Genotoxic and cytotoxic effects of thiomersal and paracetamol combination

Background: Thiomersal is used as a preservative of some vaccines or as a trace from the pathogen inactivation process in vaccine production. Prophylactic use of paracetamol upon vaccination is still common, even though paracetamol decreases immune response on some vaccines. Considering the cytotoxic and genotoxic potential of thiomersal and paracetamol and possible interaction in vivo, in vitro study was performed. Methods: The genotoxic and cytotoxic effects of thiomersal and paracetamol combination were examined on human lymphocyte cultures by using two methods: analysis of chromosomal aberrations and cytokinesis-block micronucleus cytome assay. Blood samples of three healthy donors were analyzed with the following concentrations of tested substances: thiomersal 1 µg/ml and 0.5 µg/ml, paracetamol 20 µg/ml, thiomersal 0.5 µg/ml with paracetamol 20 µg/ml and thiomersal 1.0 µg/ml with paracetamol 20 µg/ml. Results: The analysis of structural chromosomal aberrations was significantly increased in all treated cultures. In cells treated with the combination of thiomersal 1 µg/ml and 20 µg/ml of paracetamol, the number of aberrations was significantly decreased. Cytokinesis-block micronucleus cytome assay analyses showed significantly increased micronucleus frequency in lymphocytes cultivated with thiomersal 1 µg/ml compared to lymphocytes cultures exposed to thiomersal 0.5 µg/ml. Conclusions: Induction of structural chromosome aberrations and micronucleus is shown as a sign of genotoxicity for the examined concentrations of thiomersal and paracetamol. The suppressing effect of paracetamol on thiomersal genotoxicity in lymphocytes culture treated with thiomersal was shown to be indicative of further examination of paracetamol use in the prevention of genotoxicity

Chemical Composition and Antioxidant Activity of Fraxinus ornus L. and Fraxinus excelsior L.

In this work, the total content of phenolics, flavonoids, and phenolic acids in the leaves and bark extracts of Fraxinus ornus L. and Fraxinus excelsior L. was determined. In addition, the identification and quantification of phenolic acids was performed using the HPLC-DAD technique. Samples were tested for antioxidant activity using the ABTS and DPPH methods. Eight extract samples were prepared by Soxhlet and ultrasound extraction using 70 % ethanol as solvent. The content of phenolic compounds ranged from 7.59 for F. ornus to 88.93 mg GAE/g for F. excelsior in the bark extracts obtained by Soxhlet extraction. The highest content of total flavonoids in F. ornus and F. excelsior was found in the leaves extracts obtained by ultrasound extraction, which was 5.68 and 1.74 mg GAE/g extract, respectively. The results also showed that the highest content of total phenolic acids was found in F. ornus (105.33 mg CAE/g) and F. excelsior (97.97 mg CAE/g) in the bark extract obtained by Soxhlet extraction. The highest content of gallic acid (112.96 ± 1.32 mg g–1 extracts) and chlorogenic acid (246.94 ± 0.82 mg g–1 extracts) was found in the F. ornus bark extract obtained by ultrasound extraction. As for antioxidant activity, the bark extract obtained by Soxhlet extraction of F. ornus showed the best antioxidant activity by ABTS method with IC50 value of 0.062 mg ml–1. The results of the DPPH method show that the leaves extract obtained by Soxhlet extraction had the best antioxidant activity for the F. ornus sample, with an IC50 value of 0.23 mg ml–1

Determination of α-solanine content in two varieties of potatoes by the densitometric method

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\u27s t-test was used. The results showed that the use of artificial insecticides caused a very intense biosynthesis of αS in the conven-tionally 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). 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)

A New Indirect Spectrofluorimetric Method for Determination of Ascorbic Acid with 2,4,6-Tripyridyl-S-Triazine in Pharmaceutical Samples

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

A New Indirect Spectrofluorimetric Method for Determination of Ascorbic Acid with 2,4,6-Tripyridyl-S-Triazine in Pharmaceutical Samples

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

HPLC-ED Analysis of Phenolic Compounds in Three Bosnian Crataegus Species

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