Degradation of 5-hydroxymethylfurfural during yeast fermentation

5-Hydroxymethyl furfural (HMF) may occur in malt in high quantities depending on roasting conditions. However, the HMF content of different types of beers is relatively low, indicating its potential for degradation during fermentation. This study investigates the degradation kinetics of HMF in wort during fermentation by Saccharomyces cerevisiae. The results indicated that HMF decreased exponentially as fermentation progressed. The first-order degradation rate of HMF was 0.693 × 10(-2) and 1.397 × 10(-2)min(-1) for wort and sweet wort, respectively, indicating that sugar enhances the activity of yeasts. In wort, HMF was converted into hydroxymethyl furfuryl alcohol by yeasts with a high yield (79-84% conversion). Glucose and fructose were utilised more rapidly by the yeasts in dark roasted malt than in pale malt (p<0.05). The conversion of HMF into hydroxymethyl furfuryl alcohol seems to be a primary activity of yeast cells, and presence of sugars in the fermentation medium increases this activity.     

Parameters affecting 5-hydroxymethylfurfural exposure from beer

5-Hydroxymethylfurfural (HMF) is generated during food and beverage heating processes and/or storage. Its daily intake, estimated as 4–10 mg day^?1, is several orders of magnitude higher than other process contaminants. Beer can be of relevance to the evaluation of HMF exposure; however, the information concerning its occurrence in different types of beer and during product storage is scarce. Therefore, the major goal of this work was to assess the amounts of HMF in different commercial beers, as well as the impact of storage, to deepen knowledge about the contribution of beer to HMF exposure. Blonde beers presented a mean content of 4.29 ± 1.05 mg L^?1, which was significantly lower (P ≤ 0.05) than those obtained for amber (6.84 ± 0.75 mg L^?1) and dark beers (6.99 ± 0.52 mg L^?1). Additionally, to study kinetic of HMF formation, fresh pilsner beers were stored at 30, 40 and 50°C during 40 days; a zero-order reaction was observed. The dependence of the rate constant on temperature was described by the Arrhenius equation and calculated activation energy was 101.85 kJ mol^?1. Storage can increase drastically HMF content, which means higher exposure for consumers. Thus, beer contribution to HMF exposure should not be neglected, since the intake of 1 L of beer entails a consumption of 4–7 mg of HMF or even more, depending on storage time and temperature.     

Transfer of aflatoxin B1 and fumonisin B1 from naturally contaminated raw materials to beer during an industrial brewing process

The aim of this research was to determine the fate of aflatoxins (AFs) and fumonisins (FBs) naturally occurring in raw materials (maize grit and malted barley) during four industrial brewing processes. The aflatoxin B₁ (AFB₁) level in raw materials varied from 0.31 to 14.85 µg kg^?1, while the fumonisin B₁ (FB₁) level (only in maize grit) varied from 1146 to 3194 µg kg^?1. The concentration in finished beer ranged from 0.0015 to 0.022 µg l^?1 for AFB₁ and from 37 to 89 µg l^?1 for FB₁; the other aflatoxins and fumonisin B₂ were not found in beer samples. The average percentage of toxins recovered in finished beer, referring to the amounts contained in raw materials, were 1.5% ± 0.8% for AFB₁ and 50.7% ± 4.7% for FB₁. These results were mainly due to the different solubility of the two mycotoxins during the mashing process. If raw materials comply with the limits fixed by European Commission Regulations, the contribution of a moderate daily consumption of beer to AFB₁ and FB₁ intake does not contribute significantly to the exposure of the consumer.     

Dietary intake estimate for perfluorooctanesulphonic acid (PFOS) and other perfluorocompounds (PFCs) in UK retail foods following determination using standard addition LC–MS/MS

