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Related Experiment Videos

Antimony quantification in Leishmania by electrothermal atomic absorption spectroscopy

W L Roberts1, P M Rainey

  • 1Department of Laboratory Medicine, Yale University, New Haven, Connecticut 06510.

Analytical Biochemistry
|May 15, 1993
PubMed
Summary

This study quantifies trivalent (Sb(III)) and pentavalent (Sb(V)) antimony in Leishmania parasites using atomic absorption spectroscopy. The developed method accurately measures antimony uptake and release, crucial for understanding antimonial drug efficacy.

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Area of Science:

  • Parasitology
  • Analytical Chemistry
  • Biochemistry

Background:

  • Leishmania parasites are causative agents of leishmaniasis.
  • Antimonial drugs are a primary treatment, but their efficacy is hampered by resistance.
  • Understanding antimony uptake and efflux in parasites is crucial for developing new therapies.

Purpose of the Study:

  • To quantify trivalent (Sb(III)) and pentavalent (Sb(V)) antimony in Leishmania mexicana pifanoi.
  • To develop and validate an atomic absorption spectroscopy method for antimony quantification in parasitic cells.
  • To characterize the influx and efflux kinetics of Sb(III) and Sb(V) in both amastigote and promastigote forms of the parasite.

Main Methods:

  • Quantification of Sb(III) and Sb(V) using atomic absorption spectroscopy with electrothermal atomization.

Related Experiment Videos

  • Parasites (amastigotes and promastigotes) were cultured and treated with potassium antimony tartrate [Sb(III)] or sodium stibogluconate [Sb(V)].
  • Parasite samples underwent nitric acid digestion before spectroscopic analysis.

    • Main Results:

    • The atomic absorption spectroscopy method demonstrated linearity from 0 to 7 ng of antimony with a low limit of detection (95 pg).
    • Influx rates varied significantly between Sb(III) and Sb(V), and between amastigote and promastigote forms.
    • Efflux of both Sb(III) and Sb(V) from parasites followed biphasic kinetics with distinct half-lives.

    Conclusions:

    • A sensitive and reliable method for quantifying antimony in Leishmania parasites was established.
    • The study provides kinetic data on antimony influx and efflux, offering insights into drug transport mechanisms.
    • These findings are vital for understanding antimonial drug action and resistance in leishmaniasis treatment.