Assessment of organohalogenated pollutants in breast milk from the Czech Republic.

This biomonitoring survey brings new information on the occurrence of a total of 94 organohalogenated pollutants in 231 human breast milk samples collected in 2019 and 2021 from women living in two regions of the Czech Republic (Karvina and Ceske Budejovice). This study aimed to evaluate the concentrations of 6 indicator polychlorinated biphenyls (PCBs), 10 organochlorine pesticides (OCPs), 34 halogenated flame retardants (HFRs), 29 perfluoroalkyl and polyfluoroalkyl substances (PFAS) and 15 polychlorinated naphthalenes (PCNs). PCBs, OCPs, most of HFRs and PCNs were identified/quantified by gas chromatography coupled to (tandem) mass spectrometry (GC-MS(/MS)), while PFAS, hexabromocyclododecane isomers (HBCD), brominated phenols, and tetrabromobisphenol A (TBBPA) by ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). The mean value of the sum of the 6 indicator PCBs was 123.12 nanogram per gram of lipid weight (ng g-1 lw). Hexachlorobenzene (HCB), ß-hexachlorocyclohexane (ß-HCH) and p,p'-dichlorodiphenyl-dichloroethylene (p,p'-DDE) were the most abundant OCPs, detected in 100 % (mean 11.8 ng g-1 lw), 94.8 % (mean 6.1 ng g-1 lw) and 100 % (mean 101.5 ng g-1 lw) of samples, respectively. PCN congeners 20, 52 and 66 were detected in <1 % of the samples. The HFRs concentrations were relatively low compared to the levels of OCP; The detection rate of polybrominated diphenyl ethers (PBDEs, # 47, 99 and 153) ranged 21-68 % with a mean concentrations of 0.34 ng g-1 lw - 0.42 ng g-1 lw. PFAS concentrations were also low, with perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) dominant in this group (means of 22 pg ml-1 and 21 pg ml-1, respectively). Our results confirmed the long-term trend of declining levels of banned POPs in Czech mothers. The amounts of PCBs and OCPs were higher in older breastfeeding primiparous women.

Comprehensive Characterization of Micronized Wholemeal Flours: Investigating Technological Properties across Various Grains.

With a suitable milling system, it is achievable to produce wholegrain flours that match the granulation and technological properties of refined flours while maintaining a complete nutritional profile. This process also minimizes the generation of additional industrial waste. This study aimed to characterize wholemeal flours with a fine granulation size of less than 160 µm: wheat (MWF), rye (MRF), spelt (MSF), barley (MBF), buckwheat (MBWF), and sorghum (MSGF). For comparison, the plain wheat flour type 530 (T530) was analyzed. The flours were assessed in terms of their chemical compositions and alpha amylase activities (the Falling Number assay), pasting properties (amylograph and a Rapid Visco Analyser (RVA)), water absorption using a farinograph, and technological quality based on their water (WRC) and sodium carbonate solvent retention capacity (SRC) profiles. Among the micronized wholemeal flours, wheat flour (MWF) exhibited the highest nutritional value, greatest water absorption, and highest final gelatinization temperature, but had the lowest energy value, carbohydrate content, water SRC, and sodium carbonate SRC. Wholemeal rye flour (MRF) displayed the lowest nutrient content and the highest amylolytic activity, water absorption, and sodium carbonate SRC. The plain wheat flour type 530 (T530) had the lowest water absorption. Special buckwheat flour (MBWF) showed the highest energy value due to its elevated carbohydrate content, along with the lowest sugar and TDF contents, amylolytic activity, and pasting temperature.

Are fish oil-based dietary supplements a significant source of exposure to chlorinated paraffins?

