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Laboratorial quantification of serum copper by Vis spectrophotometry in comparison to flame atomic absorption spectrometry

BACKGROUND: Copper is an essential nutrient for humans and maintenance of its adequate levels is important, since its metabolism is involved with oxidative stress and patolologies, such as Wilson's disease. In this work, a visible spectrophotometric method was validated, applied in young subjects and compared to flame atomic absorption spectrometry (FAAS) in serum copper levels determination. METHODS: Serum copper concentrations were measured by FAAS and by spectrophotometry, through copper reaction with bathocuproine, l = 484 nm. Aqueous curves and spiked serum curves were realized to verify linearity, recovery and precision of the visible spectrophotometric method. Blood samples from 12 subjects (mean 22 years) were analyzed in both methodologies and compared to each other. The results were expressed by mean ± standard deviation. RESULTS: Spiked serum and aqueous curves (n = 5) showed coefficient of regression superior to 0.99 and inter and intra-day coefficient of variation lower than 15%. The serum copper values found to Vis spectrophotometric method were 1.17 ± 0.39 and 0.73 ± 0.14 mg/l to women and men, respectively. To FAAS 1.13 ± 0.43 and 0.59 ± 0.13 mg/l were found to women and men, respectively. Both results showed a significant Pearson correlation (r = 0.946; p < 0.05). Besides, serum copper levels were significantly higher in women than in men (p < 0.05). CONCLUSION: The Vis spectrophotometric method demonstrated to be linear, reproducible and applied, offering a reliable alternative to FAAS copper quantification. Moreover, Vis spectrophotometry is a simple, quickly and not expensive technique, and it could be widely used as routine in clinical laboratory.

Copper; Serum; FAAS; Vis spectrophotometry


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