The impact of thawing duration on leptin and adiponectin levels in frozen human milk samples.

Abstract

Measurements of hormone concentrations in human milk from understudied populations across the world is of growing scientific interest. Due to limited access to suitable laboratory facilities at many research sites, human milk samples are often shipped frozen to distant laboratories for analysis. Shipping and handling exposes samples to the risk of thawing and degradation as the results of delays, mishandling, or other unforeseen circumstances. Similar degradation risk factors are well known in serum samples. However, the vulnerability of hormone degradation in human milk to thawing during transportation is largely unexplored. Leptin and adiponectin are of particular importance due to their roles in feeding behavior and metabolism in infants, hence our focus on these two hormones. In addition, leptin and adiponectin have been shown to be vulnerable to pasteurization temperature degradation in human milk samples. We tested the degradation of human milk metabolic hormones - leptin and adiponectin - in a controlled environment in response to a variety of freeze/thaw conditions that are reflective of potential temperature fluctuations during transportation. Human milk samples were experimentally subjected to thaw durations ranging from 4 to 16 h and assayed for hormonal levels to observe changes from baseline. Samples were also subjected to two additional experimental conditions, re-freezing after an extended period outside freezers or remaining thawed with additional ice packs, to investigate the impact of common transportation conditions. We found the assayed levels of leptin were not significantly impacted by an extended thaw cycle of 16 h. However, leptin levels were impacted by the extended period of 40 h outside of the freezer. Adiponectin showed a decrease in concentration percentage after the initial 16-h thaw period, but the increased degradation between 0-16 h and 16-40 h was not as severe as that seen in the leptin samples. The results of this experiment can be used to inform hormone measurement consistency given different thawing lengths or freezing conditions during transportation. Additionally, this research informs decisions regarding transportation, storage/handling, and data analysis in human milk research.