TY  - CONF
AU  - Vasiljević, Sanja
AU  - Mikulić, Mira
AU  - Cvejić, Jelena
AU  - Nikolić, Zorica
AU  - Katanski, Snežana
AU  - Mamlić, Zlatica
AU  - Uhlarik, Ana
PY  - 2023
UR  - http://fiver.ifvcns.rs/handle/123456789/3759
AB  - Isoflavones are one of the best studied groups of phytoestrogens. Phytoestrogens are plant compounds that can exhibit a biological effect similar to the reproductive hormone estrogen. From nutritive and pharmaceutical aspect, red clover is one of the most important sources of these compounds, and they are often used as a raw material for production of dietary supplements used for alleviation of menopausal symptoms.
The objective of this study was to determine the concentrations of isoflavones in 11 diploid (2n) and 10 tetraploid (4n) cultivars of red clover (Trifolium pratense L.) from different origine, in a whole plant and separately: in stems, leaves and flowers at the stage of full flowering. Herbage samples from whole and different parts of plant were dried at 60˚C for about 48 hours, homogenized and grounded to a particle size of Ӕ = 0.8 mm as well as mixed with 2 ml of water on 37°C. After that, HCl and ethanol are added and mixture is heated to boiling. Obtained extract are purified by solid phase extraction on HLB cartridges and analyzed on HPLC after filtration. Zorbax SB C 18 reversed phase HPLC column is used for separation of main isoflavones present in red clover: formononetin, biochanin A, genistein and daidzein. Isoflavones are identified by comparing the retention times in HPLC chromatograms and UV spectral patterns with those of standard compounds. Isoflavone concentrations are quantified by external standard method using five-point regression curves of formononetin, biochanin A, genistein, and daidzein standard compounds.
The content of total isoflavones ranged from 3.83 (Marieta -2n cultivar) to 17.11 mg g-1 DM (Margot-4n cultivar). Red clover plant parts were significantly different according to isoflavone content. The leaf was the richest source of isoflavones (6.54 mg g-1 DM), followed by flowers (1,96 mg g-1 DM) and stem (1.16 mg g-1 DM). Biochanin A and formononetin were the predominant isoflavones in leaves, while genistein was mostly present in flowers. 
Tetraploid red clover cultivars (4n) had significantly higher isoflavones content compared to diploids (2n). On average, the content of isoflavones in tetraploids (11.0 mg g-1 DM) is statistically significantly higher than in diploids (8.44 mg g-1 DM).
PB  - Olomouc:Palacký University Olomouc
C3  - Abstracts of the 35th Meeting of the EUCARPIA Fodder Crops and Amenity Grasses Section in cooperation with the EUCARPIA Festulolium Working Group, 10-14 September 2023, Brno
T1  - Isoflavones profiles of some diploid and tetraploid red clover cultivars (Trifolium pratense L.) at flowering stage
EP  - 27
SP  - 27
DO  - 10.5507/vup.23.24463414
ER  - 

