Humic Substances in Ecosystems 8, 2009

INTERNATIONAL SCIENTIFIC CONFERENCE“

Humic substancesin ecosystems 8
Book of abstractsŠOPORŇA, Slovakia, 13 – 16. september 2009

Humic Substances in Ecosystems 8, 2009
EDITORS:
Assoc. Prof. Ing. Anton Zaujec, CSc.
Ing.Jiří Heczko
Slovak university of Agriculture, Department of pedology and geology
Tr.A.Hlinku 2, SK – 949 76 Nitra, Slovak Republic
e-mail: anton.zaujec@uniag.sk
Prof. RNDr. Pavol Bielek, DrSc.
Soil Science and Conservation Research Institute
Gagarinova 10, SK – 827 13 Bratislava, Slovak Republic
e-mail: bielek@vupop.sk
Prof. Dr.hab. Slawomir S. Gonet
Nicolaus Copernicus University, Department of Soil Science
30 Sienkiewicz St, 87-1000 Torun, Polande-mail: gonet@geo.uni.torun.pl
Prof. Dr.hab. Božena Debska
University of Technology and Agriculture, Department of Environmental Chemistry
6, Bernardynska St., PL-85-029 Bydgoszcz, Polande-mail: debska@utp.edu.pl

ISBN: 978-80-89128-60-0
Humic Substances In Ecosystems 8
Humic Substances – Organic Matter – Soils – Water – Ecosystems© 2009 Soil Science and Conservation Research Institute, Bratislava, Slovakia© 2009 Slovak University of Agriculture, Nitra, Slovakia
Proceedings of the 8th International Conference “Humic Substances in Ecosystems 8” Šoporňa,September 13-17, 2009, organized by Slovak University of Agriculture in Nitra, Soil Science
and Conservation Research Institute Bratislava and Societas Pedologica Slovaca, Slovakia incooperation with University of Technology and Agriculture Bydgoszcz and Nicolaus
Copernicus University Torun, Poland

Prited in Slovakia: Soil Science and Conservation Research Institute, Gagarinova 10, 827 13 Bratislava

Humic Substances in Ecosystems 8, 2009
List of participants:
1. BABULICOVÁ MÁRIA, Mgr. Ing. PhD., Research Institute of plant production, Piešťany, Slovakia
2. BAACH-SZOTT MAGDALEA, dr inż., University of Technology and Life Sciences, Dept. of Environmental Chemistry,
Bydgoszcz, Polska
3. BARAČÍKOVÁ GABRIELA, RNDr. CSc., Soil Science Conservation and Research Institute, Bratislava, Slovakia
4. BARTOSZEK MARIOLA, Dr., Dept. of Environmental Chemistry and Technology, Institute of Chemistry, University ofSilesia, Katowice, Poland.
5. BOGDEVITCH IOSSIFF, Prof., Research Institute for Soil Science and Agrochemistry, Minsk, Belarus
6. CIESCISKA BOGUMILA, dr inż., University of Technology and Life Sciences, Dept. of Environmental Chemistry,
Bydgoszcz, Poland
7. CIESCISKI JACEK, dr inż., University of Technology and Life Sciences, Dept. of Environmental and Civil Engineering,
Bydgoszcz, Poland
8. DEBSKA BOZEA, dr hab., University of Technology and Life Sciences, Dept. of Environmental Chemistry, Bydgoszcz,
Poland
9. DRAG MALGORZATA, dr inż., University of Technology and Life Sciences, Dept. of Environmental Chemistry, Bydgoszcz,
Poland
10. DZIAMSKI ADRZEJ, PhD., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland University ofTechnology and Life Sciences, Dept. of Botany and Ecology, Bydgoszcz, Poland
11. FASUROVÁ ADĚŽDA, Mgr., Ph.D., Brno University of Technology, Institute of Physical and Applied Chemistry, Brno,
Czech Republic
12. FIALA KAREL, Ing. PhD., Agrovýzkum Rapotín s.r.o., Česká republika
13. FEREC SADOR, Dr., AU Gödöllö, Hungary
14. FESZTEROVA MELAIA, Ing. PhD., Dept. Chemistry, Constantine Philosophy University, Nitra, Slovakia
15. GLAZEWSKI ROMA, dr., University of Warmia and Mazury in Olsztyn, Polska
16. GOET SLAWOMIR, Prof. Dr., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland
17. GÓRA RÓBERT, RNDr., PhD. Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University
18. HALAS JÁ, Ing., Soil Science Conservation and Research Institute, Bratislava, Slovakia
19. HALKO RADOLAV, RNDr., PhD. Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University
20. HECZKO JIŘÍ, Ing., Dept. Pedology and Geology, SAU Nitra, Slovakia
21. HUTTA MILA, Doc.RNDr., PhD. Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University
22. HORVÁTOVÁ MÁRIA, Ing., Dept. Pedology and Geology, SAU Nitra, Slovakia
23. CHLPÍK JURAJ, Doc. Ing. PhD., Dept. Pedology and Geology, SAU Nitra, Slovakia
24. JEDLOVSKÁ LÝDIA, Ing. CSc., Dept. Environmental and Zoology, SAU Nitra, Slovakia
25. JOCZAK JERZY, dr., Pomeranian University in Słupsk, Dept. of Geomorphology and Quaternary Geology, Poland
26. KARABCOVÁ HAA, Mgr., Agrovýzkum Rapotín s.r.o., Česká republika
27. KRHOVJÁKOVÁ JAA, Ing., Agrovýzkum Rapotín s.r.o., Česká republika
28. KULIKOVA ATALIA, Dr., Dept. of Soil Science, Lomonosov Moscow State University, Russia
29. YAKIMEKO OLGA, PhD, Soil Science Dept., Moscow State University, Moscow, Russia
30. MARKIEWICZ MACIEJ, Dr., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland
31. MIELIK LILLA, PhD., Dept. of Physics and Agrophysics, West Pomeranian University of Technology in Szczecin, Poland
32. MICHALSKI ADAM, dr inż., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland
33. MIKHAILOUSKAYA ATALLIA, PhD., Research Institute for Soil Science and Agrochemistry, Minsk, Belarus
34. PHILIPPOVA OLGA, Ing., Dept. of Soil Science Lomonosov Moscow State University, Moscow, Russia
35. POLAK JUSTYA, Dr., Dept. of Environmental Chemistry and Technology, Institute of Chemistry, University of Silesia,
Katowice, Poland.
36. POSPÍŠILOVÁ ĽUBICA, RNDr., CSc., Mendelova zemědělská a lesnická univerzita v Brně, AF, Ústav agrochemie,půdoznalství, mikrobiologie a výživy rostlin, Brno, Česká republika
37. RABAJCZYK AA, Kielce, Poland
38. SEWERIAK PIOTR, Dr., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland
39. SIEKIEWICZ MARIOLA, PhD., Dept. of Physics and Agrophysics, West Pomeranian University of Technology in
Szczecin, Poland
40. STRZELECKA JOAA, dr inż., Nicolaus Copernicus University, Dept. of Soil Science, Torun, Poland
41. SZOMBATHOVÁ ORA, Ing. PhD., Dept. Pedology and Geology, SAU Nitra, Slovakia
42. ŠEREMEŠIĆ SRDJA, Mr., Faculty of Agriculture Novi Sad, Serbia
43. ŠIMASKÝ VLADIMÍR, Ing. PhD., Dept. Pedology and Geology, SAU Nitra, Slovakia
44. TOBIAŠOVÁ ERIKA, Ing. PhD., Dept. Pedology and Geology, SAU Nitra, Slovakia
45. WOJEWODSKI PIOTR, dr inż., University of Technology and Life Sciences, Dept. of Environmental Chemistry,
Bydgoszcz, Poland
46. ZAUJEC ATO, Doc. Ing. CSc., Dept.Pedology and Geology, SAU Nitra, Slovakia

Humic Substances in Ecosystems 8, 2009

Contents
1. THE IFLUECE OF ATHROPOGEIC POLLUTIO O THE PROPERTIES OF HUMIC ACIDS OF
LIGHT SOIL MAURED WITH ORGAIC FERTILIZERS
Agnieszka Grzelakowska, Magdalena Banach-Szott................................................................................................................72.

PROPOSAL OF EVALUATIO OF SOIL ORGAIC MATTER SORPTIO CAPACITY
Gabriela Barančíková..............................................................................................................................................................83.

MR STUDIES OF HUMIC SUBSTACES EXTRACTED FROM BOTTOM SEDIMET AD WATER
Mariola Bartoszek, Justyna Polak, Wiesław W. Sułkowski....................................................................................................10

4. DYAMICS OF CHAGES THE SELEIUM COTET AD DEHYDROGEASES ACTIVITY I SOIL
DEVELOPED BY ORGAIC FERTILIZATIO
Katarzyna Borowska, Jan Koper ...........................................................................................................................................11
5. IFLUECE OF LOG- TERM FERTILIZATIO AD CROP ROTATIO O THE QUATITY AD
SELECTED PROPERTIES IDEXES OF QUALITY SOIL HUMI SUBSTACES.
Bogumiła Cieścińska, Bożena Dębska ...................................................................................................................................12
6. CARBO AD ITROGE I THE BOTTOM SEDIMETS OF THE „ŻURSKIE” LOWLAD BARRAGE
LAKE
Jacek Cieściński .....................................................................................................................................................................137.

eFECT OF THE EXTRACTIO tIME O PROPERTIES OF HUMIC SUBSTACES ALKALIE EXTRACTS
Joanna Cieslewicz..................................................................................................................................................................14

8. CHROMATOGRAPHIC CHARACTERISTICS OF HUMIC ACIDS OF SOIL FERTILIZED WITH VARIOUS
ORGAIC FERTILIZERS
Debska B., Banach-Szott M., Dziamski A., Gonet S.S3...........................................................................................................15
9. ROLE OF POST-HARVEST RESIDUE I DEVELOPIG THERMAL PROPERTIES OF HUMIC ACIDS I
THE SOILS OF DIFFERET TYPES
Bożena Debska, Małgorzata Drag, Erika Tobiashova...........................................................................................................16
10. DYAMICS OF SOIL ORGAIC CARBO: A APPROACH OF COMBIED FRACTIOATIO TOOLSFOR ISOLATIG ECOLOGICALLY RELEVAT FRACTIOS
Elke Schulz, Marc Breulmann, Tatjana Böttger ....................................................................................................................17
11. SPECTROSCOPIC CHARACTERISTICS OF HUMATES ISOLATED FROM DIFFERET SOIL TYPES;aděžda Fasurová, Lubica Popíšilová ..................................................................................................................................18
12. UV-VIS SPECTROSCOPIC PROPERTIES OF HOT WATER-EXTRACTABLE ORGAIC CARBO I SOIL
Roman Glazewski, Barbara Kalisz, Andrzej Lachacz............................................................................................................20
13. SPATIAL VARIABILITY OF SOIL ORGAIC CARBO COTET (SOC): COMPARISO BASED O
DIFFERET DATA SOURCES
Ján Halas, Martina ;ováková ...............................................................................................................................................21
14. THE STRUCTURE OF HUMIC ACIDS OF MECHAOACTIVATED PEAT AD THEIR IFLUECE O
DYAMICS OF UMBER AD DESTRUCTIVE ACTIVITY OF HYDROCARBO-OXIDIZIGMICROORGAISMS
Alexander Ivanov ...................................................................................................................................................................22
15. LEAF LITTER FALL DECOMPOSITIO I AGE-DIFFERETIATED POPLAR STADS OF HYBRID 275
Jerzy Jonczak .........................................................................................................................................................................24

Humic Substances in Ecosystems 8, 200916. SOIL ORGAIC MATTER AD PHYSICAL CHARACTERISTICS
Hana Karabcová, Marie Svozilová, Jana Krhovjáková.........................................................................................................26
17. EFFECTS OF AGROTECHICAL TREATMETS O SPECTROPHOTOMETRIC PROPERTIES OF
HUMIC ACIDS OF BROW PODZOLIC SOILS
Krystyna Kondratowicz-Maciejewska....................................................................................................................................27
18. THE EFFECT OF THE ECOLOGICAL AD ITEGRATED SYSTEM O THE SOIL PHOSPHATASE
ACTIVITY
Jan Koper, Joanna Lemanowicz.............................................................................................................................................28
19. UPTAKE OF HUMIC SUBSTACES BY WHEAT PLATS: PREFERETIAL ACCUMULATIO I LIPID
FRACTIO
;atalia A. Kulikova, Eugenia A. Tsvetkova, Gennady A, Badun, Maria G. Chernysheva, Viktor I. Korobkov, Zinaida A.
Tyasto, Irina V. Perminova3 .................................................................................................................................................29
20. THE STRUCTURE AD PROPERTIES OF HUMIC SUBSTACES FROM BROW COAL 11 YEARSAFTER ADDITIO ITO SOIL
Jolanta Kwiatkowska-Malina ................................................................................................................................................31
21. THE IFLUECE OF BROW COAL O PHYSICO-CHEMICAL PROPERTIES OF SADY SOILS
Alina Maciejewska, Jolanta Kwiatkowska-Malina ................................................................................................................33
22. RELATIOS OF EZYME ACTIVITIES WITH DIFFERET FRACTIOS OF SOIL ORGAIC MATTER;atallia Mikhailouskaya and Iossif Bogdevitch.....................................................................................................................35
23. BIOLOGICAL ACTIVITY OF HUMIC SUBSTACES AD THEIR FE-ERICHED DERIVATIVESTOWARDS CUCUMBER PLATS UDER FE-DEFICIECY CODITIO
Philippova O.I., Sorkina T.A, Kulikova ;.A, Zhelezova S.V., Lebedeva G.F., Perminova I.V. .............................................36
24. PROPERTIES OF HUMIC SUBSTACES EXTRACTED FROM BOTTOM SEDIMET AT
GOCZAŁKOWICE DAM RESERVOIR
J. Polak, M. Bartoszek, W.W. Sułkowski ................................................................................................................................38
25. CHARACTERIZATIO OF SOIL ORGAIC CARBO AD ITS FRACTIO LABILE CARBO I
ECOSYSTEMSĽubica Pospíšilová,Veronika Petrášová, Jiřina Foukalová, Eduard Pokorný ......................................................................39
26. SOIL ORGAIC CARBO AD PARTICLE SIZE DISTRIBUTIO OF ARABLE SOILS I WITER
WHEAT CROPPIG
Srdjan Seremesic, Dragisa Milosev, Maja Manojlovic..........................................................................................................40
27. THE ROLE ORGAIC MATTER AS SOLID SUPPORT OF HEAVY METALS I THE WATER
EVIROMET
Anna Rabajczyk, Elżbieta Sykała ...........................................................................................................................................41
28. QUALITY OF SOIL ORGAIC MATTER I DIFFERET ECOSYSTEMS
Tobiašová Erika- Debska Božena- Horvátová Mária- Zaujec Anton- Szombathová ;ora- Chlpík Juraj- ;ádašský Ján .....42
29. COMPARISO THE PROPERTIES OF HUMIC ACIDS FRACTIOATED FROM AGRICULTURAL RICEAD FOREST SOILS
Erh-Shuan Kuo, Tsung-Yao Tseng, Kuo Chuan Lin and Ming Kuang Wang ........................................................................43
30. STRUCTURE AD PROPERTIES OF COMMERCIAL HUMATES FROM COALIFIED MATERIALS, PEATAD SAPROPELOlga Yakimenko, Alexey Izosimov .........................................................................................................................................44

Humic Substances in Ecosystems 8, 2009
31. THE EFFECT OF GAMMA IRRADIATIO O PROPERTIES OF SOIL HUMIC ACIDS
Joanna Strzelecka and Slawomir S. Gonet.............................................................................................................................46
32. STATUS OF ORGAIC MATTER I FOREST SOILS AFTER FIRE
Slawomir S. Gonet, Helena Dziadowiec, Aleksandra Kwiatkowska ......................................................................................47
33. HUMIC SUBSTACES EFFECT O POMEGRAATE URSERY PRODUCTIO
Ferenc Sandor........................................................................................................................................................................48
34. RELATIOS BETWEE ORGAIC HORIZO PROPERTIES AD FOREST FLOOR VASCULAR PLATSCOMPOSITIO I SCOTS PIE (PIUS SYLVESTRIS L.) STADS I SOUTH WEST POLAD
Piotr Sewerniak......................................................................................................................................................................50
35. THE IFLUECE OF ORGAIC MAURIG O THE COTET OF THE SOIL ORGAIC MATTER I
THE LOG - TERM MOOCULTURE
Babulicová, M. – Kotorová, D. ..............................................................................................................................................52
36. ORGAIC MATTER COTETS AD DISTRIBUTIO I CHEROZEM-RELATED SOILS OF POLADAD SLOVAKIA
Maciej Markiewicz, Michał Jankowski, Vladimir Simansky, Sławomir Gonet ......................................................................54
37. AALYSIS AD CHARACTERIZATIO OF HUMIC SUBSTACES ISOLATED FROM PEAT AD SOIL
USIG OFF-LIE COMBIATIO OF LIQUID CHROMATOGRAPHIC METHODS
Róbert Góra, Pavol Rohárik, Milan Hutta.............................................................................................................................55
38. O-LIE FLOW-THROUGH EXTRACTIO–PRECOCETRATIO-LARGE VOLUME IJECTIO
REVERSED-PHASE LIQUID CHROMATOGRAPHY METHOD FOR TRACE DETERMIATIO OF SELECTEDPYRETHROIDS I SLOVAK SOIL MICRO-SAMPLES
Mária Chalányová, Milan Hutta, Martin Pagáč....................................................................................................................56
39. STUDY OF OFF-LIE COMBIATIOS OF HPLC AD CZE METHODS FOR AALYSIS OF LOW
MOLECULAR WEIGHT SUBSTACES POTETIALLY RELATED TO HUMIC SUBSTACES DEGRADATIOPRODUCTS
Milan Hutta, Dušan Kaniansky, Róbert Góra........................................................................................................................57
40. UTILIZATIO OF IMMOBILIZED ALUMIIUM(III) METAL IO AFFIITY CHROMATOGRAPHY FORAALYSIS OF HUMIC ACID
Radoslav Halko, Tibor ;euročný, Milan Hutta .....................................................................................................................58
41. THE CARBO AD THE FRACTIOS OF SULPHUR I ATURE RESERVE ŽITAVSKÝ LUH ISLECTED SOILS
Melánia Feszterová, Lýdia Jedlovská ....................................................................................................................................59
42. IFLUECE OF FARMIG SYSTEMS O AREA HETEROGEEITY OF TOTAL ORGAIC CARBO
COTETJiří Heczko, Anton Zaujec......................................................................................................................................................60
43. HUMIC SUBSTACES I CRYOCOITE O A SVALBARD GLACIERS
Slawomir S. Gonet, Jerzy Melke, Agnieszka Marciniak and Ewa Meller ..............................................................................61