Chlorinated paraffins (CPs) are an emerging group of environmental pollutants associated with adverse effects on human health (such as endocrine disruption and possible carcinogenicity). CPs are classified into several groups: short- (SCCPs), medium- (MCCPs) and long-chain chlorinated paraffins (LCCPs). In this study, CPs were determined in fish oil-based omega-3 dietary supplements (DS) representing widely consumed products that might be contaminated with various lipophilic contaminants including CPs. The CPs were isolated from DS (n = 85) by solid phase extraction. The SCCPs and MCCPs were determined by gas chromatography coupled with high-resolution mass spectrometry operated in a negative chemical ionisation mode. The LCCPs (up to C21) were screened using supercritical fluid chromatography coupled with high-resolution mass spectrometry with electrospray ionisation operated in negative mode. The CP concentrations varied from <0.01 to 56.48 µg/g fat for SCCPs (median 0.12 µg/g fat; limit of quantification, LOQ, for SCCPs was exceeded in 51 out of 85 samples) and from <0.03-89.08 µg/g fat for MCCPs (median 0.26 µg/g fat; LOQ for MCCPs was exceeded in 66 out of 85 samples), respectively. The LCCPs were not quantified in this study, nevertheless their limit of detection (0.5 µg/g fat) was exceeded in 7 out of 85 samples. Due to high levels of CPs, DS might pose as a significant source of CPs exposure to some population groups.

Short- and medium-chain chlorinated paraffins in human blood serum of Czech population.

Short- and medium-chain chlorinated paraffins (SCCPs; MCCPs) are widespread environmental pollutants with bioaccumulation potential and adverse effects on human health. The analysis of blood serum is an important strategy to assess the human exposure to various contaminants, including SCCPs and MCCPs. Lately, the information about the exposure of Chinese population has been reported; nevertheless, data on human exposure to SCCPs and MCCPs outside East Asia are still very limited. In this pilot study, SCCPs and MCCPs were determined in 27 serum samples obtained from Czech adults. The samples were extracted by a three-step extraction (repeated with a clean solvent) by a mixture of n-hexane:diethyl ether (9:1, v/v) with subsequent clean-up on Florisil® solid phase extraction column. Gas chromatography coupled with high resolution mass spectrometry operated in negative chemical ionisation was employed for the instrumental analysis. The method recoveries ranged from 71 to 89% with repeatabilities of <20% (expressed as relative standard deviation). In the samples, SCCP concentrations were in the range of <150-2600 ng/g lipid weight, lw (median 370 ng/g lw) and the MCCP concentrations were in the range of <200-2110 ng/g lw (median 360 ng/g lw), respectively. To the best of our knowledge, our reported results are the first data about chlorinated paraffins in human blood serum in Europe, showing exposure to these compounds with yet to be studied effects on human health.

Gentle Sterilization of Carrot-Based Purees by High-Pressure Thermal Sterilization and Ohmic Heating and Influence on Food Processing Contaminants and Quality Attributes.

Pressure-enhanced sterilization (PES) and ohmic heating (OH) are two emerging sterilization techniques, currently lacking implementation in the food industry. However, both technologies offer significant benefits in terms of spore inactivation using reduced thermal intensity in food products, as well as minimized effects on sensory and nutritional profiles. In this study, PES and OH were tested based on possible food safety process windows in comparison to thermal retorting, to optimize the food quality of carrot-based purees. The following parameters related to food quality were tested: texture, carotenoid content, color, and detectable amount of food processing contaminants (FPC) formed. Application of the innovative sterilization techniques resulted in a better retention of color, texture, and carotenoids (for PES) as well as a reduced formation of food processing contaminants. Importantly, a significant reduction in the formation of furan and its derivates was observed, compared to the retorted samples. Hence, both sterilization technologies showed promising results in the mitigation of potential toxic processing contaminants and retention of quality attributes.

Application of the GC-HRMS based method for monitoring of short- and medium-chain chlorinated paraffins in vegetable oils and fish.

The aim of this study was to develop methods for the determination of short- and medium-chain chlorinated paraffins (SCCPs; MCCPs) in vegetable oils and fish employing gas chromatography coupled with high-resolution mass spectrometry because of a lack of information on the presence of chlorinated paraffins in food consumed in Europe. For isolation of CPs from fish, an ethyl acetate extraction followed by a clean-up of the extract by gel permeation chromatography was performed. The same purification step was used for the isolation of CPs from the vegetable oils. The concentration range for SCCPs was <10-389 ng/g lipid weight (lw, mean 36 ng/g lw for the oils and 28 ng/g lw for the fish) and that for MCCPs was <20-543 ng/g lw (mean 55 ng/g lw for the oils and 59 ng/g lw for the fish). There was found a high variability in concentrations of CPs influenced by area of origin.