@conference{
author = "Vasiljević, Sanja and Mikulić, Mira and Cvejić, Jelena and Nikolić, Zorica and Katanski, Snežana and Mamlić, Zlatica and Uhlarik, Ana",
year = "2023",
abstract = "Isoflavones are one of the best studied groups of phytoestrogens. Phytoestrogens are plant compounds that can exhibit a biological effect similar to the reproductive hormone estrogen. From nutritive and pharmaceutical aspect, red clover is one of the most important sources of these compounds, and they are often used as a raw material for production of dietary supplements used for alleviation of menopausal symptoms.
The objective of this study was to determine the concentrations of isoflavones in 11 diploid (2n) and 10 tetraploid (4n) cultivars of red clover (Trifolium pratense L.) from different origine, in a whole plant and separately: in stems, leaves and flowers at the stage of full flowering. Herbage samples from whole and different parts of plant were dried at 60˚C for about 48 hours, homogenized and grounded to a particle size of Ӕ = 0.8 mm as well as mixed with 2 ml of water on 37°C. After that, HCl and ethanol are added and mixture is heated to boiling. Obtained extract are purified by solid phase extraction on HLB cartridges and analyzed on HPLC after filtration. Zorbax SB C 18 reversed phase HPLC column is used for separation of main isoflavones present in red clover: formononetin, biochanin A, genistein and daidzein. Isoflavones are identified by comparing the retention times in HPLC chromatograms and UV spectral patterns with those of standard compounds. Isoflavone concentrations are quantified by external standard method using five-point regression curves of formononetin, biochanin A, genistein, and daidzein standard compounds.
The content of total isoflavones ranged from 3.83 (Marieta -2n cultivar) to 17.11 mg g-1 DM (Margot-4n cultivar). Red clover plant parts were significantly different according to isoflavone content. The leaf was the richest source of isoflavones (6.54 mg g-1 DM), followed by flowers (1,96 mg g-1 DM) and stem (1.16 mg g-1 DM). Biochanin A and formononetin were the predominant isoflavones in leaves, while genistein was mostly present in flowers. 
Tetraploid red clover cultivars (4n) had significantly higher isoflavones content compared to diploids (2n). On average, the content of isoflavones in tetraploids (11.0 mg g-1 DM) is statistically significantly higher than in diploids (8.44 mg g-1 DM).",
publisher = "Olomouc:Palacký University Olomouc",
journal = "Abstracts of the 35th Meeting of the EUCARPIA Fodder Crops and Amenity Grasses Section in cooperation with the EUCARPIA Festulolium Working Group, 10-14 September 2023, Brno",
title = "Isoflavones profiles of some diploid and tetraploid red clover cultivars (Trifolium pratense L.) at flowering stage",
pages = "27-27",
doi = "10.5507/vup.23.24463414"
}

Vasiljević, S., Mikulić, M., Cvejić, J., Nikolić, Z., Katanski, S., Mamlić, Z.,& Uhlarik, A.. (2023). Isoflavones profiles of some diploid and tetraploid red clover cultivars (Trifolium pratense L.) at flowering stage. in Abstracts of the 35th Meeting of the EUCARPIA Fodder Crops and Amenity Grasses Section in cooperation with the EUCARPIA Festulolium Working Group, 10-14 September 2023, Brno
Olomouc:Palacký University Olomouc., 27-27.
https://doi.org/10.5507/vup.23.24463414

Vasiljević S, Mikulić M, Cvejić J, Nikolić Z, Katanski S, Mamlić Z, Uhlarik A. Isoflavones profiles of some diploid and tetraploid red clover cultivars (Trifolium pratense L.) at flowering stage. in Abstracts of the 35th Meeting of the EUCARPIA Fodder Crops and Amenity Grasses Section in cooperation with the EUCARPIA Festulolium Working Group, 10-14 September 2023, Brno. 2023;:27-27.
doi:10.5507/vup.23.24463414 .

Vasiljević, Sanja, Mikulić, Mira, Cvejić, Jelena, Nikolić, Zorica, Katanski, Snežana, Mamlić, Zlatica, Uhlarik, Ana, "Isoflavones profiles of some diploid and tetraploid red clover cultivars (Trifolium pratense L.) at flowering stage" in Abstracts of the 35th Meeting of the EUCARPIA Fodder Crops and Amenity Grasses Section in cooperation with the EUCARPIA Festulolium Working Group, 10-14 September 2023, Brno (2023):27-27,
https://doi.org/10.5507/vup.23.24463414 . .

