Sequential Elution Technique in Moss Pseudoscleropodium purum: Comparison between the Commonly Used Extracellular Extractant NiCl2 and Other New Extractants

The sequential elution technique (SET) is used to determine the distribution of elements in the different cell fractions of mosses. The extracellular extractants most commonly used in this technique are NiCl2 and EDTA, although there are certain disadvantages associated with their use. In order to avoid such problems, we searched for new extractants that displace extracellularly bound metals, either because they are present at high concentrations (Ca) or because they have a high affinity for cation exchange sites (Hg and Au). The compounds HCl, NaCl, and CaCl2 were tested as extractants of the former type, as possible alternatives for the determination of extracellular metals in the moss Pseudoscleropodium purum. Calcium chloride was finally chosen as is it potentially the most successful in terms of binding to the cation exchange sites without altering the membrane permeability. The concentration chosen was 160 mM, as this yielded maximum displacement of Zn without membrane alterations. An experiment was then carried out to test the efficiency of Ca in extracting extracellular Zn, under laboratory and field conditions. In addition, Hg and Au were tested, at different concentrations, as extractants with high affinity for cation exchange sites, as neither of these elements has previously been used in the SET, and both display electronic characteristics that suggest their potential usefulness in displacing other cations from cation exchange sites. The results obtained show that extraction of extracellularly bound metal by high concentrations of Ca should be ruled out, as total extraction of Zn was not achieved. Both Hg and Au produced membrane alterations at low concentrations and moreover, neither was more efficient at extracting Zn than the reference extractant (20 mM NiCl2).