Standard −20 °C freezer storage protocols may cause substantial plasma renin cryoactivation

Paul Bonnitcha; Mark Rigdwell; Peter Ward; Douglas Chesher

doi:10.1515/cclm-2022-1190

Published by De Gruyter February 21, 2023

Standard −20 °C freezer storage protocols may cause substantial plasma renin cryoactivation

Paul Bonnitcha , Mark Rigdwell , Peter Ward and Douglas Chesher

From the journal Clinical Chemistry and Laboratory Medicine (CCLM)

https://doi.org/10.1515/cclm-2022-1190

Showing a limited preview of this publication:

Abstract

Objectives

To assess the appropriate preanalytical process for storage of plasma for renin concentration analysis. This study was initiated due to the wide variation in preanalytical handling of samples observed within our network, particularly with respect to freezing for longer term storage.

Methods

Pooled plasma from patient samples was analysed immediately post separation for renin concentration (n=30, concentration 4.0–204 mIU/L). Aliquots from these samples were frozen in a −20 °C freezer and then analysed, with the renin concentration compared to the respective baseline concentration. Comparisons were also made to: aliquots snap frozen using a dry ice/acetone bath, aliquots stored at room temperature, and aliquots stored at 4 °C. Subsequent experiments investigated the potential sources of cryoactivation observed in these initial studies.

Results

Substantial and highly variable cryoactivation was observed in samples frozen using a −20 °C freezer, with renin concentration increasing over 300% from baseline in some samples (median 21.3%). This cryoactivation could be prevented by snap freezing samples. Subsequent experiments determined that long term storage in a −20 °C freezer could prevent cryoactivation provided samples were initially frozen rapidly in a −70 °C freezer. Rapid defrosting of samples was not required to prevent cryoactivation.

Conclusions

Standard −20 °C freezers may not be appropriate for freezing samples for renin analysis. Laboratories should consider snap freezing their samples using a −70 °C freezer or similar to avoid cryoactivation of renin.

Keywords: cryoactivation; preanalytical; renin; stability

Corresponding author: Paul Bonnitcha, Chemical Pathology Department, NSW Health Pathology, Royal North Shore Hospital, Level 5, Acute Service Building, St Leonards, NSW 2065, Australia; and School of Medical Sciences, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2006, Australia, Phone: +61 2 9926 7111, E-mail: paul.bonnitcha@health.nsw.gov.au

Acknowledgments

All authors would like to thank the Australasian Association for Clinical Biochemistry and Laboratory Medicine (AACB) ARR Harmonisation Group for helpful discussions.

Research funding: None declared.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Authors state no conflict of interest.
Informed consent: Not applicable.
Ethical approval: Research involving human subjects complied with all relevant national regulations, institutional policies and is in accordance with the tenets of the Helsinki Declaration (as revised in 2013), and has been approved by the authors’ Institutional Review Board (NSWHP Human Research Ethics Committee).

References

1. Yang, T, Xu, C. Physiology and pathophysiology of the intrarenal renin-angiotensin system: an update. J Am Soc Nephrol 2017;28:1040–9. https://doi.org/10.1681/asn.2016070734.Search in Google Scholar PubMed PubMed Central

2. Martini, AG, Danser, AHJ. Juxtaglomerular cell phenotypic plasticity. High Blood Pres Cardiovasc Prev 2017;24:231–42. https://doi.org/10.1007/s40292-017-0212-5.Search in Google Scholar PubMed PubMed Central

3. Danser, AHJ. Prorenin: back into the arena. Hypertension 2006;47:824–6. https://doi.org/10.1161/01.hyp.0000215952.91959.4a.Search in Google Scholar PubMed

4. Funder, JW, Carey, RM, Mantero, F, Murad, MH, Reincke, M, Shibata, H, et al.. The management of primary aldosteronism: case detection, diagnosis, and treatment: an endocrine society clinical practice guideline. J Clin Endocrinol Metab 2016;101:1889–916. https://doi.org/10.1210/jc.2015-4061.Search in Google Scholar PubMed

5. Libianto, R, Fuller, PJ, Young, MJ, Yang, J. Primary aldosteronism is a public health issue: challenges and opportunities. J Hum Hypertens 2020;34:478–86. https://doi.org/10.1038/s41371-020-0336-2.Search in Google Scholar PubMed

