The effect of water activity (aw) on growth and aflatoxin production by Aspergillus parasiticus NRRL 2999 was determined using submerged cultures in which the aw was adjusted by addition of glycerine, glucose, or a mixture of salts. At a sub-optimal aw aflatoxin production was low in the glycerol and glucose media while no strong inhibition of mycelial growth occurred. A similar effect was obtained in surface cultures on agar media in which the aw was adjusted by addition of glycerine or sucrose. The effect of a sub-optimal temperature was the reverse; compared to inhibition of mycelial growth in surface cultures, the effect on aflatoxin production was slight. No detectable quantities of aflatoxin B1 were formed at 0.83 aw and at 10 C nor at four combinations of higher aw and temperature. The aw was measured by a recently developed device using the dewpoint principle.

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Abstract

A collaborative study has been carried out among 20 laboratories in The Netherlands, representing governmental and industrial institutes, on the determination of aflatoxin B1 in peanut butter extracts. Blank peanut butter extracts prepared according to the proposed official Dutch method were spiked with aflatoxin B1, representing contamination levels of 0, 3, 6, and 12 μg B1/kg. Samples and standards were spotted on silica gel G TLC plates by the antidiagonal spot application technique described herein. Spotted plates were developed by 2-dimensional TLC with diethyl ether-methanol-water (94+4.5+1.5; lined tank) in the first direction and chloroform-acetone (90+10; unlined tank) in the second direction. Separated B1 spots from sample and standards developed in both directions were free from background interference. The quantity of aflatoxin B1 present in the sample was established by visual comparison of the fluorescent intensities of sample and standard B1 spots. For this procedure the variability of measurements within and between laboratories was statistically investigated: 80–90% of the complete results given by the participants were correct for the hlank and spiked extracts (contamination level of 12 μg B1/kg). For contamination levels of 3 and 6 μg B1/kg an approximate coefficient of variation of 35% was calculated for within- and between-laboratory results. Results obtained in this investigation were compared with those found by previous investigators who used the one-dimensional TLC technique. It is concluded that, with the antidiagonal procedure, small amounts of aflatoxin B1 (lowest level tested, 3 ppb) may be detected.