The prevalence of diabetes has reached alarming levels in India, making it essential to understand the concentration of nutritional-trace elements (Fe, Cu, Zn, Cr. and Se) in blood samples from diabetic adults. In this study, 208 whole blood samples from diabetic (n = 104) and non-diabetic (n = 104) adults across various age groups were analyzed using total reflection X-ray fluorescence (TXRF) spectroscopy with a sample dilution method. Statistical analysis was performed to assess descriptive statistics and determine a significant correlation between elemental concentrations in the blood samples of diabetic and non-diabetic adults. The mean concentration of nutritional-related trace elements in diabetic blood was as follows: Fe (46 ± 5) >
Zn (1.28 ± 0.14) >
Cu (0.10 ± 0.01) >
Cr (0.05 ± 0.004) >
Se (0.013 ± 0.002) in mg/L, respectively. Additionally, this study investigated the influence of nutrition-related trace element concentrations across various age groups such as 25-40 years (young adults), 41-55 years (middle-aged adults), and 56-70 years (older adults). In this investigation, Zn (p <
0.002) and Cr (p <
0.05) concentrations differed significantly between diabetic and non-diabetic adults aged 56-70 years. These findings will help us to understand age-dependent changes in element concentrations, clarify their role in diabetes, and improve risk factor management associated with diabetes.