Home Pre-analytical stability of haematinics, lactate dehydrogenase and phosphate in whole blood at room temperature up to 24 h, and refrigerated serum stability of lactate dehydrogenase, folate and vitamin B12 up to 72 h using the CRESS checklist
Article
Licensed
Unlicensed Requires Authentication

Pre-analytical stability of haematinics, lactate dehydrogenase and phosphate in whole blood at room temperature up to 24 h, and refrigerated serum stability of lactate dehydrogenase, folate and vitamin B12 up to 72 h using the CRESS checklist

  • Robert Williams , Monika Jankute , Rizwan Ifrahim , Jane Cordle and Sophie Hepburn ORCID logo EMAIL logo
Published/Copyright: December 2, 2024

Abstract

Objectives

There is a lack of analyte stability data in whole blood (WB). The aim of this study was to determine the allowable delay in WB processing for lactate dehydrogenase (LDH), folate, vitamin B12, iron and phosphate measurement. The stability of LDH, folate and vitamin B12 was also assessed in stored serum at clinically relevant time points.

Methods

Blood was taken from n=10 volunteers into Sarstedt serum gel tubes. We assessed stability in WB at room temperature up to 24 h, and stability in refrigerated serum up to 72 h. Mean percentage deviation at each time point was compared to criteria for minimum allowable bias.

Results

Results produced from one individual were removed due to discordant results, leaving n=9 specimens at each time point. Stability of folate and phosphate was variable in WB across 24 h, but was deemed to be clinically acceptable. LDH was unstable in WB, iron was stable for at least 12 h, and vitamin B12 and ferritin were acceptable for up to 24 h. Serum LDH, folate and vitamin B12 all demonstrated acceptable stability in refrigerated serum stored for up to 72 h.

Conclusions

Blood should ideally be centrifuged within 7 h for phosphate, LDH and folate, and 12 h for iron. However, for phosphate, folate and iron, there is likely to be little clinical impact if serum separation is delayed up to 24 h. Further research is needed to assess LDH stability in WB at 0–12 h, but changes are minimal at 12 h. All other analytes assessed showed acceptable stability across the time-points tested.


Corresponding author: Sophie Hepburn, Blood Sciences, NHS Highland, Raigmore Hospital, Old Perth Road, Inverness, Scotland, IV2 3UJ, UK, E-mail:

Acknowledgments

We would like to thank the volunteers that kindly donated blood samples for the study.

  1. Research ethics: Not applicable.

  2. Informed consent: Not applicable.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission. SH oversaw data analysis and interpretation. RW and SH wrote the first draft of the report, with MJ and RI contributing to the literature review. JC and RI contributed to the concept of the study and carried out laboratory sample analysis. RI performed venous blood sampling of volunteers. All authors approved the final version.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: None declared.

  7. Data availability: Not applicable.

References

1. Sikaris, KA. Enhancing the clinical value of medical laboratory testing. Clin Biochem Rev 2017;38:107–14.Search in Google Scholar

2. Lewis, T, Wood, M, Myers, M. Pathology GIRFT programme national speciality report. 2021. Available from: https://www.gettingitrightfirsttime.co.uk/wp-content/uploads/2022/03/Pathology-29Mar22i.pdf.Search in Google Scholar

3. Cornes, MP, Atherton, J, Pourmahram, G, Borthwick, H, Kyle, B, West, J, et al.. Monitoring and reporting of preanalytical errors in laboratory medicine: the UK situation. Ann Clin Biochem 2016;53:279–84. https://doi.org/10.1177/0004563215599561.Search in Google Scholar PubMed

4. Carraro, P, Plebani, M. Errors in a stat laboratory: types and frequencies 10 years later. Clin Chem 2007;53:1338–42. https://doi.org/10.1373/clinchem.2007.088344.Search in Google Scholar PubMed

5. Monneret, D, Godmer, A, Le Guen, R, Bravetti, C, Emeraud, C, Marteau, A, et al.. Stability of routine biochemical analytes in whole blood and plasma from lithium heparin gel tubes during 6-hr storage. J Clin Lab Anal 2016;30:602–9. https://doi.org/10.1002/jcla.21909.Search in Google Scholar PubMed PubMed Central

6. Plebani, M, Carraro, P. Mistakes in a stat laboratory: types and frequency. Clin Chem 1997;43:1348–51. https://doi.org/10.1093/clinchem/43.8.1348.Search in Google Scholar

