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Lejla Klepo

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Damir Suljevic, M. Mitrašinović-Brulić, L. Klepo, R. Škrijelj, M. Fočak

In the current study, we assessed the hematological/biochemical alterations, histopathological changes in the liver, and blood cell disorders in Wistar rats exposed to a toxic concentration of carbon tetrachloride (CCl4) and the potential protective effect of a 30‐day oral extract of chokeberry (Aronia melanocarpa, AM). The concentration of AM (3.38 mg/kg) obtained by quantitative purification from AM fruit showed the highest antioxidant activity (AOA) in vitro and was used for oral ingestion. In addition to high AOA, high values of total phenols (85.334 mg/g), total phenolic acid (606.95 mg/g), total flavonids (22.10 mg/g), and total anthocyanins (11.01 mg/g) were recorded in chokeberry extract. CCl4 treatment caused serious liver injury, hepatocyte and blood cell impairment. AM extract given to rats before CCl4 application had a moderate hepatoprotective effect in comparison to after CCl4 application. White blood count and leukocytes were significantly altered by CCl4, however, the protective role of AM in leukocyte disorders was not established. A high number of microcytes, stomatocytes, anisocytes, and hemolyzed erythrocytes during CCl4 exposure was reduced by AM extract. Flower erythrocytes in the AM + CCl4 group were recorded. Supplementation with chokeberry extract without CCl4 caused hyperproteinemia and hyperalbuminemia. Although the results indicate a weak protective role for AM, it is nevertheless important for improved erythropoiesis and regulation of the development of anemia. The hepatoprotective role of AM was moderate, and the immune response was not proven. Daily consumption of chokeberry extract can improve health. However, the results of our study showed that the ingestion of AM extract at this dose with the highest AOA would have more effective effects if the supplementation were significantly increased.

L. Klepo, Jelena Ostojić, Olga Borota, S. Zero, Dušan Čulum

Pyrophyllite was investigated in this study as a potential adsorbent for the removal of glyphosate, a common herbicide for the control of a wide variety of weeds. There are several publications in the literature that discuss how glyphosate interacts with clay minerals such as montmorillonite, kaolinite, and bentonite, but there is no information regarding the possibility of glyphosate removal from aqueous solution using pyrophyllite. In this study, the adsorption of glyphosate from water samples onto pyrophyllite, a mineral clay from the Parsovici, Konjic mine, was investigated. The ninhydrin reaction was used to spectrophotometrically quantify glyphosate. At 570 nm, the purple Ruhemman product's absorbance was determined. Adsorption capacity (qe), removal efficiency (R), and the partition coefficient are used to express the results (Kd). The outcomes demonstrated that distinct ionic species of glyphosate at different pHs cause the adsorption of glyphosate on pyrophyllite to be pH-dependent. The findings demonstrated that during different contact times, the basic medium can remove up to 75% of glyphosate. According to results from fitting isotherm models, the Freundlich model is slightly better fitted. According to the results of kinetic investigations, the pseudo-2nd order model is more suitable for this kind of adsorption.

A. Tahirović, Amira Čopra – Janićijević, Nedžad Bašić, L. Klepo, M. Subašić

Vitamin C or ascorbic acid content has been determinated by spectrophotometric method and titrimetric method in flowers of some Bosnian hawthorns (Crataegus L). species. Spectrophotometric method used in this study was based on the kinetic reaction between Vitamin C and methylene blue. Measurements were carried out at absorption maximum, λmax= 665 nm. We found that the lowest content of vitamin C was 617.07 mg/100 g of dry sample in flowers of the C. microphylla, and the highest level of Vitamin C was found in the C. monogyna (1104 mg/100 g of dry sample) flowers. Recoveries of the results obtained by the spectrophotometric method were 94 % 100% with relative standard deviation (RSD) values from 4.5% – 6.7 %. Obtained results shown that flowers of investigated Crataegus L. species are good source of vitamin C.

Crnkic Mirsad, L. Klepo

Tannins are polyphenolic compounds that can be divided into two groups by hydrolyzing and condensing. Tannins are produced in organisms of vascular plants called tannosomes and sequestered in vacuoles. The essential uses of tannins are in leather production, as adhesives, additives for wine, beer and fruit juices, etc. They are most present in the growing tissues of the plant as a crust, so beer that ripens in new oak barrels will contain a higher concentration of tannins. Tannins are extracted from malt during grinding and from the hops during cooking. The objective of this research is to determine the tannins content in twelve domestic beer samples (three samples of dark beer and nine samples of light beer) by spectrophotometric method. The method is based on the reduction of Fe(III) to Fe(II) by tannins. The iron(II) reacts with 1,10-phenanthroline at pH 4.4 to form a color complex. The absorbance measurements were made at 540 nm. As a standard tannic acid was used, tannin content was in range 15.49-1722.05 μg/mL. Tannins are present in all beers, above the threshold of detection. When tannins are present in excess, they negatively impact beer by causing astringency but beer completely devoid of tannins does not taste right.

