Patulin In Fruit Juices Research Articles

Long‐term investigation and analysis of patulin occurrence in juice products in China

SummaryExcessive patulin contamination in fruit juice products pose a significant risk to human health. However, there is a notable absence of a comprehensive and long‐term survey regarding the contamination of patulin in fruit juices within the Chinese market. The present study revealed a discernible decline in the patulin contamination levels, which were as follows: 60% (33/55), 18.28% (17/93), 0% (0/122), 15.58% (12/77), and 0% (0/63) at various survey time dates (2014, 2016, 2018, 2020, 2022). Among them, 18.18% (in 2014), 6.45% (in 2016), and 3.90% (in 2020) of samples exceeded the maximum limit concentration of 50 μg/L, respectively. Despite the presence of varying levels of patulin contamination, the average daily intake of patulin across all years did not present a discernible risk to human health. This investigation offers valuable insights for evaluating the extent of human exposure to patulin and establishing regulatory measures to ensure the safety of juice products.

Cellulose-based magnetic nanomaterials immobilized esterases as a reusable and effective detoxification agent for patulin in apple juice

Patulin, an important contaminant in apple juice processing, exhibits various toxicities to humans. In existing patulin detoxification methods, enzymes are desirable biological detoxification agents. Here, magnetic nanocrystalline cellulose (MNCC) was prepared and employed as support for the immobilization of porcine liver esterase (PLE) to synthesize a novel cellulose-based magnetic patulin detoxification agent (PLE@MNCC). The detoxification rate of patulin by PLE@MNCC exceeded 80%. PLE@MNCC also enabled rapid separation and recovery after application while exhibiting superior storage stability, reusability and biocompatibility. After 8 repetitive cycles of usage, PLE@MNCC retained approximately 50% of its initial activity. Moreover, the cytotoxicity evaluation demonstrated a significant decrease in patulin toxicity after the detoxification. Crucially, there were no significant adverse changes in the quality of apple juice after detoxification. PLE@MNCC as an effective, reusable, environmentally friendly and safe enzyme agent provided an efficient strategy for the detoxification of patulin in fruit juices.

Visualization detection of mycotoxin patulin in fruit juices by a small-molecule fluorescent probe.

The mycotoxin patulin is a common contaminant in rotten fruits, posing severe food safety risks and threats to human health. Developing a convenient, sensitive and reliable method for patulin detection is of utmost importance but remains challenging. In this study, we have successfully designed and synthesized a small-molecule fluorescent probe, FITC-Lys, which demonstrates good sensitivity in detecting patulin. Upon contact with patulin, the terminal Lys group of the FITC-Lys probe reacts with patulin, resulting in the formation of the fluorescein dimer that subsequently quenches fluorescence. This variation of fluorescence enables the visualization and sensitive detection of patulin. The probe exhibits good sensitivity with a low LOD of 8 ng mL-1 for the fluorescence spectrum method and a LOD of 12 ng mL-1 for the fluorescence imaging method. Moreover, we have validated the probe's capability for patulin detection in apple and pear juices, achieving good recoveries ranging from 98.60% to 103.80%. Notably, the probe FITC-Lys is the first small-molecule fluorescent probe that has proven successful in visualizing patulin in juices derived from decayed apples and pears. Consequently, this probe holds great potential as a practical tool for monitoring patulin in foodstuffs, thereby contributing to enhanced food safety standards.