6. Stowasser, M, Ahmed, AH, Pimenta, E, Taylor, PJ, Gordon, RD. Factors affecting the aldosterone/renin ratio. Horm Metab Res 2012;44:170–6. https://doi.org/10.1055/s-0031-1295460.Search in Google Scholar PubMed

7. Lonati, C, Bassani, N, Gritti, A, Biganzoli, E, Morganti, A. Measurement of plasma renin concentration instead of plasma renin activity decreases the positive aldosterone-to-renin ratio tests in treated patients with essential hypertension. J Hypertens 2014;32:627–34. https://doi.org/10.1097/hjh.0000000000000076.Search in Google Scholar

8. Ahmed, AH, Gordon, RD, Taylor, PJ, Ward, G, Pimenta, E, Stowasser, M. Are women more at risk of false-positive primary aldosteronism screening and unnecessary suppression testing than men? J Clin Endocrinol Metab 2011;96:E340. https://doi.org/10.1210/jc.2010-1355.Search in Google Scholar PubMed

9. Sealey, JE, Blumenfeld, J, Laragh, JH. Prorenin cryoactivation as a possible cause of normal renin levels in patients with primary aldosteronism. J Hypertens 2005;23:459–60. https://doi.org/10.1097/00004872-200502000-00030.Search in Google Scholar PubMed

10. Diasorin. LIAISON® direct renin, instructions for use, REF 310470. Saluggia, Italy: Diasorin; 2020.Search in Google Scholar

11. Rutledge, AC, Johnston, A, Bailey, D, Booth, RA, Edmond, P, Leung, V, et al.. Survey of renin and aldosterone testing practices by Ontario laboratories – providing insight into best practices. Pract Lab Med 2021;25:e00229. https://doi.org/10.1016/j.plabm.2021.e00229.Search in Google Scholar PubMed PubMed Central

12. Locsei, Z, Racz, K, Patocs, A, Kovacs, GL, Toldy, E. Influence of sampling and storage conditions on plasma renin activity and plasma renin concentration. Clin Chim Acta 2009;402:203–5. https://doi.org/10.1016/j.cca.2009.01.013.Search in Google Scholar PubMed

13. Alfred Health. Pathology handbook – renin: alfred health; 2022. [Online]. Available from: https://pathology.alfred.org.au/handbook/index?testid=481 [Accessed 26 Oct 2022].Search in Google Scholar

14. NSW Health Pathology. Pathology test catalogue – renin; 2022. [Online]. Available from: https://catalogue.pathology.health.nsw.gov.au/search?searchValue=renin&searchType=startsWith&page=1&pageSize=10 [Accessed 26 Oct 2022].Search in Google Scholar

15. The Royal Children’s Hospital Melbourne. Royal children hospital – specimen collection – renin direct; 2022. [Online]. Available from: https://www.rch.org.au/specimen-collection/Renin_Direct/ [Accessed 26 Oct 2022].Search in Google Scholar

16. Synnovis. Aldosterone/renin ordering info; 2015. [Online]. Available from: http://www.viapath.co.uk/our-tests/aldosteronerenin [Accessed 13 Jan 2023].Search in Google Scholar

17. ARUP Laboratories. Lab test directory, renin, direct; 2023. [Online]. Available from: https://ltd.aruplab.com/Tests/Pub/2001575 [Accessed 13 Jan 2023].Search in Google Scholar

18. Auckland District Health Board. LabPLUS test guide – renin; 2022. [Online]. Available from: https://testguide.adhb.govt.nz/EGuide/ [Accessed 13 Jan 2023].Search in Google Scholar

19. Ismail, AA. The effects of repeated freeze-thaw cycles on endocrine parameters in plasma and serum. Ann Clin Biochem 2017;54:622–3. https://doi.org/10.1177/0004563217706090.Search in Google Scholar PubMed

20. Anton, G, Wilson, R, Yu, ZH, Prehn, C, Zukunft, S, Adamski, J, et al.. Pre-analytical sample quality: metabolite ratios as an intrinsic marker for prolonged room temperature exposure of serum samples. PLoS One 2015;10:e0121495/1. https://doi.org/10.1371/journal.pone.0121495.Search in Google Scholar PubMed PubMed Central