The analysis of 252 food samples (UK-produced and imported) purchased from a variety of retail outlets in the UK was undertaken for the presence of perfluorooctanesulphonic acid (PFOS), perfluorooctanoic acid (PFOA) and nine other perfluorocompounds (PFCs). A limit of quantification (LOQ) of 1 µg/kg was achieved for all target analytes, in all samples. Standard addition was used for quantification of PFC levels. All 11 of the targeted PFCs were detected in 75 individual food items. In 70% of the samples, including all meat other than offal, none of the analytes were present above the LOD. The highest levels found were 59 µg/kg perfluorooctanesulphonic acid (PFOS) and 63 µg/kg total PFCs (ΣPFCs) in an eel sample, and 40 µg/kg PFOS (62 µg/kg ΣPFCs) in a whitebait sample. The highest level in an offal sample was 10 µg/kg, in a wild roe deer liver. There were six samples with ΣPFCs >15 µg/kg (fish, shellfish, crustaceans), a further seven samples with ΣPFCs ranging 11–15 µg/kg (including a liver), nine with ΣPFCs ranging 6–10 µg/kg (fish and livers), 31 with ΣPFCs in the range 2–5 µg/kg (including kidneys, popcorn and processed peas) and a further 22 with ΣPFCs close to the LOD of 1 µg/kg (including eggs and potatoes). These concentrations indicate that UK consumers are being exposed to a low level of PFC contamination from food. The estimated upper bound dietary intake of 10 ng/kg bodyweight (bw)/day of PFOS for average adult consumers is well below the 0.15 µg (150 ng)/kg bw tolerable daily intake (TDI) set by the European Food Safety Authority. The lower bound adult dietary intake estimate of 1 ng/kg bw/day is similar to estimates undertaken and reported in countries such as Canada, Germany and Spain.     

A novel approach to simultaneous screening and confirmation of regulated pharmaceutical compounds in dietary supplements by LC/MS/MS with an information-dependent acquisition method

The commercial success of synthetic phosphodiesterase-5 (PDE-5) inhibitors (viz. sildenafil, vardenafil and tadalafil) for erectile dysfunction (ED) has led to their widespread use as adulterants in dietary supplements (DSs). Reports on adulteration by ED drugs or their analogues in DSs suggest they may cause a serious threat to human health. The problem is becoming more complex as hidden and structurally modified analogues are continuously being reported. To analyse known drugs and their analogues, three commonly used PDE-5 inhibitors, naturally existing icariin and yohimbin, and their 19 analogues were analyzed in this study. They were identified using ion-spray liquid chromatography/tandem mass spectrometry (LC/MS/MS) using multiple reaction monitoring (MRM). This MRM procedure gave a limit of detection of less than 0.02?ng?ml^?1 for the 24 compounds, selectivity of fragmentation using MRM for 2.5?–?8.5?min in a single run and peak height repeatability of coefficient of variation of 3.9?–?31.8%. An IDA method using the MRM scans to detect the presence of known analytes was set up and added to a built-in library for screening for PDE-5 inhibitors. These MRM experiments were used to trigger product ion scans using a hybrid quadrupole-linear ion trap instrument. The product ion scan was compared and confirmed by a library search of MS/MS spectra acquired from a reference standard. To search for new analogues of PDE-5 inhibitors, a precursor ion scan of an expected ion m/z 283, which was one of the mass fragments from the analogues of sildenafil or vardenafil, was performed and fragmentation of the precursor ion, by combining a precursor ion scan with automatic confirmation using EPI spectra, was acquired. Of the 37 DSs tested, two were eventually found to be adulterated with yohimbin and vardenafil, respectively. The approach proposed in this study would be valuable in characterizing chemical constituents of drug residues and their analogues with identical chemical substructures from complex natural and synthetic sources in DSs using an information-dependent acquisition-enhanced product ion (IDA–EPI) scan.     

Melatonin is synthesised by yeast during alcoholic fermentation in wines

Melatonin (N-acetyl-5-methoxytryptamine) is a neurohormone produced in the pineal gland. Its biological properties are related to the circadian rhythm. Recently, the European Food Safety Authority (EFSA) accepted the health claim related to melatonin and the alleviation of subjective feelings of jet lag. This molecule has been detected in some foods. In this work, 13 grape varieties were studied; 7 monovarietal wines were produced in an experimental winery under strictly controlled conditions and were sampled in different steps. The grape varieties used to make the wines were: Cabernet Sauvignon, Merlot, Syrah, Tempranillo, Tintilla de Rota, Palomino Fino and Alpha red. Liquid chromatography tandem mass spectrometry (LC-MS/MS) unequivocally confirmed the presence of melatonin in wines. The main contribution of this paper is the results that clearly show that melatonin is synthesised during the winemaking process, specifically after the alcoholic fermentation. Indeed, melatonin is absent in grapes and musts and is formed during alcoholic fermentation.     