Lemon Grass Essential Oil Does not Modulate Cancer Cells Multidrug Resistance by Citral-Its Dominant and Strongly Antimicrobial Compound.

With strong antimicrobial properties, citral has been repeatedly reported to be the dominant component of lemongrass essential oil. Here, we report on a comparison of the antimicrobial and anticancer activity of citral and lemongrass essential oil. The lemongrass essential oil was prepared by the vacuum distillation of fresh Cymbopogon leaves, with a yield of 0.5% (w/w). Citral content was measured by gas chromatography/high-resolution mass spectrometry (GC-HRMS) and determined to be 63%. Antimicrobial activity was tested by the broth dilution method, showing strong activity against all tested bacteria and fungi. Citral was up to 100 times more active than the lemongrass essential oil. Similarly, both citral and essential oils inhibited bacterial communication and adhesion during P. aeruginosa and S. aureus biofilm formation; however, the biofilm prevention activity of citral was significantly higher. Both the essential oil and citral disrupted the maturated P. aeruginosa biofilm with the IC50 7.3 ± 0.4 and 0.1 ± 0.01 mL/L, respectively. Although it may seem that the citral is the main biologically active compound of lemongrass essential oil and the accompanying components have instead antagonistic effects, we determined that the lemongrass essential oil-sensitized methicillin-resistant S. aureus (MRSA) and doxorubicin-resistant ovarian carcinoma cells and that this activity was not caused by citral. A 1 mL/L dose of oil-sensitized MRSA to methicillin up to 9.6 times and a dose of 10 µL/L-sensitized ovarian carcinoma to doxorubicin up to 1.8 times. The mode of multidrug resistance modulation could be due to P-glycoprotein efflux pump inhibition. Therefore, the natural mixture of compounds present in the lemongrass essential oil provides beneficial effects and its direct use may be preferred to its use as a template for citral isolation.

Estimation of human exposure to polycyclic aromatic hydrocarbons (PAHs) based on the dietary and outdoor atmospheric monitoring in the Czech Republic.

In everyday life, humans can be exposed to various chemicals including ubiquitous polycyclic aromatic hydrocarbons (PAHs) mostly through food consumption and/or inhalation. In the presented study, we evaluated PAH concentrations in duplicate samples (n = 251). Concurrently, the outdoor concentrations of PM2.5-bound PAHs in filters (n = 179) were also monitored. The daily exposure to PAHs was subsequently estimated for the risk group of pregnant women living in two different cities (Most city and Ceske Budejovice city) in the Czech Republic. This is the first unique study in Europe to evaluate human daily exposure to 20 PAHs both from inhalation (outdoor air) and dietary intake. For the analysis of samples collected during the years 2016/2017, a gas chromatography coupled to tandem mass spectrometry was applied. Focusing on the diet samples, a slightly higher sum of detected PAHs was measured in duplicates obtained from the mothers living in the Most city (0.115-186 ng g-1) compared to the Ceske Budejovice city (0.115-97.1 ng g-1). This could be due to a higher occurrence of major analytes (pyrene, phenanthrene and fluoranthene) and the different composition of daily diet. The values of toxic and most often detected substance, namely benzo[a]pyrene (BaP), were also higher by 35% in the Most city. Regarding the outdoor air contamination (only particulate phase - PM2.5 was assessed), here the opposite situation was observed, relatively higher amounts of all PAHs were monitored in the Ceske Budejovice city (median: 2.22 ng m-3) than in the Most city (median: 1.07 ng m-3). These higher PAH concentrations in the Ceske Budejovice city are probably caused by more intense traffic, higher population and also by the occurrence of old-fashioned heating plant. Depending on a seasonal variability, especially during the cold season, the concentrations of BaP exceeded the European average emission limit (1 ng m-3) by 1.5-6 times. This highest inhalation exposure to all PAHs was observed in February. However, the dietary intake still represents the dominant contributor (>90%) to the total PAH exposure.