Humic Substances in Ecosystems 8, 2009
THE IFLUECE OF ATHROPOGEIC POLLUTIO O THE PROPERTIES OF
HUMIC ACIDS OF LIGHT SOIL MAURED WITH ORGAIC FERTILIZERS

Agnieszka Grzelakowska*, Magdalena Banach-Szott
Department of Environmental Chemistry, Faculty of Agriculture, University of Technology andLife Sciences, Bydgoszcz, Poland
E-mail:agrzelakowska@wp.pl
Sudden development of technology contributed to a phenomenon of mounting number oftechnological processes in which for many years so called environment friendly production was
not planned. Environmental pollution, local as well as global, appeared as its consequence. The
research aimed at the assessment of the influence of anthropogenic acidification, salinification
and alkalisation on the chosen parameters of humic acids, which to a large extent are responsible
for the state of soils. Complex structure of humic substances and their changeability suggest
research on new research methods, which would result in the greatest possible amount of
information on their structure and transformation. High-performance liquid chromatography(HPLC) is one of the analytical methods, which may contribute to broadening the knowledge ofhumic substances transformation processes taking place in the soils. Incubation experiment was
conducted in plastic vases of the capacity of 1l. To the ventilation of the incubated material
plastic tubes (3 tubes/each vase) were used. The incubation was conducted through one year
(from October to September) in the conditions of stable humidity (on the level of 50% of field
capacity), in the variable temperature of the region. The initial material used for incubationconsisted of light soil with/or without the addition of organic fertilisers (straw, vermicompost,
cattle manure) In all the cases, a significant difference in the parameters controlling the level ofpollution (pH- variant of acidification and alkalisation, proper conductivity- salinisation) was
obtained. After the incubation, the samples from the vases were air dried, strained through a sievewith meshes diameter of 1mm, and humic acids (HA) were extracted. Basic physical andchemical parameters of the obtained HA preparations were determined. Moreover, thesusceptibility to chemical oxidation was assessed and fractionation with the use of HPLC method
was performed. After taking the measures, the ratio of percentage content of hydrophilic fraction
to hydrophobic fraction (HIL/HOB) was calculated. On the basis of the tests conducted on thesamples of humic acids the following conclusions may be drawn:
1. The short-term action of the polluting factors did not influence significantly theproperties and transformations of humic substances described with the use of standard parameters(elementary composition), spectroscopic properties, susceptibility to chemical degradation).
2. However, the use of high-performance liquid chromatography (HPLC) proved thesignificant diversity of the examined humic acids considering their fractional composition. The
analysis of the obtained results showed obvious influence of both: the polluting substance and thekind of organic fertilizer (the addition of vermicompost to the soil material caused the greatest
difference in the examined humic acids).
KEYWORDS: humic substances, HPLC, anthropogenic pollution

7

Humic Substances in Ecosystems 8, 2009
PROPOSAL OF EVALUATIO OF SOIL ORGAIC MATTER SORPTIO CAPACITY

Gabriela Barančíková
Soil Science and Conservation Research Institute Bratislava, Regional Station Prešov,
Raymannova 1, 088 01 Prešov, Slovakia
E-mail: g.barancikova@vupop.sk
Adsorption reactions are defined as reversible and equilibrium reactions. In soil
compartment exists two main matrices where adsorption reactions can be realised. A whole a lot
is known about the concept of adsorption by inorganic constituents or clay minerals. The theoryof adsorption in soils is more concerned with the type of concentrating material at the solid-liquidinterfaces of clay minerals and not much data are available on adsorption by humic substances(HS).
However, it is well known that sorption capacity of humic substances (HS) is much higher
in comparison to clay minerals. HS can react with bases and acids, hence carry both positive andnegative charges. All these charges are developed by the ionization or dissociation of various
functional groups. The negative charges are attributed to dissociation of protons from thefunctional groups in the humic molecule. The two most important functional groups in thisrespect are the carboxyl and phenolic-OH groups. In general, these two functional groups control
the electrochemical behaviour of humic matter and they are the main reason for adsorption,
cation exchange, complex and chelation reactions. Carboxyl groups are the most important in theformation of negative charges. Since fulvic acids (FA) exhibit higher total acidity values thanhumic acids (HA), results of study on chelation and complexation analyses indicate that metalchelatation by humic acids appears more effective than by fulvic acids. The substantially larger
molecules and the more complex structures of humic acids are accepted to be the reason for morebinding sites and higher binding capacity in contrast to fulvic acids, which are smaller and lesscomplex. Humic acids are higher in aromatic structures than fulvic acids and dispose higher
chelatation capacity. Aromatic structures of HA play very important role ininteraction/incorporation of HA with organic xenobiotics.
For this reason proposal of evaluation of soil organic matter sorption capacity use basic
quantitative and qualitative humus parameters – content of soil organic carbon (SOC), ratio ofcarbon of humic and fulvic acids (Cha/Cfa) and optical parameter (Q46) but also such selectedparameters of HA chemical structure, which the most effect sorption capacity of humic acids –content of carboxylic groups and aromatization degree (α), which take into account humification
degree of HA.
High sorption humus capacity is characterized for arable soil locality with soil organiccarbon content higher or equal as 2, with ratio of Cha/Cfa > 1 and value of optical parameter Q46< 4.5. Locality with high sorption humus capacity have to dispose with high carboxylic groups(COOH>4) and degree of aromatization (α>50). In Slovak agriculture soils there are mainlysome localities of Fluvisols, Chernozems and Andosols. Low value of humus sorption capacity is
characterized for arable soils of Planosols, Rendzinas and some of Cambisols.

8Humic Substances in Ecosystems 8, 2009
Table 1. Proposal of evaluation of humus sorption capacity for agriculture soils

category
I.
II.
III.
I.
II.
III.

SOC (%) Cha/Cfa Q46
COOH (meq/1g HK)
>= 2
>1
< 4.5
>4
1.5 - 2
0.8 - 1
4.5 - 5
3-4
< 1.5
< 0.8
>5
<3
– high humus sorption capacity– medium humus sorption capacity– low humus sorption capacity

α (%)
>50
40 - 50< 40

KEYWORD: soil organic matter, sorption capacity, humic acids
This work was supported by the Slovak Research and Development Agency under thecontract No. APVV-0333-06.

9

Humic Substances in Ecosystems 8, 2009
MR STUDIES OF HUMIC SUBSTACES EXTRACTED FROM BOTTOM
SEDIMET AD WATER

Mariola Bartoszek, Justyna Polak, Wiesław W. Sułkowski
Department of Environmental Chemistry and Technology, Institute of Chemistry,
University of Silesia, Szkolna 9, 40-006 Katowice, Poland.
E-mail: mariola.bartoszek@us.edu.pl
NMR spectroscopy has been used for characterization of humic substances extracted frombottom sediment and water. The samples for studies were collected in spring and in autumn
season from eight places of the Goczałkowice Reservoir.
Fulvic acids were extracted both from bottom sediment and water whereas humic acids
were present only in bottom sediment.
The 1H NMR and 13C NMR analysis of humic substances facilitates the identification ofindividual functional groups. It was also used to estimate the relative contents of aromatic andaliphatic protons in humic substances. The HAr/HAli ratio makes possible to determine at which
stage of the humification process the water or the sediment occurs.
Solution 31P NMR spectroscopy has been used for determination the contribution ofdifferent classes of organic and inorganic phosphorus compounds in each sample.
KEYWORD: humic acids, fulvic acids, bottom sediment, 1H ;MR, 13C ;MR, 31 P ;MR10

Humic Substances in Ecosystems 8, 2009
DYAMICS OF CHAGES THE SELEIUM COTET AD DEHYDROGEASES
ACTIVITY I SOIL DEVELOPED BY ORGAIC FERTILIZATIO

Katarzyna Borowska, Jan Koper
Department of Biochemistry, University of Technology and Life Science,85-029 Bydgoszcz, 6 Bernardynska St., Poland, kborowska56@o2.pl
The concentration of selenium in plants and animals is strongly correlated with its
concentration in soils. Plants and products derived from plants transfer the selenium taken up
from the soils to humans. Selenium is one of the trace elements strongly affected by
microbiologically mediated redox processes influencing its solubility and consequently its
mobility, bioavailability and uptake in soil-plant system. Studies of enzyme activities in soil are
important as they indicate the potential of the soil to support biochemical processes which areessential for the maintenance of soil fertility. Soil samples were taken from the long-term
experiment established at the Experimental Station Grabow of the IUNG Pulawy on the soil
classified as Albic Luvisols. The experiment was conducted with crop rotation system: potato winter wheat - spring barley and red clover. The soil was affected of organic fertilization in aform of manure under potato in doses of 0, 20, 40, 60 and 80 t/ha. Total selenium content wasdetermined by the method of Watkinson with 2,3-diaminonaphtalene (DAN) using a Hitachi F2000 spectrofluorometer. Dehydrogenases activity was assayed colorimetrically using TTC as a
substrate. The application of manure resulted in the highest amounts of organic carbon in soil.
The selenium content from control plots ranged from 0.090 to 0.115 mg/kg. Statistical analyses
confirmed that application of manure resulted in the highest amounts of total selenium content insoil, with increasing doses of manure. We observed the increase of total selenium content about
40% in soil manured with the highest dose of FYM in comparison with soil from control plots.
Mean value of dehydrogenases activity in soil from control plots was 0.012 mg TPF·g-1·24h-1. We
observed the increase of enzymatic activity with increasing doses of manure. The dehydrogenases
activity in soil from plots manured with the highest dose was about 3 times higher than fromcontrol. We found a very significant correlation between total selenium content anddehydrogenases activity, and between this parameters and organic carbon in soil under study.

11

Humic Substances in Ecosystems 8, 2009IFLUECE OF LOG- TERM FERTILIZATIO AD CROP ROTATIO O THE
QUATITY AD SELECTED PROPERTIES IDEXES OF QUALITY SOIL HUMISUBSTACES.

Bogumiła Cieścińska, Bożena Dębska
Departament of Environmental Chemistry, University of Technology and Life Sciences,
Bernardynska 6, 85-029 Bydgoszcz, Poland.
E-mail: ciescinska@utp.edu.pl
In researches examined and compared composition of soil organic matter fraction andspectrofotometric properties of humic acids (HA) from soil samples in dependence on
agricultural treatments.
The research was performed on the basis of a multi-year long fertilization experimentcarried out by SGGW since 1923 in Skierniewice Research Station. The research involved theuse of soil samples from the stands under the monoculture and crop rotation of rye and potato,
considering selected fertilization combinations.
It was found that humus in the stand after rye in monoculture was characterized by higher
contribution of carbon of humic acids, higher value of humic acids carbon to fulvic acids carbon
proportion (CKH:CKF) and by lower participation of humins.
In the 5-fields rotation system comparable relations as in monoculture were found inhumus under potato. The highest values of CKH:CKF were obtained in both cultivation systems
with complete mineral-organic fertilization, lower values were noted in soil fertilized only by
NPK. Taking into consideration two fractions of humic acids – labile (FI) and strongly boundwith mineral part of soil (FIII), higher values of absorbance ratios (A2/4, A2/6, A4/6) were recordedfor HAs of FI fraction extracted from soil fertilized with participation of manure under rye inmonoculture and rotation. Absorbance ratios of HA from labile fraction were higher in soil inrotation system than in monoculture.
KEYWORD: fraction composition, UV-VIS, monoculture, crop rotation

12

Humic Substances in Ecosystems 8, 2009
CARBO AD ITROGE I THE BOTTOM SEDIMETS OF THE „ŻURSKIE”
LOWLAD BARRAGE LAKE

Jacek Cieściński
Department of Environmental and Civil Engineering, University of Technology and Life
Sciences, Sucha 9, 85-796 Bydgoszcz, Poland.
E-mail: jacek.ciescinski@wp.pl
Publication presents results of analyses of carbon and nitrogen content as well as roasting
losses from bottom sediments of barrage lake “Jezioro Żurskie”.
The samples of sediments with consideration of 3 cm layers were taken from the core of
sediments extracted by Instorf probe from central part of the lake. Obtained results indicate twolayers of sediments. In the layer of thickness 62-41 cm (the oldest sediments) there was no
significant vertical differentiation of analyzed parameters. Starting from the layer 38 cm
characterized by the lowest values of examined parameters to the layer of present sediments there
is noted systematical increase of carbon and nitrogen content as well as roasting losses. Carbon
content in dry mass of sediments reached values from 10,86 to 15,95% and nitrogen 0,59 to
1,46%.
KEYWORDS: barrage lake, bottom sediment, carbon, nitrogen13

Humic Substances in Ecosystems 8, 2009
EFECT OF THE EXTRACTIO TIME O PROPERTIES OF HUMIC SUBSTACES
ALKALIE EXTRACTS

Joanna Cieslewicz
Department of Environmental Chemistry, University of Technology and Life Sciences,2, Bernardynska St., 85-029 Bydgoszcz, Poland.
E-mail: joanna@utp.edu.pl
The aim of the present paper was to determine the effect of the extraction time on thealkaline properties of humic substances extracts from organic and mineral lake bottom sediments.
The extraction was performed with the use of 0.5 M NaOH. Both for organic and mineralsediments, the ratio during extraction were 1:10 and 1:50 (w/v). In both variants the extraction
time was, respectively, 1, 5, 12, 24 and 48 hours. In the mixtures of humic and fulvic acids
obtained, the following were determined: the content of carbon and nitrogen and thespectrometric properties for the UV-VIS range. Based on the present results, the values of
TOC/Nt, absorbance ratios, extraction efficiency and specific UV absorbance (SUVA) at the
wavelength of 280 nm were calculated.
The results demonstrate, most of all, that the extraction time has a significant effect on theproperties of extracts. It was observed that the extracts after a longer extraction time, in general,
showed a higher content of carbon and nitrogen as well as higher SUVA values. The present
research revealed that in the case of sediments, the extraction performed in the ratio of 1:50 (w/v)
is more efficient as compared with the extraction in the ratio of 1:10 (w/v).
KEYWORD: lake, humic substances, extraction

14

Humic Substances in Ecosystems 8, 2009
CHROMATOGRAPHIC CHARACTERISTICS OF HUMIC ACIDS OF SOIL
FERTILIZED WITH VARIOUS ORGAIC FERTILIZERS

Debska B.1, Banach-Szott M.1, Dziamski A.2, Gonet S.S.3
1

Department of Environmental Chemistry, University of Technology and Life Sciences,
Bernardynska 6, 85-029 Bydgoszcz, Poland.
E-mail: debska@utp.edu.pl2
Department of Botany and Ecology, University of Technology and Life Sciences,Kaliskiego 7, 85- Bydgoszcz, Poland.
E-mail: botanika@utp.edu.pl3
Department of Soil Science, ;icolaus Copernicus University, Gagarina 9, 87-100 Torun,
Poland.
E-mail:gonet@umk.pl
The aim of the present paper was to determine the effect of the kind of organic fertilisers
applied and the soil grain fraction the organic matter accumulates in on the hydrophilichydrophobic properties and the polydispersity of humic acids.
In the model incubation experiment organic fertilisers (cattle manure, vermicompost andwheat straw) were mixed with soil material (sandy texture). The soil was sampled for testing after
one year and a three-year period of incubation. Each sample was separated, according to graining,
into two fractions: grain diameter >0.1 mm (sand fraction) and <0.1 mm (silt and clay fraction)and then humic acids were isolated. For the humic acids isolated hydrophilic-hydrophobic
properties were defined using the RP-HPLC, and the division was made into low- and highmolecular-weight fractions (HPSEC).
It was demonstrated that introducing organic fertilizers to soil resulted in a decrease in theshare of the fraction of hydrophilic properties and an increase in the high-molecular fractions in
HAs molecules. However, there was shown no clear-cut effect of the kind of organic fertilizers
on the hydrophilic-hydrophobic properties and the polydispersity of humic acids. Humic acids offine-grained fraction (φ < 0.1 mm) demonstrated a higher share of hydrophilic fractions and, as aresult, higher values of the HIL/HOB ratio as compared with HAs sand fraction. The
relationships recorded show that the organic matter accumulated in finer grain fractions of soil ismore humified than the organic matter of coarser grain fractions.
KEYWORDS: organic fertilizers, humic acids, polydispersity, hydrophilic and hydrophobic
fraction

