No abstract

Multielemental analysis of whole blood can provide significant information for the evaluation of nutritional status, diagnosis of certain diseases as well as for the assessment of exposure to potentially toxic metals. However, the quantification of multiple elements in whole blood is not easy partly because of the wide variation in element concentrations (from ng•L-1 to g•L-1) and the complex matrix. The aim of this work was to develop a fast, sustainable and reliable analytical method, in combination with low power TXRF, for multielemental analysis of blood samples. Firstly, a set of experiments were carried out to select the best diluent type and dilution factor using the control material SeronormTM Trace Elements Whole Blood L-1. A critical evaluation of the parameters affecting the sample deposition on the reflector was also carried out including a study of the shape and element distribution of the deposited residue on the reflector by micro X-ray fluorescence spectrometry. Using the best analytical conditions, limits of detection estimated were in the low mg•kg-1 range and similar to those obtained using more complex sample treatments such as digestion. Accuracy and precision of the results were in most cases acceptable (Recoveries: 89-102 %, RSD: 6-8%, n=5). Only underestimated values were obtained for light elements such as potassium. To prove the applicability of the method, several blood samples from control and thyroid diseases patients were analysed. Despite the fact that more samples need to be analysed, it seems that Zn and Br contents in some of the patients are significantly higher compared to control samples.

Helichrysum italicum is a plant traditionally used for skin-related disorders that is becoming an increasingly popular ingredient in cosmetic products. In this work, a “green” ultrasound-assisted extraction method for H. italicum phenolics was developed using skin-friendly cyclodextrins (CDs). Extraction conditions needed for the greatest yield of target compounds (total phenolics, phenolic acids, and flavonoids) were calculated. The composition of the extracts was determined using LC-MS and spectrophotometric methods. Among the tested CDs, 2-hydroxylpropyl-beta-CD (HP-β-CD) was the best suited for extraction of target phenolics and used to prepare two optimized extracts, OPT 1 (the extract with the highest phenolic acid content) and OPT 2 (the extract with the highest total phenol and flavonoid content). The extracts were prepared at 80 °C, using 0.089 g of plant material/g solvent (0.6 mmol of HP-β-CD), with or without addition of 1.95% (w/w) lactic acid. The main metabolite in both extracts was 3,5-O-dicaffeoylquinic acid. It was found that the addition of lactic acid greatly contributes to the extraction of arzanol, a well-known anti-inflammatory agent. IC50 values of the anti-elastase (22.360 ± 0.125 μL extract/mL and 20.067 ± 0.975 for OPT-1 and OPT-2, respectively) and anti-collagenase (12.035 ± 1.029 μL extract/mL and 14.392 ± 0.705 μL extract/mL for OPT-1 and OPT-2, respectively) activities of the extracts surpassed those of the applied positive controls, namely ursolic and gallic acids. This activity deems the prepared extracts promising ingredients for natural cosmetics, appropriate for direct use in cosmetic products, removing the need for the evaporation of conventional solvents.

A reliable method for spectrophotometric determination of urinary malondialdehyde (MDA), according to the thiobarbituric acid (TBA) assay, is described. To account for matrix interference and differences in individual urine composition, standard addition procedure was applied. The method is adequately selective (LoQ = 0.09 μM in the presence of 0.1 M creatinine and 0.5 M urea) and reliable (within-day and between-day variability of less than 5 %). The mean level of urinary MDA was 1.52 ± 0.73 µM that is in good agreement with spectrofluorometric determination (1.20 ± 0.56 μM; p = 0.085) as well as with previous studies that used HPLC. Furthermore, it is demonstrated that MDA is stabile in urine at room temperature for 24 h and when stored at -20 °C for 6 months. The described method enables simple, rapid and cost-effective determination of urinary MDA as a relevant and non-invasive marker of "whole-body" oxidative stress.

The aim of this study was to estimate a level of citric and tartaric acids in fruit nectars (n = 17) and juices (n = 13) available on the Croatian market. For analysis reverse-phase HPLC with UV/Vis detector set at 214 nm was used. Mobile phase was phosphate buffer (50 mM, pH = 2.80) at flow rate 0.5 mL min-1. The method was linear (r 2 = 0.9999). LoD was 0.01 g L-1 , LoQ was 0.03 g L-1 and the intra-day along with the inter-day variability were up to 3 %. The level of citric and tartaric acids in fruit nectars ranged from 1.26 to 4.42 g L-1 and 0.68 to 0.86 g L-1 , respectively, and in fruit juices ranged from 3.03 to 7.67 g L-1 and 3.09 to 4.68 g L-1 , respectively. A higher level of citric acid in fruit juices than in fruit nectars was detected (p < 0.05; MannWhitney U test). Six fruit juices contained a higher level of citric acid allowed by EU regulation implying the importance of monitoring concentrations of both acids in food products.