Development of a Multicomponent Microbiological Soil Inoculant and Its Performance in Sweet Potato Cultivation.

The cultivation and consumption of sweet potato (Ipomoea batatas) are increasing globally. As the usage of chemical fertilizers and pest control agents during its cultivation may lead to soil, water and air pollution, there is an emerging need for environment-friendly, biological solutions enabling increased amounts of healthy crop and efficient disease management. Microbiological agents for agricultural purposes gained increasing importance in the past few decades. Our goal was to develop an agricultural soil inoculant from multiple microorganisms and test its application potential in sweet potato cultivation. Two Trichoderma strains were selected: Trichoderma ghanense strain SZMC 25217 based on its extracellular enzyme activities for the biodegradation of plant residues, and Trichoderma afroharzianum strain SZMC 25231 for biocontrol purposes against fungal plant pathogens. The Bacillus velezensis strain SZMC 24986 proved to be the best growth inhibitor of most of the nine tested strains of fungal species known as plant pathogens, therefore it was also selected for biocontrol purposes against fungal plant pathogens. Arthrobacter globiformis strain SZMC 25081, showing the fastest growth on nitrogen-free medium, was selected as a component with possible nitrogen-fixing potential. A Pseudomonas resinovorans strain, SZMC 25872, was selected for its ability to produce indole-3-acetic acid, which is among the important traits of potential plant growth-promoting rhizobacteria (PGPR). A series of experiments were performed to test the selected strains for their tolerance to abiotic stress factors such as pH, temperature, water activity and fungicides, influencing the survivability in agricultural environments. The selected strains were used to treat sweet potato in two separate field experiments. Yield increase was observed for the plants treated with the selected microbial consortium (synthetic community) in comparison with the control group in both cases. Our results suggest that the developed microbial inoculant has the potential to be used in sweet potato plantations. To the best of our knowledge, this is the first report about the successful application of a fungal-bacterial consortium in sweet potato cultivation.

Advantages of Multiplexing Ability of the Orbitrap Mass Analyzer in the Multi-Mycotoxin Analysis.

In routine measurements, the length of the analysis time and nfumber of samples analysed during a time unit are crucial parameters, which are especially important for the food analysis, particularly in the case of mycotoxin determinations. High-resolution equipment, including time-of-flight or Orbitrap analyzators, can provide stable instrumental background for high-throughput analyses. In this report, a short, 1 min MS-based multi-mycotoxin method was developed with the application of a short column as a reduced chromatographic separation, taking advantages of the multiplexing and high-resolution capability of the QExactive Orbitrap MS possessing sub-1 ppm mass accuracy. The performance of the method was evaluated regarding selectivity, LOD, LOQ, linearity, matrix effect, and recovery, and compared to a UHPLC-MS/MS method. The final multiplexing method was able to quantify 11 mycotoxins in defined ranges (aflatoxins (corn, 2.8-600 µg/kg; wheat, 1.5-350 µg/kg), deoxynivalenol (corn, 640-9600 µg/kg; wheat, 128-3500 µg/kg), fumonisins (corn, 20-1500 µg/kg; wheat, 30-3500 µg/kg), HT-2 (corn, 64-5200 µg/kg; wheat, 61-3500 µg/kg), T-2 (corn, 10-800 µg/kg; wheat, 4-250 µg/kg), ochratoxin (corn, 4.7-600 µg/kg; wheat, 1-1000 µg/kg), zearalenone (corn, 64-4800 µg/kg; wheat, 4-500 µg/kg)) within one minute in corn and wheat matrices at the MRL levels stated by the European Union.

Comprehensive assessment of heavy elements and evaluation of potential human health risk in the urban environment: a case study from Novi Sad, Serbia.

