The Effects of a Single Freeze-Thaw Cycle on Concentrations of Nutritional, Noncommunicable Disease, and Inflammatory Biomarkers in Serum Samples

Ransi Ann Abraham; Garima Rana; Praween K Agrawal; Robert Johnston; Avina Sarna; Sowmya Ramesh; Rajib Acharya; Nizamuddin Khan; Akash Porwal; Sucheta Banerjee Kurundkar; Arvind Pandey; Raghu Pullakhandam; Krishnapillai Madhavan Nair; Geeta Trilok Kumar; Hps Sachdev; Umesh Kapil; Sila Deb; Arjan de Wagt; Ajay Khera; Lakshmy Ramakrishnan

doi:10.1055/s-0041-1726575

The Effects of a Single Freeze-Thaw Cycle on Concentrations of Nutritional, Noncommunicable Disease, and Inflammatory Biomarkers in Serum Samples

J Lab Physicians. 2021 Mar;13(1):6-13. doi: 10.1055/s-0041-1726575. Epub 2021 May 24.

Authors

Ransi Ann Abraham¹, Garima Rana¹, Praween K Agrawal², Robert Johnston², Avina Sarna³, Sowmya Ramesh³, Rajib Acharya³, Nizamuddin Khan³, Akash Porwal³, Sucheta Banerjee Kurundkar⁴, Arvind Pandey⁵, Raghu Pullakhandam⁶, Krishnapillai Madhavan Nair⁶, Geeta Trilok Kumar⁷, Hps Sachdev⁸, Umesh Kapil⁹, Sila Deb¹⁰, Arjan de Wagt², Ajay Khera¹⁰, Lakshmy Ramakrishnan¹

Affiliations

¹ Department of Cardiac Biochemistry, All India Institute of Medical Sciences, Delhi, India.
² UNICEF, Delhi, India.
³ Population Council, Delhi, India.
⁴ Clinical Development Services Agency, Translational Health Science & Technology Institute, Faridabad, Haryana.
⁵ National Institute of Medical Statistics, Indian Council of Medical Research, Delhi, India.
⁶ National Institute of Nutrition, Hyderabad, Telangana, India.
⁷ Institute of Home economics, Delhi, India.
⁸ Paediatrics and Clinical Epidemiology, B-16 Qutab Institutional Area, New Delhi, India.
⁹ Department of Human Nutrition, All India Institute of Medical Sciences, Delhi, India.
¹⁰ Ministry of Health and Family Welfare, Delhi, India.

Abstract

Background The stability of biological samples is vital for reliable measurements of biomarkers in large-scale survey settings, which may be affected by freeze-thaw procedures. We examined the effect of a single freeze-thaw cycle on 13 nutritional, noncommunicable diseases (NCD), and inflammatory bioanalytes in serum samples. Method Blood samples were collected from 70 subjects centrifuged after 30 minutes and aliquoted immediately. After a baseline analysis of the analytes, the samples were stored at - 70°C for 1 month and reanalyzed for all the parameters. Mean percentage differences between baseline (fresh blood) and freeze-thaw concentrations were calculated using paired sample t -tests and evaluated according to total allowable error (TEa) limits (desirable bias). Results Freeze-thaw concentrations differed significantly ( p < 0.05) from baseline concentrations for soluble transferrin receptor (sTfR) (- 5.49%), vitamin D (- 12.51%), vitamin B12 (- 3.74%), plasma glucose (1.93%), C-reactive protein (CRP) (3.45%), high-density lipoprotein (HDL) (7.98%), and cholesterol (9.76%), but they were within respective TEa limits. Low-density lipoprotein (LDL) (- 0.67%), creatinine (0.94%), albumin (0.87%), total protein (1.00%), ferritin (- 0.58%), and triglycerides (TAG) (2.82%) concentrations remained stable following the freeze-thaw cycle. In conclusion, single freeze-thaw cycle of the biomarkers in serum/plasma samples after storage at - 70°C for 1 month had minimal effect on stability of the studied analytes, and the changes in concentration were within acceptable limit for all analytes.

Keywords: analysis; bioanalytes; freeze-thaw; nutrition and noncommunicable diseases biomarkers; serum sample; stability; storage.

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