Yeast cell based feed additives: studies on aflatoxin B1 and zearalenone

Thirty commercially available yeast cell wall products and two reference bentonites were tested for their ability to bind aflatoxin B₁ (AFB1) and zearalenone (ZON) in buffer solutions at pH 3 and pH 6.5 as well as in real gastric juice. For most products, the binding efficacy of AFB1 correlated with the ash content, which was between 2.6 and 89%, and constituted the inorganic non-volatile components, such as mineral clays, of the samples. Samples with smectite as the main ash component showed the highest binding efficacy; yet, a correlation with the content of mannanooligosaccharides (MOS) and β-glucans from yeast cell walls was not observed. Products containing >30% ash showed AFB1 adsorption values >90% at least in one of the investigated media whereas most products with <10% ash did not exceed adsorption rates of 20%. In the case of ZON, adsorption efficiency ranged between 10 and 60%. It tended to be lowest for products with MOS and β-glucan contents <10% and greatest for products with MOS and β-glucan contents >50%. However, there was no general correlation between the adsorption of ZON and the concentration of MOS and β-glucans. Different products of one brand sold in different countries were observed to bind AFB1 to different degrees, which was explained by the difference in ash contents and mineral composition. In the case of ZON, differences in adsorption between products of the same brand were less pronounced.     

Bioaccumulation of cyanuric acid in edible tissues of shrimp following experimental feeding

Due to concerns that cyanuric acid (CYA)-contaminated feed had been used in aquaculture and could enter the human food chain, a method to quantify CYA residues in the edible tissues of fish and shrimp was previously developed and validated. This paper provides further data on the deliberate feeding of CYA to shrimp to determine the extent of residue accumulation in edible tissue. Liquid chromatography–tandem mass spectrometry (LC–MS/MS) was employed for the analysis of CYA in shrimp tissue. Edible tissue of shrimp fed 1666 or 3333?mg?kg^?1 CYA in their diet (approximately 55 and 124?mg?kg^?1 body weight) contained 0.767 and 0.406?mg?kg^?1 CYA, respectively. The residue levels are below the World Health Organization (WHO) tolerable daily intake level for CYA and are generally considered unlikely to pose a human health risk.     

Risk assessment for furan contamination through the food chain in Belgian children

Young, old, pregnant and immuno-compromised persons are of great concern for risk assessors as they represent the sub-populations most at risk. The present paper focuses on risk assessment linked to furan exposure in children. Only the Belgian population was considered because individual contamination and consumption data that are required for accurate risk assessment were available for Belgian children only. Two risk assessment approaches, the so-called deterministic and probabilistic, were applied and the results were compared for the estimation of daily intake. A significant difference between the average Estimated Daily Intake (EDI) was underlined between the deterministic (419?ng?kg^?1 body weight (bw) day^?1) and the probabilistic (583?ng?kg^?1 bw day^?1) approaches, which results from the mathematical treatment of the null consumption and contamination data. The risk was characterised by two ways: (1) the classical approach by comparison of the EDI to a reference dose (RfD_chronic-oral) and (2) the most recent approach, namely the Margin of Exposure (MoE) approach. Both reached similar conclusions: the risk level is not of a major concern, but is neither negligible. In the first approach, only 2.7 or 6.6% (respectively in the deterministic and in the probabilistic way) of the studied population presented an EDI above the RfD_chronic-oral. In the second approach, the percentage of children displaying a MoE above 10,000 and below 100 is 3–0% and 20–0.01% in the deterministic and probabilistic modes, respectively. In addition, children were compared to adults and significant differences between the contamination patterns were highlighted. While major contamination was linked to coffee consumption in adults (55%), no item predominantly contributed to the contamination in children. The most important were soups (19%), dairy products (17%), pasta and rice (11%), fruit and potatoes (9% each).     

Multi-residue method for the confirmation of four avermectin residues in food products of animal origin by ultra-performance liquid chromatography–tandem mass spectrometry

A confirmatory method was developed for the rapid determination of abamectin, ivermectin, doramectin and eprinomectin residues in various food products of animal origin, such as pork muscle, pork liver, fish and milk. Samples were homogenized, extracted and de-proteinized by acetonitrile, cleaned via two-step cleaning procedure using Bond Elut C₁₈ SPE columns and then alumina-N cartridges. All the four avermectin residues in different animal-food products were simultaneously separated and determined by ultra-performance liquid chromatography–electrospray ionization tandem mass spectrometry (UPLC–ESI–MS/MS) within 3.5?min. Data acquisition under positive ESI–MS/MS was performed by applying multiple reaction monitoring (MRM) for both identification and quantification, and mass spectrometric conditions were optimized to increase selectivity and sensitivity. The matrix-matched calibration curves for different matrices, such as pork muscle, pork liver, fish and milk, were constructed and the interference effect of different sample matrices on the ionization was effectively eliminated. The UPLC–MS/MS method was validated with satisfactory linearity, recovery, precision and stability. Matrix-matched calibration curves of abamectin, ivermectin, doramectin and eprinomectin in four different matrices were linear (r^{2 ?}≥?0.990, goodness-of-fit coefficients ≤12.8%) in the range 2.5–200?µg?kg^?1. The limits of detection and quantification for the four avermectins were in the range 0.05–0.68 and 0.17–2.27?µg?kg^?1, respectively. Recoveries were 62.4–104.5% with good intra- and inter-day precision. The method was rapid, sensitive and reliable, and can be applied to the quantitative analysis of avermectin residues in different animal-food products.     