This study aimed to determine seasonal (summer vs. winter) and spatial distribution of the selected heavy elements (HEs) (As, Cd, Co, Cr, Cu, Ni, and Pb) in soil samples collected from a typical Central European town, Novi Sad, Serbia. The highest Pb concentrations were observed in summer because of intensive ground-flushing, whereas Cu had its highest concentration in winter, which may be attributed to traffic emissions. Source characterization and spatial distribution were carried out using cluster analysis (CA), principal component analysis (PCA), variogram calculation and theoretical model fitting, GIS-based geostatistical methods, and positive matrix factorization (PMF) data processing tools. Environmental impact of HEs found in different functional areas demonstrated that the quantified concentrations of Pb, As, Co, and Cu ranked soil as unpolluted to moderately polluted, while the presence of Co, Ni, and Cr classified urban soil as moderately polluted. Pollution load index (PLI) suggested a significant HEs enrichment while the new modified approach to Nemerow integrated risk index (NIRI) indicated high risk, being extreme for some selected locations. Machine learning classifiers were used for the first time to identify the differences between urban soil and dust samples in situations when simultaneous analysis of both matrices was carried out, as well as for temporal distribution (summer versus winter), based on the obtained concentration of HEs. Variogram calculation suggested that the pattern in spatial variability within the system emerged from the combined action of key structural factors (e.g., the parent soil material, landforms and topography, and climate) and random factors related to human activities. The estimated human health risk for two segments of the population revealed that ingestion is the primary route of exposure to HEs for children and adults.

Comprehensive characterization of stress tolerant bacteria with plant growth-promoting potential isolated from glyphosate-treated environment.

The application of plant growth-promoting bacteria in agricultural systems is an efficient and environment-friendly strategy to improve crop yields and maintain soil quality. However, as different soils have diverse and specific ecological characteristics and may represent adverse abiotic conditions, in vivo application requires the careful selection of the desired beneficial microorganisms. In this study we report Ensifer adhaerens SZMC 25856 and Pseudomonas resinovorans SZMC 25875 isolates recovered from glyphosate-treated soil to possess yet undiscovered plant growth-enhancing potential. The strains were found to promote the growth of tomato seedlings significantly, to have the ability of synthesizing indole-3-acetic acid and siderophores, to tolerate pH in the range of 6.59-7.96, salinity up to 12.5 g L-1 NaCl and drought up to 125 g L-1 polyethylene glycol 6000, as well as to survive in the presence of various pesticides including glyphosate, diuron, chlorotoluron, carbendazim and thiabendazole, and heavy metals such as Al, Fe, Mn, Zn, Pb and Cu. The plant growth-promoting traits of the examined E. adhaerens and P. resinovorans isolates and their tolerance to numerous abiotic stress factors make them promising candidates for application in different agricultural environments, including soils polluted with glyphosate.

Comprehensive characterization of PAHs profile in Serbian soils for conventional and organic production: potential sources and risk assessment.

This study presents a comprehensive characterization of occurrence and levels of 16 polycyclic aromatic hydrocarbons (PAHs) in arable soils used for conventional and organic production in northern and central part of Serbia as well as cross-border region with Hungary. Furthermore, this study includes a characterization of PAH sources and carcinogenic/non-carcinogenic human health risk for PAHs accumulated in analysed arable soils. The total concentration of 16 PAHs varied between 55 and 4584 µg kg-1 in agricultural soil used for conventional production and between 90 and 523 µg kg-1 in agricultural soil used for organic production. High molecular weight (HMW) PAHs were dominant compounds with similar contribution in both soil types (86% and 80% in conventional and in organic soil, respectively). Principal component analysis and diagnostic ratios of selected PAHs were used for identification of PAH sources in the analysed soils. Additionally, positive matrix factorization was applied for quantitative assessment. The results indicated that the major sources of PAHs were vehicle emissions, biomass and wood combustion, accounting for ~ 93% of PAHs. Exposure of farmers assessed through carcinogenic (TCR) and non-carcinogenic (THQ) risk did not exceed the acceptable threshold (TCR < 10-6 and THQ < 1). Oral ingestion was the main exposure route which accounted for 57% of TCR and 80% of THQ. It was followed by dermal contact. This investigation gives a valuable data insight into the PAHs presence in arable soils and reveals the absence of environmental and health risk. It also acknowledges the importance of comprehensive monitoring of these persistent pollutants.

Pollution status and health risk caused by heavy elements in the flooded soil and vegetables from typical agricultural region in Vojvodina Province, Serbia.

