Biochemical Analytes Stability in Refrigerated (2 - 8oc) and Frozen (-20oc) Serum Samples at Kericho County Hospital Laboratory, Kenya
DOI:
https://doi.org/10.51867/scimundi.5.1.6Keywords:
Biochemical Analytes, Freezing, Refrigeration, Serum Samples, StabilityAbstract
Pre-analytical variables, including samples storage, can adversely affect the reliability of medical laboratory results. Add-on tests, alongside unprecedented delays in testing, require that samples are stored in a manner that maintain their integrity and optimally preserve the various analytes. Refrigeration and freezing, are well-documented preservation methods used by medical laboratories for short and long term storage of retained specimen. Different analytes however vary considerably in stability when samples are stored over a period of time. This study aimed at evaluated the stability of selected biochemical analytes in refrigerated (2 – 8oC) and frozen (-20oC) serum, to determine the maximum storage period at which accurate and reliable results can still be obtained at the Kericho County Hospital laboratory, guiding specific analytes delayed and add-on testing storage requirement and timelines. Samples drawn from twenty healthy volunteering adult participants, randomly picked from the local population, were processed and analysed at baseline, then a set of aliquots refrigerated and another frozen, in batches marked for analysis on the 7th, 14th, 21st and 28th days, on a well maintained, accurately calibrated, and quality checked HumaStar 100, an automated biochemistry analyser. Fridge and freezer temperatures were maintained relatively constant at 2 – 8oC and -20oC respectively, with routine monitoring throughout the period. The data was coded and analysed on SPSS version 22. The statistical significance of the difference in the determined analyte concentrations in comparison to baseline levels was evaluated using student paired T-test and Wilcoxon rank test. From the findings, alkaline phosphate, creatinine, and aspartate aminotransferase were found to be stable up to 28 days in the refrigerated and frozen serum, with mean percentage differences less than calculated reference change value of 16.48, 35.97 and 42.25 respectively. Sodium, total and direct bilirubin, were however unstable, as their mean percentage difference was more than the calculated reference change value of 2.16, 0.657 and 1.14. Urea and glucose showed stability at 2-8℃ for up to the 14th day, while potassium and chloride were only stable in refrigerated serum up to the 7th day analysis. Freezing improved the stability of all the analytes except bilirubin, ALT and sodium. This study recommends immediate analysis of serum for sodium, bilirubin, and ALT, which should never be part of add-on tests, and freezing of the samples when analysis is delayed for the other analytes.
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References
Alavi, N., Khan, S. H., Saadia, A., & Naeem, T. (2020). Challenges in preanalytical phase of laboratory medicine: Rate of blood sample nonconformity in a tertiary care hospital. EJIFCC, 31(1), 21.
Asmelash, D., Worede, A., & Teshome, M. (2020). Extra-analytical clinical laboratory errors in Africa: A systematic review and meta-analysis. EJIFCC, 31(3), 208.
Bonini, P., Plebani, M., Ceriotti, F., & Rubboli, F. (2002). Errors in laboratory medicine. Clinical Chemistry, 48(5), 691-698.
https://doi.org/10.1093/clinchem/48.5.691
PMid:11978595
Chua, C., Tifigiu, E., Boroujeni, A. M., Lin, B., Laskar, D., Shafique, K., Zuretti, A., Michl, J., & Pincus, M. R. (2018). Stability of values for the activities of critical enzymes assayed in serum frozen for prolonged time periods. Annals of Clinical and Laboratory Science, 48(5), 618-626.
Cray, C., Rodriguez, M., Zaias, J., & Altman, N. H. (2009). Effects of storage temperature and time on clinical and biochemical parameters from rat serum. Journal of the American Association for Laboratory Animal Science, 48, 202-204.
Divya, P. D., & Jayavardhanan, K. K. (2010). Effect of temperature and storage time on hepatobiliary enzyme activities in goat serum. Veterinary World, 3, 277-279.
Emre, H. O., Karpuzoglu, F. H., Coskun, C., Sezer, E. D., Ozturk, O. G., Ucar, F., & Coskun, A. (2021). Utilization of biological variation data in the interpretation of laboratory test results: Survey about clinicians' opinion and knowledge. Biochemia Medica, 31(1), 93-102. https://doi.org/10.11613/BM.2021.010705
PMid:33380892 PMCid:PMC7745156
Fauziah, A. N., Martsiningsih, M. A., & Setiawan, B. (2021). Electrolyte levels (Na, K, Cl) in serum stored at 4°C temperature. Indonesian Journal of Medical Laboratory Science and Technology, 3(2), 90-98. https://doi.org/10.33086/ijmlst.v3i2.1870
Flores, C. F. Y., Pineda, Á. D. L. M. H., Bonilla, V. M. C., & Sáenz-Flor, K. (2020). Sample management: Stability of plasma and serum on different storage conditions. EJIFCC, 31(1), 46.