Liquid chromatography tandem mass spectrometry determination of maduramycin residues in the tissues of broiler chickens

Maduramycin is a polyether ionophoric coccidiostat used to prevent coccidiosis in poultry at a prescribed concentration over a certain time interval. Due to public health concerns about the presence of coccidiostat residues in poultry, the aim of the present study was to determine the level of maduramycin residues in the tissues of broiler chickens fed commercially produced feed containing 5 mg kg^?1 of maduramycin in complete feed throughout the 5-day withdrawal period (WP). The residues were investigated by liquid chromatography (LC) coupled with electrospray ionisation (ESI) tandem mass spectrometry (MS/MS). The limit of detection (LOD) and limit of quantification (LOQ) of the method were 0.3 and 0.8 µg kg^?1, respectively. The average recovery based on matrix-fortified calibrations for chicken tissues was 90%. Maduramycin was found to be rapidly distributed in all tissues. The highest concentrations of maduramycin residues were found in the heart followed by the skin, liver, gizzard, kidneys and, finally, muscle (thigh and breast). On day 5 of the WP, residue concentrations of maduramycin did not decline below the LOQ of the method. Our results emphasize the need to establish a maximum residue limit (MRL) for maduramycin to control its residue levels in edible tissues from chickens before slaughter.     

Furan in coffee: pilot studies on formation during roasting and losses during production steps and consumer handling

The occurrence of furan in some food products has already been known for a few decades, and it has been reconfirmed in more recent investigations that furan is present in a variety of foodstuffs. This list of products includes roasted coffee, which has been shown to generate furan as a result of the heat treatment at roasting which is applied to achieve the desired aroma and flavour profile of a roasted coffee. The objective of this study is to provide data to allow a better understanding of the available data of furan in coffee, the kinetics of furan generated during roasting, and to estimate the reduction of furan levels afterwards due to subsequent processing steps and consumer handling. Finally, the study is meant as a contribution to establish exposure data on the basis of scientific data at the stage of coffee consumption. This paper shows that the formation of furan during roasting is dependent on roasting conditions and is, therefore, directly linked to achieving targeted flavour profiles. Furthermore, it is demonstrated that modifications in process conditions potentially to reduce furan levels may have the opposite effect on other undesired reaction products of the roasting chemistry such as, for example, acrylamide. Due to the high volatility of furan, any subsequent processing step or consumer handling has an impact on the level of furan. As a guidance from this study and in consideration of the identified losses of each process and handling step on the basis of the trial conditions, it is estimated that only approximately 10% of the initially generated furan during roasting gets into the cup of coffee for consumption.     

Evaluation of mepiquat in malted barley and beer using LC‐MS/MS

Following recent studies that showed that the agrochemical mepiquat (1,1‐dimethylpiperidinium) forms during the roasting of coffee beans and barley, this work investigates the presence of mepiquat in malted barley and commercially available beers. Liquid chromatography–tandem mass spectrometry was used to develop a sensitive and precise analytical method, with detection limits of 0.031 ng/g in malted barley and 0.014 ng/g in beer. Mepiquat was detected in nine out of 10 malted barley samples, with all results under the Canadian maximum residue limit (100 ng/g). The data suggest a relationship between perceived malted barley colour and mepiquat concentration. The concentration of mepiquat in the beers analysed was also below the maximum residue limits in Canada (100 ng/g) and in the EU (600 ng/g), suggesting that mepiquat is not a regulatory concern in finished beers. Copyright © 2015 The Institute of Brewing & Distilling     

酵母发酵对杏原料中氰化物的降解作用

杏中含有微量的氢氰酸，使其食用受到一定限制。在杏酒的开发过程中，对酵母酒精发酵是否能减少氢氰酸的含量作了探索研究。在采用的4种葡萄酒活性干酵母中，1号酵母菌和3号酒精发酵完成后，氰化物含量分别减少了56．7％和32．2％；2号酵母菌和4号酵母菌降解率低于10％。采用1号酵母菌的研究表明，随着发酵的进行氰化物含量逐步减少，在第6d降至最低。研究结果可能对利用微生物发酵解除或降低食品原料中有毒物质有所启发。     

