Distribution of selected metals among cytosolic biomolecules in gills and liver of chub (Squalius cephalus (Linnaeus) and Squalius vardarensis Karaman; Actinopterygii, Cyprinidae) from metal contaminated natural waters

Citosolske raspodjele nekoliko odabranih elemenata (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se i Zn) u jetrima i ńkrgama dvaju slatkovodnih bioindikatorskih organizama, klena (Squalius cephalus) iz rijeke Sutle u Hrvatskoj i vardarskog klena (Squalius vardarensis) iz triju rijeka sjeveroistoĉne Sjeverne Makedonije, izuĉavane su primjenom tekućinske kromatografije visoke djelotvornosti s iskljuĉenjem po veliĉini (SEC-HPLC) i spektrometrije masa visoke rezolucije s induktivno spregnutom plazmom (HR ICP-MS). OdreĊene su raspodjele metala/nemetala meĊu citosolskim biomolekulama razliĉitih molekulskih masa svojstvene uvjetima niske izloņenosti metalima u rijeĉnoj vodi, kao i promjene u raspodjelama koje su javljaju kao posljedica povińenih razina izloņenosti. Primjena anionsko izmjenjivaĉke kromatografije (AEC-HPLC) kao drugog stupnja razdvajanja i proĉińćavanja citosolskih biomolekula te dviju tehnika spektrometrije masa (MALDI-TOF-MS i LC-MS/MS) omogućila je identifikaciju ili precizno odreĊivanje molekulskih masa nekoliko citosolskih biomolekula koje veņu metale u jetrima i ńkrgama vardarskog klena, poput izoformi metalotioneina koje veņu Cd, Cu i Zn, hemoglobinskih podjedinica koje veņu Fe te toplinski stabilnih biomolekula koje veņu Mo. Primijenjeni analitiĉki pristup pokazao se korisnim u istraņivanju unutarstaniĉne sudbine i ponańanja metala u organima riba te otkrivanju potencijalnih novih biomarkera izloņenosti i uĉinaka metala.The cytosolic distributions of several selected elements (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se, and Zn) in the liver and gills of two freshwater bioindicators, European chub (Squalius cephalus) from the Sutla River in Croatia and Vardar chub (Squalius vardarensis) from three rivers of north-eastern Macedonia, were studied using size-exclusion high performance liquid chromatography (SEC-HPLC) and high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Distributions of metals/non-metal among the biomolecules of different molecular masses which are characteristic for low metal exposure in the river water were determined, as well as the changes in distribution which occur as a consequence of increased exposure to metals. The use of anion-exchange high performance liquid chromatography (AEC-HPLC) as the second step of separation and purification of cytosolic biomolecules, as well as the use of two mass spectrometry techniques (MALDI-TOF-MS and LC-MS/MS) have enabled the identification or exact determination of molecular masses of several cytosolic biomolecules that bind metals in the liver and gills of Vardar chub, such as the metallothionein isoforms that bind Cd, Cu and Zn, the hemoglobin subunits that bind Fe and heat stable biomolecules that bind Mo. Applied analytical approach has proven to be useful in the study of intracellular fate and behavior of metals in organs of fish, as well as in the process of discovery of potential new biomarkers of metal exposure and effects

Distribution of selected metals among cytosolic biomolecules in gills and liver of chub (Squalius cephalus (Linnaeus) and Squalius vardarensis Karaman; Actinopterygii, Cyprinidae) from metal contaminated natural waters

