MPŠ MP&Scaron MP&Scaron MP&Scaron Avtorji

Jožef Stefan
Postgraduate School

Jamova 39
SI-1000 Ljubljana

Phone: +386 1 477 31 00
Fax: +386 1 477 31 10


Course Description

Speciation of Elements in Biological Systems and Environment


Ecotechnologies, third-level study programme


prof. dr. Radmila Milačič
prof. dr. Milena Horvat
prof. dr. Janez Ščančar
prof. dr. Vekoslava Stibilj


To introduce the students the importance of speciation analysis in investigations of cycling of elements in environment and biologicaly systems. Students will gain basic knowledge on chemistry of numerous elements that in various chemical forms exhibit different toxicity or are even essencial.
The basic tools of speciation analysis that enable reliable determination of different chemical species of elements in matrices of biological and environmental samples will be introduced.
The role of chemical speciation will be presented for particular elements in the field of health, nutrition, uptake of particula chemical species to plants and in the investigations of biogeochemicaly cycling.


1. Introduction to the field of chemical speciation analysis. (Definition of chemical speciation regarding the oxidation state of element and speciation of organic and inorganic species of elements).
2. The role of speciation of elements regarding the toxicity and essenciality. (Some elements e.g. chromium exhibit different toxicity in different chemical forms. Hexavalent chromium is extremly toxic species, while trivalent chromiumn species are far less toxic and are even essential).
3. Basic analytical methods for speciation analysis. (Co-precipitation, selective extraction, spectrophotometry, chromatographic methods (HPLC, GC, electrophoresis…) coupled on-line and off-line with element specific detectors (AAS, ICP-MS, ICP-OES, AFS, INAA) in combination
with mass spectrometry.
4. Cycling and transformation of elements in environment. (Biogeochemical cycling of element species in system soil, water, air, the chemical species of elements in living organisms and foodstufs).
5. Speciation analysis of selected elements (Mercury, chromium, tin, selenium, srsenic, antimony, aluminium, cadmium, zinc, halogen elements…).

Course literature:

Caroli, S., Element Speciation in Bioinorganic Chemistry, Willey and sons, Inc. 1996.

Cornelis, R., Handbook of Elemental Speciation, Techniques and Methodology, Willey and sons, Inc. 2003.

Cornelis, R., Handbook of Elemental Speciation, Species in the Environment, Food Medicien and Occupational Health, Willey and sons, Inc. 2005.

Szpunar, J., Lobinski, R., Hyphenated Techniques in Speciation Analysis, Athenaeum Press Ltd, Gateshead, 2003

Significant publications and references:

• MILAČIČ, Radmila, ŠTUPAR, Janez. Simultaneous determination of chromium(III) complexes and
chromium(VI) by fast protein anion-exchange liquid chromatography - atomic absorption spectrometry.
Analyst (Lond.), 1994, vol. 119, str. 627-632.

• MILAČIČ, Radmila, ŠTUPAR, Janez. Fractionation and oxidation of chromium in tannery waste-and
sewage sludge-amended soils. Environ. sci. technol.. [Print ed.], 1995, vol. 29, str. 506-514.
MITROVIĆ, Bojan, MILAČIČ, Radmila, PIHLAR, Boris. Speciation of aluminium in soil extracts by
employing cation-exchange fast protein liquid chromatography-inductively coupled plasma atomic
emission spectrometry. Analyst (Lond.), 1996, vol. 121, str. 627-634.

• BANTAN POLAK, Tjaša, MILAČIČ, Radmila, PIHLAR, Boris. Quantitative determination of trace
amounts of Al-citrate by anion-exchange FPLC-ETAAS. Talanta (Oxford). [Print ed.], 1998, vol. 47, str.

• BANTAN POLAK, Tjaša, MILAČIČ, Radmila, MITROVIĆ, Bojan, PIHLAR, Boris. Combination of
various analytical techniques for speciation of low molecular weight aluminium complexes in plant sap.
Fresenius' j. anal. chem., 1999, vol. 365, str. 545-552.

