![]() ![]() The deposited phase was completely transferred to another test tube and evaporated to dryness. The extraction solvent was rapidly injected into the sample solution by a glass syringe.Īfter forming a cloudy solution, the extraction was centrifuged at 4,000 rpm for 3 min subsequently, chloroform and ethyl acetate were collected from the bottom of a conical test tube. The extraction solvent contained a mixture of 1.0 mL acetonitrile as the disperser and 250 μL ethyl acetate: chloroform (190 μL: 60 μL). The separation of the dispersive phase was performed using a centrifuge model, namely Tlettich universal 320.įor the implementation of DLLME, under optimum conditions, 5.00 mL blank apple juice was spiked with 5 μL of 50 ppb patulin in a 10-mL glass test tube. The detection was performed at a wavelength of 276 nm. A mixture of water and acetonitrile (97:3) at a flow rate of 1 mL/min was utilized as the mobile phase in isocratic elution mode. A chromolith HPLC column (15 cm, Agilent) was selected for the measurement. The HPLC system equipped with an auto-sampler (Waters 717), an HPLC pump (Waters 1525), and a dual λ absorbance ultraviolet detector (Waters 2487) were used for the analysis. Afterwards, the filtrate was diluted at 1:4 ratios with deionized water and used for dispersive liquid-liquid microextraction (DLLME) procedure. The fresh juice was centrifuged at 3,500 rpm for 15 min, the supernatant was then filtered through a 0.45-μm membrane filter. Once opened, they were stored in specific food containers at 4☌ and analyzed within 5 days. The samples were stored at ambient temperature before further use. The apple juice samples were collected from different supermarkets (Shiraz, Iran). Patulin and standard solutions were prepared by dilution in ethanol. The stock solution of patulin was 10 mg/L. Water and acetonitrile supplied of high-performance liquid chromatography (HPLC) grade and other chemicals, such as acetone, ethanol, methanol, chloroform, carbon tetrachloride, dichloromethane, carbon disulfide, ethyl acetate, and sodium chloride, were obtained from the Merck Company (Darmstadt, Germany). Patulin (with purity of ≥ 99%) and hydroxymethylfurfural standards were supplied from the Sigma-Aldrich Corporation (USA). These samples were produced by 15 Iranian manufacturers. This study was conducted on a total of 75 samples, including 38 and 37 samples of apple juice and apple cans, respectively, which were randomly purchased from the retail markets during 2016. Regarding this and considering the toxic effects of patulin on people’s health, especially children, the present study was conducted with the aim of estimating the level of patulin in apple juice and cans of different brands. There are limited number of studies investigating this mycotoxin in Iran. observed the concentration level of ≥ 50 µg/L patulin in 10% of the samples. In some other studies performed in Spain and Japan, the apple products were reported to be contaminated with patulin. ![]() In a study conducted in Tunisia, the concentration of patulin in 35% of the samples was over 50 µg/L. The level of patulin in apple products has been reported in several countries. Īccordingly, the European Union in the Commission Regulation 1425/2003 has recommended the patulin levels of < 50, 25, and 10 µg/L for apple juice, beverage containing apple juice, and solid apple products, respectively. ![]() Currently, the Alimentarius and Food and Drug Administration have recommended a maximum level of 50 µg/L patulin for apple products. Patulin is reported more frequently in apple products among other different fruit products. Patulin has been recognized as a health-threatening substance in several countries. Although this heat-resistance toxin was initially studied as a potential new antibiotic, different studies have demonstrated its immunotoxic, genotoxic, embryotoxic, and neurotoxic properties. Patulin is mainly found in the rotten parts of fruits and vegetables, such as apples, pears, peaches, apricots, and grapes. Patulin (4-hydroxy-4H-furopyran-2(6H)-one), is one of the important mycotoxins, produced by a wide range of molds, in particular Penicillium, Aspergillus, and Bysochlamys, among which Penicillium expansum is the most isolated species. The toxins may form on fields or during the storage or even processing of the foods. ![]() Mycotoxins are secondary abiotic hazard metabolite produced by fungi in the contaminated foods. ![]()
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