Citosolske raspodjele nekoliko odabranih elemenata (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se i Zn) u jetrima i ńkrgama dvaju slatkovodnih bioindikatorskih organizama, klena (Squalius cephalus) iz rijeke Sutle u Hrvatskoj i vardarskog klena (Squalius vardarensis) iz triju rijeka sjeveroistoĉne Sjeverne Makedonije, izuĉavane su primjenom tekućinske kromatografije visoke djelotvornosti s iskljuĉenjem po veliĉini (SEC-HPLC) i spektrometrije masa visoke rezolucije s induktivno spregnutom plazmom (HR ICP-MS). OdreĊene su raspodjele metala/nemetala meĊu citosolskim biomolekulama razliĉitih molekulskih masa svojstvene uvjetima niske izloņenosti metalima u rijeĉnoj vodi, kao i promjene u raspodjelama koje su javljaju kao posljedica povińenih razina izloņenosti. Primjena anionsko izmjenjivaĉke kromatografije (AEC-HPLC) kao drugog stupnja razdvajanja i proĉińćavanja citosolskih biomolekula te dviju tehnika spektrometrije masa (MALDI-TOF-MS i LC-MS/MS) omogućila je identifikaciju ili precizno odreĊivanje molekulskih masa nekoliko citosolskih biomolekula koje veņu metale u jetrima i ńkrgama vardarskog klena, poput izoformi metalotioneina koje veņu Cd, Cu i Zn, hemoglobinskih podjedinica koje veņu Fe te toplinski stabilnih biomolekula koje veņu Mo. Primijenjeni analitiĉki pristup pokazao se korisnim u istraņivanju unutarstaniĉne sudbine i ponańanja metala u organima riba te otkrivanju potencijalnih novih biomarkera izloņenosti i uĉinaka metala.The cytosolic distributions of several selected elements (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se, and Zn) in the liver and gills of two freshwater bioindicators, European chub (Squalius cephalus) from the Sutla River in Croatia and Vardar chub (Squalius vardarensis) from three rivers of north-eastern Macedonia, were studied using size-exclusion high performance liquid chromatography (SEC-HPLC) and high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Distributions of metals/non-metal among the biomolecules of different molecular masses which are characteristic for low metal exposure in the river water were determined, as well as the changes in distribution which occur as a consequence of increased exposure to metals. The use of anion-exchange high performance liquid chromatography (AEC-HPLC) as the second step of separation and purification of cytosolic biomolecules, as well as the use of two mass spectrometry techniques (MALDI-TOF-MS and LC-MS/MS) have enabled the identification or exact determination of molecular masses of several cytosolic biomolecules that bind metals in the liver and gills of Vardar chub, such as the metallothionein isoforms that bind Cd, Cu and Zn, the hemoglobin subunits that bind Fe and heat stable biomolecules that bind Mo. Applied analytical approach has proven to be useful in the study of intracellular fate and behavior of metals in organs of fish, as well as in the process of discovery of potential new biomarkers of metal exposure and effects

Distribution of selected metals among cytosolic biomolecules in gills and liver of chub (Squalius cephalus (Linnaeus) and Squalius vardarensis Karaman; Actinopterygii, Cyprinidae) from metal contaminated natural waters

Citosolske raspodjele nekoliko odabranih elemenata (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se i Zn) u jetrima i ńkrgama dvaju slatkovodnih bioindikatorskih organizama, klena (Squalius cephalus) iz rijeke Sutle u Hrvatskoj i vardarskog klena (Squalius vardarensis) iz triju rijeka sjeveroistoĉne Sjeverne Makedonije, izuĉavane su primjenom tekućinske kromatografije visoke djelotvornosti s iskljuĉenjem po veliĉini (SEC-HPLC) i spektrometrije masa visoke rezolucije s induktivno spregnutom plazmom (HR ICP-MS). OdreĊene su raspodjele metala/nemetala meĊu citosolskim biomolekulama razliĉitih molekulskih masa svojstvene uvjetima niske izloņenosti metalima u rijeĉnoj vodi, kao i promjene u raspodjelama koje su javljaju kao posljedica povińenih razina izloņenosti. Primjena anionsko izmjenjivaĉke kromatografije (AEC-HPLC) kao drugog stupnja razdvajanja i proĉińćavanja citosolskih biomolekula te dviju tehnika spektrometrije masa (MALDI-TOF-MS i LC-MS/MS) omogućila je identifikaciju ili precizno odreĊivanje molekulskih masa nekoliko citosolskih biomolekula koje veņu metale u jetrima i ńkrgama vardarskog klena, poput izoformi metalotioneina koje veņu Cd, Cu i Zn, hemoglobinskih podjedinica koje veņu Fe te toplinski stabilnih biomolekula koje veņu Mo. Primijenjeni analitiĉki pristup pokazao se korisnim u istraņivanju unutarstaniĉne sudbine i ponańanja metala u organima riba te otkrivanju potencijalnih novih biomarkera izloņenosti i uĉinaka metala.The cytosolic distributions of several selected elements (Cd, Co, Cu, Fe, Mn, Mo, Pb, Se, and Zn) in the liver and gills of two freshwater bioindicators, European chub (Squalius cephalus) from the Sutla River in Croatia and Vardar chub (Squalius vardarensis) from three rivers of north-eastern Macedonia, were studied using size-exclusion high performance liquid chromatography (SEC-HPLC) and high resolution inductively coupled plasma mass spectrometry (HR ICP-MS). Distributions of metals/non-metal among the biomolecules of different molecular masses which are characteristic for low metal exposure in the river water were determined, as well as the changes in distribution which occur as a consequence of increased exposure to metals. The use of anion-exchange high performance liquid chromatography (AEC-HPLC) as the second step of separation and purification of cytosolic biomolecules, as well as the use of two mass spectrometry techniques (MALDI-TOF-MS and LC-MS/MS) have enabled the identification or exact determination of molecular masses of several cytosolic biomolecules that bind metals in the liver and gills of Vardar chub, such as the metallothionein isoforms that bind Cd, Cu and Zn, the hemoglobin subunits that bind Fe and heat stable biomolecules that bind Mo. Applied analytical approach has proven to be useful in the study of intracellular fate and behavior of metals in organs of fish, as well as in the process of discovery of potential new biomarkers of metal exposure and effects