Evaluation of the Burdening on the Czech Population by Brominated Flame Retardants.

The completed environmental study was concerned with assessing the exposure of the Czech population to polybrominated diphenyl ethers (PBDEs). Simultaneously, the levels of polychlorinated pollutants such as polychlorinated biphenyls (PCBs) and chlorinated diphenyl ethanes (DDTs) were also monitored. The pollutant levels were newly measured in solid fat tissue removed during plastic surgery. A total of 107 samples of fat were taken from 19-76-year-old volunteers. A total of 16 PBDE congeners were determined, of which only six occur in more than 38% of fat tissue samples. The total PBDE level attains an average value of 3.31 ng/g, which is 25% less than was measured in 2009. On the other hand, there was an increase in the levels of two PCB congeners, which was caused by an increase of the total PCB concentration from level of 625.5 ng/g, published in 2009, to the current level of 776 ng/g. The level of DDTs decreased and currently has a value of 467.4 ng/g, which is about 24% lower than in 2009. The contamination of obese middle-aged women in Czechia by more modern types of pollutants, such as PBDEs, is incomparably lower than that by PCBs and DDTs and is also decreasing in time.

Integration of five groups of POPs into one multi-analyte method for human blood serum analysis: An innovative approach within biomonitoring studies.

Within this study, a new analytical strategy was developed and validated for the simultaneous determination of 78 organohalogenated contaminants in human blood serum, namely 40 flame retardants (FRs) including 7 "novel" brominated and chlorinated FRs (novel FRs), 19 perfluoroalkylated substances (PFASs), 11 organochlorine pesticides (OCPs) and 8 polychlorinated biphenyls (PCBs). The integral sample preparation procedure was implemented for the isolation of non-polar compounds, based on three-step solvent extraction using a mixture of n-hexane:diethylether (9:1, v/v), followed by purification using a solid-phase extraction (SPE) on a Florisil® column. For isolation of more polar and lipophobic analytes, the remaining fraction from the first extraction step was further processed, using a modified QuEChERS method. Depending on the polarity and volatility of target compounds, either gas chromatography coupled to (tandem) mass spectrometry (GC-MS/(MS)), or ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (UHPLC-MS/MS), was employed for their identification/quantification. Within the subsequent pilot study, the new validated procedure was successfully applied to the monitoring of organohalogenated contaminants in 38 samples of human blood serum obtained from Prague, Czech Republic. From 78 targeted analytes, 10 PFASs, 10 OCPs, 8 PCBs and 6 BFRs were detected in serum at concentrations above method quantification limits (MQLs). In the serum samples, the amounts of determined PFASs were in the range<0.01-8.97ngmL-1 (mean 0.631ngmL-1), OCPs and PCBs ranged from <0.1-1626ngg-1 lw (mean 40.0ngg-1 lw) and<0.1-481ngg-1 lw (mean 63.3ngg-1 lw), respectively.

A novel approach to assess the quality and authenticity of Scotch Whisky based on gas chromatography coupled to high resolution mass spectrometry.

Whisky is one of the most popular spirit drinks in the world. Unfortunately, this highly valued commodity is vulnerable to fraud. To detect fraudulent practices and document quality parameters, a number of laboratory tests based on various principles including chromatography and spectroscopy have been developed. In most cases, the analytical methods are based on targeted screening strategies. Non-targeted screening (metabolomics fingerprinting) of (semi)volatile substances was used in our study. Following the pre-concentration of these compounds, either by solid phase microextraction (SPME) or by ethyl acetate extraction, gas chromatography (GC) coupled to tandem mass spectrometry (Q-TOF mass analyser) was employed. Unsupervised principal component analysis (PCA) and supervised partial least squares discriminant analysis (PLS-DA) were used for evaluation of data obtained by analysis of a unique set of 171 authentic whisky samples provided by the Scotch Whisky Research Institute. Very good separation of malt whiskies according to the type of cask in which they were matured (bourbon versus bourbon and wine) was achieved, and significant ´markers' for bourbon and wine cask maturation, such as N-(3-methylbutyl) acetamide and 5-oxooxolane-2-carboxylic acid, were identified. Subsequently, the unique sample set was used to construct a statistical model for distinguishing malt and blended whiskies. In the final phase, 20 fake samples were analysed and the data processed in the same way. Some differences could be observed in the (semi)volatile profiles of authentic and fake samples. Employing the statistical model developed by PLS-DA for this purpose, marker compounds that positively distinguish fake samples were identified.