15

Humic Substances in Ecosystems 8, 2009
ROLE OF POST-HARVEST RESIDUE I DEVELOPIG THERMAL PROPERTIESOF HUMIC ACIDS I THE SOILS OF DIFFERET TYPES

Bożena Debska1, Małgorzata Drag1, Erika Tobiashova21

Department of Environmental Chemistry, University of Technology and Life Sciences,
Bernardynska 6, 85-029 Bydgoszcz, Poland.
E-mail: debska@utp.edu.pl2
Department of Geology and Pedology, Slovak Agricultural University, Tr.A. Hlinku 2,
949 01, ;itra, Slovakia.
E-mail: Erika.Tobiasova@uniag.sk
The aim of the present paper was to determine parameters of thermal decomposition of
humic acids of the soils of different types with no additives and when mixed with post-harvest
residue before and after 360 days of incubation.
The research was based on a model incubation experiment carried out at the Departmentof Pedology and Geology of the Slovak Agricultural University in Nitra. Plant residues (mixturesof aerial parts and roots) of corn, oilseed rape, sunflower, potato and rye were mixed in the ratio
10:1 with the following soil samples: luvisol, chernozem, arenosols and gleic phaeosems and the
mixtures were incubated at 25oC and 60% of field water capacity. The plant-soil mixtures wereanalysed prior the experiment and after 360 days of incubation. Humic acids were separated fromthe control soil (without plant residue), and plant-soil mixtures before and after incubation withthe standard Schnitzer method. Thermal properties were determined with the use of the
Derivatograph C (MOM, Hungary). Samples of 40 mg humic acids were mixed with Al2O3 (1:9)
and heated in the air at the rate of 3.3 °C. min -1.
The results of thermal analysis showed that parameters of thermal decomposition of
humic acids were determined both by the soil type and by the properties of post-harvest residue
introduced into soil. The process of decomposition of post-harvest residue was connected with,e.g. a decreased value of the parameter corresponding to the burning heat of humic acids. Low
values of parameter ‘Z’ were due to the presence of non-decomposed lignin and cellulose
structures in the molecules of humic acids
KEYWORDS: post-harvest residue, humic acids, TG, DTG, DTA

16

Humic Substances in Ecosystems 8, 2009
DYAMICS OF SOIL ORGAIC CARBO: A APPROACH OF COMBIED
FRACTIOATIO TOOLS FOR ISOLATIG ECOLOGICALLY RELEVAT
FRACTIOS

Elke Schulz, Marc Breulmann, Tatjana Böttger
Helmholtz Centre for Environmental Research – UFZ,
Theodor-Lieser-Str. 4, D-06120 Halle, Germany
E-mail: elke.schulz@ufz.de
Up to now the most commonly used parameter for soil quality is the total organic carbon(TOC) content in soil. Changes of TOC as well as its distribution in differently stabilized soilorganic matter (SOM) pools are very slowly running processes.
SOM research is connected with questions of SOM extractability, structure, turnover,stability and function. However, actually no applicable methods and practical solutions for
managing carbon content in soil could be developed.
Studying SOM dynamics we have to consider at least two pools: a stable / stabilized(relative “inert”) pool and a decomposable pool of SOM. The “inert” fraction has almost no
considerable share in carbon transformation processes having residence times of some hundredyears. Only the decomposable pool will be transformed in relevant time intervals and may be
influenced by human activities. This part is strongly correlated with transformation processes oforganic substances in soil (transformation of nutrients as well as pollutants).
Recently a wide range of methods are used to isolate and characterize SOM pools for
estimating both their amount and suitability as indicators for soil quality.
Depending on the scientific question several methods and techniques can be used fordiscrimination of different SOM pools:•
Hot water extraction (HWC)•
Cold water extraction (WEOM)•
Potassium permanganate oxidization (labile C)•
Clay associated organic matter (stable carbon)•
Specific light organic matter (different stability of organic matter).
Our approach for assessing of SOM quality is a combination of physical and chemical
separation techniques, including size-density fractionation and isolation of a readily
decomposable organic carbon pool by hot-water extraction. C content in hot water extractablefraction is a simple determinable parameter for estimating decomposable SOM supply andnutrient release. However, management (fertilization) also contributes to C pool associated toclay minerals of soil matrix.
The objectives of here presented research are: (1) to give a short overview of selected
methods of SOM pool discrimination, (2) to show relations between selected pools, (3) todemonstrate the suitability of hot water extractable C for estimating and calculating
decomposable SOM pool.
Results from long-term field experiments support hypotheses that impact of management
practices on SOM could be discriminated with the hot water extractable SOM fraction whileorganic matter in size-density fractions represent more stabilized OM with special importance forsustainability and C accumulation.
KEYWORD: hot water extraction, particle size and density fractionation, stability of SOM

17

Humic Substances in Ecosystems 8, 2009
SPECTROSCOPIC CHARACTERISTICS OF HUMATES ISOLATED FROMDIFFERET SOIL TYPES

Naděžda Fasurová1, Lubica Popíšilová2
1

University of Technology, Faculty of Chemistry, Institute of Physical and Applied Chemistry,
Purkyňova 118, 612 00 Brno, Czech Republic.
E-mail: fasurova@fch.vutbr.cz
2
Mendel University of Agriculture and Forestry, Institute of Agrochemistry, Soil Science,
Microbiology and Plant ;utrition, Zemědělská 1, 613 00 Brno, Czech Republic.

The aim of our work was to compare spectroscopic properties of soil humates isolated
from different soil matrices (Haplic Cambisol, Leptic Cambisol, Eutric Cambisol, Luvi-Haplic
Chernozem). The optical methods (VIS and SFS) on this project have been used. Synchronous
fluorescence spectra (SFS) were measured by Spectrofluorimeter Aminco Bowman Series 2,
Thermospectronics. Bandpass of both monochromators was set to 4 nm, temperature was 20°C
and constant difference between excitation and emission monochromators was (∆λ = (λem. - λex.)= 55 nm). VIS spectra were measured by spectrometer Varian Cary 50 Probe within the range
300 - 700 nm.
Isolation of HA preparation was made according to the IHSS method. Elemental analysis
of isolated HA was kindly made in Engineering Test Institute Brno. We followed total carbon
content, fractional composition, and humification degree.
The highest humic acids content had Eutric Cambisol and the lowest in Leptic Cambisol.
The highest humification degree had sample Luvi-Haplic Chernozem and the lowest Leptic
Cambisol. This result is equal to humic substances maturity in different soil types.
The highest content of organic carbon and humus was in Eutric Cambisol(grassland), the lowest values was in Haplic Cambisol. The highest humic acids content was in
Eutric Cambisol.
SFS spectra (Figure 1) measured in range from 200 to 600 nm (at ∆λ=55 nm) contained 5
or 6 peaks at λex./λem.=413/468, 450/505, 467/522, 482/537, 492/547 (303/358). Last one wasobserved in two Eutric Cambisols samples and in Haplic Cambisols. The maximum relative
fluorescence intensity at 522 nm gave Haplic Chernozem. The lowest relative fluorescenceintensity gave Leptic Cambisol at the same wavelength. Higher value of relative fluorescenceintensity gave the sample of Eutric Cambisol isolated from grassland to compare with the sample
isolated from arable soil. We have founded out that SFS spectra gave to main fluoprophores withrespect of changing of ∆λ difference between both monochromators at about: 450/480, 467/500530 nm. The relative fluorescence indexes were calculated from SFS spectra (as the ratio
I468/I522). Colour indexes (Q4/6) were calculated as ratio A465/A665 from VIS spectra. The highest
fluorescence index (F) was obtained in Eutric Cambisol (arable soil) and the lowest in EutricCambisol (grassland). The highest Q4/6 index had Leptic Cambisol and the lowest had LuviHaplic Chernozem. Linear correlation between organic carbon content and fluorescence indexes
was found (R2=0.92).

18

Humic Substances in Ecosystems 8, 2009
Fig.1. SFS spectra of soil humates at ∆λ=55 nm

KEYWORD: soil humic substances, Synchronous fluorescence spectroscopy
ACK;OWLEDGEME;T.
This work has been supported by Grant Agency of the Czech Republic ProjectNo.104/03/D135, by the Project NAZVA No. QH72039 and by the Research plan No.
MSM6215648905.

19

Humic Substances in Ecosystems 8, 2009
UV-VIS SPECTROSCOPIC PROPERTIES OF HOT WATER-EXTRACTABLE
ORGAIC CARBO I SOIL

Roman Glazewski1, Barbara Kalisz2, Andrzej Lachacz2
1) Department of Environment Protection Engineering, University of Warmia and Mazury in
Olsztyn, ul. Prawochenskiego 1/38, 10-975 Olsztyn, Poland.
E-mail: rom@uwm.edu.pl2) Department of Soil Science and Soil Protection, University of Warmia and Mazury in Olsztyn,
pl. Lodzki 3/109, 10-727 Olsztyn, Poland
E-mail: andrzej.lachacz@uwm.edu.pl, bsam@O2.pl
Surface organic soil layers are the main source of allochthonous organic matter for water
reservoirs. This kind of soil formations after dewatering undergoes the processes of rapidtransformation of humic substances, which are transformed and mineralised. This leads to themodification of organic compounds composition. Their water repellency and lability in soil
solution changes. The study comprises three groups of soil formations – with an ongoing processof organic matter accumulation, dewatered organic formations containing more than 12% of OC,
and dewatered soil formations containing less than 12% of OC which are considered degraded.
UV-VIS spectroscopic properties of soil organic matter soluble in hot water were studied.
Soil samples were extracted in a closed system for 18 hours and then filtered through 0.45 µm
filters. The spectra were taken out in the range of 200-700 nm in 1 cm quartz cuvette using
double beam scanning spectrophotometer Shimadzu 1601PC. Quantitative measurements ofdissolved organic carbon using analyser Shimadzu TOC5000 were made as well. The results
enabled us to compare the dynamics of absorbance Aλ and SUVAλ as well as the ratio forvarious values of λ. It was estimated that the organic compounds of the analysed soil formations
had different sizes, degree of condensation (humification), lability and solublity in water. The
degraded soil formations (<12% of OC) contain more hydrophobic condensed aromatic cores,
which was proved by the highest values of SUVA280.
KEYWORD: organic soil, hot water solubility, UV-Vis spectra, HWC, DOC

20

Humic Substances in Ecosystems 8, 2009SPATIAL VARIABILITY OF SOIL ORGAIC CARBO COTET (SOC):
COMPARISO BASED O DIFFERET DATA SOURCES

Ján Halas1, Martina Nováková2
1

2

Soil Science and Conservation Research Institute, Raymannova 1, 080 01 Prešov
Soil Science and Conservation Research Institute, Gagarinova 10, 827 13 Bratislava
E-mail: j.halas@vupop.sk, m.novakova@vupop.sk

Common crop management is based on the assumption according which any field being abasic spatial unit of agricultural production is considered to be homogenous for its natural
conditions. Uniform agricultural technologies such as soil agrotechnology, fertilization, and plant
protection are applied over the whole area of individual fields. Moreover, any field is considered
to be a basic spatial and statistical unit for specifying the relevant soil and landscape properties.
On the other hand, there is a significant spatial variability in soil organic carbon content which
does not respect artificial land parcels. Organic carbon represents a key soil component withsignificant impact on soil physical, chemical and biological properties of soil (including the soil
production potential).
The aim of this paper is: i) to demonstrate the existence of spatial within-field variability
in relation to the content of soil organic carbon (SOC) and map this variability; ii) to delineate
homogenous areas based on SOC content classification and to quantify their spatial extent; iii) tocompare historical and the most recently measured data on SOC content at the same areas, andiv) to demonstrate vertical variability of SOC content at selected soil profiles which are tocharacterize some of the identified homogenous areas.
The experiment was carried out on pilot area consisting of 7 different plots of arable landwith the total area of 720 ha. Model area is located at Podunajská nížina lowland which ischaracterized as both the warmest and very dry climate region in Slovakia (with average yearlytemperature 10 °C and average yearly rainfall about 550 mm). The soil sampling oriented todetermine SOC content was performed by auger sampling in regular triangular network (with aside of 131,5 m) and with a density of approximately 1 sampling point per 1,446 ha. Totally, 521
soil samples were taken from depth of 0-0,2m and analyzed for SOC content (Cox) by standard
laboratory methods. Position (longitude, latitude) of all sampling points was recorded by GPS.
The results of experiment confirm the existence of within-field spatial variability related
to the SOC content, as well as significant differences in SOC content has been proven among thestudied plots (the SOC content difference is from very low to very high depending on the SOC
content classification used). Relatively homogenous zones delimited on the basis of applied SOC
content classification have a potential to explain the SOC balance in soil within the individual
plots. The comparison of historical and our measured data have shown an increase in SOC
content from 8 to 104%. Moreover, the delimited zones homogenous as for its SOC content after
they are further characterized by additional soil and landscape data can provide the spatial units
for the crop production modeling in detailed scales.
KEYWORD: spatial variability, soil organic carbon
ACK;OWLEDGEME;T:This work was supported by the Slovak Research and Development Agency under thecontract No. APVV-0333-06.

21

Humic Substances in Ecosystems 8, 2009
THE STRUCTURE OF HUMIC ACIDS OF MECHAOACTIVATED PEAT AD
THEIR IFLUECE O DYAMICS OF UMBER AD DESTRUCTIVE ACTIVITYOF HYDROCARBO-OXIDIZIG MICROORGAISMS

Alexander Ivanov
Institute of Petroleum Chemistry SB RAS, Akademichesky Avenue 3, 634021 Tomsk, Russia.
E-mail: ivanov@ipc.tsc.ru
Due to development of nature-protecting biotechnologies the interest to the study ofprocesses with participation of aerobic hydrocarbon-oxidizing microorganisms (HOM) has
increased. First of all it is concerned with the pollution of soil, surface and underground waters
by oil and oil products. The intensity and character of decomposition of oil hydrocarbons in soil
basically is defined by a functional activity of HOM, capable to assimilate of oil as a uniquesource of carbon [1]. Processes of soil self-cleaning under conditions of environment proceed
extremely slowly and require stimulation. As hydrocarbon-oxidizing microorganisms areconstant components of soil biocoenosis, there was an aspiration to use them catabolic activity
for restoration of petropolluted soils by the addition of preparations on a basis of humic acids(HA) of peat into polluted soil [2].
The purpose of this investigation is to study the influence of mechanoactivation (MA) of
peat on changes of structure of HA and to investigate the influence of HA on dynamics ofnumber and destructive activity of HOM of petropolluted soil.
A high-moor moss peat of «Temnoe» field of the Tomsk Oblast (Russia) with a low
degree of decomposition of 5 % was the subject of the present study.
MA was carried out in the AGO–2 planetary mill-activator without additives (HA2) and in
the presence of solid alkali 3 % NaOH (HA3). HA1 from initial nonmechanoactivated peat
served as the control.
MA of peat results in the changes in yields and quantitative characteristics of main organic
components [3]. The efficiency of HA extraction after MA raises – their yield has increased in 7
times. The increase of the HA yield may be explained by the destruction of the hard-to-hydrolyse
substances. Moreover, results obtained with use of gel-permeation chromatography and 13C
NMR– spectroscopy testify about the decrease in molecular weight and a structural
reconstruction of macromolecules of HA. The presence of aromatic skeleton in HA provides theirability to binding organic compounds that are result of hydrophobic interactions. Therefore, theaffinity of HA with hydrophobic organic compounds increases due to increase of contribution ofaromatic skeleton in their structure. The greatest degree of aromaticity and the least degree ofoxidability have observed for HA after mechanoactivation of peat with alkali.
It is known, that the first stage of oil degradation begins its physicochemical andmicrobiological destruction [4]. In this period a relative content of alkanes decreases and aquantity of resinous substances increases in residual oil. This stage of recultivation corresponds
to the most toxic geochemical situation, a maximum inhibition of biocoenosises and on this stage
it is expediently to carry out actions for the intensification of microbiological processes andphotochemical and physical processes of decomposition of oil.
The analysis of a chemical composition of petropolluted soil in 30 days after introductionof HA has shown that the quantity of extracted bitumen-like compounds decreases in 40 %. In
structure of bitumen-like compounds the share of paraffin-naphthene fractions has decreased, inparticular, n-alkanes and the quantity of resins has increased. It testifies about a destructive
oxidizing activity of microorganisms stimulated by HA.
22