7. Plebani, M. Quality indicators to detect pre-analytical errors in laboratory testing. Clin Biochem Rev 2012;33:85–8.Search in Google Scholar

8. Vermeersch, P, Frans, G, Meyer, A, Costelloe, S, Lippi, G, Simundic, A. How to meet ISO15189:2012 pre-analytical requirements in clinical laboratories? A consensus document by the EFLM WG-PRE. Clin Chem Lab Med 2021;59:1047–61. https://doi.org/10.1515/cclm-2020-1859.Search in Google Scholar PubMed

9. Lippi, G, Cornes, MP, Grankvist, K, Nybo, M, Simundic, A. EFLM WG-Preanalytical phase opinion paper: local validation of blood collection tubes in clinical laboratories. Clin Chem Lab Med 2016;54:755–60. https://doi.org/10.1515/cclm-2015-1274.Search in Google Scholar PubMed

10. ISO 15189:2022. Medical laboratories – requirements for quality and competence. Geneva: International Organization for Standardization (ISO), 2022.Search in Google Scholar

11. Vercruysse, K, Lambrecht, S, Oyaert, M. Total lab automation: sample stability of clinical chemistry parameters in an automated storage and retrieval module. Clin Chem Lab Med 2021;60:52–9. https://doi.org/10.1515/cclm-2021-0866.Search in Google Scholar PubMed

12. Fantz, CR, Greene, DN. Where are the preanalytical stability standards? J Appl Lab Med 2018;2:830–2. https://doi.org/10.1373/jalm.2018.026062.Search in Google Scholar PubMed

13. Tanner, M, Kent, N, Smith, B, Fletcher, S, Lewer, M. Stability of common biochemical analytes in serum gel tubes subjected to various storage temperature and time pre-centrifugation. Ann Clin Biochem 2008;45:375–9. https://doi.org/10.1258/acb.2007.007183.Search in Google Scholar PubMed

14. van Balveren, JA, Huijskens, MJ, Gemen, EF, Péquériaux, NC, Kusters, R. Effects of time and temperature on 48 routine chemistry, haematology and coagulation analytes in whole blood samples. Ann Clin Biochem 2017;54:448–62. https://doi.org/10.1177/0004563216665868.Search in Google Scholar PubMed

15. Henriksen, LO, Faber, NR, Moller, MF, Nexo, E, Hansen, AB. Stability of 35 biochemical and immunological routine tests after 10 hours storage and transport of human whole blood at 21 °C. Scand J Clin Lab Invest 2014;74:603–10. https://doi.org/10.3109/00365513.2014.928940.Search in Google Scholar PubMed PubMed Central

16. Gómez-Rioja, R, Espartosa, DM, Segovia, M, Ibarz, M, Llopis, MA, Bauça, JM, et al.. Laboratory sample stability. Is it possible to define a consensus stability function? An example of five blood magnitudes. Clin Chem Lab Med 2018;56:1806–18. https://doi.org/10.1515/cclm-2017-1189.Search in Google Scholar PubMed

17. Gómez-Rioja, R, Amaro, MS, Diaz-Garzón, J, Bauçà, JM, Espartosa, DM, Fernández-Calle, P, et al.. A protocol for testing the stability of biochemical analytes. Technical document. Clin Chem Lab Med 2019;57:1829–36. https://doi.org/10.1515/cclm-2019-0586.Search in Google Scholar PubMed

18. Cornes, M, Simundic, AM, Cadamuro, J, Costelloe, SJ, Baird, G, Kristensen, GBB, et al.. The CRESS checklist for reporting stability studies: on behalf of the European federation of clinical chemistry and laboratory medicine (EFLM) working group for the preanalytical phase (WG-PRE). Clin Chem Lab Med 2020;59:59–69. https://doi.org/10.1515/cclm-2020-0061.Search in Google Scholar PubMed

19. Gómez-Rioja, R, Von Meyer, A, Cornes, M, Costelloe, S, Vermeersch, P, Simundic, AM, et al.. Recommendation for the design of stability studies on clinical specimens. Clin Chem Lab Med 2023;61:1708–18. https://doi.org/10.1515/cclm-2023-0221.Search in Google Scholar PubMed

