Determination of Cu/Zn and Fe in human serum of patients with sickle cell anemia using radiation synchrotron

doi:10.1016/j.apradiso.2011.12.028

Applied Radiation and Isotopes

Volume 70, Issue 7, July 2012, Pages 1277-1280

https://doi.org/10.1016/j.apradiso.2011.12.028 Get rights and content

Abstract

In this work we analyzed serum samples from patients with Sickle Cell Anemia (SCA) using Total Reflection X-Ray Fluorescence using Synchrotron Radiation (SRTXRF). The SRTXRF measurements were performed at the X-Ray Fluorescence Beamline at the Brazilian National Synchrotron Light Laboratory (LNLS). We studied forty-three patients aged 18–50 suffering from SCA and sixty healthy volunteers aged 18–60. It was possible to determine the concentrations of the following elements: P, S, Cl, K, Ca, Fe, Cu, Zn, Br and Rb. Moreover, there are evidences of an association among Fe, Cu, Zn and Cu/Zn in the SCA pathogenesis process. The concentrations of Fe and Cu in the serum samples of patients with SCA were larger, 120% and 20%, respectively, when compared with the CG. The serum level Cu/Zn ratio was significantly higher (60%) in the serum samples from patients suffering from SCA than from the CG. Therefore, the Cu/Zn ratio can be used as an adjuvant index in enhancement for diagnosis of SCA.

Highlights

► Serum samples from patients with Sickle Cell Anemia (SCA) were analyzed by SRTXRF. ► It was possible to determine the concentrations of the P, S, Cl, K, Ca, Fe, Cu, Zn, Br and Rb. ► There are evidences of an association among Fe, Cu, Zn and Cu/Zn in the SCA process. ► The results indicate that the Cu/Zn ratio can be used as an adjuvant index for diagnosis of SCA.

Introduction

Sickle Cell Anemia (SCA) is a blood disorder that affects hemoglobin, the protein found in red blood cells that help carry oxygen throughout the body. It is a disease that originated in Africa and is now the most prevalent hereditary disease in the whole world because of the vast racial mixing that affects millions of people. SCA occurs when a person inherits two genes for sickle hemoglobin (one from each parent) or homozygous sickle cell anemia (HbSS) (Yuditskaya et al., 2009). Instead of being flexible and disk-shaped, these cells are stiffer and curved in the shape of the old farm tool known as a sickle. These irregular-shaped blood cells die prematurely, resulting in a chronic shortage of red blood cells. In addition, they can get stuck when traveling through small blood vessels, which can slow or block blood flow and oxygen to certain parts of the body. Moreover, this causes pain and can lead to serious complications: anemia, delayed growth, infections and acute chest syndrome. Therefore, the determination of trace element levels in human serum is of interest for the biomedical area since several elements take part in all metabolic processes. Thus, it has been established that trace element levels in human serum can be utilized as indicators for several pathological conditions. In this way, the simultaneous detection of certain elements in the serum offers a very interesting approach in the diagnosis and treatment of various diseases including SCA (JongHwa et al., 2007).

The total reflection X-ray fluorescence (TXRF) is a multielemental technique widely used in the analysis of low concentrations (ng g⁻¹) in environmental, medical and biological samples. TXRF analysis is a well established analytical technique for the detection of major, minor and trace elements, especially suited for samples, whenever only small specimen mass is available. On the other hand, synchrotron radiation is an excellent source for exciting total reflection X-ray fluorescence (Wobrauschek, 2007).

In this study, the quantitative analysis of Fe, Cu. Zn and Cu/Zn in the serum from patients with Sickle Cell Anemia (SCA) and healthy subjects was performed by Total Reflection X-Ray Fluorescence using Synchrotron Radiation. Moreover, the results can be used to analyze the role of serum trace elements as an adjuvant index in enhancement for diagnosis of SCA.

Section snippets

Sampling serum

Forty-three patients (15 males and 28 females) aged 18–50, all of them suffering from Sickle Cell Anemia, and sixty healthy volunteers (41 males and 19 females) aged 18–60 were studied. The diagnosis of the sickling condition was established by medical history, physical examination and hematological studies. In addition, medical history was evaluated for each patient to exclude other pathologies. All the serum samples provided by “Instituto Estadual de Hematologia Arthur de Siqueira Cavalcanti”

Results and discussion

Fig. 1 shows a typical X-ray fluorescence spectrum of a serum sample using SR-TXRF. It was possible to detect the presence of ten elements: Phosphorus, Sulfur, Chlorine, Potassium, Calcium, Iron, Copper, Zinc, Bromine and Rubidium. In order to confirm the accuracy of the multi-elemental analysis of serum samples by means of SR-TXRF, we carried out elemental analyses of the CertiPUR^® reference material manufactured by Merck. The results are shown in Table 1. The results obtained are in agreement

Conclusions

SRTXRF is recognized as a powerful tool for the determination of multielemental concentrations in human serum samples. The elements P, S, Cl, K, Ca, Fe, Cu, Zn, Br and Rb were identified in serum samples of healthy subjects (CG) and patients with SCA. The values found in this study for the mean concentrations of Cu and Zn in CG are close to the values found in literature. The increase of Fe, in the SCA group, may be connected to folic acid ingestion, which helps in the maintenance of red blood

Acknowledgments

This study was partially developed at the Brazilian National Synchrotron Light Laboratory (LNLS-Project XRF-6753/07) with financial support by FAPERJ, CNPq and CAPES.

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Determination of Cu/Zn and Fe in human serum of patients with sickle cell anemia using radiation synchrotron

Abstract

Highlights

Introduction

Section snippets

Sampling serum

Results and discussion

Conclusions

Acknowledgments

Am. J. Clin. Nutr.

J. Am. Diet. Assoc.

Am. J. Clin. Nutr.

Blood

Reference ranges of copper and zinc and prevalence on their deficiencies in an Arab population aged 15–80 years

Biol. Trace Elem. Res.

Discontinuing prophylactic transfusions used to prevent stroke in sickle cell disease

N. Engl. J. Med.

Plasma malondialdehyde and serum trace element concentrations in patients with active pulmonary tuberculosis

Biol. Trace. Elem. Res.

Increased serum copper and decreased serum zinc levels in children with iron deficiency anemia

Biol. Trace Elem. Res.