Humic Substances in Ecosystems 8, 2009
It is known, that the oil pollution increases of activity of HOM which play the importantrole in the decomposition of oil and detoxication of soils [5]. The growth of bacteria andactinomicetes – basic destructors of oil, has noted in the end of experiment (in 30 days ofgrowth). The additions of HA in pure soil were intensifying the growth of both groups, and ingreat degree of bacteria. The greatest stimulating effect of studied groups of microorganisms has
been observed after the addition into soil of HA3 solutions. The essential increase in number ofactinomicetes and bacteria was noted for petropolluted soil after the addition these HA. Obtained
data can be explained that a soil microflora uses peripheral parts of HA molecules in an initialperiod of growth.
The change of fermentative activity of soil microflora is one of indicators of oil
biodestruction. The growth of microorganisms’ number was accompanied by the increase theirfermentative activity which provides processes of oil hydrocarbon destruction.
A stimulating action of HA3 on a destructive activity of soil microflora has confirmed by
the method of IR-spectroscopy. Spectral coefficients, calculated according to optical density ofabsorption bands in a certain area of spectrum and which characterize biodegradation processes,
testified about the decomposition of paraffin hydrocarbons, increase in degree of branching and,as a whole, about a deeper processes of oil biodegradation with the use of HA3.
Thus, results of researches have shown that the addition of HA (0.005 mass % from soil
weight) isolated from mechanoactivated peat (in the presence of solid alkali 3 % NaOH), in
petropolluted soil increases the number of heterotrophic bacteria in 8 times, actinomicetes in 5
times and fermentative activity of microorganisms in 3 times. The content of extracted bitumenlike compounds decreases to 40 %. A biochemical essence of stimulating effect of HA wascaused by the activation of cell fermentative activity which depends on a growth rate and biomass
accumulation and mechanochemical activation of peat allows to improve the efficiency of HA
extraction and can cause the increase in availability of their separate components and parts ofmolecules, including micro-and macroelements which are used as sources of nutrition and energy
by a microbic cell.
Obtained data will make it possible to extend our knowledge about the influence of HA on
the development of hydrocarbon-oxidizing soil microflora and can represent a practical interestfor the development of technology of new humic preparations production from peat which can beused for biological recultivation of petropolluted soils.
KEYWORD: peat, mechanochemical activation, humic acids, petropolluted soil, hydrocarbonoxidizing microorganisms

23

Humic Substances in Ecosystems 8, 2009
LEAF LITTER FALL DECOMPOSITIO I AGE-DIFFERETIATED POPLAR
STADS OF HYBRID 275

Jerzy Jonczak
Pomeranian University in Słupsk, Department of Geomorphology and Quaternary Geology,
Partyzantów str. 27, 76-200 Słupsk, Poland
E-mail: jonczak@apsl.edu.pl
Plant litter fall decomposition is a critical stage in nutrients turnover in ecosystems. Habitat
conditions such as temperature, humidity and the biological activity of the soil, as well as thechemical composition of the initial material, are decisive factors determining the intensity of thisprocess. The contents of such substances like lignin, waxes, tannins and resins greatly influence
the rate of decomposition. These substances resist decomposition and make organic remains lessattractive to microorganisms. Chemical composition, and in particular the presence of diverse
substances resistant to decomposition, is a feature of each plant species and each fraction of thelitter fall. This feature changes over time. As plants age the chemical composition of their organs
might change, each to a different extent, which is then reflected in the chemical properties of thelitter fall. Considering this, it might be expected that the decomposition rate of fractions of thelitter fall would be connected with the age of the contributing plants. The aim of this study is tocompare the decomposition of leaves, the main component of the plant litter fall, among
plantations of Black Poplar (Populus nigra L.) clones Hybrid 275 of different ages.
The study was conducted in 2000-2002 at three plantations of Hybrid 275 Black Poplar ofdifferent ages. These plantations are in northern Poland in Wierzbiczany (17 years old), Gronowo(28 years old) and Ostromecko (54 years old) forest districts. All these plantations have similar
total precipitation, temperature ranges and soil nutrients abundance. Studies of the decomposition
were conducted using 30 cm x 30 cm litter bags made of 1 mm mesh nylon netting. Each bag was
filled with 15 g of dry leaves collected in the autumn of 2000 when litter fall was at its maximum.
Ten bags were located at each of 5 points in each study plot. Every two months one bag wascollected from each point and analyzed (content of C, N, P, K, Mg, Ca). The content of lignin in
the initial material was measured by weight after extraction with 72% H2SO4.
The process of decomposition occurred at a different rate at each plantation. It was fastest in
the 54-year-old stand (54Y), slightly slower in the 28-year-old stand (28Y) and slowest in the 17year-old stand (17Y). After 12 months in the 54Y stand almost 80% of the initial mass was
decomposed, in the youngest stand it was slightly more than 40%. The coefficients ofdecomposition, based on Olson’s model, were 1.49 for 54-year-old plantation, 0.97 for 28-yearold plantation and 0.61 for 17-year-old plantation. Poplar leaves decomposed quickly incomparison with other broadleaved trees. The index of decomposition was closely related to theage of the trees, with a correlation coefficient of 0.99. Because the temperature and humidity
conditions were similar at all locations, the different rate of decomposition can be attributed tothe different chemical composition of the initial material. The content of phosphorus, calcium andmagnesium varied greatly in the initial material, and the content of the nitrogen, lignin and theratio C/N varied less. However, only the lignin content was correlated with the age of poplars,with their proportion decreasing with the age of the trees. A high correlation coefficient between
the content of lignin and the intensity of decomposition of leaves (-0.97) was obtained. The
important role these compounds play in shaping the rate of decomposition of plant remains has
been pointed out by many authors. Other important factors that affect the rate of decomposition
are the content of nitrogen and the ratio C/N in the initial material. These are limiting factors in24

Humic Substances in Ecosystems 8, 2009
the development of the soil microorganisms responsible for the decomposition. The content of
nitrogen, both in the initial material and in the humic horizon of soils (0.082 – 0.087 g·kg-1), wassimilar at all stands. So this was not the differentiating factor. The ratio C/N in the initial material
was high and similar at all the plantations. During decomposition at all the locations the ratio C/Nnarrowed to values of about 25:1, which was linked to the immobilisation of the nitrogen.
Nitrogen concentration had a constantly increasing trend. On the one hand this was an effect ofslower rate nitrogen was released compared with other elements, and on the other hand it
reflected the absolute accumulation of nitrogen, due to its influx with rainwater, the binding of
atmospheric nitrogen by microorganisms, and also the result of mycelium filaments penetrating
the litter bags. After the first 10 months of the experiment the store of nitrogen in thedecomposed material increased, despite the loss of leaf mass. Immobilization of nitrogen occurs
with a high ratio of C/N. A strong deficit of nitrogen in the relation to carbon takes place whenmicroorganisms decompose material with a high C/N ratio and all the nitrogen released in thisprocess is used by the microorganisms to build their own proteins. An actual release of nitrogenat all the poplar stands was observed only in the second year. Potassium was leached out quickly
during decomposition. At all the plantations most of the potassium store had already beenreleased during first two months. This tendency has often been observed by other researchers.
Potassium occurs in organic remains as ions, which are easily leached out by rainwater. The
concentration of phosphorus and magnesium remained relatively constant during decomposition,
which differed at each plantation. The stores of these elements decreased with the loss of leaf
mass. The concentration of calcium tended to rise at all the plantations, but its supply decreased
gradually.
Despite the differences in the chemical composition of the initial material at the poplar
plantations analysed, the dynamics of the concentrations of each nutrient and of their supplies
showed similar trends.
The study showed a high intensity of leaves decomposition at plantations of poplar clone
Hybrid 275 in comparison with other species of broadleaved trees. A high correlation coefficient
between the age of poplars and the rate of decomposition was found. Leaves of old trees
decomposed quicker than those of younger trees. The observed differences most probably result
from different chemical composition of the initial material. Distinct differences were noted
between the plantations in the content of phosphorus, potassium and magnesium. Only for lignin
were high negative correlations obtained between their concentration and the age of the poplars
from which the leaves originated, and the index of decomposition. Only small differences wereobserved in the nitrogen content and the C/N ratio, which excludes these as differentiating
factors.
KEYWORDS: poplar, Hybrid 275, leaf litter fall, decomposition, nutrient turnover25

Humic Substances in Ecosystems 8, 2009
SOIL ORGAIC MATTER AD PHYSICAL CHARACTERISTICS
1*

Hana Karabcová, 2Marie Svozilová, 1Jana Krhovjáková

1

2

Agroresearch Rapotín, Ltd., Výzkumníků 267, 788 13 Vikýřovice, Czech Republic
Research Institute for Cattle Breeding, Ltd., Rapotín, Výzkumníků 267, 788 13 Vikýřovice, Czech
Republic*Corresponding author E-mail: hana.karabcova@vuchs.cz

The paper deals with soil organic matter and its influence on physical soil parameters on
the permanent grassland. In 2005 – 2007 the effect of these attributes were observed in conditionsof the Hrubý Jeseník Mountains. The experiment is situated in 400 m above sea level. The soil issandy-loam of the cambisol type. We used organic fertilisation with three graduated systems ofload. The soil bulk density, total porosity, maximum capillary water capacity from physicalproperties and organic soil carbon were determined. Soil samples for determination of physicaland chemical properties were taken from depth 0.02 – 0.15 m and 0.15 – 0.30 m in spring. The
aim of this work was to show relationship between organic soil carbon and selected physicalcharacteristics in these conditions. Statistically significant relationship was found between
organic soil carbon and soil bulk density. No statistically significant impact was found between
organic soil carbon and porosity and maximum capillary water capacity. Obtained data were
tested by GLM statistics.
KEYWORDS: soil organic matter, bulk density, porosity, maximum capillary water capacity

26

Humic Substances in Ecosystems 8, 2009
EFFECTS OF AGROTECHICAL TREATMETS O SPECTROPHOTOMETRICPROPERTIES OF HUMIC ACIDS OF BROW PODZOLIC SOILS

Krystyna Kondratowicz-Maciejewska
University of Technology and Life Sciences, Department of Environmental Chemistry,
6 Bernardynska St., 85-029 Bydgoszcz, Poland.
e-mail: chem@utp.edu.pl
Characteristics of humic substances is usually done by investigations of the properties ofsoil extracts isolated with the use if selective solvents and by the analysis of some groups of
selected compounds, such as humic and fulvic acids. One of the criterions used in thecharacteristics of a complex nature of humic substances are the values of absorbance of theirsolutions in the range of ultraviolet and visible light (UV and VIS). Also infrared spectroscopy(IR) is an important method of such studies of humic substances. The objective of theinvestigations was to evaluate effects of agrotechnical treatments on physicochemical propertiesof humic acids of brown podzolic soils. The investigations were carried out as a part of a longterm static experiment of the Puławy ITFSS. There were three experimental factors: 1 – crop
rotation (A-potato, wheat, spring barley, corn; B-potato, winter wheat + charlock as aftercrop forploughing, barley with companion crop, mixture of clover with grasses), 2 – fertilization withmanure, 3 – fertilization with mineral nitrogen. Humic caids separated by according to Schnitzer
were analysed for optical properties in the UV-VIS range and their IR spectra in the range 4004000 cm-1 were recorded. Crop rotation appeared to be the basic factor affecting the opticaldensity properties of humic acids. Rotation of crops enriching the soil in organic matter caused
higher absorbance values of humic acids solutions than in the one depleting the soil. Higher
values of the A2/4 and ∆log K parameters in the crop rotation B indicated that humic acids
isolated from the soil suplemented with organic matter had a higher content of humus in thestarting point of decomposition than humic acids of the variant depleting the soil. The kind ofcrop rotation modified spectroscopic parameters of humic acids. Humic acids of the soil underrotation of crops introducing less organic matter had a higher percentage of lignin structures thanthose of other one crop rotation. As compared with mineral fertilization manuring generated thedevelopment of humic acids of a smaller molecular weight. A clear presence of absorption bands
in the range 1460-1000 cm-1 in the IR spectra of humic acids separated from the combinationenriching the soil in organic matter showed their “chemical youth”.
KEYWORDS: humic acids, fertilization, crop rotation, IR, UV-VIS

27

Humic Substances in Ecosystems 8, 2009
THE EFFECT OF THE ECOLOGICAL AD ITEGRATED SYSTEM O THE SOIL
PHOSPHATASE ACTIVITY

Jan Koper, Joanna Lemanowicz
Department of Biochemistry, Faculty of Agriculture, University of Technology and Life Sciences,
6 Bernardyńska St., 85-029 Bydgoszcz, Poland, tel. (052) 374-95-59,
E-mail: lemanowicz@utp.edu.pl
The aim of the present paper was to determine the effect of organic and integrated farmingwith varied fertilization on the phosphatase activity of soil and how it affects the content ofphosphorus available for plants. In the organic system there was used only organic fertilization in
a form of cattle manure, while in the integrated system – organic fertilization combined withmineral fertilization (NPK). The soil was sampled twice during the vegetation period of crops: inspring – in April and in autumn – in September . It was found that both the organic system andthe integrated system affect the changes in the parameters researched. There was demonstrated anincrease in the content of available phosphorus in the soil sampled in autumn. There wasobserved a significant effect of the farming system on the activity of alkaline and acidphosphatases.
KEYWORD: available phosphorus, activity alkaline and acid phosphatase, soil

28

Humic Substances in Ecosystems 8, 2009UPTAKE OF HUMIC SUBSTACES BY WHEAT PLATS:
PREFERETIAL ACCUMULATIO I LIPID FRACTIO

Natalia A. Kulikova1, Eugenia A. Tsvetkova2, Gennady A, Badun3, Maria G.
Chernysheva3, Viktor I. Korobkov3, Zinaida A. Tyasto3, Irina V. Perminova3
1

Department of Soil Science, Lomonosov Moscow State University, 119991 Moscow, Russia
2;.D. Zelinskii Institute of Organic Chemistry of RAS, 119991 Moscow, Russia
3
Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia
E-mail: knat@darvodgeo.ru

Humic substances (HS) have been the subject of numerous scientific studies due to theirbeneficial effects of HS on living organisms. The principal effects exhibited by HS onto livingorganisms include an increase in biomass accumulation, nutrient uptake, biosynthesis, antiviral
activity, and others [1]. In spite of numerous studies on the biological effects of HS, themechanism of their action remains unclear. The main reason seems to be the stochastic nature of
HS. In contrast to common biological macromolecules, which are synthesized by a livingorganism according to the information encoded in DNA (nuclear acids, proteins, enzymes,
antibodies etc.), HS are the products of stochastic synthesis. They are characterized aspolydisperse materials having elemental compositions that are non-stoichiometric, and structures,
which are irregular and heterogeneous. The above features hamper a use of common biologicalapproaches to study biological activity of HS.
Several studies on the uptake of HS by higher plants have been performed. Earlier work
relied on the color changes in the plant organs as an indication of HS uptake. The more recent
study used 14C-labeled humic-like substances or 3H-labeled HS [2, 3]. Those studies alsoestablished that both humic-like substances and HS and were taken by the plants. Therefore, next
goal in going deeper inside understanding mode of action of HS is a study distribution of HS inplant tissues. This study was aimed to find out uptake and distribution of HS in wheat seedlings.
For this study humic acids (HA) derived from leonardite and IHSS standard fulvic acids(FA) were used. To synthesize isotope-labeled HA, an approach developed in [4] for the
preparation of tritium-labeled HS was used. The obtained 3H-HA sample was dissolved in aphosphate buffer (0.028 M, pH 6.8) and purified by dialysis during a month at 4ºC. It allowed
eliminating exchangeable tritium of OH–, COOH–, and NHn groups of HS. Seeds of wheat
Triticum aestivum L. were germinated at 24°C in the dark for 72 h followed by transferring
seedlings into the 0.5 l tanks containing Knopp nutrition solution. After another 72 h plants weretransferred into the vials containing HS at concentration 50 mg l–1 with specific radioactivity 0.10.2 Curie l–1. Then lipid fraction was extracted according to [5] followed by radioactivity
measurements.
Obtained results are presented in the Table 1.
Table 1. Contents of HS in lipid fraction of wheat plantHS
Coal HA
Aquatic FA

Contents of HS in lipid fractions, % of taken up
Roots
Shoots
Total
82±6
16±2
98±8
49±10
16±8
64±5

29

Humic Substances in Ecosystems 8, 2009
As it can be seen from the Table, almost entire amount of HA that were taken up by plants
were found in lipid fraction (98%); for FA that value decreased to 64%. Taking into account that
contents of lipid fraction in plants were about 3-5%, one could conclude on preferential
accumulation of HS in lipid fraction. Accumulation coefficient which was calculated as ratio of
HS concentration in plant to concentration in lipids was as 43±2 and 25±2 for HA and FA
respectively. On the other hand, lipid contents in HS usually vary in the range 3-8%. Therefore,
preferential HS accumulation in lipid fraction was evident for preliminary transformation of HS
on the root surface or nearby it before HS entrance into the plants. That finding could be also
confirmed by equal amounts of HA and FA in lipid fraction of shoots (16%). The latter indicatedthat only specific fragments of HS, probably hydrophobic, were able to enter plants vascularsystem.
So, our experiments demonstrated that HS taken up by plants were preferentiallyaccumulated in lipid fraction. Before entrance HS were seemingly to be partly transformed andmainly their hydrophobic fragments were adsorbed by plants. However, lipid fraction in plants iswell known to be a wide group of natural compounds distinguishing greatly in both structure andfunctions. They are presented by several different compounds including pigments, fats, fatty
acids, waxes, steroids, cutin, suberin and others. Therefore, further study of distribution of HS inlipid fraction of plants is required.
KEYWORDS: wheat, uptake, lipid, tritium, label

30

Humic Substances in Ecosystems 8, 2009
THE STRUCTURE AD PROPERTIES OF HUMIC SUBSTACES FROM BROW
COAL 11 YEARS AFTER ADDITIO ITO SOIL