20. Hepburn, S, Ifrahim, R, Cordle, J. Stability of anti-thyroid stimulating hormone receptor antibody in whole blood and serum: caution required for reflective and batch testing. Ann Clin Biochem 2022;59:373–6. https://doi.org/10.1177/00045632221111342.Search in Google Scholar PubMed

21. Thirkettle, S, Blaszczyk, P, Evans, R, Wheatley, M, Abbas, M, Russell, J, et al.. Stability assessment of serum tumour markers: calcitonin, chromogranin A, thyroglobulin and anti-thyroglobulin antibodies. Ann Clin Biochem 2023;60:212–6. https://doi.org/10.1177/00045632231160371.Search in Google Scholar PubMed

22. Williams, RD, Duff, CJ, Yates, A. Determination of in vitro stability of routine haematinics tests using EFLM standards and the CRESS checklist. Ann Clin Biochem 2023;60:367–73. https://doi.org/10.1177/00045632231177247.Search in Google Scholar PubMed

23. Šupak-Smolčić, V, Franin, L, Horvat, M, Šonjić, P, Svatić, R, Antončić, D, et al.. Serum bicarbonate stability study at room temperature – influence of time to centrifugation and air exposure on bicarbonate measurement reported according to the CRESS checklist. Clin Chem Lab Med 2023;61:1046–53. https://doi.org/10.1515/cclm-2022-1182.Search in Google Scholar PubMed

24. Guder, WG, da Fonseca-Wollheim, F, Heil, W, Schmitt, Y, Töpfer, G, Wisser, H, et al.. Quality of diagnostic samples. Recommendations of the working group on preanalytical quality of the German society for clinical chemistry and laboratory medicine, 3rd ed. BD Diagnostics; 2010. Available from: https://scribd.com/document/515629757/Guder-et-al-Recommends-of-the-Working-Group-on-Prenalytical-Quality-of-the-German-Society-for-Clinical-chemistry-and-lab-Medecine-english [Accessed 26 Feb 2024].Search in Google Scholar

25. Roche IFU for LDH: Roche Diagnostics instructions for use (IFU). LDHI2, Lactate Dehydrogenase acc. to IFCC ver.2. Published Aug 2019, version 7.0 English. Document ID: 0005169330190c701V7.0.Search in Google Scholar

26. Oddoze, C, Lombard, E, Portugal, H. Stability study of 81 analytes in human whole blood, in serum and in plasma. Clin Biochem 2012;45:464–9.10.1016/j.clinbiochem.2012.01.012Search in Google Scholar PubMed

27. Illana, FJ, García-Osuna, Á, Sospedra, M, Ferrer, R, Martínez-Brú, C, Guiñón, L. Quality assurance of add-on testing in plasma samples: stability limit for 29 biochemical analytes. Biochem Med 2024;34:020704.10.11613/BM.2024.020704Search in Google Scholar PubMed PubMed Central

28. Ono, T, Kitaguchi, K, Takehara, M, Shiiba, M, Hayami, K. Serum-constituents analyses: effect of duration and temperature of storage of clotted blood. Clin Chem 1981;27:35–8.10.1093/clinchem/27.1.35Search in Google Scholar

29. Boyanton, BLJr, 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

30. Zhang, DJ, Elswick, RK, Miller, WG, Bailey, JL. Effect of serum-clot contact time on clinical chemistry laboratory results. Clin Chem 1998;44:1325–33.10.1093/clinchem/44.6.1325Search in Google Scholar

31. Fazili, Z, Sternberg, MR, Paladugula, N, Whitehead, RDJr, Chen, H, Pfeiffer, CM. The loss of 5-methyltetrahydrofolate in human serum under suboptimal preanalytical conditions can only partially be recovered by an oxidation product. J Nutr 2014;144:1873–9.10.3945/jn.114.198358Search in Google Scholar PubMed PubMed Central

32. Cuhadar, S, Atay, A, Koseoglu, M, Dirican, A, Hur, A. Stability studies of common biochemical analytes in serum separator tubes with or without gel barrier subjected to various storage conditions. Biochem Med 2012;22:202–14.10.11613/BM.2012.023Search in Google Scholar PubMed PubMed Central

33. Heins, M, Heil, W, Withold, W. Storage of serum or whole blood samples? Effects of time and temperature on 22 serum analytes. Eur J Clin Chem Clin Biochem 1995;33:231–8.10.1515/cclm.1995.33.4.231Search in Google Scholar PubMed