TY  - JOUR
AU  - Vasiljević, Sanja
AU  - Mikulić, Mira
AU  - Aćimović, Milica
AU  - Kiprovski, Biljana
AU  - Krstić, Sanja
AU  - Katanski, Snežana
AU  - Mamlić, Zlatica
PY  - 2020
UR  - http://fiver.ifvcns.rs/handle/123456789/2662
AB  - Phytoestrogens are plant compounds that can exhibit estrogen-like activity. Isoflavones are one of 
the best studied groups of phytoestrogens known for their human health benefits. Red clover (Trifolium 
pratense L.) is one of the leading forage legumes in northern and south-eastern Europe, the United States 
and Canada. In recent years, the use of natural antioxidants found in red clover has attracted interest 
due to their presumed nutritional and therapeutic values (Vlaisavljevic et al., 2014; Vlaisavljevic et al., 2017). Studies of utilization of red clover by the pharmaceutical industry have pointed that dietary phytoestrogens play an important role in the prevention of menopausal symptoms (Lipovac et al., 2010), osteoporosis (Atkinson et al., 2004a), estrogen-related cancers like breast cancer, prostate cancer (Atkinson et al., 2004b; Velentzis et al., 2008) and heart disease (Cano et al., 2010; Dixon, 2004). The isoflavones content in red clover is controlled by genetic and environmental factors (Boot et al., 2006; Sivesind and Seguin, 2005; Tsao et al., 2006; Visnevschi-Necrasov et al., 2013).
PB  - Skopje : Ss Cyril and Methodius University in Skopje, Faculty of Pharmacy
PB  - Skopje : Macedonian Pharmaceutical Association
T2  - Macedonian Pharmaceutical Bulletin
T1  - Isoflavones profiles of red clover (Trifolium pratense L.) at different growth stages
EP  - 20
SP  - 19
VL  - 66 (Suppl 2)
DO  - 10.33320/maced.pharm.bull.2020.66.04.009
ER  - 

@article{
author = "Vasiljević, Sanja and Mikulić, Mira and Aćimović, Milica and Kiprovski, Biljana and Krstić, Sanja and Katanski, Snežana and Mamlić, Zlatica",
year = "2020",
abstract = "Phytoestrogens are plant compounds that can exhibit estrogen-like activity. Isoflavones are one of 
the best studied groups of phytoestrogens known for their human health benefits. Red clover (Trifolium 
pratense L.) is one of the leading forage legumes in northern and south-eastern Europe, the United States 
and Canada. In recent years, the use of natural antioxidants found in red clover has attracted interest 
due to their presumed nutritional and therapeutic values (Vlaisavljevic et al., 2014; Vlaisavljevic et al., 2017). Studies of utilization of red clover by the pharmaceutical industry have pointed that dietary phytoestrogens play an important role in the prevention of menopausal symptoms (Lipovac et al., 2010), osteoporosis (Atkinson et al., 2004a), estrogen-related cancers like breast cancer, prostate cancer (Atkinson et al., 2004b; Velentzis et al., 2008) and heart disease (Cano et al., 2010; Dixon, 2004). The isoflavones content in red clover is controlled by genetic and environmental factors (Boot et al., 2006; Sivesind and Seguin, 2005; Tsao et al., 2006; Visnevschi-Necrasov et al., 2013).",
publisher = "Skopje : Ss Cyril and Methodius University in Skopje, Faculty of Pharmacy, Skopje : Macedonian Pharmaceutical Association",
journal = "Macedonian Pharmaceutical Bulletin",
title = "Isoflavones profiles of red clover (Trifolium pratense L.) at different growth stages",
pages = "20-19",
volume = "66 (Suppl 2)",
doi = "10.33320/maced.pharm.bull.2020.66.04.009"
}

Vasiljević, S., Mikulić, M., Aćimović, M., Kiprovski, B., Krstić, S., Katanski, S.,& Mamlić, Z.. (2020). Isoflavones profiles of red clover (Trifolium pratense L.) at different growth stages. in Macedonian Pharmaceutical Bulletin
Skopje : Ss Cyril and Methodius University in Skopje, Faculty of Pharmacy., 66 (Suppl 2), 19-20.
https://doi.org/10.33320/maced.pharm.bull.2020.66.04.009

Vasiljević S, Mikulić M, Aćimović M, Kiprovski B, Krstić S, Katanski S, Mamlić Z. Isoflavones profiles of red clover (Trifolium pratense L.) at different growth stages. in Macedonian Pharmaceutical Bulletin. 2020;66 (Suppl 2):19-20.
doi:10.33320/maced.pharm.bull.2020.66.04.009 .

