Biological Trace Element Research

, Volume 183, Issue 1, pp 173–181 | Cite as

Speciation of As(ΙΙΙ)/As(V) and Total Inorganic Arsenic in Biological Fluids Using New Mode of Liquid-Phase Microextraction and Electrothermal Atomic Absorption Spectrometry

  • Lida Haghnazari
  • Nezam Mirzaei
  • Hossein Arfaeinia
  • Kamaladdin Karimyan
  • Hooshmand Sharafi
  • Nazir Fattahi


In this paper, a new extraction method based on countercurrent liquid–liquid microextraction (CLLME) has been developed for the extraction and preconcentration of inorganic arsenic (iAs) in plasma and urine samples prior to their analysis by electrothermal atomic absorption spectrometry (ETAAS). In this method, firstly, 5 ml of water is added to the extraction vessel. Then 30.0 μl of the extracting solvent is added to it in order for the extracting solvent to be placed in the narrow-necked vessel. In total, 10 ml of a standard solution or a pretreated real sample is added to the sample container and it is connected to the extraction vessel via a connector. While opening the embedded valve at the bottom of the sample container and the one in the extraction vessel, the sample solution flows into the extracting solvent with the same flow rate, leading to the successful extraction of metal ligand into the extracting organic solvent. Under the optimum conditions, calibration curves are linear in the range of 0.1–50 μg l−1, and limit of detections (LODs) are in the range of 0.03–0.05 μg l−1. The enhancement factor and enrichment factor were in the range of 220–240 and 198–212, respectively. Repeatability (intra-day) and reproducibility (inter-day) of method based on seven replicate measurements of 5.0 μg l−1 of arsenic were in the range of 2.3–3.5% and 4.0–5.7%, respectively. The applicability of the proposed CLLME and ETAAS methods was demonstrated by analyzing the iAs in spiked urine and plasma samples. The obtained recoveries of the arsenic in the range of 92–107% indicated the excellent capability of the developed method for speciation of arsenic from plasma and urine samples.

Graphical Abstract


Arsenic speciation Countercurrent liquid–liquid microextraction Extraction solvent lighter than water Biological analysis 



The authors gratefully acknowledge the Research Council of Kermanshah University of Medical Sciences (Grant Number: 96422) for the financial support.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.

Informed Consent

All the participants were informed about the purpose of the study and gave informed consent.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Department of Clinical Biochemistry, School of MedicineKermanshah University of Medical SciencesKermanshahIran
  2. 2.Department of Environmental Health Engineering, Faculty of HealthKashan University of Medical SciencesKashanIran
  3. 3.Department of Environmental Health Engineering, School of Public HealthTehran University of Medical SciencesTehranIran
  4. 4.Department of Environmental Health Engineering, School of Public HealthBushehr University of Medical SciencesBushehrIran
  5. 5.Environmental Health Research CenterKurdistan University of Medical SciencesSanandajIran
  6. 6.Research Center for Environmental Determinants of Health (RCEDH)Kermanshah University of Medical SciencesKermanshahIran

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