Jolanta Kwiatkowska-Malina
Department of Spatial Planning and Environmental Sciences, Warsaw University of Technology,
Pl. Politechniki 1, 00-661 Warsaw, Poland.
E-mail: j.kwiatkowska@gik.pw.edu.pl
Transformations (mineralization and humification) of soil organic substances lead toformation of humic substances (HS), the most spread in nature group of organic compounds.
Diversity of organic matter and the environment create varying structures and compositions, andthus differences in properties of HS. Soils in many areas of Poland are characterized by low
contents of organic matter, which greatly contributes to their limited fertility and productivity,and higher vulnerability to contamination and degradation. Soil degradation in some areas causes
a dramatic need to improve the structure and contents of organic matter.
The requirements of EU legislation are key factors controlling soil protection. Although
the EU has no soil directive yet, 2 documents have an impact on the requirements for national
legislation and policy on soil protection. In the Communication “Towards a Thematic Strategy
for Soil Protection” (COM 2002, 179), depletion of soil organic matter is mentioned as one of 8
main hazards for soils. According to the Directive establishing a framework for the protection ofsoil (COM 2006, 232, final), soil can be considered a non-renewable resource as degradation
processes are usually faster that soil formation and remediation processes.
Recently, the use of organic amendments other than traditional manure showed to beefficient for the improvement and/or restoration of soil organic matter. One of the potential
sources of organic matter in soil is an organic-mineral preparation obtained from brown coal, theso-called Rekulter (Kwiatkowska et al 2005). Organic matter from brown coal added to soil may
be transformed into HS in (micro)biological and abiotic ways.
The objective of this study was the qualitative evaluation of structure and physicalchemical properties of HS in soil 11 years after application of brown coal in the form of Rekulter(a dose of 40 t ha-1) as a source of organic matter. Structural analyses were carried out for humic
acids (HAs) extracted from the Haplic Luvisol, and they include: solide-state 13C NMR
spectroscopy, electron paramagnetic resonance (EPR), infrared and UV-VIS spectra, share ofhydrophilic and hydrophobic fractions by High Performance Liquid Chromatography (HPLC), as
well as elemental composition and parameters of thermal decomposition by differential scanningcalorimetry (DSC) measurement.
Application of organic matter from brown coal resulted after 11 years in an increases of
the carbon contents of HAs extracted from soil, and consequently decreased values of the H:C
ratio. HAs of amended soil showed higher absorbance values in the UV-VIS region, higher
contents of oxygen functional groups, and were more resistant to thermal decomposition
compared with non-amended ones (control). HAs molecules were also richer in aromatic
structures and resistant to decomposition with a high potential of sorption properties. For FT-IR
spectra of HAs an increase of the relative intensity of peaks at 1720 cm-1 with respect to that at
1670 cm-1 was observed, thus providing spectroscopic evidence of higher contents of carboxylic
groups in aromatic rings, as compared to HAs extracted from non-amended soils.
Organic matter from brown coal introduced to soil in the form of Rekulter relatively
slowly undergoes transformation and causes permanent soil enrichment with HS of properties

31

Humic Substances in Ecosystems 8, 2009
similar to these originating from traditional sources. It also demonstrates long-term positiveeffects on physical-chemical parameters, composition and structure of SH and soil.
Table 1. Elemental composition (in atomic percentage), atomic ratios and degree of internal oxidation (ω)ω) of
HA extracted from soil

ObjectCH
N
O„0” - control
36,1 43,4 2,7
17,8-1
36,6 45,0 2,6
15,8
Rekulter (40 t ⋅ ha )notation: ω - internal of oxidation degreee

H:C
1,20
1,23

O:C
0,49
0,43

O:H
0,41
0,35

N:C
ω
0,074 0,002
0,072 0,150

Fig. 1. FTIR spectra of HAs extracted from soil

Table 2. Spectral properties of HAs extracted from soil

Object„0” control
Rekulter (40 t ⋅ ha-1)

A280
5,2
6,9

A465
0,99
1,13

A600 A665 A2/4
0,362 0,201 5,20
0,403 0,226 6,09

A2/6
25,7
30,3

A4/6
4,95
4,98

∆logK
0,680
0,690

Table 3. Percentages of some carbon links in HAs molecules calculated from 13C MR spectra

ObjectCal
Car
Clig
C-COOH P„0” - control
54,5
30,4 5,3
9,7
14,9
.
-1
Rekulter (40 t ha )49,532,9 7,0
10,6
17,5
notation: Cal, Car, Clig – C in aliphatic, aromatic and lignite structures
KEYWORD: structure, properties, soil, humic substances, brown coal32

Cal/Car1,79
1,50

Humic Substances in Ecosystems 8, 2009
THE IFLUECE OF BROW COAL O PHYSICO-CHEMICAL PROPERTIES OFSADY SOILS

Alina Maciejewska, Jolanta Kwiatkowska-Malina
Department of Spatial Planning and Environmental Sciences, Warsaw University of Technology,
Pl. Politechniki 1, 00-661 Warsaw, Poland
E-mail: j.kwiatkowska@gik.pw.edu.pl
Soil organic matter plays an important role in stability and fluxes of trace greenhousegases between land surface and atmosphere. A steady decline of organic matter in soils in Poland
is observed, therefore the pending problem and new challenge for soil and environmental
scientists is to increase its level, particularly in sandy soils. A growing interest was observed inlast decades in different sources of organic matter, for example from brown coal, its derivatives
and composts that are considered as an important source of enriching the soil with organic matter(Dębska et al. 2002; Kwiatkowska et al. 2005).
The long-term experiment was carried out in a test field in Klon on rusty soil originatedfrom light sands. The Rekulter – a brown coal fertilizer was introduced 20 cm deep into the soil
in autumn 1987 at 2 doses of 80 and 160 tons/ha. After 1 (1988) and 20 (2008) years soil samples
were taken at the depth of 20 cm, and the following parameters were determined: pHH20 and
pHKCl, hydrolitic acidity (Hh) and total exchange basic (TEB) cations by the Kappen’s method,
the total organic carbon (TOC)–by the Tiurin method, and the total nitrogen (Nt) content–by the
Kjeldahl method. Additionally, cations exchange capacity (CEC) and base saturation (BS) were
calculated.
Applying the Rekulter resulted in changes of physicochemical properties of the soil (Table
1). One year after application, the highest increase in soil reaction pH (pHH2O=7.5; pHKCl=7.4)
was found in the case of higher dose of Rekulter, and the same behavior (pHH2O=7.2; pHKCl=6.4)
was observed 20 years after its application. In objects with higher dose of the Rekulter, the TOC
decreased from about 20 g.kg-1 after 1 year to 15 g.kg-1 after 20 years of its application. The
highest growth of the Nt content of 1.1 g.kg-1 (1988) and 0.9 g.kg-1 (2008) occurred at higher doseof the Rekulter. The highest value of TOC to Nt ratio of 20 obtained for soil sample with higher
dose of the Rekulter was due to the highest carbon content in this object. In other experimentswith the Rekulter and brown coal amendments similar results were obtained (Maciejewska et al.
2001).
Table 1. The effect of the Rekulter on basic properties of soil samples

SampleYear

Dose(t ha-1)“0” (control)

1988
2008
1998
2008
1988
2008

0
0
80
80
160
160

pH in H2O

pH in KCl
(g.kg-1)

TOC(g.kg-1)

Nt

TOC:N

5.6
4.7
7.6
0.6
12.7
5.9
4.7
6.3
0.4
14.6
Rekulter
7.0
6.8
14.5
0.9
16.1
7.1
6.3
11.5
0.7
16.2
Rekulter
7.5
7.4
22.1
1.1
20.1
7.2
6.4
15.5
0.9
19.5
LSD = 0.05
0.2
0.4
0.7
0.08
Selected physical-chemical properties of soil after application of the Rekulter arepresented in Table 2. A decrease of Hh was observed in the case of both doses applied, with the33

Humic Substances in Ecosystems 8, 2009
lowest value of 0.7 cmol (+) .kg found in the case of higher dose of the Rekulter. The highsorption capacity of the Rekulter significantly influenced TEB, CEC and BS of amended soil.
The Rekulter applied to soil demonstrates long-term positive effects on basic properties
and physical-chemical parameters of the sandy soil.
Table 2. The effect of the Rekulter on hydrolitic acidity (Hh), total exchange basic (TEB) cations, cationexchange capacity (CEC) and base saturation (BS) of soil

SampleYear“0” (control) 1988
2008
Rekulter
1988
2008
Rekulter
1988
2008
LSD = 0.05

Dose of amendment Hh TEB(t ha-1)
cmol(+) .kg-1
0
4.1 2.60
3.8 3.680
0.9 13.980
1.9 19.7160
0.4 32.0160
0.7 36.6
0.1 2.1

CEC
6.7
7.4
14.7
21.6
32.4
37.3
3.3

BS
%
39.3
48.6
93.9
91.2
98.9
98.1
4.2

KEYWORD: organic matter, physico-chemical properties, soil, brown coal, Rekulter

34

Humic Substances in Ecosystems 8, 2009
RELATIOS OF EZYME ACTIVITIES WITH DIFFERET FRACTIOS OF SOIL
ORGAIC MATTERNatallia Mikhailouskaya and Iossif Bogdevitch
Research Institute for Soil Science and Agrochemistrye, Kazintsa, 62, 220108 Minsk,Belarus.
E-mail: bionf@yandex.ru, brissa5@mail.belpak.by
Important ecological function of humic substances is their capability of stabilization of
extracellular enzyme activities that are the primary means by which soil microorganisms
decompose complex organic compounds into assimilable subunits. Humus–enzyme complexes
have a functional capacity under unfavorable conditions; they are resistant to proteolysis as wellas to chemical or physical stresses. Stable humus pool and humus–enzyme complexes play anessential role in the functioning of soil systems.
The objective was to establish the relations of enzyme activities with the contents of stable
and active SOM fractions. Cellulase and invertase were chosen for their critical role in the C
cycle in soil. Urease was studied because of its role in releasing inorganic nitrogen in the N cycle.
Long-term organic amendments (8 and 16 t ha-1 of FYM in rotation) positively affected
the contents of stable and active SOM fractions in Luvisol loamy sand soil. Close correlations
were found between stable humus pool and hydrolase’s activities in soil (R2; P < 0.05): forcellulase – 0.71 – 0.91; for invertase – 0.88 – 0.92; for urease – 0.73 – 0.76. Regression analysis
revealed reliable but not so close relationships of hydrolase’s activities with decomposable
fractions of SOM (R2; P < 0.05): for cellulase – 0.63 – 0.68; for invertase – 0.57 – 0.69; forurease – 0.66 – 0.71.
KEYWORD: SOM stable and active fractions, cellulase, invertase, urease35

Humic Substances in Ecosystems 8, 2009
BIOLOGICAL ACTIVITY OF HUMIC SUBSTACESAD THEIR FE-ERICHED DERIVATIVES TOWARDS CUCUMBER PLATSUDER FE-DEFICIECY CODITIO

Philippova O.I.1, Sorkina T.A.2, Kulikova N.A.1, Zhelezova S.V.1, Lebedeva G.F.1,
Perminova I.V.2
1

Department of Soil Science, Lomonosov Moscow State University, 119991 Moscow, Russia
2
Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia
E-mail: philolga@mail.ru

Humic substances (HS) are generally considered to increase availability of micronutrientssuch as Fe, Zn, Mg, Mn, Ca and others, low levels of which cause leaf yellowing called chlorosis.
So, some HS preparation enriched with micronutrients are available in the agricultural market.
The observed beneficial effect of HS is generally attributed to the chelating activity of HS thatprovides plants with micronutrients in easily assimilated form. However, HS are known to beuniversal bio-activators providing plant adaptation to stress of various nature. Therefore HS
could be expected to mitigate negative effects of chlorosis due to their own biological activity.
This study was aimed to compare influence of HS, their Fe-enriched derivative and commerciallyavailable iron supplier on cucumber plant growth under Fe-deficiency conditions.
Following preparation were used: potassium humate derived from coal (Solntsevskoe
deposit, Sakhalin Island, Russia) assigned as K-Sh, its Fe-enriched derivative assigned as Fe-HS,and commercially available agricultural iron additive ferrous chelate of ethylenediaminedi(ohydroxyphenylacetic) acid (Fe-EDDHA). The content of iron was measured using ophenantroline method after oxidative digestion and was as 1%, 9%, and 6% in K-HS, Fe-HS and
Fe-EDDHA respectively.
To estimate biological activity of HS under iron deficiency conditions we appliedbioassay technique. Plants of cucumbers Cucumis sativus L. were used as a target object.
Cucumber seeds were grown in distilled water for 120 hours at 25°C in the dark followed bytransferring seedlings into 0.05 mM solution of CaSO4 for 24 hours. Then seedlings weretransferred to plastic pots containing 0.1 l of Hoagland’s nutrition solution supplied with
25 µmol/l of iron and 15 mg/l of HS. Values of pH of all the solutions were adjusted to 8. The
cucumber plants were grown with 12-hours light day. Hoagland’s nutrition solution without Fe
was used as blank.
After 32 days of growing plant photosynthesis efficiency were estimated in terms of the
maximum activity of PSII (Fv/Fm) using pulse amplitude modulation (PAM) fluorometer (PAM2000, Walz, Germany). Then cucumber plants were harvested and subjected to measuring shoots
and roots length and weight and chlorophyll content. The content of iron in plant tissue wasmeasured using o-phenantroline method after oxidative digestion.
The obtained data showed that in the lack of iron in the nutrition media poor cucumber
growth was observed. Plants demonstrated sings of chlorosis such as yellow or yellow-whiteleaves, poor photosynthesis efficiency (Fv/Fm = 0.45), and low chlorophyll a/b ratio (1.8). Wheniron was added in the form Fe-Sh or Fe-EDDHA no chlorosis symptoms was observed, andvalues of Fv/Fm and chlorophyll a/b ratio increased to 0.77-0.78 and 2.4-2.6. As a consequence,
the biometric parameters of the treated plants – length and weight – were substantially increased
as well (Figure 1).

36

Humic Substances in Ecosystems 8, 2009
% of blankweightlengtha600% of blank

600

500

500

400

400

300

300

200

200

100

100

0

0
blank

FeEDDHA

Fe-Sh

weightlength

b

blank

K-Sh

FeEDDHA

Fe-Sh

K-Sh

,

Figure 1. The influence of commercial iron chelate Fe-EDDHA parent humic material K-Sh and its Feenriched derivative Fe-Sh on cucumber shoots’(a) and roots’ (b) weight and length increase.

As it can be seen from the Figure 1, plants treated with Fe-EDDHA or Fe-Sh possessedhigher weight and length of both shoots and roots as compared to blank. However, efficiency ofstudied iron suppliers did not differ one form another if shoots parameters were used as a targetfunction, whereas substantial differences were observed for both length and weight of roots.
Of interest was that addition of K-Sh solely also led to a partial recovery of the Fedeficient plants. In particular, the photosynthesis efficiency (Fv/Fm = 0.77) and chlorophyll a/b
ratio (2.4) were significantly increased in humus-treated plants. At that, length and weight of
cucumbers’ shoots and roots were similar to those of plants treated with Fe suppliers.
Fe content in the leaves of treated plants were estimated as 508, 158, and 93 mg/kg of dry
weight for Fe-EDDHA, Fe-Sh, and K-Sh respectively. The highest iron content was found in FeEDDHA treated plants whereas humics treated plants possessed much lower iron. The latter was
evident for the fact that both Fe availability in the nutrition media and beneficial effect of HS
were responsible for plants’ recovery from Fe-deficiency chlorosis. So, mitigating activity ofhumic-based Fe additives under Fe-deficiency conditions resulted not only from the fact thatplants could use Fe, but from protective activity of HS in relation to photosystem II either.

37

Humic Substances in Ecosystems 8, 2009

PROPERTIES OF HUMIC SUBSTACES EXTRACTED FROM BOTTOM SEDIMET
AT GOCZAŁKOWICE DAM RESERVOIR

J. Polak, M. Bartoszek, W.W. Sułkowski
Department of Environmental Chemistry and Technology, Institute of Chemistry,
University of Silesia, Szkolna 9, 40-006 Katowice, Poland, wieslaw.sulkowski@us.edu.pl
Studies of humification processes of bottom sediments collected from various points of
the Goczałkowice dam were conducted. The humic and fulvic acids were extracted from thecollected sediment. For study of physico-chemical properties of humic and fulvic acids EPR andIR spectroscopy was used. For extracted humic and fulvic acids, concentration of free radicals
and the g factor were determined with EPR. The presence of characteristic functional groups was
confirmed with IR spectroscopy.
It was found that humic and fulvic acids extracted from bottom sediment at 8 places of the
Goczałkowice Reservoir are characterized by similar physico-chemical properties, which points
to a similar composition of bottom sediment in the whole reservoir. Humic acids are relatively
high aromatised, have a low free radicals content and a low value of the g factor. They havecondensed aromatic rings with side-aliphatic chains. However, the extracted fulvic acids are lessaromatised than humic acids. They have a weakly formed aromatic core with the dominance ofside- aliphatic chains.
The influence of the depth of the lake on the content of oxygen functional groups in theextracted humic and fulvic acids was also observed.
KEYWORD: humic acids, fulvic acids, bottom sediment, EPR, IR

38

Humic Substances in Ecosystems 8, 2009
CHARACTERIZATIO OF SOIL ORGAIC CARBO AD ITS FRACTIO LABILE
CARBO I ECOSYSTEMSĽubica Pospíšilová,Veronika Petrášová, Jiřina Foukalová, Eduard Pokorný
Institute of Agrochemistry, Soil Science, Microbiology and Plant ;utrition, Mendel University ofAgriculture and Forestry, Zemědělská 1, 613 00 Brno, Czech Republic.
E-mail: lposp@mendelu.cz
Parameters evaluating humus content and quality were determined in soils under different
management systems. Our work represents a part of long - term field experiments that are carriedout in the Czech Moravian Upland. Soil samples were selected from the topsoil of the following
soil types: Leptic Cambisol (Ocmanice, grassy soil), Haplic Cambisol (Náměšť n/Oslavou,
arable soil), Eutric Cambisol (Vatín, grassy soil), Eutric Cambisol (Vatín, arable soil). Among theall parameters labile soil carbon and basal respiration activity were measured. Oxidimetrictitration for total organic carbon content determination was used. Humic substances (HS) sum,humic acids (HA) sum and fulvic acids (FA) sum were determined by short fractionation method.
Basal respiration was determined under laboratory conditions using Vaisala GMT 222 apparatus.
Hot water extraction for labile organic carbon determination was used. UV-VIS spectra were
measured in the mixture of 0.1 M sodium pyrophosphate and 0.1 M NaOH.
Results showed that total carbon content varied from 1.3 – 1.8 %. Higher content wasdetermined in grassy soils. Labile carbon content was higher in grassy soils to compare witharable soils (varied from 540 – 620 mg/kg). HS sum was the lowest in grassy soil (locality
Ocmanice) about 6.5 mg/kg. Quality of humus was low and HA/FA ratio was less than 1 for all
samples. UV-VIS spectral characteristics showed low absorbance and high color indexes Q4/6 forall samples. Humification degree was low (less than 20%). Basal respiration was higher in arable
soils to compare with grassy soils. Production of CO2 varied from 0.31 – 3.8mg CO2/100g/h.
Complex factor (Fig.1) is connected with C and N microbial utilization after both elements
application to the soil samples. Results showed higher average values in grassland. We canconclude that total carbon content, labile carbon content, basal respiration and humic substances
quality were mainly affected by land use.
KEYWORD: labile soil carbon, basal respiration activity, humus fractionation, UV-VIS
spectroscopy
This study was supported by the Research plan No. MSM6215648905 “Biological and Technological
Aspects of Sustainability of Controlled Ecosystems and their Adaptability to Climate Change“, which is financed by
the Ministry of Education, Youth and Sports of the Czech Republic and by the project NAZVA QH 81200.