Vasiljević, Sanja, Mikulić, Mira, Aćimović, Milica, Kiprovski, Biljana, Krstić, Sanja, Katanski, Snežana, Mamlić, Zlatica, "Isoflavones profiles of red clover (Trifolium pratense L.) at different growth stages" in Macedonian Pharmaceutical Bulletin, 66 (Suppl 2) (2020):19-20,
https://doi.org/10.33320/maced.pharm.bull.2020.66.04.009 . .

TY  - CONF
AU  - Vasiljević, Sanja
AU  - Radinović, Irena
AU  - Vlaisavljević, Sanja
AU  - Mikulić, Mira
AU  - Milošević, Branko
AU  - Katanski, Snežana
AU  - Dolapčev, Anja
AU  - Dragić, Vasiljka
PY  - 2019
UR  - http://fiver.ifvcns.rs/handle/123456789/2614
AB  - Saglasno trendovima održive i organske poljoprivrede u proizvodnji zdravstveno-bezbedne stočne hrane, noviji pravci u oplemenjivanju crvene deteline imaju za cilj stvaranje dugovečnijih sorti, poboljšanog kvaliteta. Istovremeno brojni su primeri alternativnog korišćenja crvene deteline u proizvodnji funkcionalne hrane bogate izoflavonima. Rizomska forma podzemnog stabla nije svojstvena crvenoj detelini, međutim neke srodne vrste (Trifolium medium) poseduju takvu formu. Kulturom embriona u skorije vreme je dobijen (Jakešova et al, 2011, 2014) i komercijalizovan prvi hibrid između Treifolium pretense i Trifolium medium, pod nazivom „Pramedi“ sa rizomskom formom podzemnog stabla čime je pre svega povećana perzistentnost.
PB  - Beograd : Društvo za krmno bilje Republike Srbije
C3  - Zbornik apstrakata, 14. Simpozijum o krmnom bilju "Značaj i uloga krmnih biljaka i održivoj poljoprivredi Srbije", Zemun, 18-19. april 2019.
T1  - Novi izazovi u oplemenjivanju crvene deteline (Trifolium pretense L.) u održivoj poljoprivredi
EP  - 22
SP  - 21
UR  - https://hdl.handle.net/21.15107/rcub_fiver_2614
ER  - 

@conference{
author = "Vasiljević, Sanja and Radinović, Irena and Vlaisavljević, Sanja and Mikulić, Mira and Milošević, Branko and Katanski, Snežana and Dolapčev, Anja and Dragić, Vasiljka",
year = "2019",
abstract = "Saglasno trendovima održive i organske poljoprivrede u proizvodnji zdravstveno-bezbedne stočne hrane, noviji pravci u oplemenjivanju crvene deteline imaju za cilj stvaranje dugovečnijih sorti, poboljšanog kvaliteta. Istovremeno brojni su primeri alternativnog korišćenja crvene deteline u proizvodnji funkcionalne hrane bogate izoflavonima. Rizomska forma podzemnog stabla nije svojstvena crvenoj detelini, međutim neke srodne vrste (Trifolium medium) poseduju takvu formu. Kulturom embriona u skorije vreme je dobijen (Jakešova et al, 2011, 2014) i komercijalizovan prvi hibrid između Treifolium pretense i Trifolium medium, pod nazivom „Pramedi“ sa rizomskom formom podzemnog stabla čime je pre svega povećana perzistentnost.",
publisher = "Beograd : Društvo za krmno bilje Republike Srbije",
journal = "Zbornik apstrakata, 14. Simpozijum o krmnom bilju "Značaj i uloga krmnih biljaka i održivoj poljoprivredi Srbije", Zemun, 18-19. april 2019.",
title = "Novi izazovi u oplemenjivanju crvene deteline (Trifolium pretense L.) u održivoj poljoprivredi",
pages = "22-21",
url = "https://hdl.handle.net/21.15107/rcub_fiver_2614"
}