Gómez-Rioja, R., Amaro, M. S., Diaz-Garzón, J., Bauçà, J. M., Espartosa, D. M., & Fernández-Calle, P. (2019). A protocol for testing the stability of biochemical analytes: Technical document. Clinical Chemistry and Laboratory Medicine, 57(12), 1829-1836.
https://doi.org/10.1515/cclm-2019-0586
PMid:31348756
Hedayati, M., Razavi, S. A., Boroomand, S., & Kheradmand, K. S. (2020). The impact of pre‐analytical variations on biochemical analytes stability: A systematic review. Journal of Clinical Laboratory Analysis, 34(12), e23551. https://doi.org/10.1002/jcla.23551
PMid:32869910 PMCid:PMC7755813
Hirigo, A. T. (2020). Effect of storage time and temperature on the stability of serum analytes. Health Science Journal, 15(7).
Ikeda, K., Ichihara, K., Hashiguchi, T., Hidaka, Y., Kang, D., Maekawa, M., Matsumoto, H., Matsushita, K., Okubo, S., Tsuchiya, T., Furuta, K., & Committee for Standardization, The Japanese Society of Laboratory Medicine (JSLM). (2015). Evaluation of the short-term stability of specimens for clinical laboratory testing. Biopreservation and Biobanking, 13(2), 135-143.
https://doi.org/10.1089/bio.2014.0072
PMid:25880474
Kachhawa, K., Kachhawa, P., Varma, M., Behera, R., Agrawal, D., & Kumar, S. (2017). Study of the stability of various biochemical analytes in samples stored at different predefined storage conditions at an accredited laboratory of India. Journal of Laboratory Physicians, 9(1), 11-15.
https://doi.org/10.4103/0974-2727.187928
PMid:28042210 PMCid:PMC5015491
Mrazek, C., Lippi, G., Keppel, M. H., Felder, T. K., Oberkofler, H., Haschke-Becher, E., & Cadamuro, J. (2020). Errors within the total laboratory testing process, from test selection to medical decision-making: A review of causes, consequences, surveillance, and solutions. Biochemia Medica, 30(2), 215-233.
https://doi.org/10.11613/BM.2020.020502
PMid:32550813 PMCid:PMC7271754
Najat, D. (2017). Prevalence of pre-analytical errors in clinical chemistry diagnostic labs in Sulaimani city of Iraqi Kurdistan. PLOS ONE, 12(1), e0170211. https://doi.org/10.1371/journal.pone.0170211
PMid:28107395 PMCid:PMC5249186
Pleus, S., Freckmann, G., Baumstark, A., & Haug, C. (2022). Stability of glucose concentrations in frozen plasma. Journal of Diabetes Science and Technology, 16(5), 1096-1100.
https://doi.org/10.1177/1932296820963657
PMid:33034207 PMCid:PMC9445330
Razi, F., Bandarian, F., Niazpour, F., Shafiee, G., Heshmat, R., Abdollahi, M., Seiedzadeh, M., Forouzanfar, K., Homayouni, M., Rambod, C., & Nasli-Esfahani, E. (2020). Stability of thirty-four analytes in blood samples of diabetic patients. Clinical Laboratory, 66(1). https://doi.org/10.7754/Clin.Lab.2019.190514
PMid:32013343
Shimizu, Y., & Ichihara, K. (2019). Elucidation of stability profiles of common chemistry analytes in serum stored at six graded temperatures. Clinical Chemistry and Laboratory Medicine, 57(9), 1388-1396. https://doi.org/10.1515/cclm-2018-1109
PMid:30860975
Van Vrancken, M. J., Briscoe, D., Anderson, K. M., & Wians Jr, F. H. (2012). Time-dependent stability of 22 analytes in lithium-plasma specimens stored at refrigerator temperature for up to 4 days. Laboratory Medicine, 43(6), 268-275.
https://doi.org/10.1309/LM53SBZ4VQGWVQOM
Widell, K. N., Dhakal, P. R., Thakur, M., & Hafner, A. (2019). Refrigeration to prevent food losses. International Institute of Refrigeration. https://doi.org/10.18462/iir.icr.2019.424
Zemlin, A. E. (2018). Errors in the extra-analytical phases of clinical chemistry laboratory testing. Indian Journal of Clinical Biochemistry, 33(2), 154-162. https://doi.org/10.1007/s12291-017-0657-2
PMid:29651205 PMCid:PMC5891449
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