Non-transferrin bound iron measurement is influenced by chelator concentration

Kolb, A.M.; Smit, Nico P.M.; Lentz-Ljuboje, R.; Osanto, Susanne; Pelt, J. van

doi:10.1016/j.ab.2008.11.003

Cited by 24 publications

(28 citation statements)

References 17 publications

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“…In addition to the type of chelator used, its concentration is also found to play a critical role. It has been observed that with the increase in the concentration of NTA there is also a parallel increase in the concentration of NTBI estimated [63]. In the method described by Gosriwatana et al [19,62,64,65] 80 mM NTA was suggested as compared to the other methods where lower concentrations of NTA was used.…”

Section: Chelation Of Ntbi With Chelator Followed By Its Separation Amentioning

confidence: 99%

“…In the method described by Gosriwatana et al [19,62,64,65] 80 mM NTA was suggested as compared to the other methods where lower concentrations of NTA was used. However Kolb et al [63] suggested that 4 mM NTA is the best suitable concentration for chelation of NTBI. This chelated fraction of iron is separated from plasma protein with ultra filter of cut off of 30,000 [64].…”

Section: Chelation Of Ntbi With Chelator Followed By Its Separation Amentioning

confidence: 99%

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Non Transferrin Bound Iron: Nature, Manifestations and Analytical Approaches for Estimation

Patel

Ramavataram

2012

Ind J Clin Biochem

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Section: Chelation Of Ntbi With Chelator Followed By Its Separation Amentioning

confidence: 99%

Section: Chelation Of Ntbi With Chelator Followed By Its Separation Amentioning

confidence: 99%

Non Transferrin Bound Iron: Nature, Manifestations and Analytical Approaches for Estimation

Patel

Ramavataram

2012

Ind J Clin Biochem

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“…However, NTA must be used judiciously not to mobilize TBI [15], as extraction of even a small fraction (1-5%) of iron from holoTf or from iron-chelates (in chelated patients) can lead to unwarranted overestimation of NTBI [14]. This is particularly the case when traces of chelators might be present in circulation.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, unlike other studies [11,14,15], the LPI assay does not depend on supplementation of strong iron-mobilizing agents, or high-affinity chelators [16,20]. Thus, the assay avoids the inadvertent detection of TBI [15] or labile iron-chelates [19,[21][22][23]. The detected LPI reflects the most labile or redox-active forms of NTBI in native plasma or serum, but not putative forms bound to non-labile ligands (e.g., citrate, albumin).…”

Section: Rationale Of the Studymentioning

confidence: 99%

“…This led to simple and sensitive assays that are performed under nearly physiological conditions, with 40-lM ascorbate added to initiate redox cycling of the iron and ensuing generation of reactive oxygen species (ROS) that are quantified by a fluorescence assay (Table I). Furthermore, unlike other studies [11,14,15], the LPI assay does not depend on supplementation of strong iron-mobilizing agents, or high-affinity chelators [16,20]. Thus, the assay avoids the inadvertent detection of TBI [15] or labile iron-chelates [19,[21][22][23].…”

Section: Rationale Of the Studymentioning

confidence: 99%

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Non‐transferrin bound iron in Thalassemia: Differential detection of redox active forms in children and older patients

Breuer¹,

Ghoti

Shattat³

et al. 2011

American J Hematol

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Non-transferrin bound iron (NTBI) is commonly detected in patients with systemic iron overload whose serum iron-binding capacity has been surpassed. It has been perceived as an indicator of iron overload, impending organ damage and a chelation target in poly-transfused thalassemia patients. However, NTBI is a heterogeneous entity comprising various iron complexes, including a significant redox-active and readily chelatable fraction, which we have designated as ''labile plasma iron'' (LPI). We found that LPI levels can be affected by plasma components such as citrate, uric acid, and albumin. However, the inclusion of a mild metal mobilizing agent in the LPI assay (designated here as ''eLPI''), at concentrations that do not affect transferrin-bound iron, largely overcomes such effects and provides a measure of the full NTBI content. We analyzed three distinct groups of poly-transfused, iron overloaded thalassemia patients: non-chelated children (3-13 yrs, Gaza, Palestine), chelated adolescents-young adults (13-28 yrs, Israel), and chelated adults (27-61 yrs, Israel) for LPI and eLPI. The eLPI levels in all three groups were roughly commensurate (r 2 5 0.61-0.75) with deferrioxamine-detectable NTBI, i.e., DCI. In older chelated patients, eLPI levels approximated those of LPI, but in poly-transfused unchelated children eLPI was notably higher than LPI, a difference attributed to plasma properties affected by labile iron due to lack of chelation, possibly reflecting age-dependent attrition of plasma components. We propose that the two formats of NTBI measurement presented here are complementary and used together could provide more comprehensive information on the forms of NTBI in patients and their response to chelation. Am. J. Hematol. 87:55-61, 2012. V

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Ability to determine the desferrioxamine‐chelatable iron fractions of nontransferrin‐bound iron using HPLC

Koba

Słomka

Bączek

et al. 2013

J of Separation Science

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Iron is an essential element in human development. It is imperative for oxygen and electron transport and also for DNA and neurotransmitters synthesis. On the other hand, this metal is able to participate in Fenton's reaction that in turn leads to free radical damage. The most toxic fraction of iron - nontransferrin-bound iron and its part desferrioxamine-chelatable iron - can serve as an exquisite biomarker in the identification of iron imbalance. The goal of the present study was to devise a simple, repeatable, and inexpensive method for the determination of desferrioxamine-chelatable iron in serum blood samples. The assay procedure is based on desferrioxamine complex formation with iron ions followed to ferrioxamine and its quantitative measurement using RP-HPLC method. The desferrioxamine-chelatable iron was extracted from blood by centrifugation and SPE method. Chromatographic separation was performed at 40°C by step-form gradient elution using Cadenza CD-C18 column (150 × 4.6 mm id, particle size of 3.0 μm) connected with precolumn for contaminants removal. Gradient HPLC elution has been carried out with solvent A (10 mM Tris-HCl, pH 5.5) and solvent B (ACN). The flow rate was 1.2 mL/min, and the total separation time was 5 min. The linear quantitation range was 2.5-500 μM (r = 0.9973), and the LOD and LOQ were 0.42 and 1.29 μM, respectively. Proposed HPLC method allowed for the determination of desferrioxamine-chelatable iron fraction's of nontransferrin-bound iron, both in the buffer and the serum supplemented with iron ions as well as in the patients' serum samples with good results of precision and recovery. The developed method found to be sufficiently precise and reproducible for established conditions and after validation and may be used for routine assay of desferrioxamine-chelatable iron in biological samples.

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Non-transferrin bound iron measurement is influenced by chelator concentration

Cited by 24 publications

References 17 publications

Non Transferrin Bound Iron: Nature, Manifestations and Analytical Approaches for Estimation

Non Transferrin Bound Iron: Nature, Manifestations and Analytical Approaches for Estimation

Non‐transferrin bound iron in Thalassemia: Differential detection of redox active forms in children and older patients

Ability to determine the desferrioxamine‐chelatable iron fractions of nontransferrin‐bound iron using HPLC

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