34. Hedayati, M, Razavi, SA, Boroomand, S, Kia, SK. The impact of pre-analytical variations on biochemical analytes stability: a systematic review. J Clin Lab Anal 2020;34:e23551.10.1002/jcla.23551Search in Google Scholar PubMed PubMed Central

35. Drammeh, BS, Schleicher, RL, Pfeiffer, CM, Jain, RB, Zhang, M, Nguyen, PH. Effects of delayed sample processing and freezing on serum concentrations of selected nutritional indicators. Clin Chem 2008;54:1883–91.10.1373/clinchem.2008.108761Search in Google Scholar PubMed

36. Zwart, SR, Wolf, M, Rogers, A, Rodgers, S, Gillman, PL, Hitchcox, K, et al.. Stability of analytes related to clinical chemistry and bone metabolism in blood specimens after delayed processing. Clin Biochem 2009;42:907–10.10.1016/j.clinbiochem.2009.02.010Search in Google Scholar PubMed

37. van Eijsden, M, van der wal, MF, Hornstra, G, Bonsel, GJ. Can whole-blood samples be stored over 24 hours without compromising stability of C-reactive protein, retinol, ferritin, folic acid, and fatty acids in epidemiologic research? Clin Chem 2005;51:230–2.10.1373/clinchem.2004.042234Search in Google Scholar PubMed

38. Cuerq, C, Peretti, N, Chikh, K, Mialon, A, Guillaumont, M, Drai, J, et al.. Overview of the in vitro stability of commonly measured vitamins and carotenoids in whole blood. Ann Clin Biochem 2015;52:259–69.10.1177/0004563214542471Search in Google Scholar PubMed

39. Jansen, EHJM, Beekhof, PK, Cremers, JWJM, Schenk, E. Long-term (in)stability of folate and vitamin B12 in human serum. Clin Chem Lab Med 2012;50:1761–3.10.1515/cclm-2012-0108Search in Google Scholar PubMed

40. Roche Diagnostics instructions for use (IFU). Elecsys folate III. Folate serum/plasma application. Published Dec 2019, version 5.0 English. Document ID: 07027290500v5.0.Search in Google Scholar

41. Roche Diagnostics instructions for use (IFU). Elecsys Vitamin B12 II. Published Nov 2021, version 5.0 English. Document ID: ms_07212771190V5.0.Search in Google Scholar

42. Abraham, RA, Agrawal, PK, Acharya, R, Sarna, A, Ramesh, S, Johnston, R, et al.. Effect of temperature and time delay in centrifugation on stability of select biomarkers of nutrition and non-communicable diseases in blood samples. Biochem Med 2019;29:020708. https://doi.org/10.11613/bm.2019.020708.Search in Google Scholar PubMed PubMed Central

43. Roche Diagnostics instructions for use (IFU). IRON2. Iron Gen.2. Published Dec 2019, version 11.0 English. Document ID: 0105169291190c701V11.0.Search in Google Scholar

44. Stahl, M, Brandslund, I. Controlled storage conditions prolong stability of biochemical components in whole blood. Clin Chem Lab Med 2005;43:210–5.10.1515/CCLM.2005.036Search in Google Scholar PubMed

45. Simundic, AM, Bölenius, K, Cadamuro, J, Church, S, Cornes, MP, van Dongen-Lases, EC, et al.. Joint EFLM-COLABIOCLI recommendation for venous blood sampling. Clin Chem Lab Med 2018;56:2015–38.10.1515/cclm-2018-0602Search in Google Scholar PubMed

46. Sarstedt blood collection systems. Available from: 20_563_0000_200_be_systeme_1117.pdf (sarstedt.com) [Accessed 11 Jan 2024].Search in Google Scholar

47. European federation of clinical chemistry and laboratory medicine (EFLM) Biological Variation Database, 2024 [Online]. Available from: https://biologicalvariation.eu/ [Accessed 11 Jan 2024].Search in Google Scholar

48. Park, H, Park, J, Park, K, Chang, P. Effects of freezing rate on structural changes in L-lactate dehydrogenase during the freezing process. Sci Rep 2021;11:13643.10.1038/s41598-021-93127-6Search in Google Scholar PubMed PubMed Central

49. Clement, NF, Kendall, BS. Effect of light on vitamin B12 and folate. Lab Med 2009;40:657–9.10.1309/LMWD82YZ7QZPSNQPSearch in Google Scholar