The investigation conducted in the Vojvodina Province, as a typical European and one of the biggest agricultural regions in the Balkans, offers the research methodology that could be used for any non/flooded agricultural region. The flood impact on heavy elements (HE) content in the flooded arable soil (n = 16) in relation to the control soil (n = 16) was examined, as well as their accumulation in the most often cultivated vegetables (n = 96) in the studied area. Results revealed that the flood did not significantly change the pseudo total HEs concentration in the soil as well as their amounts accumulated in different soil fractions. In both soils, only the average content of Ni exceeded the maximum permissible values set by Serbian soil quality standard which is in line with the Dutch standard. In comparison with the background values, notable enrichment is found for most of analyzed elements in both soils. Soil pollution status was assessed through several indices indicating that contamination range was in the domain from a moderate to a highly polluted. The principal component analysis demonstrated that soil contamination was probably originated from agricultural/anthropogenic activities (Cd, Cu, As, Pb), apart from Ni, Cr, and Co which came from natural weathering of the parent material. Carcinogenic and non-carcinogenic risks of selected HEs for Serbian population in the investigated region were below the threshold values. The average levels of Pb in investigated potato and carrot samples were higher than the maximum allowable concentrations established by EU/Serbian regulation. The total hazard quotients (THQ) of HEs through intake of analyzed vegetables were below the safe threshold (˂ 1), suggesting the absence of adverse health effects.

Characterization of the Plant Growth-Promoting Activities of Endophytic Fungi Isolated from Sophora flavescens.

Endophytic fungi in symbiotic association with their host plant are well known to improve plant growth and reduce the adverse effects of both biotic and abiotic stresses. Therefore, fungal endophytes are beginning to receive increased attention in an effort to find growth-promoting strains that could be applied to enhance crop yield and quality. In our study, the plant growth-promoting activities of endophytic fungi isolated from various parts of Sophora flavescens (a medicinally important plant in Mongolia and China) have been revealed and investigated. Fungal isolates were identified using molecular taxonomical methods, while their plant growth-promoting abilities were evaluated in plate assays. Altogether, 15 strains were isolated, representing the genera Alternaria, Didymella, Fusarium and Xylogone. Five of the isolates possessed phosphate solubilization activities and twelve secreted siderophores, while all of them were able to produce indoleacetic acid (IAA) in the presence or absence of tryptophan. The endogenous and exogenous accumulation of IAA were also monitored in liquid cultures using the HPLC-MS/MS technique to refine the plate assay results. Furthermore, for the highest IAA producer fungi, the effects of their extracts were also examined in plant bioassays. In these tests, the primary root lengths of the model Arabidopsis thaliana were increased in several cases, while the biomasses were significantly lower than the control IAA treatment. Significant alterations have also been detected in the photosynthetic pigment (chlorophyll-a, -b and carotenoids) content due to the fungal extract treatments, but these changes did not show any specific trends.

Occurrence and assessment of environmental risks of endocrine disrupting compounds in drinking, surface and wastewaters in Serbia.

The present study is the first comprehensive monitoring of 13 selected endocrine disrupting compounds (EDCs) in untreated urban and industrial wastewater in Serbia to assess their impact on the Danube River basin and associated freshwaters used as sources for drinking water production in the area. Results showed that natural and synthetic estrogens were present in surface and wastewater at concentrations ranging from 0.1 to 64.8 ng L-1. Nevertheless, they were not detected in drinking water. For alkylphenols concentrations ranged from 1.1 to 78.3 ng L-1 in wastewater and from 0.1 to 37.2 ng L-1 in surface water, while in drinking water concentrations varied from 0.4 to 7.9 ng L-1. Bisphenol A (BPA) was the most abundant compound in all water types, with frequencies of detection ranging from 57% in drinking water, to 70% in surface and 84% in wastewater. Potential environmental risks were characterized by calculating the risk quotients (RQs) and the estrogenic activity of EDCs in waste, surface and drinking water samples, as an indicator of their potential detrimental effects. RQ values of estrone (E1) and estradiol (E2) were the highest, exceeding the threshold value of 1 in 60% of wastewater samples, while in surface water E1 displayed potential risks in only two samples. Total estrogenic activity (EEQt) surpassed the threshold of 1 ng E2 L-1 in about 67% of wastewater samples, and in 3 surface water samples. In drinking water, EEQt was below 1 ng L-1 in all samples.

Does the application of human waste as a fertilization material in agricultural production pose adverse effects on human health attributable to contaminants of emerging concern?