Influence of thermal treatment of rapeseed on the canolol content

4-Vinylsyringol, also referred to as canolol, is a highly active antioxidant and potent lipidperoxyl radical scavenger found in rapeseed. The canolol content of rapeseed can be increased through the decarboxylation of sinapic acid via roasting treatments. Different roasting conditions were tested and compared and an optimum for the canolol formation was found at 160 °C. The canolol content of the rapeseed samples with optimal roasting increased by a factor of 120 in relation to the unroasted sample. The rapeseed was ground, extracted and analysed by normal-phase HPLC/UV. The structure of canolol was confirmed by NMR and MS techniques. Several rapeseed oils were purchased in German food stores and analysed. No differences in canolol content were observed in both cold-pressed and rape kernel oil samples tested. Dehulled rapeseed samples demonstrated no significant difference in canolol content when compared to unpeeled rapeseed samples.     

Determination of cyanidin-3-glucoside (red kernel food colour) in beverages by high performance liquid chromatography and a study of its degradation by quadruple time-of-flight mass spectrometry

A method for the determination of cyanidin-3-glucoside (red kernel colour, RKC) in various matrices, including carbonated soft drinks, fruit-based soft drinks, sugar confectionery and milk-based drinks, by high performance liquid chromatography (HPLC) based on UV–Vis detection (at 520?nm) have been developed. Pre-treatment procedures for various matrices have been optimised. For soft drinks, the pH values were adjusted to 2.0, and sugar confectionery and milk-based drinks were extracted with 30 and 50% methanol in 0.1% formic acid (pH?～?2.5) aqueous solutions, respectively. The limits of detection for the established methods ranged from 0.2?mg/kg for soft drinks to 2.0?mg/kg for the sugar confectionery and milk-based drinks with the relative standard deviations lower than 7%, which satisfy the method performance requirements for quality control for the beverages. The stability of the colourant RKC under various thermal and pH conditions was investigated by quadruple time-of-flight mass spectrometry (Q-TOF-MS). The main colourant component of RKC, cyanidin-3-glucoside (Cyd-3-Glu), lost the glucose entity under thermal treatment. A new component was identified when RKC degraded under various pH conditions and a degradation pathway is proposed. The results will assist in understanding the degradation mechanism of the colourant Cyd-3-Glu.     

Mass-defect filtering of isotope signatures to reveal the source of chlorinated palm oil contaminants

This paper reports new insights at the molecular level into the route of a worldwide problem of the food industry: the occurrence of monochloro-propanediol (MCPD) esters. The application of mass defect-driven workflows is described to generate a hypothesis on the identity and occurrence of those thermally labile, chlorinated contaminant precursors that may act as chlorine donors during the formation of MCPD esters. For the first time, holistic mass-defect filtering of isotope signatures is used to pinpoint completely unknown and unexpected chlorine-containing substances naturally present in various extracts of palm fruit and partially and fully refined oils. Supervised multivariate analysis showed the effective classification of samples from various stages of industrial processing, suggesting that these steps strongly impact a complex and dynamic pool of chlorinated substances. In-vitro experiments confirmed that several of these naturally occurring chlorinated plant constituents decompose upon heat treatment, thus potentially being a source of chlorine for further reactions with palm oil lipids in a subsequent chlorination cascade. It is hypothesised that during oil refining the organochlorines naturally present in palm fruits act as a ‘chlorine source’ for the generation MCPD diesters. This discovery implies that industrial efforts targeting the mitigation of chlorinated substances must intervene at the earliest possible production stage or preferably even prior to oil processing. Current performance and limitations of mass-defect filtering are discussed and future developments are outlined.     