Relationship between atmospheric pollution in the residential area and concentrations of polycyclic aromatic hydrocarbons (PAHs) in human breast milk.

Human milk is an important source of beneficial nutrients and antibodies for newborns and infants and, under certain circumstances, its analysis may provide information on mothers' and infants' exposure to various contaminants. In the presented study, we have introduced the new analytical approach for analysis of 24 highly occurring polycyclic aromatic hydrocarbons (PAHs) in this indicator matrix. The sample preparation procedure is based on an ethyl acetate extraction of milk; the transfer of analytes into an organic layer is enhanced by addition of inorganic salts, i.e. sodium chloride and magnesium sulphate. Following the clean-up of a crude extract on silica SPE columns, gas chromatography coupled to triple quadrupole mass spectrometry is used for PAH identification and quantitation. The average recoveries of targeted PAHs from spiked samples were in the range of 68-110% with repeatabilities below 30% and method quantitation limits ranging from 0.03 to 0.3ng/g lipid weight. This newly validated method was successfully applied for analyses of 324 human milk samples collected from nonsmoking women during two sampling periods (summer and winter) in two residential areas in the Czech Republic differing in atmospheric pollution by PAHs. From 24 targeted analytes 17 were detected at least in one sample. Phenantherene, fluoranthrene, pyrene and fluorene were the most abundant compounds found at average concentration of 13.81, 1.80, 0.86, and 2.01ng/g lipid weight respectively. Comparing the data from two sampling periods, in both areas higher concentrations were measured in samples collected during winter. Also in the highly industrialized locality with heavily contaminated air PAH amounts in milk were higher than in the control locality. These first data on PAH concentrations in human milk collected in the Czech Republic are comparable with measurements for nonsmoking women reported earlier in the United States but significantly lower than results from China, Turkey or Italy.

Gas chromatography-triple quadrupole tandem mass spectrometry: a powerful tool for the (ultra)trace analysis of multiclass environmental contaminants in fish and fish feed.

A new method for rapid determination of 73 target organic environmental contaminants including 18 polychlorinated biphenyls, 16 organochlorinated pesticides, 14 brominated flame retardants and 25 polycyclic aromatic hydrocarbons in fish and fish feed using gas chromatography coupled with triple quadrupole tandem mass spectrometry (GC-MS/MS) was developed and validated. GC-MS/MS in electron ionization mode was shown to be a powerful tool for the (ultra)trace analysis of multiclass environmental contaminants in complex matrices, providing measurements with high selectivity and sensitivity. Another positive aspect characterizing the newly developed method is a substantial simplification of the sample preparation, which was achieved by an ethyl acetate QuEChERS (quick, easy, cheap, effective, rugged and safe) based extraction followed by silica minicolumn clean-up. With use of this sample preparation approach the sample laboratory throughput was increased not only because six samples may be prepared in approximately 1 h, but also because all the above-mentioned groups of contaminants can be determined in a single GC-MS/MS run. Under the optimized conditions, the recoveries of all target analytes in both matrices were within the range from 70 to 120% and the repeatabilities were 20% or less. The method quantification limits were in the range from 0.005 to 1 µg kg(-1) and from 0.05 to 10 µg kg(-1) for fish muscle tissue and fish feed, respectively. The developed method was successfully applied to the determination of halogenated persistent organic pollutants and polycyclic aromatic hydrocarbons in fish and fish feed samples.