The QuEChERS (quick, easy, cheap, effective, rugged, and safe) ultrasound-assisted method was successfully used to determine the presence of contaminants of emerging concern (CECs) in both the growing medium (i.e. soil) and vegetable samples (i.e. potato, onion, celery, parsnip and carrot) cultivated in household gardens under field conditions impacted by the application of livestock manure mixed with human waste. CECs with a broad range of physico-chemical properties including pharmaceuticals, plasticizers, herbicides, personal care products, and biocides were investigated. Among all studied CECs, diclofenac (DCL), carbamazepine, bisphenol A (BPA) and estrogenic hormones (estrone (E1), 17α-ethinyl estradiol (EE2)) were detected and quantified in vegetable samples with a concentration range from 0.114 to 13.3 ng/g fresh weight (f.w.), while BPA, E1, and EE2 were detected in soil samples with concentration ranges from 0.526 to 0.830 ng/g f.w., 0.121-0.199 ng/g f.w. and 0.118-0.333 ng/g f.w., respectively. DCL was only quantified above the limit of quantification in one soil sample (0.151 ng/g f.w.). The human health risk was estimated using the threshold of toxicological concern approach and a standard diet for two segments of the population (i.e. adult and child). The consumption of investigated vegetables contaminated with CECs poses no risk for the Serbian population.

Occurrence of heavy elements in street dust from sub/urban zone of Tianjin: pollution characteristics and health risk assessment.

Main purpose of this study was to determine the concentrations of selected heavy elements (As, Cd, Pb, Cu, Co, Cr and Ni) in the street dust samples (n = 49) collected from seven districts located in suburban/urban zone of Tianjin in order to estimate their possible sources and degree of environmental pollution as well as human health risk. Mean concentrations (mg kg-1) of As (19.3), Cd (0.60), Pb (28.4) and Cu (62.7) were above their corresponding soil background values. According to the results of multivariate statistical analysis, the accumulation of As, Cd, Pb, Cu and Cr in street dust was affected by anthropogenic activities, while the contents of Ni and Co were associated with natural sources. Pollution degree by geo-accumulation index had the following trend: Cd > Cu > As > Pb > Cr > Ni > Co. Dust contamination with Cd ranged from unpolluted to highly polluted. Potential ecological risk indicated low (Pb, Cu, Cr, Co and Ni) to high (Cd) risk, while potential risk index showed moderate and very high risks. Non-carcinogenic risk of the studied elements was below safe level (<1). Data obtained in this investigation gave the additional values to the knowledge needed for future monitoring and risk assessment, relating the presence of heavy elements studied in suburban/urban areas.

The extracellular matrix proteoglycan lumican improves survival and counteracts cardiac dilatation and failure in mice subjected to pressure overload.

Left ventricular (LV) dilatation is a key step in transition to heart failure (HF) in response to pressure overload. Cardiac extracellular matrix (ECM) contains fibrillar collagens and proteoglycans, important for maintaining tissue integrity. Alterations in collagen production and cross-linking are associated with cardiac LV dilatation and HF. Lumican (LUM) is a collagen binding proteoglycan with increased expression in hearts of patients and mice with HF, however, its role in cardiac function remains poorly understood. To examine the role of LUM in pressure overload induced cardiac remodeling, we subjected LUM knock-out (LUMKO) mice to aortic banding (AB) and treated cultured cardiac fibroblasts (CFB) with LUM. LUMKO mice exhibited increased mortality 1-14 days post-AB. Echocardiography revealed increased LV dilatation, altered hypertrophic remodeling and exacerbated contractile dysfunction in surviving LUMKO 1-10w post-AB. LUMKO hearts showed reduced collagen expression and cross-linking post-AB. Transcriptional profiling of LUMKO hearts by RNA sequencing revealed 714 differentially expressed transcripts, with enrichment of cardiotoxicity, ECM and inflammatory pathways. CFB treated with LUM showed increased mRNAs for markers of myofibroblast differentiation, proliferation and expression of ECM molecules important for fibrosis, including collagens and collagen cross-linking enzyme lysyl oxidase. In conclusion, we report the novel finding that lack of LUM attenuates collagen cross-linking in the pressure-overloaded heart, leading to increased mortality, dilatation and contractile dysfunction in mice.

Separation and Purification of Aflatoxins by Centrifugal Partition Chromatography.

Aflatoxins are mycotoxins that are produced by several species of filamentous fungi. In the European Union, the concentration limits for this group of mycotoxins in food and feed products are very low (on the order of parts per billion). Thus, relatively high amounts of these substances in their pure forms are required as reference standards. Chromatographic techniques based on solid stationary phases are generally used to purify these molecules; however, liquid-liquid chromatographic separations may be a promising alternative. Therefore, this study proposes a liquid-liquid chromatographic method for the separation of four aflatoxins and impurities. To optimise the method, numerous biphasic solvent systems (chloroform-, acetone- and acetic acid-based systems) were tested and evaluated in terms of their effectiveness at partitioning aflatoxins; the toluene/acetic acid/water (30:24:50, v/v/v/%) system was found to be the most efficient for application in centrifugal partition chromatographic instrument. Using liquid-liquid instrumental separation, the four aflatoxins, namely B1 (400 mg), B2 (34 mg), G1 (817 mg) and G2 (100 mg), were successfully isolated with 96.3%-98.2% purity from 4.5 L of Aspergillus parasiticus fermented material in a 250 mL centrifugal partition chromatography column. The identities and purities of the purified components were confirmed, and the performance parameters of each separation step and the whole procedure was determined. The developed method could be effectively used to purify aflatoxins for analytical applications.