39

Humic Substances in Ecosystems 8, 2009
SOIL ORGAIC CARBO AD PARTICLE SIZE DISTRIBUTIO OF ARABLE
SOILS I WITER WHEAT CROPPIG

Srdjan Seremesic, Dragisa Milosev, Maja Manojlovic
Department of Field and Vegetable Crops, Faculty of Agriculture ;ovi Sad, Sq. D. Obradovica
8, 21000 ;ovi Sad, Serbia
Email: srdjan@polj.ns.ac.yu
In order to investigate effects of cropping systems on content of soil organic carbon (SOC)and distribution of particle size fractions of soil, different cropping systems of winter wheat wereanalyzed. Soil samples were collected from the long-term experiments carried out at the Rimski
Sancevi experimental station, Novi Sad. The following treatments were analysed:
1. Three–year crop rotation (maize-soybean-wheat), mineral fertilizer 100 kg N ha-1 + crop
residues
2. Two–year crop rotation (maize-wheat), mineral fertilizer 100 kg N ha-1 + crop residues
3. Monoculture (wheat-wheat), mineral fertilizer 100 kg N ha-1 + crop residues
4. Two–year rotation (maize-wheat), without fertilizers + crop residues
5. Three–year rotation (maize-soybean-wheat), without fertilizers + crop residues
6. Four–year crop rotation (sugar beet-spring barley-maize-wheat), 40 t ha-1 manure without
mineral fertilizers
7. Four–year crop rotation (sugar beet-spring barley-maize-wheat), 40 t ha-1 manure + mineral
fertilizer 100 kg N ha-1
8. Four–year crop rotation (sugar beet-spring barley-maize-wheat), mineral fertilizer 200 kg
N ha-1without crop residue
9. Four–year crop rotation (sugar beet-spring barley-maize-wheat), mineral fertilizer 200 kg
N ha-1 + crop residues
10. Control, native vegetation with short grasses (the adjacent land which has never beencultivated)
Three soil depths were analyzed (0-20, 20-40 and 40-60 cm). Organic carbon content wasdetermined by CHNS analyzer. Particle size fractions (>2000 µm, 2000-250 µm, 250-53µm and<53µm) were obtained by dispersion in water with series of sieves.
The use of organic and mineral fertilizers influenced SOC content and particle size
fractions. Obtained results showed deterioration of soil structure in arable soil compared with
control as a result of applied management practice. Significantly higher content ofmacroaggregates in 0-20 cm soil layer was found in soil samples from control plots comparedwith wheat cropping systems. Addition of manure without crop residue incorporation was not
sufficient for soil structure preservation. In arable soil higher content of microaggregates (<250µm) was found in 0-20 and 20-40 cm soil layers, whereas 40-60 cm depth had equal proportionof macro- and microaggregates as clay fraction predominates.
SOC content decreased with the depth and differences in SOC among depths wereassociated with the below-ground biomass of crops in rotation. Higher content of SOC in 0-20
cm and 20-40 cm depths of different size fractions was observed with addition of mineral
fertilizer (200 kg N ha-1) and incorporation of crop residues. An increase of SOC content withincreased particle size was observed. Accordingly, macroaggregates turnover was crucial in theprocesses of soil organic matter stabilization.
KEYWORD: soil organic carbon, particle size fractions, cropping systems

40

Humic Substances in Ecosystems 8, 2009
THE ROLE ORGAIC MATTER AS SOLID SUPPORT OF HEAVY METALS I THE
WATER EVIROMET

Anna Rabajczyk, Elżbieta Sykała
Samodzielny Zakład Ochrony i Kształtowania Środowiska, Uniwersytet HumanistycznoPrzyrodniczy Jana Kochanowskiego w Kielcach, ul. Świętokrzyska 15G, 25-406 Kielce, Polska
E-mail: chromium@ujk.kielce.pl, sykala@ujk.kielce.pl
Metals are perceived as pollution but they are also natural substances in environment.
Metals get into the water environment from atmosphere as a result heavy rainfall or snowfall.
They are often leaches from rocks or soils. Contents of heavy metals are very often exceeded
because of emission from industrial land (Stögbauer et al., 2008). Among pollution the mostdurable are heavy metals. Their contents in water environment components is very divers, but thebiggest of heavy metals contents are in suspension (about 80%), while the littlest are in water(Adamiec and Helios-Rybicka, 2002; Aleksander - Kwaterczak and Helios - Rybicka, 2004;
Helios - Rybicka et al., 2005; Luck et al., 2008). The surface water environment is under the
influence of continuous industrial pressure.
Pollution of surface water environment which find to the water ecosystem direct orindirect occur various forms. They are solution as well as suspension. Suspense Particular Matter(SPM) is very essential component of surface water. SPM plays essential role in metals pollution
transport (Blo et al., 2003; Cenci, Martin, 2004). Solid phase of SPM consists of mainly frominorganic compounds but they are also organic compounds. Most trace elements, especially ofheavy metals don’t stay a long time in dissolve form. Heavy metals as few from inorganic
compounds have strong adsorptive properties. They are stopped on phase line among othersuspension and other solid body occurred in water. Heavy metals consist in surface watercomponents decrease in rows: suspension > deposit > water (Luck et al., 2008).Heavy metals
ratio in industrials components are very divers because of outside factor and biogeochemical
processes.
The aim of researches was determined of heavy metals contents in the water in form
dissolve and suspension. The samples of water were taken from Bobrza River, from source of ariver to Sitkówka - Nowiny. Chosen length a river encompasses different lands about various
degrees degradation of the environment. They are: source-naturals, Białgon - sour, Sitkówka –Nowiny - alkaline. In the samples of water was determining: pH, conduction, suspension andconcentration of heavy metals in the water in the form dissolve and suspension.
KEYWORD: heavy metals, organic matter, solution, suspension

41

Humic Substances in Ecosystems 8, 2009QUALITY OF SOIL ORGAIC MATTER I DIFFERET ECOSYSTEMS

Tobiašová Erika- Debska Božena- Horvátová Mária- Zaujec Anton- Szombathová NoraChlpík Juraj- Nádašský Ján
Department of Pedology and Geology, Slovak Agricultural University in ;itra (Slovak Republic).
E-mail: Erika.Tobiasova@uniag.sk
This study deals about quality of soil organic matter in natural ecosystems and arable land.
Sampling places were located in Nitra, Komjatice, Močenok and Šaľa on soil types Eutric
Fluvisols and Haplic Chernozems. There were four different ecosystems- agri-ecosystem (arable
land), forest ecosystem, meadow ecosystem and anthropic soil included. The highest qualityof soil organic matter, evaluated according to extracted carbon of humic acids, was in agriecosystem 19 %, next it was in forest ecosystem 18 %, meadow ecosystem 16 % and with thelowest value 13 % it was in anthropic soil. The highest soil organic matter quality in agriecosystem on Haplic Chernozems was in variant CM-03, with share of cereals 72 % (maize
42 %), root crops 14 % and oil crops 14 %. Just in this variant the highest carbon and nitrogencontents were determined. In Eutric Fluvisols the highest soil organic matter quality was invariant FM-04, with share of cereals 72 % (maize 14 %), root crops 14 % and with the highest
share of oil crops 14 % just in this crop rotation. In this variant also in one year spring cereal andmaize for grain were growing. In other variants maize was growing for silage.
KEYWORD: ecosystem, crop rotation, humic acids, soil organic matter

42

Humic Substances in Ecosystems 8, 2009COMPARISO THE PROPERTIES OF HUMIC ACIDS FRACTIOATED FROMAGRICULTURAL RICE AD FOREST SOILS

Erh-Shuan Kuo1, Tsung-Yao Tseng2, Kuo Chuan Lin3 and Ming Kuang Wang1*
1. Department of Agricultural Chemistry, ;ational Taiwan University, Taipei, Taiwan, 106; 2.
School of Forestry and Resource Conservation, ;ational Taiwan University, Taipei, Taiwan; 3.
Forestry Research Institute, Taipei, Taiwan.
This study aimed to evaluate the properties of humic acids (HA) fractionated byconventional humic acids extraction methods. The humic acids from agricultural rice and forest
soils were fractionated by the International Humic acids Society (IHSS) methods subjected toelemental, infrared and E4/E6 ratio analyses, and compared for their differences of physical andchemical properties, the characteristics and structures of humic acids. The degree of humificationof the Chi-Shan (CS) agricultural rice soils was lower than that of the Ta-Ta-Chia (TTC) forest
soils. It was caused by the tillage or fertilization in the agricultural rice soils and showed thelower degree of humification. Under the different climate and vegetative conditions, the forest
soils are more favor to the degree of humification than that of agricultural rice soils.
KEY WORDS: C/; ratio. E4/E6,humic substance, humification, infrared analysis

43

Humic Substances in Ecosystems 8, 2009
STRUCTURE AD PROPERTIES OF COMMERCIAL HUMATES FROM COALIFIED
MATERIALS, PEAT AD SAPROPELOlga Yakimenko, Alexey Izosimov
Department of Soil Science, Moscow State University, Leninskie Gori, Moscow 119992, Russia
E-mail: iakim@soil.msu.ru
Since the beneficial role of humic substances (HS) in the environment is well known,
today many companies all over the world produce commercial humates (HUM) from organic raw
materials and promote them as soil conditioners, plant growth stimulators, for remediation of
degraded soils, and sorbtion of toxicants. Industrial companies manufacture HUM using a big
variety of raw source materials and different industrial processes, and therefore the products havehighly variable properties and there are not enough data about their structure and actions. Since
the mechanism of HS-action is apparently correlated with their certain chemical parameters, theeffect of HS will be different depending on their origin. In this study a possible connection
between the origin of organic matter, and the ability of HUM to stimulate plant growth wasinvestigated. A number of HUM produced from coalified materials (brown coal BC, leonardite
Le, lignite Li and humalite Hu) and plant-originated resources peat (Pe), sapropel (Sa) andorganic waste material (OW) were examined in respect to their chemical composition andstructure, content of HS and auxin-like activity.
Chemically HUM are mostly Na or K humates with different additives – microelements,
silica, or nutrients. A detailed chemical analysis was conducted for HUM-products. Content of C
total varies between 30-45%, being higher for products from Pe, BC and Le and lower for ones
from Sa, Hu, Li and OW (Fig.1). HUM from coalified materials contain minor amount of N,
whereas products from Pe and Sa are significantly higher in N most probably due to presence ofproteins in raw materials (Fig.2). Some individual samples contain up to 4-7% of S.
55

3,0
Mean

2,5

Mean±SD

Mean±1,96*SD

Mean±SD

Mean±1,96*SD

45
Ctot, %

2,0
N tot, %

Mean

50

1,5

40

1,0

35

0,5

30

0,0

Pe

Sa

BC Hu

Le

Li

OW

25
Pe

Fig.2. Box plot of N tot

Sa

BC

Hu

Le

Li

Ow

Fig.1. Box plot of C total

Active ingredients in HUM-products are considered to be HS: HA or probably FA. For all
the samples total extractable C achieved 20-40%, which is approximately equal to 16-75% from
TOC. The distribution pattern demonstrates that HUM from Sa, Hu and Li content the lowest
amounts of extractable C most probably due to links with mineral impurities (Fig.3). Amount of
HA and fulvic acid fraction in individual samples varies greatly, although ratio CHA/CFA shows
uniform distribution except of BC-products rich in HA and poor in FA (Fig.4).
44

Humic Substances in Ecosystems 8, 2009

8

50
Mean

Mean±SD

Mean

Mean±1,96*SD

Mean±SD

Mean±1,96*SD

45

TEC, %

40

6

35
30

4

25
20

2

15
10

Sa

Pe

BC

Hu

Le

Li

OW

0
Pe

Sa

BC

Hu

Le

Li

OW

Fig.4. Box & Whisker Plot of CHA:CFA

Fig.3. Box & Whisker Plot of TEC

Advanced investigations of molecular structure of HA extracted from selected HUM were
performed; results are discussed in presentation. In brief, molecular structure HUM from plantoriginated material (Pe, Sa and OW) were enriched in O-containing functional groups, whereas
HUM from coalified materials in aromatic moieties.
In vitro experiments were conducted with plants treated with and without HUM. Reddish
seeds (Raphanus sativis L.) have been soaked in solutions of HUM at concentrations 625, 125 и
12,5 mg/L, and germinated at dark at 27 o C for 72 h. The physiological activity of HUM wasestimated as a change in primary root length of seedlings. Not all of the HUM tested
demonstrated auxin-like activity. The level of growth-stimulating activity varied greatly for
different HUM-brands, depending on their chemical structure and organic matter origin. FAfraction of HUM acted more actively then HA-fraction, whereas the highest effect was revealed
for unfractionated HUM-products.
Statistical evaluation of possible correlations between structural parameters and auxin-like
activity have been performed.
KEYWORD: humic substances, humic fertilizers, commercial humate,auxin-like activity
This work was supported by Russian Foundation for Basic Research, project 07-04-01510.

45

Humic Substances in Ecosystems 8, 2009
THE EFFECT OF GAMMA IRRADIATIO O PROPERTIES OF SOIL HUMICACIDSJoanna Strzelecka and Slawomir S. Gonet
Department of Soil Science, Institute of Geography, ;icolaus Copernicus University
9 Gagarina St., 87-100 Torun, Poland
E-mail: gonet@umk.pl
Humic substances are naturally occurring, biogenic, heterogeneous organic substances thatcan generally be described as yellow to black in colour, of high molecular weight and refractory.
Humic acids (HA) are the fraction of humic substances that is insoluble in water under acidic
conditions (below pH=2), but is soluble at higher pH values. HAs play an important role increation of many soil properties profitable for plant development but the processes of theirformation have not been entirely discovered so far. The HAs are resistant to chemical andmicrobial decomposition, therefore their mean residence time in soil is evaluate on about 1000
years.
The aim of this work was to identify if humic acids are relatively resistant not only tochemical and microbiological factors but to gamma irradiation too.
HA samples were extracted from soil samples (LAS – from O-horizon of Albic Aresonol
under pine forest; PUL – from Ap horizon of Haplic Luvisol used for agriculture; TOR – from 030 layer of Eutri-Fibric Histosol) and purified by the standard Schnitzer’s methods. Samples of
HA were divided into those that have been exposed to gamma radiation and for those that havenot been exposed to radiation. In the experiment “cobalt bomb” was used with 60Co as the sourceof gamma rays. Time of irradiation of samples was 1.5 months, the radiation dose was 160 kGy.
Irradiated and not irradiated HA samples were analysed for: elemental composition, infrared, 13CNMR CP/MAS solid state and UV-VIS spectra.
Obtained results showed that humic acids are resistant to gamma irradiation at the dose of
160 kGy. Elemental composition (see table), UV-VIS, IR and NMR analysis of irradiated HAsamples did not differ from comparable HA samples not irradiated. Studies have shown clearly
the differentiation in structure and properties of HA depending on their source of origin (type ofsoil).
Table 1. Elemental analysis (in atomic percentage) of HA samples
atomic %SampleVariant
H:C N:C O:C
C
H
N
O
0 kGy
35.4 40.7 2.0 21.9 1.15 0.06 0.62
LAS
160 kGy 35.5 40.5 1.9 22.1 1.14 0.05 0.62
0 kGy
32.8 43.7 2.8 20.7 1.33 0.09 0.63
PUL
160 kGy 32.9 43.4 2.8 20.8 1.32 0.09 0.63
0 kGy
35.0 42.3 1.8 20.9 1.21 0.05 0.59
TOR
160 kGy 34.9 42.3 1.8 21.0 1.21 0.05 0.60

KEYWORDS: humic acids, gamma radiation

46

O:H

ω

0.54
0.54
0.47
0.47
0.49
0.50

0.257
0.265
0.186
0.201
0.140
0.146

Humic Substances in Ecosystems 8, 2009STATUS OF ORGAIC MATTER I FOREST SOILS AFTER FIRE

Slawomir S. Gonet, Helena Dziadowiec, Aleksandra Kwiatkowska
Department of Soil Science, Institute of Geography, ;icolaus Copernicus University
9 Gagarina St., 87-100 Torun, Poland
E-mail: gonet@umk.pl
Samples were taken 13 years after the fire (2900 ha, surface fire) from the following sites:
control (area not burned, 90-years old Pinus sylvestris L. stand) and burned (area after surface
fire, stand as in control). Morphology of the soil profiles (Brunic Albic Arenosol, WRB 2006),
soil properties and stocks of organic carbon were determined.
The organic horizon in control soil consists of two sub-horizons: Ol and Ofh. In soil onarea after surface fire, the organic horizon consists of the sub-horizon Ol (“fresh build after fire)
and the sub-horizon named Os (“old” litter, burned during the fire). Thickness of the Os varied
from 0.3 to 2.5 cm. Os sub-horizon was characterized by black colour (N 2/0 when moist, 10YR
2/1 when dry), 8-22% of organic matter content and 10-13 of C:N ratio. Degree of aromaticity of
the Os horizon organic matter was 1.5 (after 13-C NMR CPMAS spectra), higher than Ofhhorizon – 0.7. Organic matter stocks in organic horizon of buried area (2.66 kg/m2) were lower
than in soil organic horizon of non-buried area (4.55 kg/m2). Lower organic matter stocks inhumus horizon were noted also, respectively.