Assessment of the acrylamide intake of the Belgian population and the effect of mitigation strategies

The acrylamide (AA) intake of the Belgian consumer was calculated based on AA monitoring data of the Belgian Federal Agency for the Safety of the Food Chain (FASFC) and consumption data of the Belgian food consumption survey coordinated by the Scientific Institute for Public Health (3214 participants of 15 years or older). The average AA exposure, calculated probabilistically, was 0.4 µg kg^?1 body weight (bw) day^?1 (P97.5 = 1.6 µg kg^?1 bw day^?1), the main contributors to the average intake being chips (23%), coffee (19%), biscuits (13%), and bread (12%). Additionally, the impact of a number of AA mitigation scenarios was evaluated (German minimization concept, scenarios for mitigation from the literature, signal values), which is an important issue for public health as well as for policy-makers. Specific actions in cooperation with the food industry to reduce the AA content of foods seems to be a more efficient strategy than mere implementation of signal values. Considering that an important share of the AA intake is due to prepared meals, the catering industry as well as consumers need to be better informed on the various possibilities for keeping the AA content of meals as low as possible.     

The chemistry and analysis of annatto food colouring: a review

Annatto food colouring (E160b) has a long history of use in the food industry for the colouring of a wide range of food commodities. The principal colouring component of annatto is the oil-soluble diapo carotenoid bixin, which is the methyl ester of the dicarboxylic acid norbixin and soluble in aqueous alkali. Bixin and norbixin, therefore, exhibit not only physicochemical properties normally associated with carotenoids, but also certain anomalous properties that have an impact on the stability, food colouring applications, and importantly the analysis of annatto. This review summarizes the key aspects of the structural determination of bixin (and norbixin) with special attention to cis-trans isomerization and how this links with its chemical structure, spectroscopic properties, and stability. The oxidative, thermal, and photo-stability of annatto and the subsequent implications for its use in the colouring of foods, food processing, and the analysis of foods and beverages are discussed along with important mechanistic, thermodynamic and kinetic aspects. The main analytical techniques used for the chemical characterization of annatto, i.e. spectrophotometry, nuclear magnetic resonance (NMR), chromatography (particularly high-performance liquid chromatography (HPLC)) and mass spectrometry are reviewed in detail and other methods are discussed. This links in with a review of the methods available for the detection and measurement of annatto in colour formulations and foods and beverages, which highlights the importance of the need for a good understanding and knowledge of the chemistry of bixin and norbixin in order to meet new analytical challenges.     

Identification and quantification of the migration of chemicals from plastic baby bottles used as substitutes for polycarbonate

The results of a study on the analytical identification and quantification of migration of chemicals from plastics baby bottles found in the European Union market made of materials that are now present as substitutes for polycarbonate (PC) are reported. A total of 449 baby bottles with a focus on first age or sets of bottles were purchased from 26 European Union countries, Canada, Switzerland and the USA. From this collection, which contained several duplicates, a total of 277 baby bottles were analysed. The materials included different types of plastic such as PC, polyamide (PA), polyethersulphone (PES), polypropylene (PP), but also silicone, and from the United States a co-polyester marketed under the trade name Tritan?. The bottles were subjected to the conventional migration test for hot fill conditions, i.e. 2?h at 70°C. The simulant used was that specified in European Union legislation (2007/19/EC) for milk, i.e. 50% ethanol. In a first phase 1, migration was conducted since the scope of this investigation was a screening rather than a true compliance testing check. Second and third migrations were performed on selected articles when migrated substances exceeded limits specified in the legislation. In order to verify some materials, a portion of the bottle was cut to run an FT-IR fingerprint to confirm the nature of the polymer. The migration solutions in general showed a low release of substances. Results showed that bottles made of PP and silicones showed a greater number of substances in the migration solutions and in greater quantity. Chemicals from PP included alkanes, which could be found in >65% of the bottles at levels up to 3500?µg?kg^?1; and benzene derivatives in 17% of the baby bottles and found at levels up to 113?µg?kg^?1. Some substances were found on a regular basis such as plasticisers, esters and antioxidants (e.g. tris(2,4-di-tert-butylphenyl)phosphate, known as Irgafos 168. Some substances found were not included in the Community positive list, which means that those should not be found even in the first migration. Such substances included 2,6-di-isopropylnaphthalene (DIPN), found in 4% of the bottles at levels up to 25?µg?kg^?1, 2,4-di-tert-butyl phenol (in 90% of the bottles at levels up 400?µg?kg^?1). Moreover, bisphenol A (BPA) was detected and quantified in baby bottles made of PA, but limited to one brand and model specific (but labelled BPA free). Results for baby bottles made of silicone also indicated the presence of components, e.g. potentially coming from inks (benzophenone, diisopropyl naphtahalene – DIPN, which could come for example from the presence of instruction leaflets in the bottles). In the case of silicone, phthalates were also found in relevant concentrations, with levels for DiBP and DBP from the first migration test of 50–150?µg?kg^?1 and DEHP at levels 25–50?µg?kg^?1.