Determination of Flavonoids in Pulp and Peel of Mandarin Fruits

The aim of this study was to determine total flavonoids and individually flavanon glycosides as well as antioxidant capacity in pulp and peel of two mandarin groups, namely Satsuma (Citrus unshiu Marcovitch) cv. Saigon and Clementine (Citrus reticulate var. clementine) cv. Corsica SRA 63. Total flavonoids content was measured using colorimetric method, whereas HPLC-PDA detection was used for the analysis of individual flavanone glycosides (narirutin, naringin and hesperidin). In addition FRAP method was used to determine the antioxidant capacity. The results of colorimetric method showed that there was high concentration of flavonoids in all investigated samples, especially in peels (1156 mg/100 g in Satsuma peel and 804 mg/100 g in Clementine peel). Among the flavonone glycosides, hesperidin, was determined in the highest concentration in both investigated pulps. In peels flavanon glycosides were present in decreasing sequence as follow: narirutin>hesperidin>naringin. According to FRAP method, all samples possess an evident antioxidant capacity especially peels. Correlation between total flavonoids, hesperidin, naringin, narirutin and antioxidant capacity was very high with correlation coefficiants between 0.81 and 0.98

Accumulation of metals relevant for agricultural contamination in gills of European chub (Squalius cephalus)

The study of metal bioaccumulation in the gills of European chub (Squalius cephalus) was conducted in September 2009 at medium-size rural river Sutla, characterized by agricultural and municipal type of water contamination. The concentration ranges were established for the first time in the soluble, metabolically available fractions of chub gills for 12 metals, which are environmentally extremely relevant and yet only seldom studied, as follows in a decreasing order: K, 225-895 mg L-1 ; Na, 78-366 mg L-1 ; Ca, 19-62 mg L-1 ; Mg, 13-47 mg L-1 ; Rb, 164-1762 µg L-1 ; Sr, 24-81 µg L-1 ; Ba, 13-67 µg L-1 ; Mo, 1.3-16 µg L-1 ; Co, 0.7-2.7 µg L- 1 ; Li, 0.4-2.2 µg L-1 ; Cs, 0.2-1.9 µg L-1 ; and V, 0.1-1.8 µg L-1. The concentrations of Fe (1.6-6.4 mg L-1) and Mn (16-69 µg L-1) were also determined and were in the agreement with previous reports. By application of general linear modeling, the influence of different abiotic (metal exposure level) and biotic parameters (fish sex, age, size and condition) on metal bioaccumulation was tested. It was established that bioaccumulation of many metals in fish depended on various physiological conditions, wherein Ba could be singled out as metal exhibiting the strongest association with one of biotic parameters, being significantly higher in smaller fish. However, it was also undoubtedly demonstrated that the concentrations of three metals can be applied as reliable indicators of metal exposure even in the conditions of low or moderate water contamination, such as observed in the Sutla River, and those were nonessential elements Li and Cs, and essential element Fe. The results of our study present an important contribution to maintenance of high ecological status of European freshwaters, through enrichment of knowledge on the bioaccumulation of various metals in gills of European chub as frequently applied bioindicator species in monitoring of water pollution