M. Radić, L. Klepo, L. Karic, H. Džudžević-Čančar, J. Toromanović, A. Ajanović, A. Uzunović

Solanine is a glycoalkaloid found in the Solanaceae family, such as the potato. It is very poisonous even in small quantities because it has pesticide and fungicide effects and represents a natural plant defense mechanism. Its concentration increases when the plant is exposed to the agents that can cause plant stress (fertilization, insecticide use, etc.). This paper aims to examine the influence of three cultivation systems (conventionally, organically and naturally) on the biosynthesis of α-solanine (αS) through his quantification in young potatoes using densitometry. Two varieties of potatoes were analyzed: Aladdin (Ala) and Mona Lisa (MoL). For statistical analysis, the Student's t-test was used. The results showed that the use of artificial insecticides caused a very intense biosynthesis of αS in the conventionally grown Ala variety (1.19 mg/100 g of fresh tubers (f.t.)) in comparison to the average α-solanine content (AαSC) by the organically grown Ala (0.62 mg/100 gf.t.) (it is close to the statistical significance, (p=0.08)). It is difficult to explain the very high AαSC of natural Ala cultivation (1.62 mg/100 gf.t.). Analysis of potatoes of the MoL variety showed that the AαSC of conventionally grown potatoes (1.35 mg/100 gf.t.) was statistically higher than the AαSC of naturally grown potatoes (0.59 mg/100 g of f.t.) (p<0.05). Also, AαSC of the organically grown MoL (1.40 mg/100 gf.t.) was higher than the AαSC of naturally grown MoL, but without statistical significance (p>0.05). Concentrations of αS founded in the case of conventionally, organically and naturally grown potatoes are considered safe and such potatoes are suitable for consumption. However, because of a slight reduction in toxic αS, it is recommended to consume organically grown potatoes (Ala variety), and naturally grown potatoes (MoL variety).

Dušan Čulum, A. Čopra-Janićijević, Danijela Vidic, L. Klepo, A. Tahirović, N. Bašić, M. Maksimović

The aim of this work was the qualitative and quantitative determination of selected phenolic compounds in three Crataegus species grown in Bosnia. Crataegus plants are consumed for medicinal purposes and as foodstuff in the form of canned fruit, jam, jelly, tea, and wine. Two samples of plant material, dry leaves with flowers, and berries of three Crataegus species—Crataegus rhipidophylla Gand., Crataegus x subsphaericea Gand., and Crataegus x macrocarpa Hegetschw.—were analyzed. Twelve ethanolic extracts were isolated from the selected plant material using Soxhlet and ultrasound extraction, respectively. Soxhlet extraction proved to be more effective than ultrasound extraction. A simple and sensitive method, high-performance liquid chromatography with electrochemical detection, HPLC-ED, was used for the simultaneous determination of phenolic acids and flavonoids in Crataegus species. The content of gallic acid in the extracts ranged from 0.001 to 0.082 mg/g dry weight (DW), chlorogenic acid from 0.19 to 8.70 mg/g DW, and rutin from 0.03 to 13.49 mg/g DW. Two flavonoids, vitexin and hyperoside, commonly found in chemotaxonomic investigations of Crataegus species, were not detected in the examined extracts. In general, leaves with flowers samples are richer in gallic acid and rutin, whereas the berries samples are richer in chlorogenic acid. Distinct similarities were found in the relative distribution of gallic acid among the three species. Extracts of C. x macrocarpa had the highest content of all detected compounds, while significant differences were found in rutin content, depending on the plant organ. To the best of our knowledge, this is the first study reporting content of phenolic compounds in Crataegus rhipidophylla Gand., Crataegus x subsphaericea, and Crataegus x macrocarpa from Bosnia.

L. Klepo, A. Čopra-Janićijević, L. Kukoc-Modun

Ascorbic acid (AA) is a water-soluble vitamin which shows no fluorescence. However, in reaction with iron(III), AA is oxidised to dehydroascorbic acid and iron(III) is reduced to iron(II) which forms a complex with 2,4,6-tripyridyl-S-triazine (TPTZ) in buffered medium. The relative fluorescence intensity of the resulting Fe(TPTZ)22+ complex can be measured at excitation and emission wavelengths of 393 and 790 nm, respectively. Based on this data, a new indirect spectrofluorimetric method for the determination of AA in pharmaceutical samples was proposed. Influence of the reaction conditions, such as acidity of acetic buffer, concentration of TPTZ and iron(III), reaction time and instrumental parameters were investigated in detail. The linear range was from 5.4 × 10−4 to 5.4 × 10−6 mol·L−1 (R = 0.9971). The LOD was 7.7 × 10−7 mol·L−1 and LOQ was 2.3 × 10−4 mol·L−1. Fourteen pharmaceutical samples containing various amounts of AA were analysed. Influences of potential interfering substances were also examined. Analysis of commercial pharmaceutical formulations showed good correlation with the nominal values given by the manufacturers and with the results obtained by a titration method. The proposed method can be applied in routine quality control in the pharmaceutical industry due to its sensitivity, simplicity, selectivity and low cost.

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