Determination of patulin in apple juice by liquid chromatography-electrospray tandem mass spectrometry

Patulin is a mycotoxin produced by several fungi, (Penicillium, Aspergillus, Byssochlamys). The main sources of patulin intake in human diet are apples, apple juice and apple nectar, and for this reason, apple based foods are monitored for the presence of this mycotoxin. Commission Regulation EC No 1881/2006 lays down maximum residue limits (MRLs) of 50 µg/kg in apple juice and cider, 25 µg/kg in solid apple products, and 10 µg/kg in products for infants and young children. In Serbia, maximum permitted amounts of patulin in fruit juices, reconstituted concentrated fruit juices and fruit nectars, as well as in solid apple products, including apple compote and apple puree, intended for direct human consumption are prescribed in the Regulation on maximum concentrations of certain contaminants in foodstuffs. This paper presents the LC-MS/MS method for quantitative determination of patulin in apple juice. Criteria for method validation were taken from Commission Decision 2002/657/EC. Linearity was confirmed in the concentration ranges of 0-100 µg/kg, with the limit of detection (LoD) of 9.85 µg/kg. The performance of the method was successfully verified by participating in a proficiency study.

Optimization of green and rapid analytical procedure for the extraction of patulin in fruit juice and dried fruit samples by air-assisted natural deep eutectic solvent-based solidified homogeneous liquid phase microextraction using experimental design and computational chemistry approach

In this paper, a green and inexpensive air-assisted natural deep eutectic solvent-based solidified homogeneous liquid phase microextraction procedure was optimized for extraction of patulin in fruit juice and dried fruit samples using experimental design prior to its spectrophotometric determination. Four different natural deep eutectic solvent were prepared and applied to ensure efficient, and selective extraction of patulin. The significant variables including Zn(II) amount, cooling time, pH and amount of natural deep eutectic solvent were optimized by using central composite design. Under optimized conditions, working range was 10–750 μg L−1 with 0.9996 of correlation coefficient. Detection limit and preconcentration factor were 3.5 μg L−1 and 150, respectively. The repeatability and reproducibility precision were in the range of 3.2–4.6% and 4.3–5.6% respectively. Recoveries ranging from 94% to 104% proved the accuracy of the method. The optimized method was successfully applied to the extraction and identification of patulin in the selected samples.

Leuconostoc mesenteroides subsp. mesenteroides LB7 isolated from apple surface inhibits P. expansum in vitro and reduces patulin in fruit juices

This research aimed to isolate lactic acid bacteria (LAB) from apple surface and to reveal their potential to inhibit the growth of Penicillium expansum. Besides, their ability to detoxify fruit juices contaminated with mycotoxin patulin, produced by this fungi, was also studied. The isolation was performed on a typical MRS medium under ambient conditions. The molecular identification of the strains was done by sequencing the 16S rRNA genes. Antifungal activities of the isolated strains have been evaluated using dual agar plate assay protocol. A total of 11 LAB isolates was obtained from apples. These isolates showed phenotypic traits consistent with the genera of LAB. They have been identified as Leuconostoc mesenteroides subsp. mesenteroides and Weissella paramesenteroides. Among them, the strain LB7 showed exciting inhibitory activities in vitro against P. expansum. LB7 also successfully detoxified homemade and commercial fruit juices contaminated with patulin. Further research will bring the application prospects of these LABs in food biocontrol and biopreservation strategies.

Degradation of patulin in fruit juice by a lactic acid bacteria strain Lactobacillus casei YZU01

Patulin, a secondary metabolite produced by fungi, such as Aspergillus, Penicillium, and Byssochlamys, is highly toxic to bacteria and plants and exerts serious threat to human and animal health. The use of beneficial microbes shows good effect and application prospects in removing mycotoxins. As a widely used biological control agent, lactic acid bacteria (LAB) can remove mycotoxins from foodstuff. In this research, a LAB strain that can degrade patulin, namely, Lactobacillus casei YZU01, was screened. The physiological mechanism of patulin degradation by L. casei YZU01 was investigated. L. casei YZU01 mainly degraded patulin by secreting extracellular metabolites, and its cell wall also absorbed patulin. Furthermore, L. casei YZU01 was applied for the removal of patulin in raw apple juice (RAJ) or raw pear juice (RPJ). Patulin (10 μg/mL) in RAJ or RPJ was completely degraded by L. casei YZU01 after 36 h of incubation, and patulin in commercialized apple or pear juice products was degraded after 48 h of incubation after treatment with L. casei YZU01. Results indicated the potential of L. casei YZU01 in dealing with patulin contamination in food industries.