• BANTAN POLAK, Tjaša, MILAČIČ, Radmila, MITROVIĆ, Bojan, PIHLAR, Boris. Investigation of low
molecular weight Al complexes in human serum by fast protein liquid chromatography (FPLC)-ETAAS
and electrospray (ES)-MS-Ms techniques. J. anal. at. spectrom., 1999, vol. 14, str. 1743-1748.

• SVETE, Peter, MILAČIČ, Radmila, MITROVIĆ, Bojan, PIHLAR, Boris. Potential for the speciation of Zn
using fast protein liquid chromatography (FPLC) and convective interaction media (CIM) fast monolithic
chromatography with FAAS and electrospray (ES)-MS-MS detection. Analyst (Lond.), 2001, vol. 126,
str. 1346-1354.

• BANTAN POLAK, Tjaša, MILAČIČ, Radmila, PIHLAR, Boris, MITROVIĆ, Bojan. The uptake and
speciation of various Al species in the Brassica rapa pekinensis. Phytochemistry. [Print ed.], 2001, vol.
57, str. 189-198.

• SANZ-MEDEL, Alfredo, SOLDADO CABEZUELO, Ana B., MILAČIČ, Radmila, BANTAN POLAK, Tjaša. The chemical speciation of aluminium in human serum. Coord. chem. rev., 2002, vol. 228, str. 373-383.

• KRALJ, Blaž, ŠČANČAR, Janez, KRIŽAJ, Igor, BENEDIK, Miha, BUKOVEC, Peter, MILAČIČ, Radmila. Determination of high molecular mass Al species in serum and spent CAPD fluids of dialysis patients combining SEC and anion-exchange FPLC with ETAAS detection. J. anal. at. spectrom., 2004, vol. 19, str. 101-106.

• BANTAN POLAK, Tjaša, MITROVIĆ, Bojan, MILAČIČ, Radmila. The use of fast protein liquid chromatography with ICP-OES and ES-MS-MS detection for the determination of various forms of aluminium in the roots of Chinese cabbage. Anal. chim. acta. [Print ed.], 2005, vol. 540, no. 1, str. 83-89.

• KRALJ, Blaž, KRIŽAJ, Igor, BUKOVEC, Peter, SLEJKO, Simon, MILAČIČ, Radmila. Speciation of aluminium in tea infusions by use of SEC and FPLC with ICP-OES and ES-MS-MS detection. Anal. bioanal. chem., 2005, vol. 383, str. 467-475.

• ZULIANI, Tea, LESPES, Gaëtane, MILAČIČ, Radmila, ŠČANČAR, Janez, POTIN-GAUTIER, Martine. Influence of the soil matrices on the analytical performance of headspace solid-phase microextraction for organotin analysis by gas chromatography-pulsed flame photometric detection. J. chromatogr A., 2006, vol. 1132, str. 234-240.

• MURKO, Simona, MILAČIČ, Radmila, ŠČANČAR, Janez. Speciation of Al in human serum by convective-interaction media fast-monolithic chromatography with inductively coupled plasma mass spectrometric detection. J. inorg. biochem.. [Print ed.], 2007, issue 9, vol. 101, str. 1234-1241

• ŠČANČAR, Janez, ZUPANČIČ, Marija, MILAČIČ, Radmila. Development of analytical procedure for the determination of exchangeable Cr(VI) in soils by anion-exchange fast protein liquid chromatography with electrothermal atomic absorption spectrometry detection. Water air soil pollut.. [Print ed.], 2007, vol. 185, str. 121-129.

• ZULIANI, Tea, LESPES, Gaëtane, MILAČIČ, Radmila, ŠČANČAR, Janez, POTIN-GAUTIER, Martine. Comprehensive study of the parameters influencing the detection of organotin compounds by a pulsed flame photometric detector in sewage sludge. J. chromatogr., 2008, vol. 1188, no. 2, str. 281-285.

• ŠČANČAR, Janez, MILAČIČ, Radmila. Aluminium speciation in environmental samples : a review. Anal. bioanal. chem., 2006, vol. 386, str. 999-1012.


Seminar and oral exam.

Students obligations:

Seminar and oral exam.