We planned this study to research the effect of freeze-thaw number and storage temperatures on the tests of thiol-disulfide homeostasis. Dynamic thiol-disulfide homeostasis has a crucial role in antioxidant defense reactions. Thiol disulfide homeostasis is detected in serum samples. To gauge the steadiness at different storage temperatures, sera stored at -20°C and -80°C for 1, 2, 3, 6, and 12 months. To work out the effect of the freeze-thaw cycle, sera were frozen at -80°C and subjected to ten freeze-thaw cycles at 24-hour intervals. According to our results, native thiol, total thiol, and disulfide levels in human serum samples were stable at -20°C for three months and at -80°C for 12 months. After the third month at -20°C, native thiol and total thiol levels decreased while disulfide levels increased. Additionally, as the number of freeze-thaw cycles increased, native thiol and total thiol levels decreased, and disulfide levels increased. Our results show that storage at -20°C and an increasing number of freeze-thaw processes affect thiols levels. To get the most accurate results, repeated freeze-thaw cycles should be avoided the maximum amount possible. Also, samples should be frozen at -80°C if long storage times are required.
Key words: Thiols, freezing-thawing, stability
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