A novel, green and safe ultrasound-assisted emulsification liquid phase microextraction based on alcohol-based deep eutectic solvent for determination of patulin in fruit juices by spectrophotometry

In the study, a novel, green and safe microextraction procedure was developed for the determination and extraction of patulin, α, β-unsaturated lactone (4-hydroxy-4H-furo[3,2-c] pyran-2-(6H)-one), in fruit juice samples by using ultrasound-assisted emulsification liquid phase microextraction based on alcohol-based deep eutectic solvent coupled to micro-volume ultraviolet-visible spectrophotometry. The method is based on formation of chelate complex or nucleophilic adduct formation between patulin and Mg(II) in presence of nucleophilic metal chelating tris-buffer at pH 8.0, and then extraction of the formed complex into the micro-drops of deep eutectic solvent by donor-acceptor mechanism. The optimum conditions for the effective extraction of patulin from fruit juices were 500 μL of deep eutectic solvent, 75 μmol L−1 of Mg(II) solution, pH 8.0, ultrasound time of 12 min, respectively. The patulin in enriched phase was detected by micro-volume ultraviolet-visible spectrophotometry at 304 nm. Under optimal conditions, the linear response range of the method was from 7.5 to 420 μg L−1 with a detection limit of 2.2 μg L−1. After the validation study, the applicability of the method were tested by analysis of the fruit juice samples. The average recovery for spiked fruit juices was ranged from 90.2% to 106.9%, with relative standard deviations lower than 4.2%.

Patulin in fruit juices: occurrence, bioaccessibility, and risk assessment for Serbian population

ABSTRACTThis paper reports the first study of patulin occurrence in fruit juices and risk assessment related to patulin intake by children, adolescents and adults in Serbia. During 3 consecutive years (2013–2015), 142 fruit (apple or multi-fruit) juices were collected from the market and analysed using HPLC-UV. Patulin was found in 51.4% of juices with 0.7% of the samples in excess of the legal limit of 50 μg kg−1 (mean 4.3 μg kg−1). Apple juices showed significantly higher percentage of contaminated samples (74.0% vs 27.5%), as well as higher mean patulin content (6.4 vs 2.1 μg kg−1) when compared with the multi-fruit ones. Bioaccessibility of patulin in fruit juices was studied using the standardised in vitro digestion method. A mean of 21.6% of the initial patulin amount reached the end of the intestinal phase showing a significant reduction of this toxin during the human digestion process. Risk assessment of patulin intake by Serbian children, adolescents and adults, conducted by deterministic and probabilistic approaches and including the bioaccessibility results, revealed no health concern. Although patulin alone does not represent risk, further research should consider its co-occurrence with other toxic substances in food and potential adverse effects of their mixtures.

A novel molecularly imprinted electrochemical sensor modified with carbon dots, chitosan, gold nanoparticles for the determination of patulin

In this paper, molecular imprinting technique was applied to the electrochemical sensor. We used 2-oxindole as dummy template, ρ-Aminothiophenol (ρ-ATP) as functional monomers, combined with the high sensitivity of electrochemical detection, to achieve a specific and efficient detection of patulin in fruit juice. In addition, carbon dots and chitosan were used as the modifying material to improve electron-transfer rate, expand the electroactive surface of glassy carbon electrode and enhance strength of the signal. The Au–S bond and hydrogen bond were employed to complete the assembly of the ρ-ATP and 2-oxindole on the surface of the electrode. Then, polymer membranes were formed by electropolymerization in a polymer solution containing ρ-ATP, HAuCl4, tetrabutylammonium perchlorate (TBAP) and the template molecule 2- oxindole. After elution, the specific cavity can adsorb the target patulin. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) measurements were performed to monitor the electropolymerization process and its optimization. Transmission electron microscopy (TEM), Scanning electron microscopy (SEM) and Atomic force microscopy (AFM) analyses were used for characterization. This was the first time that the molecularly imprinted polymer (MIP) technology combined with carbon dots, chitosan and Au nanoparticles modification and was applied in the electrochemical detection of patulin. The linear response range of the MIP sensor was from 1 × 10–12 to 1 × 10−9molL−1 and the limit of detection (LOD) was 7.57 × 10–13molL−1. The sensor had a high-speed real-time detection capability, low sample consumption, high sensitivity, low interference, good stability and could become a new promising method for the detection of patulin.