Vasiljević, S., Radinović, I., Vlaisavljević, S., Mikulić, M., Milošević, B., Katanski, S., Dolapčev, A.,& Dragić, V.. (2019). Novi izazovi u oplemenjivanju crvene deteline (Trifolium pretense L.) u održivoj poljoprivredi. in Zbornik apstrakata, 14. Simpozijum o krmnom bilju "Značaj i uloga krmnih biljaka i održivoj poljoprivredi Srbije", Zemun, 18-19. april 2019.
Beograd : Društvo za krmno bilje Republike Srbije., 21-22.
https://hdl.handle.net/21.15107/rcub_fiver_2614

Vasiljević S, Radinović I, Vlaisavljević S, Mikulić M, Milošević B, Katanski S, Dolapčev A, Dragić V. Novi izazovi u oplemenjivanju crvene deteline (Trifolium pretense L.) u održivoj poljoprivredi. in Zbornik apstrakata, 14. Simpozijum o krmnom bilju "Značaj i uloga krmnih biljaka i održivoj poljoprivredi Srbije", Zemun, 18-19. april 2019.. 2019;:21-22.
https://hdl.handle.net/21.15107/rcub_fiver_2614 .

Vasiljević, Sanja, Radinović, Irena, Vlaisavljević, Sanja, Mikulić, Mira, Milošević, Branko, Katanski, Snežana, Dolapčev, Anja, Dragić, Vasiljka, "Novi izazovi u oplemenjivanju crvene deteline (Trifolium pretense L.) u održivoj poljoprivredi" in Zbornik apstrakata, 14. Simpozijum o krmnom bilju "Značaj i uloga krmnih biljaka i održivoj poljoprivredi Srbije", Zemun, 18-19. april 2019. (2019):21-22,
https://hdl.handle.net/21.15107/rcub_fiver_2614 .

TY  - CONF
AU  - Malenčić, Đorđe
AU  - Miladinović, Jegor
AU  - Kiprovski, Biljana
AU  - Mikulić, Mira
AU  - Cvejic Hogervorst, Jelena Helene
AU  - Prvulović, Dejan
PY  - 2019
UR  - http://fiver.ifvcns.rs/handle/123456789/2228
AB  - Food and feed of plant origin containing phytochemicals of great antioxidant
potential are called nowadays “functional”, which means that they beside nutritive traits,
feature also some medicinal properties. Plant phenolics show useful characteristics in
human and animal metabolism, such as antiinflamatory and even anticarcinogenic. They
posses antibacterial, antimutagenic and vasodilatory activities, as well. Acting as natural
antioxidants they lower the risk of atherosclerosis and cardiac illness. Thus, soybean based
feed enriched with polyphenols may be classified as functional feed in animal diet.
Content of total polyphenols, tannins, flavonoids and anthocyanins were evaluated in the
seeds of seven soybean varieties of different seed color. One yellow seed cultivar
represented a reference genotype. Isoflavone (phytoestrogen) content and composition was
determined using HPLC analysis on an Agilent model 1100, equipped with diode array
detector (DAD). Aglycons were quantified from three five-
0.9998) obtained using the corresponding standards (daidzein, glycitein and genistein). In
addition, antioxidant activity of seed extracts was evaluated by the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) free radical scavenging activity assay.
A positive linear correlation between antioxidant activity and contents of total polyphenols
and anthocyanins was established. The highest antioxidant activity was observed in the
extracts of black and brown varieties, which also showed high levels of all polyphenol
classes examined. Yellow seed had the highest total isoflavone content (3.62 mg/g of dry
material). The highest concentration of total daidzein was determined in black seeds (>2.0
mg/g d.m.), and the highest total glycitein and genistein contents occurred in the yellow
cultivar (0.53 and 1.49 mg/g d.m., respectively). Varieties of black and brown seeds could
be of special interest due to large content of polyphenols (4.94-6.22 mg/g d.m.), especially
from the class of anthocyanins. Obtained data should enable the selection of genotypes rich
in natural antioxidants that could further be processed into functional feed in animal diet.
PB  - Novi Sad : University of Novi Sad, Faculty of Agriculture
C3  - Book of Abstracts, International symposium on animal science (ISAS), 03-08.06.2019., Herceg Novi, Montenegro
T1  - Colored soybean seeds: a new source of functional feed in animal nutrition
EP  - 25
SP  - 25
UR  - https://hdl.handle.net/21.15107/rcub_fiver_2228
ER  - 