Host metabolite producing endophytic fungi isolated from Hypericum perforatum.

In the present study, endophytic fungi have been isolated from various parts of the medicinal herb Hypericum perforatum (St. John's Wort), which is known as a source of medically important metabolites. The isolated strains were cultured in liquid media and their ability to synthesize hypericin, the secondary metabolite of the host and its suspected precursor, emodin was tested analyzing the extracts of the fermentation broth and the mycelia. The HPLC-UV analysis of the chloroform/methanol extracts of the mycelia revealed that three isolates were able to produce emodin (SZMC 23771, 19.9 ng/mg; SZMC 23772, 20.8 ng/mg; SZMC 23769, 427.9 ng/mg) and one of them also could synthesize hypericin (SZMC 23769, 320.4 ng/mg). These results were also confirmed via UHPLC-HRMS technique both in full scan and MS/MS mode. The strains producing only emodin belong to the section Alternata of the genus Alternaria, while the isolate producing both metabolites was identified as Epicoccum nigrum. The mycelial extracts of E. nigrum and the Alternaria sp. SZMC 23772 showed higher inhibitory activities in the antimicrobial tests against the six selected bacteria compared to the hypericin and emodin standards in the applied concentration (100 µg/mL), while in case of the Alternaria sp. SZMC 23771 lower inhibition activities were observed on Staphylococcus aureus and Streptomyces albus than the pure compounds.

Occurrence, seasonal variety of organochlorine compounds in street dust of Novi Sad, Serbia, and its implication for risk assessment.

The paper presents a study of organochlorine compounds in street dust, an often forgotten aspect of urban environmental pollution. Six different polychlorinated biphenyls (PCBs) and eighteen organochlorine pesticides (OCPs) were monitored in terms of levels, distribution, seasonal variation and health risks. Street dust samples were collected from fifteen urban locations in Novi Sad including schools, recreational areas, residential and industrial zones. Street dust concentration of PCBs ranged from less than the limit of detection (

Seasonal occurrence and cancer risk assessment of polycyclic aromatic hydrocarbons in street dust from the Novi Sad city, Serbia.

This is the first investigation that identified seasonal occurrence, distribution and sources of 16 polycyclic aromatic hydrocarbons (PAHs) in 60 street dust samples collected within urban zone of Novi Sad, the second largest city in Serbia. The obtained results were further used for comprehensive assessment of carcinogenic risk of Serbian inhabitants exposed to PAHs present in street dust by the incremental lifetime cancer risk method. The total level of 16 PAHs ranged between 35 µg kg-1 and 2422 µg kg-1 in samples taken in summer and between 35 µg kg-1 and 587 µg kg-1 in samples taken in winter. In both seasons, 4-ring PAHs were the most dominant compounds and high molecular weight (HMW) PAHs had similar contribution (55% in summer and 65% in winter). The highest content was determined for fluoranthene (Fly) in both seasons (597 µg kg-1 in winter, 301 µg kg-1 in summer). The PAHs source apportionment was analyzed by principal component analysis (PCA) and diagnostic ratios, and combustion of petroleum seemed to be the main sources of the PAHs in street dust. The cancer risk level for children and adult were comparable for dermal contact and by ingestion, and ranged from 10-6 to 10-4 indicating a potential risk. Additionally, the total incremental life time cancer risk (ILCR) was assessed for children and adult population taking into account three possible exposure routs and the median total cancer risk was ˃10-5, with 7% of the samples having the risk ˃10-4 that should be considered of high concern with potential health problem. These results are the first of this kind for the whole Serbia and the Western Balkan region and can be considered as the base line for future research.

Survey on the micro-pollutants presence in surface water system of northern Serbia and environmental and health risk assessment.