Immobilization of inactivated microbial cells on magnetic Fe3O4@CTS nanoparticles for constructing a new biosorbent for removal of patulin in fruit juice

Patulin contamination of juice is an important food safety issue throughout the world. In this paper, magnetic Fe3O4@CTS nanoparticles were coated with inactivated C. utilis CICC1769 cells and used as a new biosorbent for the removal of patulin in fruit juice. Prepared nanoparticles were characterized by X-ray diffraction and FT-IR spectra analyses and the results indicated that inactivated C. utilis CICC1769 cells were successfully immobilized on the surface of magnetic Fe3O4@CTS. Furthermore, this new biosorbent was found to reduce patulin by over 90% in the orange juice without any significant negative impacts on the quality parameters such as Brix, Vitamin C, and titratable acidity. This study shows that there are promising industrial applications of the as-prepared biosorbent for juice-detoxification.

Effect of Yeast Cell Morphology, Cell Wall Physical Structure and Chemical Composition on Patulin Adsorption.

The capability of yeast to adsorb patulin in fruit juice can aid in substantially reducing the patulin toxic effect on human health. This study aimed to investigate the capability of yeast cell morphology and cell wall internal structure and composition to adsorb patulin. To compare different yeast cell morphologies, cell wall internal structure and composition, scanning electron microscope, transmission electron microscope and ion chromatography were used. The results indicated that patulin adsorption capability of yeast was influenced by cell surface areas, volume, and cell wall thickness, as well as 1,3-β-glucan content. Among these factors, cell wall thickness and 1,3-β-glucan content serve significant functions. The investigation revealed that patulin adsorption capability was mainly affected by the three-dimensional network structure of the cell wall composed of 1,3-β-glucan. Finally, patulin adsorption in commercial kiwi fruit juice was investigated, and the results indicated that yeast cells could adsorb patulin from commercial kiwi fruit juice efficiently. This study can potentially simulate in vitro cell walls to enhance patulin adsorption capability and successfully apply to fruit juice industry.

Vortex-assisted liquid–liquid microextraction coupled with high performance liquid chromatography for the determination of furfurals and patulin in fruit juices

A fast and simple solvent microextraction technique using salting out-vortex-assisted liquid-liquid microextraction (salting out-VALLME) was developed for the extraction of furfurals (2-furfural (2-F), 3-furfural (3-F), 5-methylfurfural (5-MF) and 5-hydroxymethylfurfural (5-HMF)) and patulin (PAT) in fruit juice samples. The optimum extraction conditions for 5 mL sample were: extraction solvent, 1-hexanol; volume of extractant, 200 µL; vortex time, 45 s; salt addition, 20%. The simultaneous determination of the furfurals and PAT were investigated using high performance liquid chromatography coupled with diode array detector (HPLC-DAD). The separation was performed using ODS Hypersil C18 column (4.6 mm i.d × 250 mm, 5 μm) under gradient elution. The detection wavelengths used for all compounds were 280 nm except for 3-F (210 nm). The furfurals and PAT were successfully separated in less than 9 min. Good linearities (r(2)>0.99) were obtained within the range 1-5000 μg L(-1) for all compounds except for 3-F (10-5000 µg L(-1)) and PAT (0.5-100 μg L(-1)). The limits of detection (0.28-3.2 µg L(-1)) were estimated at S/N ratio of 3. The validated salting out-VALLME-HPLC method was applied for the analysis of furfurals and PAT in fruit juice samples (apple, mango and grape).

