Patulin in apple juice

Takino, M., Daishima, S., and Nakahara, T. (2003). Liquid chromatography/mass spectrometric determination of patulin in apple juice using atmospheric pressure photoionization. Rapid Commun. Mass Spectrom. 17, 1965—1972. 

Within the Codex system, the contaminants considered in this section are mainly dealt with by the Codex Committee on Food Additives and Contaminants (CCFAC), which is hosted by the Netherlands. Many of the Member States of the European Union are very active in CCFAC. For example, Denmark and the Netherlands have been instrumental in developing the Codex General Standard on Contaminants and Toxins and draft limits for lead in various foods. Sweden has developed a proposal for a limit for ochratoxin A in cereals and cereal products and France has proposed a maximum level for patulin in apple juice. 

Apples and pears are the fruits most frequently contaminated with patulin. As a consequence, most scientific publications on mycotoxin contamination of fruit juices deal with the occurrence of patulin in apple juice. Table V shows the most important surveys on the occurrence of patulin in apple juice for the last five years. The frequency of contamination ranged from 13 to 81%. Apart from one Turkish study with a mean of 140 pg/L (Yurdun et al., 2001), mean patulin concentrations were rather low and usually below 50 pg/ L. The highest patulin concentration was 733 pg/L, but concentrations up to 1150 pg/kg have occasionally been reported for commercially available apple juice (Beretta et al., 2000). 

Certain groups of a population may be at risk for elevated exposure to mycotoxins if certain fruit products significantly contribute or even dominate the daily diet. This has especially been shown for infants and young children for patulin in apple juice and ochratoxin A in dried vine fruits. Furthermore, wine and cider may significantly contribute to ochratoxin A intake of adults. As a consequence, maximum mycotoxin concentrations for certain fruit products have been established in several countries. 

Food and Drug Administration 2001. Patulin in apple juice, apple juice concentrates and apple juice products, http //vm.cfsan.fda.gov/ dms/patubckg.html April 14, 2004. 

Leggott, N.L., Shephard, G.S., Stockenstrom, S., Staal, E., and van Schalkwyk, D.J. 2001. The reduction of patulin in apple juice by three different types of activated carbon. Food Addit. 

Yurdun, T., Omurtag, G.Z., and Ersoy, O. 2001. Incidence of patulin in apple juices marketed in Turkey. J. Food Prot. 64, 1851-1853. 

The assoeiation of fungi with mammalian diseases came to the fore in 1960 with the diseovery of the aflatoxins (1.36). The death of turkeys from liver damage having been fed on groundnuts eontaminated with Aspergillus flavus led to the isolation of the highly carcinogenic aflatoxins. These developments are discussed in Chapter 9. It led to the awareness of the potential human health hazards from microbial metabolites and the implications of the presence of other mycotoxins in foodstuffs such as patulin in apple juice and the tri-chothecenes on corn. The development of analytical methods for the detection of mycotoxins has become an important aspect of food science. 

Methods exist for many matrix/mycotoxin combinations including patulin in apple juice using atmospheric pressure photoionization, zearalenone and its metabolites in fish tissue and porcine urine, muscle, and liver tissues using electrospray ionization, trichothecenes (T-2 toxin, HT-2 toxin, acetyl T-2 toxin, diacetoxyscirpenol, neosolaniol, and mono-acetoxyscirpenol) in grain using atmospheric pressure chemical ionization, and trichothecenes (T-2 toxin, HT-2 toxin, deoxynivalenol, nivalenol), zearalenone and its metabolites (a- and jS-zearalenol and a- and jS-zearalanol) in eggs. 

Patulin in apple juice 50 European Commission and Codex Alimentarius... 

Patulin in apple juice and puree Ethylacetate Pectinase treatment for puree LC coupled to UV detector LC coupled to UV detector 15pgkg 70-100... 

Scott, P. M., and B. P. C. Kennedy Improved Method for the Thin-layer chromatographic determination of Patulin in Apple Juice. J. Ass. Off. Anal. Chem. 56,813 (1973). 

Owing to the varied structures of various food dyes, they can often be differentiated from one another by their characteristic ultraviolet/visible absorbance spectra. Using HPLC coupled with a diode array detector (HPLC-DAD) it is possible to collect a compound s absorbance spectrum as it elutes from the HPLC column, which greatly assists in identification. At Reading Scientific Services Ltd (RSSL) this type of detector is routinely used in a range of analyses of such substances as patulin, a mycotoxin found in apple juice, and in the analysis of colours and vitamins, which allows a more certain assignment of a particular peak to a specific compound to be made. 

DATA FROM RECENT SURVEYS ON PATULIN CONTAMINATION IN APPLE JUICE... 