This study demonstrates the occurrence of 940 organic micro-pollutants in surface water of four rivers, one irrigation canal system, and two lakes in Vojvodina Province, the northern part of Serbia, summing in total eighteen samples. The number of detected chemicals ranged from 22 to 84, with 127 micro-pollutants detected at least once, representing 13% of the studied substances. The targeted compounds include n-alkanes, sterols, polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides polychlorinated biphenyls, pesticides, pharmaceutical active compounds, industrial chemicals, plasticizers, etc. Among the analysed compounds, sterols were the most dominant with maximum quantified concentrations. The substances which were quantified with frequency over 50% were two PAHs (2-methylnaphthalene, benzo(ghi)perylene), five sterols (cholesterol, cholestanol, stigmasterol, fucosterol, beta-sitosterol), three pharmaceuticals and personal care products (L-menthol, diethyltoluamide, caffeine), and ten household chemicals (4-tert-octylphenol, dimethyl phthalate, methyl palmitate, phenylethyl alcohol, 1-nonanol, alpha-terpineol, 2-phenoxy-ethanol, methyl myristate, acetophenone, and 2-ethyl-1-hexanol). The list of priority substances under the European Union Directive 2013/39/EU includes 49 priority substances (PSs) out of which 34 were analysed. Among these, eleven PSs were quantified, and only two compounds (fluoranthene and benzo (a) pyrene) exceeded EU Environmental Quality Standards targeted values. The obtained results were compared with the previously published data that dealt with the same targeted number of micro-pollutants in sediment samples. This revealed connections between the same sampling locations. Environmental risk assessment showed the existence of potential ecological risk as 72% of the obtained values for the ecological hazard index (HI) at investigated locations were higher that the targeted value (HI > 1). Estimated values for hazard quotient (HQ) and hazard index (HI) for non-carcinogenic risk were lower than the targeted value, indicating no non-carcinogenic risk through dermal contact and non-intentional ingestion of water. Estimated values for cancer risk were all below 1 × 10-6, which is not considered to pose significant human health risk.

Seasonal, spatial variations and risk assessment of heavy elements in street dust from Novi Sad, Serbia.

This is the first study carried out to determine the levels, spatial and temporal variations and risk indices of street dust in Serbia. Sixty street dust samples representing four functional areas: school, recreational, residential and industrial were collected from Novi Sad during summer and winter. The purpose was to obtain the concentration of seven heavy elements, evaluate the possible sources and degree of environmental pollution and human health risks. The average concentration of As, Cd, Co, Cr, Cu, Ni and Pb (mg/kg) during winter was: 7.96, 0.40, 7.73, 12.1, 57.6, 14.0 and 70.6, while in summer it was: 1.88, 0.54, 4.31, 60.1, 42.7, 28.1, and 62.5, respectively. Interpolation method together with the correlation matrix and the results of multivariate statistics supported anthropogenic origin of Pb, Cr and Cu, natural origin of Ni, Cd and As and mix origin of Co. Pollution indices such as geoaccumulation index (Igeo) indicated existence of unpolluted to extreme risk regarding to environmental pollution. Moreover, the potential pollution index (Er) showed low (Pb) to moderate (Ni) risk while potential risk index (RI) and modified potential risk index (mRI) indicated considerable and very high risks. Health risk assessment showed no non-carcinogenic risk of heavy elements for adults and children.

Ranking and similarity of conventional, microwave and ultrasound element sequential extraction methods.

This study aims to compare three extraction techniques of four sequential element extraction steps from soil and sediment samples that were taken from the location of the Pancevo petrochemical industry (Serbia). Elements were extracted using three different techniques: conventional, microwave and ultrasound extraction. A novel procedure - sum of the ranking differences (SRD) - was able to rank the techniques and elements, to see whether this method is a suitable tool to reveal the similarities and dissimilarities in element extraction techniques, provided that a proper ranking reference is available. The concentrations of the following elements Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, Ni, Pb, Si, Sn, Sr, V and Zn were determined through ICP OES. The different efficiencies and recovery values of element concentrations using each of the three extraction techniques were examined by the CRM BCR-701. By using SRD, we obtained a better separation between the different extraction techniques and steps when we rank their differences among the samples while lower separation was obtained according to analysed elements. Appling this method for ordering the elements could be useful for three purposes: (i) to find possible associations among the elements; (ii) to find possible elements that have outlier concentrations or (iii) detect differences in geochemical origin or behaviour of elements. Cross-validation of the SRD values in combination with cluster and principal component analysis revealed the same groups of extraction steps and techniques.