A comparison of the selectivity of nano-HPTLC systems used for determination of patulin in fruit juices from the internet stores and pharmacies

The new nano-HPTLC method was proposed for improved separation of patulin from the admixture of the three naturally occurring natural components of fruit juices, which in last years could be often obtained from the internet functional food stores and pharmacies. Separation of patulin (P) from quercetin (Q), ascorbic acid (A) and 5-hydroxymethylfurfural (H) was performed using three different ready-for-use plates NanoAdamant (NA), Nano-SIL-20 (NSIL) and Nano-Durasil-20 (ND) covered with the non-modified silica gel layer. After development on the 8.0 cm distance, the nano-HPTLC plates were sprayed with 0.5% aqueous solution of the 3- methyl-benzothiazolinone hydrazone hydrochloride hydrate (MBTH). Most satisfactory separation of patulin from the each mentioned compounds was obtained using the Nano-SIL-20 plates and toluene-ethyl acetate-98% formic acid (25:20:5, v/v/v) as the mobile phase. In this best nano-HPLTC method the limit of detection of patulin was near 2.5 times lower (4 ng/spot) in comparison to other used here chromatographic systems. Thus, this optimized nano-HPTLC mode could be used for analysis of the solid-phase extracts of patulin from the fresh apples, clarified apple juices and other fruit products.

Survey of the presence of patulin in fruit juices

Patulin is a mycotoxin produced by fungi belonging to the Penicillium and Aspergillus genera. The occurrence of patulin in fruit juices marketed in Italy in 2008 and purchased from supermarkets and retail shops has been measured. The aim of this study was to assess the presence of patulin in order to evaluate the potential risk for the consumer and, at the end of this food chain, to determine the quality of the raw material used. One hundred and five fruit juices (35 apple juice, 35 mixed-taste juices, 35 pear juices), produced by various Italian and European companies, were analysed using a previously published method. The analytical investigation showed that apple juices had a concentration of patulin ranging between 6 and 30 µg l−1, with a mean of 18 µg l−1; mixed fruit juices had a concentration ranging between 1 and 45 µg l−1, with a mean of 23 µg l−1. Instead, pear juices had a concentration ranging between 5 and 92 µg l−1, with a mean of 43 µg l−1, and 14 samples of the 35 analysed juices showed a patulin level above the highest regulated limit of 50 µg l−1, imposed by European Commission Regulation 1881/06.

Occurrence of patulin in various fruit juices from South Korea: An exposure assessment

To determine patulin in various fruit juices, high performance liquid chromatography (HPLC) method was optimized and validated. For validation of HPLC method, a linearity, accuracy, precision, detection limit, and quantification limit were determined. Linearity (R2 = 0.99995), accuracy (96.1–115.7%), precision (3.31–9.52), detection limit (6 ng/mL), and quantification limit (8 ng/mL) were in agreement with performance criteria for patulin as set by the European Commission hence proved that HPLC can be used to detect patulin in fruit juices. After validation, the method was applied to estimate the prevalence of patulin in fruit juices (apple, grape, and orange juices). Nine samples (12.5%, 3 apple, 2 orange, and 4 grape juices) of 72 samples were positive for patulin in the range 2.8 to 30.9 ng/mL. According to the monitoring results, daily intake was estimated to be 0.17 ng/kg BW/day which was lower than the provisional maximum tolerable daily intake (0.4 μg/kg) established by Joint Expert Committee on Food Additives. These results indicate that the detection method coincides with the performance criteria and is appropriate for analysis of patulin, and continuous monitoring of patulin in various fruit juices from Korea is necessary.

