Trace Level Analysis of Per- and Polyfluorinated Substances in Fish from Various Regions in Switzerland.

Per- and polyfluorinated substances (PFASs) are persistent man-made chemicals which can end up in the food chain. In this study, the concentrations of 15 PFASs in various wild fish species from different regions in Switzerland were determined excluding hot spots of contamination. After clean-up with SPE, the samples were analyzed by HPLC-MS/MS. PFASs were detected in all but 1 of the 83 fish samples (0.07 to 40.7 µg/kg fish muscle meat). The most abundant compound in fish from subalpine lakes was perfluorooctane sulfonic acid (PFOS), comprising more than 80% of the total contamination while perfluorononanoic (PFNA), -decanoic (PFDA) and -undecanoic (PFUnDA) acid dominated in high alpine fish. PFAS levels were more elevated in subalpine lakes (median PFASs 11.1-19.0 µg/kg) than in the high alpine Lake Sils (median PFASs 0.66-2.67 µg/kg) or streams and canals in Valais (median PFASs 0.56 µg/kg). Our results indicate that wild fish may be one of the PFAS sources in human diet.

Comparison of nutritional composition between plant-based drinks and cow's milk.

The high decline in liquid milk consumption in Western countries has been compensated by the increased consumption of processed dairy products and the rapidly increasing number of new plant-based beverages constantly introduced in the market, advertised as milk substitutes and placed on shelves near milk products. To provide better understanding about the nutritional value of these drinks compared with cow's milk, 27 plant-based drinks of 8 different species and two milk samples were purchased from two big retailers in Switzerland, and their composition regarding protein, carbohydrate, fat, vitamin, and mineral contents and residue load [glyphosate, aminomethylphosphonic acid (AMPA), and arsenic] was analyzed quantitatively and qualitatively. Energy and nutrient intakes were calculated and compared with the dietary reference values for Germany, Austria and Switzerland (D-A-CH). In addition, the digestible indispensable amino acid score (DIAAS) was calculated to estimate the quality of the proteins. Milk contained more energy; fat; carbohydrate; vitamins C, B2, B12, and A; biotin; pantothenic acid; calcium; phosphorus; and iodine than most plant-based drinks. Soy drinks provided slightly more protein and markedly more vitamins B1 and B6, folic acid, and vitamins E and D2 (with supplemented vitamin D2) and K1, magnesium, manganese, iron, and copper than milk and the other plant-based drinks. However, with the exception of cow's milk and soy drinks, which had > 3% protein, most milk alternatives contained ≤ 1% protein; therefore, they cannot be considered good protein sources. In regard to protein quality, milk was outstanding compared with all plant-based drinks and exhibited higher calculated DIAASs. Our results show that the analyzed plant-based drinks are not real alternatives to milk in terms of nutrient composition, even if the actual fortification is taken into account. Improved fortification is still an issue and can be optimized using the most bioavailable and soluble derivatives. Complete replacement of milk with plant-based drinks without adjusting the overall diet can lead to deficiencies of certain important nutrients in the long term.

Biomonitoring of ochratoxin A, 2'R-ochratoxin A and citrinin in human blood serum from Switzerland.

Biomonitoring of mycotoxins and their metabolites in biological fluids is increasingly used to assess human exposure. In this study, biomarkers of ochratoxin A (OTA) and citrinin (CIT) exposure were determined in a large number of serum samples from healthy blood donors in Switzerland. In 2019, 700 samples from different regions were obtained. From 240 donors, a second sample (taken 2-9 months later) was available for analysis. Moreover, 355 blood donor samples from 2005 from all regions in Switzerland and 151 additional samples from the southern Swiss region of Ticino from 2005 could be analysed.OTA, 2'R-ochratoxin A (2'R-OTA), ochratoxin alpha (OTα), CIT and dihydrocitrinone (DH-CIT) were analysed using validated targeted methods including precipitation and online SPE clean-up.OTA and 2'R-OTA were frequently detected (OTA in 99%; 2'R-OTA in 51% of the tested samples). The mean concentration in all positive samples was 0.4 ng/mL for OTA and 0.2 ng/mL for 2'R-OTA. OTα was not detected in any sample above the limit of quantification (LOQ). In contrast to OTA, CIT and DH-CIT were only quantifiable in 2% and 0.1% of the samples, respectively. No significant trend was observed between the samples from 2005 and the more recent samples, but OTA concentrations were usually higher in serum samples from the southern Swiss region of Ticino and in males compared to females.Our extensive data fit well within the framework of previously published values for the healthy adult European population.

Urine glyphosate level as a quantitative biomarker of oral exposure.

BACKGROUND: Since the classification of glyphosate as a Group 2A substance "probably carcinogenic to humans" by the IARC in 2015, human health concerns have been raised regarding the exposure of operators, bystanders, and consumers. Urine measurement studies have been conducted, but since toxicokinetic data on glyphosate in humans is lacking, a meaningful interpretation of this data regarding exposure is not possible. OBJECTIVE: This study aims to determine the fraction of glyphosate and AMPA excretion in urine after consuming ordinary food with glyphosate residue, to estimate dietary glyphosate intake. METHODS: Twelve participants consumed a test meal with a known amount of glyphosate residue and a small amount of AMPA. Urinary excretion was examined for the next 48 h. RESULTS: Only 1% of the glyphosate dose was excreted in urine. The urinary data indicated the elimination half-life was 9 h. For AMPA, 23% of the dose was excreted in urine, assuming that no metabolism of glyphosate to AMPA occurred. If all of the excreted AMPA was a glyphosate metabolite, this corresponds to 0.3% of the glyphosate dose on a molar basis. CONCLUSION: This study provides a basis for estimating oral glyphosate intake using urinary biomonitoring data.