@conference{
author = "Malenčić, Đorđe and Miladinović, Jegor and Kiprovski, Biljana and Mikulić, Mira and Cvejic Hogervorst, Jelena Helene and Prvulović, Dejan",
year = "2019",
abstract = "Food and feed of plant origin containing phytochemicals of great antioxidant
potential are called nowadays “functional”, which means that they beside nutritive traits,
feature also some medicinal properties. Plant phenolics show useful characteristics in
human and animal metabolism, such as antiinflamatory and even anticarcinogenic. They
posses antibacterial, antimutagenic and vasodilatory activities, as well. Acting as natural
antioxidants they lower the risk of atherosclerosis and cardiac illness. Thus, soybean based
feed enriched with polyphenols may be classified as functional feed in animal diet.
Content of total polyphenols, tannins, flavonoids and anthocyanins were evaluated in the
seeds of seven soybean varieties of different seed color. One yellow seed cultivar
represented a reference genotype. Isoflavone (phytoestrogen) content and composition was
determined using HPLC analysis on an Agilent model 1100, equipped with diode array
detector (DAD). Aglycons were quantified from three five-
0.9998) obtained using the corresponding standards (daidzein, glycitein and genistein). In
addition, antioxidant activity of seed extracts was evaluated by the 2,2-diphenyl-1-
picrylhydrazyl (DPPH) free radical scavenging activity assay.
A positive linear correlation between antioxidant activity and contents of total polyphenols
and anthocyanins was established. The highest antioxidant activity was observed in the
extracts of black and brown varieties, which also showed high levels of all polyphenol
classes examined. Yellow seed had the highest total isoflavone content (3.62 mg/g of dry
material). The highest concentration of total daidzein was determined in black seeds (>2.0
mg/g d.m.), and the highest total glycitein and genistein contents occurred in the yellow
cultivar (0.53 and 1.49 mg/g d.m., respectively). Varieties of black and brown seeds could
be of special interest due to large content of polyphenols (4.94-6.22 mg/g d.m.), especially
from the class of anthocyanins. Obtained data should enable the selection of genotypes rich
in natural antioxidants that could further be processed into functional feed in animal diet.",
publisher = "Novi Sad : University of Novi Sad, Faculty of Agriculture",
journal = "Book of Abstracts, International symposium on animal science (ISAS), 03-08.06.2019., Herceg Novi, Montenegro",
title = "Colored soybean seeds: a new source of functional feed in animal nutrition",
pages = "25-25",
url = "https://hdl.handle.net/21.15107/rcub_fiver_2228"
}

Malenčić, Đ., Miladinović, J., Kiprovski, B., Mikulić, M., Cvejic Hogervorst, J. H.,& Prvulović, D.. (2019). Colored soybean seeds: a new source of functional feed in animal nutrition. in Book of Abstracts, International symposium on animal science (ISAS), 03-08.06.2019., Herceg Novi, Montenegro
Novi Sad : University of Novi Sad, Faculty of Agriculture., 25-25.
https://hdl.handle.net/21.15107/rcub_fiver_2228

Malenčić Đ, Miladinović J, Kiprovski B, Mikulić M, Cvejic Hogervorst JH, Prvulović D. Colored soybean seeds: a new source of functional feed in animal nutrition. in Book of Abstracts, International symposium on animal science (ISAS), 03-08.06.2019., Herceg Novi, Montenegro. 2019;:25-25.
https://hdl.handle.net/21.15107/rcub_fiver_2228 .

Malenčić, Đorđe, Miladinović, Jegor, Kiprovski, Biljana, Mikulić, Mira, Cvejic Hogervorst, Jelena Helene, Prvulović, Dejan, "Colored soybean seeds: a new source of functional feed in animal nutrition" in Book of Abstracts, International symposium on animal science (ISAS), 03-08.06.2019., Herceg Novi, Montenegro (2019):25-25,
https://hdl.handle.net/21.15107/rcub_fiver_2228 .