation quantification using liquid biopsies
- Cardiovascular Diseases
- Evolution of the slopes of ST2 and galectin-3 during marathon and ultratrail running compared to a control group
- Letters to the Editor
- Biological variation of two serum markers for preeclampsia prediction
- Daily monitoring of a control material with a concentration near the limit of detection improves the measurement accuracy of highly sensitive troponin assays
- Are patients adequately informed about procedures for 24-h urine collection?
- Interferences in free thyroxine concentration using the Roche analytical platform: improvement of the third generation?
- Procedures for the diagnosis of macro-follicle stimulating hormone (FSH) in a patient with high serum FSH concentrations
- False-positive result of immunochromatographic (IC) strip test for the diagnosis of α-thalassemia in samples with autoantibodies
- Sample dilution for soluble interleukin-2 receptor α measurement: comparison of two different matrices
- The growing problem of predatory publishing: a case report
- Cerebrospinal fluid lactate levels according to the site of puncture