21. Gordon, AJ, Ford, RA. Chemist’s companion – a handbook of practical data, techniques, and references. New York: John Wiley & Sons; 1972:451 p.Search in Google Scholar

22. Australian Standards. AS 3864 – medical refrigeration equipment – for the storage of blood and blood products manufacturing requirements. 2012.Search in Google Scholar

23. 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 2021;59:59–69. https://doi.org/10.1515/cclm-2020-0061.Search in Google Scholar PubMed

24. Analyse-it for Microsoft Excel, Analyse-it Software, Ltd.; 2022.Search in Google Scholar

25. Danser, AHJ, Derkx, FHM, Schalekamp, MADH, Hense, HW, Riegger, GAJ, Schunkert, H. Determinants of interindividual variation of renin and prorenin concentrations: evidence for a sexual dimorphism of (pro)renin levels in humans. J Hypertens 1998;16:853–61. https://doi.org/10.1097/00004872-199816060-00017.Search in Google Scholar PubMed

26. Hsueh, WA, Baxter, JD. Human prorenin. Hypertension (Dallas) 1991;17:469. https://doi.org/10.1161/01.hyp.17.4.469.Search in Google Scholar PubMed

27. Chakera, AJ, McDonald, TJ, Knight, BA, Vaidya, B, Jones, AG. Current laboratory requirements for adrenocorticotropic hormone and renin/aldosterone sample handling are unnecessarily restrictive. Clin Med 2017;17:18–21. https://doi.org/10.7861/clinmedicine.17-1-18.Search in Google Scholar PubMed PubMed Central

28. Hepburn, S, Munday, C, Taylor, K, Halsall, DJ. Stability of direct renin concentration and plasma renin activity in EDTA whole blood and plasma at ambient and refrigerated temperatures from 0 to 72 hours. Clin Chem Lab Med 2022;60:1384–92. https://doi.org/10.1515/cclm-2022-0375.Search in Google Scholar PubMed

29. Sandberg, S, Fraser, CG, Horvath, AR, Jansen, R, Jones, G, Oosterhuis, W, et al.. Defining analytical performance specifications: consensus statement from the 1st strategic conference of the European federation of clinical chemistry and laboratory medicine. Clin Chem Lab Med 2015;53:833–5. https://doi.org/10.1515/cclm-2015-0067.Search in Google Scholar PubMed

30. Meijers, WC, van der Velde, AR, Kobold, ACM, Dijck-Brouwer, J, Wu, AH, Jaffe, A, et al.. Variability of biomarkers in patients with chronic heart failure and healthy controls. Eur J Heart Fail 2017;19:357–65. https://doi.org/10.1002/ejhf.669.Search in Google Scholar PubMed PubMed Central

31. Bais, R. Use of capability index to improve laboratory analytical performance. Clin Biochem Rev 2008;29(1 Suppl):S27–31.Search in Google Scholar

32. Gruson, D, Maisin, D, Lison, P, Maiter, D, Persu, A. Two-site automated chemiluminescent assay for measurement of immunoreactive renin. Biomarkers 2011;16:605–9. https://doi.org/10.3109/1354750x.2011.614015.Search in Google Scholar

33. Salib, MM, Yu, C, Apostoloska, S, McDonald, C, Moore, E, O’Shea, C, et al.. Is renin really stable when stored at −20°C? (Poster). In: AACB conference; 2019.Search in Google Scholar

34. Danser, AHJ, Batenburg, WW, van Esch, JHM. Prorenin and the (pro)renin receptor-an update. Nephrol Dial Transplant 2007;22:1288–92. https://doi.org/10.1093/ndt/gfl846.Search in Google Scholar PubMed

35. Tu, W, Eckert, GJ, Pratt, JH, Danser, AHJ. Plasma levels of prorenin and renin in blacks and whites: their relative abundance and associations with plasma aldosterone concentration. Am J Hypertens 2012;25:1030–4. https://doi.org/10.1038/ajh.2012.83.Search in Google Scholar PubMed

Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2022-1190).

Received: 2022-11-21

Accepted: 2023-02-08

Published Online: 2023-02-21

Published in Print: 2023-07-26