Glyphosate residues in Swiss market foods: monitoring and risk evaluation.

A total of 243 samples of diverse foodstuffs were analysed for glyphosate and aminomethylphosphonic acid (AMPA) using a liquid chromatography triple quadrupole mass spectrometry (LC/MS/MS) method with a relatively low limit of quantification in the range of 0.0005-0.0025 mg kg-1. Main contributors for dietary glyphosate and AMPA intake were cereals and pulses. The results suggest that pasta is a very important foodstuff for dietary glyphosate residue intake in Switzerland. Interestingly all samples of wine, fruit juice and nearly all samples of honey tested positive for glyphosate although at very low levels. A dietary risk assessment was conducted. Food products for analysis were not selected purely at random, rather products were selected for which high levels of glyphosate residues were suspected. However, even in samples where high residue levels were expected, no exceedances of maximum residue levels were found. Consequently, human exposure did not exceed neither acceptable daily intake nor acute reference dose. Therefore, glyphosate residues found in the sampled foodstuffs from the Swiss market were of no concern for human health.

Natural occurrence of bisphenol F in mustard.

Bisphenol F (BPF) was found in mustard up to a concentration of around 8 mg kg(-1). Contamination of the raw products or caused by the packaging could be ruled out. Also, the fact that only the 4,4'-isomer of BPF was detected spoke against contamination from epoxy resin or other sources where technical BPF is used. Only mild mustard made of the seeds of Sinapis alba contained BPF. In all probability BPF is a reaction product from the breakdown of the glucosinolate glucosinalbin with 4-hydroxybenzyl alcohol as an important intermediate. Hot mustard made only from brown mustard seeds (Brassica juncea) or black mustard seeds (Brassica nigra) contained no BPF. BPF is structurally very similar to bisphenol A and has a similar weak estrogenic activity. The consumption of a portion of 20 g of mustard can lead to an intake of 100-200 µg of BPF. According to a preliminary risk assessment, the risk of BPF in mustard for the health of consumers is considered to be low, but available toxicological data are insufficient for a conclusive evaluation. It is a new and surprising finding that BPF is a natural food ingredient and that this is the main uptake route. This insight sheds new light on the risk linked to the family of bisphenols.

A quantitiative LC-MS/MS method for the measurement of arachidonic acid, prostanoids, endocannabinoids, N-acylethanolamines and steroids in human plasma.

Free arachidonic acid is functionally interlinked with different lipid signaling networks including those involving prostanoid pathways, the endocannabinoid system, N-acylethanolamines, as well as steroids. A sensitive and specific LC-MS/MS method for the quantification of arachidonic acid, prostaglandin E2, thromboxane B2, anandamide, 2-arachidonoylglycerol, noladin ether, lineoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, steroyl ethanolamide, aldosterone, cortisol, dehydroepiandrosterone, progesterone, and testosterone in human plasma was developed and validated. Analytes were extracted using acetonitrile precipitation followed by solid phase extraction. Separations were performed by UFLC using a C18 column and analyzed on a triple quadrupole MS with electron spray ionization. Analytes were run first in negative mode and, subsequently, in positive mode in two independent LC-MS/MS runs. For each analyte, two MRM transitions were collected in order to confirm identity. All analytes showed good linearity over the investigated concentration range (r>0.98). Validated LLOQs ranged from 0.1 to 190ng/mL and LODs ranged from 0.04 to 12.3ng/mL. Our data show that this LC-MS/MS method is suitable for the quantification of a diverse set of bioactive lipids in plasma from human donors (n=32). The determined plasma levels are in agreement with the literature, thus providing a versatile method to explore pathophysiological processes in which changes of these lipids are implicated.

LC-MS/MS method for simultaneous analysis of uracil, 5,6-dihydrouracil, 5-fluorouracil and 5-fluoro-5,6-dihydrouracil in human plasma for therapeutic drug monitoring and toxicity prediction in cancer patients.

The chemotherapeutic drug 5-fluorouracil (5-FU) is widely used for treating solid tumors. Response to 5-FU treatment is variable with 10-30% of patients experiencing serious toxicity partly explained by reduced activity of dihydropyrimidine dehydrogenase (DPD). DPD converts endogenous uracil (U) into 5,6-dihydrouracil (UH(2) ), and analogously, 5-FU into 5-fluoro-5,6-dihydrouracil (5-FUH(2) ). Combined quantification of U and UH(2) with 5-FU and 5-FUH(2) may provide a pre-therapeutic assessment of DPD activity and further guide drug dosing during therapy. Here, we report the development of a liquid chromatography-tandem mass spectrometry assay for simultaneous quantification of U, UH(2) , 5-FU and 5-FUH(2) in human plasma. Samples were prepared by liquid-liquid extraction with 10:1 ethyl acetate-2-propanol (v/v). The evaporated samples were reconstituted in 0.1% formic acid and 10 µL aliquots were injected into the HPLC system. Analyte separation was achieved on an Atlantis dC(18) column with a mobile phase consisting of 1.0 mm ammonium acetate, 0.5 mm formic acid and 3.3% methanol. Positively ionized analytes were detected by multiple reaction monitoring. The analytical response was linear in the range 0.01-10 µm for U, 0.1-10 µm for UH(2) , 0.1-75 µm for 5-FU and 0.75-75 µm for 5-FUH(2) , covering the expected concentration ranges in plasma. The method was validated following the FDA guidelines and applied to clinical samples obtained from ten 5-FU-treated colorectal cancer patients. The present method merges the analysis of 5-FU pharmacokinetics and DPD activity into a single assay representing a valuable tool to improve the efficacy and safety of 5-FU-based chemotherapy.