Fig. 1 Typical soil profile on burned area with characteristic Os sub-horizon

KEYWORDS: organic matter stock, forest soils, forest fire, humus

47

Humic Substances in Ecosystems 8, 2009
HUMIC SUBSTACES EFFECT O POMEGRAATE URSERY PRODUCTIO

Ferenc Sandor(PhD Student, SZIE Godollo-Hungary) RoP Horticulture Director, USAID-DAI Horticultureand Livelihood Program Eastern Region, Afghanistan
E-mail: francisco@rootsofpeace.org
In the Eastern Region of Afghanistan is well known about its large scale horticulture
production. But it is also common knowledge, that climatic and soil conditions are very difficult.
In this region, where the annual precipitation is very low, the heat is very high, the soil shows
very weak development. The tested soil is a young soil in early stage of weathering. Its originatedfrom sedimentary material, which was deposited in the area. The little difference between thehorizons and the absence of E horizon also indicates the young age and poorly structured soil.
The high content of clay size particles makes the soil compact and poorly ventilated. The water
penetration is also limited into the soil.
The chemical property tests show, that the soil strongly alkaline and the nutrient content,
except the Phosphorus and Potassium, is low. The nutrient availability for plant is difficult in thealkaline context. Iron deficiency is a common problem in the production. The clayey structure,high pH, the reducing environment and low nutrient content makes difficult to work with thissoil. This land is the less suitable for high quality nursery production, which is a crucial factor forthe production. Therefore, the use of biostimulants, such as humic substances, can improve theproduction radically.
The aim of the research had been to increase pomegranate cutting survival rate and thenumber of quality saplings in nursery production through humic and fulvic acid application
during the growing period. The nursery production has been working since February 2007. The
survey study in 2007 indicated, that approximately 44-45% of the saplings were acceptable fortransplant. It is also established, that the climatic and soil conditions affected the growth of 55 %of the total cuttings.
One of the objectives of the experiment was to establish the effect of the humic substanceson the quality of the saplings in term of length, diameter, root system development and vigor.
The research established the changes of the soil physical and chemical properties andnutrient availability in the soil comparing them with the results of the original soil test.
The experiment had been conducted in four repitions. Each plot contained 100 rows.
Every 5th row functioned as control row. The production technology followed agro-ecologicalconcepts. Controlled furrow irrigation had been used according to calculations. The plant
nutrition method added animal manure to the DAP and Urea basal application. Humic acid had
been applied twice as side application and fulvic acid once as foliar application.
After 9 months the trial showed the following results:
1. The height and diameter of the test plants increased with an average of 30-35% more thanthe diameter and height of the control plants.
2. While 90% of the test plants achieved the required development stage by the end of the
production season, only 55% of the control plants could qualify under the same standard.
3. The root system analysis established that the root weight of the test plants were 2,62 times
heavier than the root weight of the control plants. The number of roots with a diameter of
2-4 mm, 4-7 mm and over 8 mm were 4-5 times larger in case of the test plants.
48

Humic Substances in Ecosystems 8, 2009
4. The soil properties showed the following results:
a. EC in 0-15 cm depth: Increased
b. EC in 15-30 cm depth: Slightly decreasedc. EC in 30-45 cm depth: Decresed
d. pH in 0-15 cm depth: Decreased
e. pH in 15-30 cm depth: Decreased
f. pH in 30-45 cm depth: Increased
g. Non-salinity in all depth
h. Available NPK content: Radically increased
i. Iron deficiency symptoms: Disappeared
The conclusion of the trial confirms that the main importance of humic substances inagricultural production lies in their capacity to:- Improve the soil physical properties- Stimulate plant growth- Enhance the effect of fertilizer
Substantial fraction of the mass of the humic acids is in carboxylic acid functional groups,
which endow these molecules with the ability to chelate positively charged multivalent ions
(Mg2+, Ca2+ Fe2+, most other "trace elements" of value to plants. By chelating the ions, theyfacilitate the uptake of these ions.
The trial clearly showed that the usually present iron deficiency did not develop in the test
plants. The mechanism how the insoluble iron oxide is absorbed by the plants requires further
tests. However, since plants have been shown to absorb and translocate the complex organic
molecules of systemic insecticides, we can no longer discredit the idea that plants may be able toabsorb the soluble forms of humus; this may in fact be an essential process for the uptake ofotherwise insoluble iron oxides.

49

Humic Substances in Ecosystems 8, 2009
RELATIOS BETWEE ORGAIC HORIZO PROPERTIES AD FOREST FLOORVASCULAR PLATS COMPOSITIO I SCOTS PIE
(PIUS SYLVESTRIS L.) STADS I SOUTH WEST POLAD

Piotr Sewerniak
Department of Soil Science, Institute of Geography, ;icolaus Copernicus University, Toruń,Poland;E-mail: sewern@uni.torun.pl
Properties of organic horizon are strongly related to the species composition ofphytocoenosis. Relations between organic horizon properties and vegetation composition are
double-sided. Plants influence on upper soil horizons properties, but also the properties determine
forest floor plant compositions. The second relation results from different plant species soil
requirements. Scots pine (Pinus sylvestris L.) is species that strongly influences soil properties
mainly by acidification upper soil horizons [e.g. Augusto i in. 1998]. In forest floor of pine stands
species that tolerate acid conditions mostly prevail. Such situation takes place even on sites ofdeciduous forests where pine was planted in silviculture what was common in south west Poland[Olaczek 1976]. The aim of the paper was to evaluate the relations between organic horizon
properties and forest floor vascular plants composition of Scots pine stands in selected plots insouth west Poland.
The researches were conducted in 100 plots located in Bolesławiec, Głogów and Oława
forest division (Fig. 1). In each plot phytosociological relevé was taken and organic horizon ofsoil was sampled. In descriptions of phytocoenosis Braun-Blanquet method was used [BraunBlanquet 1964]. Nomenclature of plants follows Rutkowski [1998]. Soil samples were taken andanalyzed by methods according to the instruction of polish forest survey [Instrukcja 2003].
Relations between vegetation and organic horizon properties were examined using CanonicalCorrespondence Analysis (CCA) in CANOCO 4,5 package [ter Braak, Šmilauer 2002].

Fig. 1. Location of Bolesławiec, Głogów and Oława forest division

The pattern of species distribution shown in figure 2 suggests that the first axis of the
diagram refer to soil fertility that increases from the left to the right part of the ordination
diagram. In its left part oligotrophic species are dominant (e.g. Vaccinium vitis-idaea, Ledum
palustre, Calluna vulgaris) while in the right part species of high soil trophy preferences prevail

50

Humic Substances in Ecosystems 8, 2009
(e.g. Alliaria petiolata, Brachypodium sylvaticum, Dactylis aschersoniana). It is confirmed byanalytical determinations of organic horizons. The gradient of characteristics positively correlated
to soil fertility (base saturation - BS, pHKCl, Nt and basic cations content) increase to the rightwhile gradient of acid cations (H+ + Al3+) content and C:N ratio increases to the left side of the
diagram. Results of CCA, forward selection and Monte Carlo permutation test identified BS, C:N
ratio, pHKCl, Mg2+, Ca2+, K+ and acid cations content as statistically important (p < 0,05) in thepattern of species distribution.

Fig. 2. CCA ordination diagram showing correlation between species composition andorganic horizon properties

The species distribution shown in figure 2 suggests also that the second axis of the
diagram refers to soil humidity. It is indicated by location of wet sites species mainly in a lower
part of the diagram (e.g. Calamagrostis canescens, Vaccinium oxycoccos, Erica tetralix) and of
fresh soils in middle and upper part of the diagram (Fig. 2). The occurance of species typical of
deciduous forests (e.g. Anamone nemorosa, Poa nemoralis) indicate that some of investigatedpine stands were planted on originally deciduous forests sites.

51

Humic Substances in Ecosystems 8, 2009
THE IFLUECE OF ORGAIC MAURIG O THE COTET OF THE SOIL
ORGAIC MATTER I THE LOG - TERM MOOCULTURE

Babulicová, M. – Kotorová, D.
Department Agrosystems, Plant Production Research Centre – Research Institute of Plant
Production Piešťany
Department Productive ecology,Plant Production Research Centre Piešťany – Institute ofAgroecology Michalovce
E-mail: babulicova@vurv.sk; kotorova@scpv-ua.sk
Soil organic matter is the source of microorganism nutrition, it is very important in thetillage of soil, in the infiltration of air and water by promoting water retention and reducing
erosion. The content and quality of soil organic matter positively affect physical, chemical andbiological soil properties. The soil fertility depends on the humus content, which is the result ofcrop rotation and organic fertilization. Nowadays the market economy is the most importantargument for the option of agricultural crops. This situation in many cases results in increasing
the concentration of one crop and limited crop rotations. With regard to the threats brought about
by continuous cropping the Research Institute of Plant Production Piešťany were carried out afield investigation in 1974 – 2009 concerning the questions: To what degree is it respectable toincrease the share of cereals in crop rotations? What are the possibilities of the decreasing thenegative impacts of the continuous cropping of cereals?
Stationary trial was established in 1974 in the experimental station Borovce. The field
experiment is located on a medium heavy textured soil (Luvi-Haplic Chernozem on loess). The
trial was situated in the area of continental climate. The trial had two parts. In the first part, thecrop rotations were with 40 %, 60 % and 80 % share of the cereals. In the second part of our trial
winter wheat and spring barley were grown in monoculture. In the first sequence there is growing
winter wheat in “pure” monoculture. Three variants of fertilization were used. At the control
variants are used only fertilizers. At the further variant straw of cereals was ploughed in the soil.
At the third variant straw and organic manuring Veget were ploughed in the soil. In the second
sequence there are rotated winter wheat and spring barley. In the third and fourth sequences: theintroduction of compensating crops (maize on silage, grain maize, oat) were used. In the years
2007 – 2008 the effect of mineral and organic fertilization on the physical and chemical
properties of winter wheat and spring barley in continuous cropping was investigated.
The winter wheat monoculture growing
During the period of 2007-2008 at the autumn sampling, the lower bulk density wasdetermined at the variant with ploughed in straw (B: 1.34 Mg.m-3) and at the variant withploughed in straw and Veget (C: 1.35 Mg.m-3) in comparison with the control variant withmineral fertilizers (A: 1.40 Mg.m-3). At the same time the total porosity observed at the variants
B (49.82 %) and C (48.15 %) was higher than at the variant A (45.16%).
The maximum water capillary capacity at the variants B (36.17%) and C (35.99%) withorganic manuring were lower than the one at the control variant A (36.99). On the contrary, thenon-capillary porosity was higher at the B (14.49 %) and C (11.90 %) variant than at the
A variant (6.29 %). The variants B and C were characterized by higher average humus contents(B: 1.836 %; C: 1.737 %) than the control variant A (1.664 %) all the time (at the spring andautumn sampling in the years 2007 and 2008). The average phosphorus content was higher at thevariants with organic manuring (B: 76 mg.kg-1; C: 78 mg.kg-1) than at the variant A (72 mg.kg-1)
in spring of the years 2007 and 2008. On the contrary, the average potassium content was lower
at the variants B and C (B: 267 mg.kg-1; C: 258 mg.kg-1) than at the control variant (275 mg.kg-1).
52

Humic Substances in Ecosystems 8, 2009
It can be supposed that the supplied organic matter in the form of ploughed in cereal straw or in
the form of ploughed in cereal straw and organic Veget contributed to a higher total porosity,
increasing of maximum water capillary capacity and increasing of content of humus. It is notpossible to exclude that the differences can bring about the heterogeneity of soil, too.
The spring barley monoculture growing
In the spring of the years 2007 and 2008 the lower bulk density was recorded at thevariant with ploughed in straw (E: 1.53 Mg.m-3) and at the variant with ploughed in straw andVeget (F: 1.51 Mg.m-3) than at the control variant with mineral fertilizers (D: 1.55 Mg.m-3).
mg.kg-1) At the same time the higher total porosity was observed at the variants E and F (E: 42.24%; F: 43.65 %) in comparison with the control variant D (41.34 %). The total porosity wasslightly higher at the variant with ploughed in straw and Veget (E: 45.53 %) than at the control
variant (D: 45.29 %) at the autumn sampling too. The maximum capillary water capacity was
higher at the variant with ploughed in straw (E: 35.71 %) in comparison with the control variant(D: 34.87 %) at the spring sampling. In autumn the non-capillary porosity was lower at bothvariants with organic manuring (E: 9.28 %; F: 9.85 %) than at the control variant (D: 10.05 %).
The variant with ploughed in straw and Veget was characterized by higher average humus
content (F:1.655 %) than the control variant (D:1.622 %) all the time (at the spring and autumn
sampling in the years 2007 and 2008). The soil reaction was at all the variants slightly acid(variant D: 6.49; E: 6.52; F: 6.57). At the variant with ploughed in straw and Veget (F: 76 mg.kg1) the content of phosphorus was higher than at the control variant (D: 68 mg.kg-1). Averagecontent of potassium was lower at the variants with organic manuring (variant E: 250 mg.kg-1 andvariant F: 267 mg.kg-1) than at the control variant (D: 275 mg.kg-1). The introduced results
suggested that the incorporation of straw by the continuous cropping is not sufficient for the
maintenance of seasonable physical and chemical soil properties. The decomposition of straw inthe soil lasts very long time and in the dry years the decomposition is especially decelerated.
Therefore, the combination of straw ploughed in with the organic manuring Veget, which has apositive impact on the soil moisture maintenance, the air content in soil conditions, seems much
more suitable.
By the continuous cropping of spring barley at the variant with ploughed in straw (variantE) the lowest content of organic carbon, the lowest content of potassium and phosphorus and thelowest total porosity was found in this variant. The present results are showing that thecontinuous growing of spring barley contributed to a stronger decline of soil physical andchemical properties than the continuous cropping of winter wheat. By the continuous cropping of
winter wheat the average bulk density was lower by 7.14 % than by the continuous cropping ofspring barley. The total porosity was higher by 7.16 %, the average maximum capillary capacity
by 6.64 %, the average non-capillary porosity by 9.15 % and the humus content by 8.4 % by thecontinuous cropping of winter than by the continuous cropping of spring barley. These findings
can be explains by the different root systems of winter wheat and spring barley. There is also
another important factor: winter wheat, its radical system together the root system secretion
influence the soil profile within almost 10 months, whereas spring barley only 3.5 – 5 months.
This result in soil moisture retention, aeration of soil profile and it influences the soil structure.

KEYWORD: winter wheat, spring barley, long- term monoculture, organic manuring, physicaland chemical soil properties, organic matter
This work was promoted by Agency for Supporting of Science and Research in Slovak
Republic on the agreement APVV-0645-06.