Determination of patulin in fruit juice and dried fruit samples by in-tube solid-phase microextraction coupled with liquid chromatography–mass spectrometry

A simple and sensitive method for the determination of patulin in fruit juice and dried fruit samples was developed using a fully automated method consisting of in-tube solid-phase microextraction (SPME) coupled with liquid chromatography–mass spectrometry (LC–MS). Patulin was separated within 5 min by high-performance liquid chromatography using a Synergi MAX-RP 80A column and water/acetonitrile (80/20, v/v) as the mobile phase. Electrospray ionization conditions in the negative ion mode were optimized for MS detection of patulin. The pseudo-molecular ion [M−H] − was used to detect patulin in selected ion monitoring (SIM) mode. The optimum in-tube SPME conditions were 25 draw/eject cycles of 40 μL of sample using a Carboxen 1006 PLOT capillary column as an extraction device. The extracted patulin was readily desorbed from the capillary by passage of the mobile phase, and no carry-over was observed. Using the in-tube SPME LC–MS with SIM method, good linearity of the calibration curve ( r = 0.9996) was obtained in the concentration range of 0.5–20 ng/mL using 13C 3-patulin as an internal standard, and the detection limit (S/N = 3) of patulin was 23.5 pg/mL. The in-tube SPME method showed >83-fold higher sensitivity than the direct injection method (10 μL injection volume). The within-day and between-day precision (relative standard deviations) were below 0.8% and 5.0% ( n = 6), respectively. This method was applied successfully for the analysis of fruit juice and dried fruit samples without interference peaks. The recoveries of patulin spiked into apple juice were >92%, and the relative standard deviations were <4.5%. Patulin was detected at ng/mL levels in various commercial apple juice samples.

Determination of patulin in fruit juices using HPLC-DAD and GC-MSD techniques

A high performance liquid chromatography with a diode-array detector (HPLC-DAD) and a gas chromatography with a mass spectrometer (GC-MSD) are described for the determination of patulin (PAT) in apple juice. The limits of detection (DL) and quantification (QL) for the HPLC-DAD and GC-MSD method were found to be (DL=0.23μgkg−1 QL=1.2μgkg−1) and (DL=5.8μgkg−1 and QL=13.8μgkg−1), respectively. The recovery factors for HPLC-DAD and GC-MSD were found to be 99.5% (RSD%=0.73) and 41% (RSD%=10.03), respectively. The HPLC-DAD method was used to determine the occurrence of PAT in 90 samples of fruit juices.Results revealed the presence of PAT in 100% of the samples examined. The mean values of PAT in concentrated fruit juices and in the commercial fruit juices collected from the Greek market were found to be 10.54μg PAT kg−1 and 5.57μg PAT kg−1 juice, respectively. The most contaminated samples were four concentrated juices ranging from 18.10μg PAT kg−1 to 36.8μg PAT kg−1 juice. The daily exposure to patulin for the consumers of all ages in Greece, is ranging from 0.008μg PAT kg−1 bw to 0.1μg PAT kg−1 bw if the daily intake of fruit juices is from 0.1 to 0.5kg. With the exception to the most contaminated sample, the daily exposure due to the samples examined, is below the provisional maximum tolerable daily intake for PAT (0.4μg PAT kg−1 bw).

Neue bestimmungsmethode des mykotoxins patulin mit hilfe der dünnschicht- und hochdruckflüssigchromatographie

The quantitative analysis of the mycotoxin patulin in fruit juices by high-pressure liquid or thin-layer chromatography A new method of determining patulin in apple and grape juice is proposed. The juice is extracted with an Extrelut ® column and the quantitative determination is made by high-pressure liquid or thin-layer chromatography with lower limits of 5 and 20 ppb (10 9), respectively. The recovery is more than 90%. Compared with other methods the proposed procedure is faster, needs less solvents and is therefore suitable for serial investigations.