Kadakal and Nas (2002b) used apples, classified by the decay proportion on the fruit surface as sound, 30, 60, or 100% decayed, in the production of apple juice, to determine the effect of apple decay proportion on the patulin content of apple juice. Patulin increased in apple juice samples as the decay proportion increased. Patulin in juice samples produced with apples that were sound, 30, 60, and 100% decayed, were 0-15.9, 47.1-500.3, 156.4-2257.5, and 54.9-2508.6 pg/kg. Similar results have been reported by Jackson et al. (2003). Patulin was not detected in juice pressed from fresh tree-picked apples but was found at levels of 40.2-374 pg/L in juice pressed from fresh ground-harvested (dropped) apples. Another possible source of patulin contamination may be contamination of apple juice with P. expansum. McCallum et al. (2002) observed extensive fungal growth and high patulin levels after inoculation of apple cider with different isolates of P. expansum. Concentrations of 538-1822 pg/ml in apple ciders were associated with incubation at room temperature (25°C), and potentially toxic patulin levels of 75-396 pg/ml also were found in refrigerated ciders (4°C) inoculated with P. expansum. 

Table VI also shows that the use of activated charcoal seems to be another effective step in reducing patulin contamination in apple juice. Leggott et al. (2000) observed a reduction of patulin from 110 to 75 pg/L by a combination of depectinization, charcoal treatment, and ultrafiltration, which was probably because of the adsorption of patulin on the activated charcoal. No further removal of patulin occurred during the remainder of the juicing process. Leggott et al. (2001) reported that the type of charcoal...

Codex Alimentarius Commission 2002. Proposed draft code of practice for the prevention of patulin contamination in apple juice and apple juice ingredients in other beverages, ftp //ftp.fao.org/... 

Huebner, H.J., Mayura, K., Pallaroni, L., Ake, C.L., Lemke, S.L., Herrera, P., and Phillips, T.D. 2000. Development and characterization of a carbon-based composite material for deducing patulin levels in apple juice. J. Food Prot. 63, 106-110. 

Sydenham, E.W., Vismer, H.F., Marasas, W.F.O., Brown, N.L., Schlechter, M., and Rheeder, J.P. 1997. The influence of deck storage and initial processing on patulin levels in apple juice. Food Addit. Contam. 14, 429-434. 

Dialysis is a cleanup method in which the analyte passes from one solution to another through a membrane while impurities do not transfer. The process can be set up to remove either low-molecular-weight or high-molecular-weight interferences. As examples, dialysis was used to extract the mycotoxin patulin from apple juice using ethyl acetate in a diphasic system, and to remove... 

In comparison with the usual HPLC/UV method, it has heen shown in the analysis of mycotoxins using patulin (TV in Rg. 9.1) that the detection limit decreases hy a factor of 1(X) if an isotopic dilution assay (cf. 5.2.6) is carried out with [ C2] patulin as the internal standard. After sily-lation and gas chromatographic/mass spectromet-ric measurement of analyte and isotopomer, 5.7-26.0 pg/1 of patulin were found in apple juices. 

During processing, affected fruit is generally removed or pressurized-water jets are utilized to remove damaged portions of fruit and contaminations (or both). These processes effectively eliminate patulin. However, if these procedures are not done properly, patulin may remain in the processed apple juice and apple products, where it is very stable. Pasteurization at 90°C only causes a reduction of 10%, however patulin is not stable in the presence of sulfur dioxide or sulfydryl compounds. The fermentation process for cider eliminates 99% of patulin. 

Patulin (C6H704), is a compound, dangerous to health in humans which is found in the juice of damaged apples or grapefruits. The current method of measuring this compound resembles that of a immunoenzymatic ELISA test. A standard solution of pure patulin was prepared in water at a concentration of 1.54 g/1 just prior to use. 

Patulin-producing strains are widespread as contaminants in food. Patulin has been found in mouldy bread, fermented sausage, commercial apple juice and animal food. The compound inhibits the growth of Gram-negative and Gram-positive bacteria however, it cannot be used as an antibiotic, for its has been shown that patulin is very toxic and teratogenic. The molecular basis for the toxicity is still unknown. 

Mycotoxins are toxic secondary metabolites produced by fungi growing within or on foods. They can be a serious threat to human and animal health (Nagler el al., 2001). Table 11.4 details mycotoxins associated with soft drinks and fruit juice manufacture and raw materials. Patulin is the most common mycotoxin associated with fruit juice, particular ly apple juice (Pitt Hocking, 1997). It commonly occurs if juice is produced from stored apples. Mould growth in infected apples increases with time, raising levels of patulin. The use of windfall apples for juice is also a factor. Avoidance of windfall apples, filtration of juice and pressing quickly after harvest are all methods to reduce the incidence of patulin in juice. Patulin can be destroyed by fermentation to cider or by the addition of ascorbic acid (Marth, 1992). Within Europe, the European Union has set a limit of 50 ig/kg for patulin in both apple juice and cider. A recent survey of apple products in Chile found that 28% of samples of juice and concentrate exceeded this limit (Canas Aranda, 1996). 

Patulin was originally considered desirable for its antibacterial properties. However, its toxicity to mammals precluded its use as an antibiotic. Patulin (fig. 4), a metabolite of many species of Penicillium and Aspergillus, has been detected in damaged apples, apple juice and apple cider made from partially decaying apples and in other fruit juices as well as in wheat. A heat-resistant fungus Byssochlamys nivea, frequently found in foods also produces patulin [109]. Due to highly reactive double bonds that readily react with sulfhydryl... 

---