50. Leino, A, Koivula, MK. Stability of chemical and immunochemical analytes in uncentrifuged plasma samples. Ann Clin Biochem 2009;46:159–61.10.1258/acb.2008.008212Search in Google Scholar PubMed

Received: 2024-09-17
Accepted: 2024-11-02
Published Online: 2024-12-02
Published in Print: 2025-03-26

© 2024 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Beyond test results: the strategic importance of metadata for the integration of AI in laboratory medicine
  4. Reviews
  5. Reference, calibration and referral laboratories – a look at current European provisions and beyond
  6. How has the external quality assessment/proficiency testing of semen analysis been developed in the past 34 years: a review
  7. Opinion Papers
  8. Data flow in clinical laboratories: could metadata and peridata bridge the gap to new AI-based applications?
  9. A comprehensive survey of artificial intelligence adoption in European laboratory medicine: current utilization and prospects
  10. Guidelines and Recommendations
  11. Guidelines for the correct use of the nomenclature of biochemical indices of bone status: a position statement of the Joint IOF Working Group and IFCC Committee on Bone Metabolism
  12. Candidate Reference Measurement Procedures and Materials
  13. Absolute quantitation of human serum cystatin C: candidate reference method by 15N-labeled recombinant protein isotope dilution UPLC-MS/MS
  14. General Clinical Chemistry and Laboratory Medicine
  15. Performance evaluation of the introduction of full sample traceability system within the specimen collection process
  16. Pre-analytical stability of haematinics, lactate dehydrogenase and phosphate in whole blood at room temperature up to 24 h, and refrigerated serum stability of lactate dehydrogenase, folate and vitamin B12 up to 72 h using the CRESS checklist
  17. Comparison of capillary finger stick and venous blood sampling for 34 routine chemistry analytes: potential for in hospital and remote blood sampling
  18. Performance evaluation of enzymatic total bile acid (TBA) routine assays: systematic comparison of five fifth-generation TBA cycling methods and their individual bile acid recovery from HPLC-MS/MS reference
  19. Clinical performance of a new lateral flow immunoassay for xylazine detection
  20. Evaluation of revised UK-NEQAS CSF-xanthochromia method for subarachnoid hemorrhage: outcome data provide evidence for clinical value
  21. Strategies to verify equimolar peptide release in mass spectrometry-based protein quantification exemplified for apolipoprotein(a)
  22. Evaluation of the clinical performance of anti-mutated citrullinated vimentin antibody and 14-3-3 eta testing in rheumatoid arthritis
  23. Diagnostic performance of specific biomarkers for interstitial lung disease: a single center study
  24. Reference Values and Biological Variations
  25. Neonatal reference intervals for serum steroid hormone concentrations measured by LC-MS/MS
  26. Paediatric reference intervals for haematology parameters analysed on Sysmex XN-9000: a comparison of methods in the framework of indirect sampling
  27. Cardiovascular Diseases
  28. Analytical characteristics and performance of a new hs-cTnI method: a multicenter-study
  29. Diabetes
  30. Use of labile HbA1c as a screening tool to minimize clinical misinterpration of HbA1c
  31. Letters to the Editor
  32. Current trends and future projections in the clinical laboratory test market: implications for resource management and strategic planning
  33. Particulate matter in water: an overlooked source of preanalytical error producing erroneous chemistry test results
  34. “Activation” of macro-AST by pyridoxal-5-phosphate in the assay for aspartate aminotransferase
  35. The correlation of albumin with total protein concentrations in cerebrospinal fluid across three automated analysers – relevance to the diagnosis of subarachnoid haemorrhage in clinical chemistry practice
  36. Adult reference intervals for serum thyroid‐stimulating hormone using Abbott Alinity i measuring system
  37. Cell population data in venous thrombo-embolism and erysipelas: a potential diagnostic tool?
  38. Diagnostic performances and cut-off verification of blood pTau 217 on the Lumipulse platform for amyloid deposition in Alzheimer’s disease
  39. The first case of Teclistamab interference with serum electrophoresis and immunofixation
  40. Congress Abstracts
  41. Annual meeting of the Royal Belgian Society of Laboratory Medicine (RBSLM): “A Neurological Journey: Brain Teasers for Laboratory Medicine”
Downloaded on 12.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/cclm-2024-1098/html
Scroll to top button