53

Humic Substances in Ecosystems 8, 2009
ORGAIC MATTER COTETS AD DISTRIBUTIO I CHEROZEM-RELATEDSOILS OF POLAD AD SLOVAKIA

Maciej Markiewicz1, Michał Jankowski1, Vladimir Simansky2, Sławomir Gonet1
1

Department of Soil Science, ;icolaus Copernicus University,
ul. Gagarina 9, 87-100 Toruń, Poland.
2
Department of Pedology and Geology, Slovak University of agriculture,
Tr. A. Hlinku 2, 949 01 ;itra, Slovakia.
E-mail: mawicz@umk.pl
Soils containing relatively deep, black, humic horizons [A] of the mollic type in the top
part of the profile, are situated among the most fertile and the most productive arable soils of theworld. There are some types of such soils occurring on the territory of Poland and Slovakia invarious geomorphological, lithological and hydrological conditions. Black color of these horizons
is often marked in their names, according to local soil systematics, for example: černozem,čiernica, czarna ziemia, smolnica.
The aim of this study is to present basic properties of humic horizons of four exemplary
profiles representing different soil types in that group of soils. Profiles are located in various
landscapes, three in Poland: smolnica (Gniew, the Starogard Lakeland, terminoglacial lake
deposits), czarna ziemia (Ostrówek, the Gniezno Lakeland, glacial till), czarnoziem (Witów,
Poland, the Proszowice Plateau, loess) and one in Slovakia: černozem (L’udovitovà, the Nitra
river valley, silt deposits). All the soils under study are in agricultural land use. They aredeveloped in medium to heavy texture deposits (silt, loam or clay), containing 1 to 24% of
calcium carbonate.
The humic horizons [A] are 40 to 70 cm deep and they are underlain by horizons
transitional to parent material [AC], spread 70 - 110 cm down the profile. Organic matter
contents vary from 1,7 to 3,7% (1.0-2.2% OC) in A horizons and from 0.7 to 1.7 (0.4-1.0% OC)
in AC horizons. In general, they have narrow C:N ratio values (9-11) and neutral or alkaline
reaction. Only one profile, czarnoziem from Southern Poland has wider C:N ratio (14) and acidreaction.
KEYWORD: Chernozems, mollic horizon, organic matter

Fig. 1. Location of the study soil profiles

54

Humic Substances in Ecosystems 8, 2009
AALYSIS AD CHARACTERIZATIO OF HUMIC SUBSTACES ISOLATEDFROM PEAT AD SOIL USIG OFF-LIE COMBIATIO OF LIQUID
CHROMATOGRAPHIC METHODS

Róbert Góra, Pavol Rohárik, Milan Hutta
Department of Analytical Chemistry, Faculty of ;atural Sciences, Comenius University in
Bratislava, Mlynská dolina CH-2, 842 15, Bratislava, Slovakia,
E-mail: gora@fns.uniba.sk
Humic Substances (HSs) belong to the most spread enviro-bio-polymers and they havedirect influence to various processes playing significant role in an environment. Characteristicfeature of this analytes group is created by diffuse non-distinct analytical signal that is producedby many detection principles. This fact is a consequence of their great complexity expressed also
by their chemical, structural and physical poly-dispersity, which further translates itself in a great
uncertainty of their analytical signal measured in almost all analytical methods, that are applied toinvestigation and measurement of HSs.
Liquid chromatography method arranged around RP HPLC using stepwise gradients ofdimethylformamide (DMF) in buffered aqueous mobile phase and a wide-pore (30nm)
octadecylsilica column had been applied to the analysis of soil and peat HSs, respectively, inorder to demonstrate the usefullness of the approach for their characterization even at trace
concentration levels.
Combination of acceptable DMF solvating and aggregation disturbing properties for HSs
together with wide pore RP sorbent improves surface interactions of the analytes and suppresses
influence of size exclusion effects in RP-HPLC. Thus it provides reproducibility ofcharacterization profiles and robustness of the method. Very good reproducibility of retention
times (± 0.5 % RSD) of peaks enforced by the step gradient shape supports well defined
characterization and/or fractionation of HSs. This creates good conditions for its combination
with the LC method based on totally different separation mechanism to create two-dimensional
separation schema. Thus, individual fractions obtained by the described RP-HPLC method were
off-line analyzed by size-exclusion chromatographic method (SEC) working on independent
separation principle and also by the described RP-HPLC method.
In SEC we achieved relatively precise and accurate determination of relative molecularmass distribution under the assumption and condition that separation or fractionation is based
entirely on separation according to size of molecule. Choice of a proper molecular mass
calibration standards mimicking elution behavior of HSs in a given chromatographic system isvery important with this respect and is discussed in more detail in poster presentation.

KEYWORDS: humic substances, RP-HPLC, step gradient, SEC, combination
This work was supported by the financial support of projects VEGA 1/4474/07 provided
by Slovak Grant Agency, APVV-0595-07 and VVCE-0070-07

55

Humic Substances in Ecosystems 8, 2009
O-LIE FLOW-THROUGH EXTRACTIO–PRECOCETRATIO-LARGE
VOLUME IJECTIO REVERSED-PHASE LIQUID CHROMATOGRAPHY METHODFOR TRACE DETERMIATIO OF SELECTED PYRETHROIDS I SLOVAK SOIL
MICRO-SAMPLES

Mária Chalányová, Milan Hutta, Martin Pagáč
Department of Analytical Chemistry, Faculty of ;atural Sciences, Comenius University, Mlynská
dolina CH-2, 842 15 Bratislava, Slovakia
E-mail: chalanyova@fns.uniba.sk
Analytical chemistry is frequently confronted with the need to measure many substancespresent in a complex matrix either at low concentrations (multicomponent trace analysis) or ina small sample mass or volume (multicomponent micro-analysis). In some cases, as are e.g.
forensic analysis or analysis of xenobiotics in the soil of rhizosphere of individual plant, we are
faced with the problem of multicomponent trace or ultra-trace micro-analysis. In the last
mentioned example the mass of available soil sample representing the problem is usually
restrcted to several ten to hundred miligrams. However, this is contradicting our general sense forrepresentativness of field soil sample, sampling plan and analytical or laboratory sample mass as
is defined in numerous textbooks. We believe that the progress must be done also in thementioned direction and so we decided to solve the problem of HPLC trace analysis of selectedpesticides in soil micro-samples by proper design, selection, testing and study of quantitative
manipulation with microsamples.
Α rapid micro-analytical multiresidue method has been developed for the analysis ofpyrethroids (kadethrin, cypermethrin and permethrin) in a micro-sample of soil (mass around 200mg). The method is based on the on-line flow-through extraction of soil micro-samples (packed
into a short glass column) with a methanol-aqueous citric acid buffer mixture, successive on-line
solid-phase extraction preconcentration (SPE) of analytes from the extract and on-line RP-HPLC
analysis with UV photometric detection.
The separation of pyrethroids was performed on a Purospher RP-18e column withmethanol-water as mobile phase. Effects of sorbent placed at bottom of short column holding thesoil sample and different kinds of on-line SPE columns were tested. Beside the influence ofvolume of the effluent on the pyrethroids recovery was also studied. Calibration curves werelinear over the range assayed from 0.01 to 0.2 µg/mL with correlation coefficients of linear
regression (least-squares method) in the range 0.998-0.999. Recovery studies were carried out at
0.25-1.00 µg/g dry soil fortification level and obtained recoveries were for kadethrin 81-84%,
cypermethrin 56-59% and for permethrin 58-63%. Achieved detection limits of studied synthetic
pyrethroids were for kadethrin 4.5 ng/g dry soil, 3.7 ng/g of cypermethrin and 3.6 ng/g ofpermethrin, respectively.

KEYWORDS: soil micro-sample, pyrethroids, on-line extraction, RP-HPLC
This work was supported by grant provided by Slovak Grant Agency VEGA 1/0870/09,
VEGA 1/4474/07, grant APVV-0595-07, and VVCE-0070-07.

56

Humic Substances in Ecosystems 8, 2009
STUDY OF OFF-LIE COMBIATIOS OF HPLC AD CZE METHODS FORAALYSIS OF LOW MOLECULAR WEIGHT SUBSTACES POTETIALLYRELATED TO HUMIC SUBSTACES DEGRADATIO PRODUCTS

Milan Hutta, Dušan Kaniansky, Róbert Góra
Department of Analytical Chemistry, Faculty of ;atural Sciences, Comenius University, Mlynská
dolina CH-2, 842 15 Bratislava, Slovakia
E-mail: hutta@fns.uniba.sk
Complexity of problem called humic substances invokes new way of thinking also inanalytical chemistry and provokes to various activities leading to analytical systems of greater
complexity just to get more general, or vice-versa more detailed information on theircomposition, concentration in the environment or adverse effects to chemical analyses of various
substances in environmental compartments.
Design and development of novel systems for characterization of humic substances and/or
their degradation products in solutions is still challenging.
Two-dimensional system devised in this work was created by off-line combination ofgradient elution reversed-phase high performance liquid chromatography and capillary zone
electrophoresis in an open system. Electrophoretic system was in our case used as a selective
detector for collected liquid chromatographic fractions. We tested orthogonality of liquid
chromatography and zone electrophoresis for group of 20 cationogenic and 28 anionogenic
compounds under conditions of high compatibility of both separation systems (identical mobile
phase and carrier electrolyte pH buffering components). The choice of the model substances was
based on criteria of their potential presence in solutions of degradation products of humic
substances (photolysis, hydrolysis, oxidation, oxidative photolysis, biodegradation etc.).
Migration and retention characteristics of analytes were evaluated both by Spearman rankcorrelation coefficients and Pearsons correlation coefficients.
We measured migration times in electrophoresis and retention times in chromatography,
respectively, of 20 basic compounds by use of phosphate buffers of varying pH values. Similar
experiments were done also with the set of 28 acidic compounds.
Combination of reversed - phase, or aqueous normal phase liquid chromatography,
respectively, and capillary electrophoresis gives us results, that are interpreted as non-existenceof proof of data sets correlations in studied combinations of CZE and HPLC systems. That
practically means formulation of separation systems orthogonality. Similar combinations oforthogonal separation systems will be used also for the characterization of treated solutions ofhumic substances.

KEYWORDS: Liquid chromatography, RP-HPLC, capillary electrophoresis, CZE, combination,orthogonal, ionogenic substancesThis work was supported by grant provided by Slovak Grant Agency VEGA 1/0870/09,
VEGA 1/4474/07, grant APVV-0595-07, and VVCE-0070-07.

57

Humic Substances in Ecosystems 8, 2009
UTILIZATIO OF IMMOBILIZED ALUMIIUM(III) METAL IO AFFIITY
CHROMATOGRAPHY FOR AALYSIS OF HUMIC ACID

Radoslav Halko, Tibor Neuročný, Milan Hutta
Department of Analytical Chemistry, Faculty of ;atural Sciences, Comenius University,
Mlynská dolina CH-2, 842 15 Bratislava, Slovakia
E-mail: halko@fns.uniba.sk
Immobilized metal ion affinity chromatography (IMAC) method is based on the ability ofmacromolecules to form coordination complexes with metal ions, which have been chelated to a
multidentate ligand immobilized onto a support material. In the present work, IMAC wasdeveloped for the fractionation of humic acid (HA).
In our case, Iontosorb Salicyl (Iontosorb, Czech Republic) was used as the chelating
sorbent containing salicylic acid bound via azo group in side chains of modified bead-form
cellulose. Sorption characteristics of Al(III) ions for this chelating sorbent were determined at
different values of pH. Simple photometric detection at 590 nm was used for monitoringaluminium as SPADNS-Al(III) chelate in eluent during breakthrough experiments. Resulting
sorption capacity is highest at pH 5.5 giving value 36 µmol of aluminium per 1 g of the sorbent.
Furher study was focused on the fractionation of the group of aromatic acids or HA by Alloaded Salicyl IMAC technique. The aromatic acids were selected as potential constituents of the
HA. The effect of various pH values (3.9, 6.7 and 8.9, resp.) on the retention of model sample ofacids was investigated. The IMAC eluent fractions of acids were separated and determined byreverse-phase high performance liquid chromatography (RP-HPLC) with UV-VIS detection.
Finally, IMAC technique was developed for the fractionation HA applying a buffer-based pH
gradient (between pH 8.9 and 2.0) for their gradual elution. The HA fractions retained on the
Al(III)-IMAC were eluted with eluent of decreasing pH value, and the fractions collected werecharacterized by UV–VIS detection. Based on the results obtained, IMAC appears to be a
promising tool for chemical separation of HA. The results will help in the understanding of thenature of HA and their metal binding characterization, and in modeling natural environments.

KEYWORDS: Immobilized aluminum (III) affinity liquid chromatography, soil acids, humic
substancesThis work was financially supported by grants VEGA-1/4474/07, VEGA-1/0870/09,
APVV-0595-07 and VVCE-0070-07.

58

Humic Substances in Ecosystems 8, 2009
THE CARBO AD THE FRACTIOS OF SULPHUR I ATURE RESERVEŽITAVSKÝ LUH I SLECTED SOILS

Melánia Feszterová1, Lýdia Jedlovská2
1

Katedra chémie, Fakulta prírodných vied, Univerzita Konštantína Filozofa, Tr. A. Hlinku 1, 949
74 ;itra, Slovensko, mfeszterova@ukf.sk
2
Katedra environmentalistiky a zoológie, Fakulta agrobiológie a potravinových zdrojov,
Slovenská poľnohospodárska univerzity, Tr. A. Hlinku 2, 949 74 ;itra

The aim of our work is to determine oxidizable carbon (Corg), selected fractions of sulphur
in soil samples from Nature Reserve Žitavský luh. This area is located on the borders of theŽitava River, on foothill of hilly country of Hronska. The wetlands are interesting in not only
from industrial point of view but they also play very important role in vegetation andcomposition of forest and plant communities by their position, climatic and soil conditions. We
compared examined indicators in samples from different soil types, which were taken from thesoils in Natural Reserve Žitavský luh in different depths, in years 2004 - 2006. From soil types
there are Mollic Fluvisol and Eutric Fluvisol in the reserve. The environment protection andecological measures oriented to decrease of discharged sulphur compounds caused the fall of the
supply of sulphur. One of the limiting factors for vegetal production is the deficit of sulphur.
Sulphur is one of the essential, irrecoverable nutrients in cultivating agricultural plants; it is a
mobile element and in environment it is reliable to very important changes because of the variety
of its chemical forms.

KEYWORDS: ;ature Reserve, Žitavský luh, environment, soil type, fractions of sulphur, sulphur
dynamics

59

Humic Substances in Ecosystems 8, 2009IFLUECE OF FARMIG SYSTEMS O AREA HETEROGEEITY OF TOTALORGAIC CARBO COTETJiří Heczko, Anton Zaujec
Department of Pedology and Geology, Slovak Agricultural University in ;itra (Slovak Republic).
E-mail: Jiri.Heczko@uniag.sk
The aim of paper was study the influence of different farming systems on area
heterogeneity of total organic carbon content. Soil samples were taken in spring 1997 and 2009
from top layer 0-0.2 m in long time field trial. The integrated and ecological farming systems
were established at the Slovak Agricultural University Research Station Dolná Malanta on Haplic
Luvisol clay-loamy soil in the fall of 1990. We measured total organic carbon content (Corg) in
the fertilized and unfertilized variants of integrated and ecological farming systems.
In spring 2009 was on ecological farming system in the fertilized variant (with farmyard
manure) average total organic carbon content 1.24% and in the unfertilized variant 1.09%. We
note in comparing with the year 1997 higher content of total organic carbon by 0.06% in thefertilized variant and lower content by 0.31% in the unfertilized variant. In spring 2009 was on
integrated farming system in the fertilized variant (with farmyard manure and NPK) average totalorganic carbon content 0.95% and in the unfertilized variant 0.82%. Compared with the year
1997 the total organic carbon content declined by 0.5% in the fertilized variant and in theunfertilized variant declined upon 0.82%. We can state, from results obtained in our study, that
ecological farming systems is from long-term duration (12 years) more sustainable and stabile fortotal organic carbon content as integrated farming systems.

KEYWORDS: Soil organic carbon, arable systems, area heterogeneity60

Humic Substances in Ecosystems 8, 2009
HUMIC SUBSTACES I CRYOCOITE O A SVALBARD GLACIERS

Slawomir S. Gonet1, Jerzy Melke2, Agnieszka Marciniak1 and Ewa Meller1
1

Department of Soil Science, ;icolaus Copernicus University, 9, Gagarina St., 87-100 Torun,
Poland2
Department of Soil Science, Maria Curie-Sklodowska University, 19 Akademicka St.,
20-033 Lublin,Poland
E-mail: gonet@umk.pl

Cryoconite is biogenic surface dust on the glacial surface, consists of mineral particles,organic matter and microbial communities (phototrophic microorganisms, cyanobacteria,
hetrotrophic bacteria) and microalgae. Biogenic impurities and dark colored their products havebeen shown to affect the surface albedo of snow and ice. Cryoconite can thereby contribute toacceleration of melting the glacier surface leading to creation of cryoconite holes – small
cylindrical depressions which occur on the surface ice of gleciers. Cryoconite holes are refugia
for life in this extreme environment.
The high light absorbency of cryoconite is due to humic substances produced frommicrobial materials in cryoconite holes and/or to organic matter of soil origin. The aim of thiswork was to investigate the properties of humus material, especially humic acids, in cryoconites
collected in a Svalbard glaciers.
Cryoconite samples from six glaciers (Aavatsmark, Waldemar, Irene, Blomli, Renard,
Scott) in the Kaffiora and Bellsund Regions in the Svalbard were analysed. Organic carbon(Corg), total nitronen (Nt) and fractional composition of organic matter (fulvic acids, humic acids
and humins fractions) in cryoconites were determined. Humic acids were extracted with 0.5M
NaOH method and then analysed for: elemental composition, UV-VIS, IR and 13C-NMR
CP/MAS spectra.
Corg content in cryoconte materials ranged from 1.77% to 3.87%, Nt content from
0.199% to 0.413%. The average value of Corg:Nt ratio was 9.1, what suggests the microbial
origin of organic material. Fractional composition of cryconite organic matter was characterized
by very low humic acids to fulvic acids ratio (average 0.07), not existing for soil organic matter.
Participation of fulvic acids carbon in total Corg was 40%, humic acids carbon – 2.5%, humins –58%. Extracted humic acids showed high aliphatic character. Atomic H:C ratio calculated fromelemental composition was 1.50. Very high value of the N:C ratio of 0.11 indicate a relatively
high content of nitrogen in humic acids structure. These results were confirmed by the infrared
spectra. Integration of 13C-NMR CP/MAS spectra showed very high content of aliphatic carbons(41% in 0-45ppm chemical shift) and very low content of aromatic carbons (14% in 110-165ppm
chemical shift). Calculated degree of humic acids aliphaticity was about 4.0 and 2-4 times higher
than for soil humic acids.

KEYWORD: cryoconite, organic matter, humic acids

61