Inter-laboratory evaluation studies for establishment of notified ELISA methods for allergic substances (peanuts)

Inter-laboratory evaluation studies were conducted for ELISA methods for allergic substances (peanuts). Extracts of biscuit, sauce, chocolate and butter spiked with peanut standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of the ELISA methods using a Peanut Protein ELISA Kit (Peanut kit) and a FASTKIT Peanut ELISA kit (Peanut ELISA kit) were mostly below 10%. Mean recoveries of the peanut standard protein from the food extracts were over 40% in the two ELISA methods. Repeatability relative standard deviations of peanut standard protein in four food extracts were in the ranges of 15.2-49.7% and 3.0-28.3% for the Peanut kit and the Peanut ELISA kit, respectively. Reproducibility relative standard deviations of peanut standard protein in four food extracts were 23.5-44.4%, 9.6-28.4% for the Peanut kit and the Peanut ELISA kit, respectively. The detection limits of both ELISA methods were 2-2.5 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of peanut protein levels in extracts of biscuit, sauce, chocolate and butter.     

Fate of coliphage in a wastewater treatment process

The fate of coliphage in a wastewater treatment plant in the central part of Japan was investigated from March to December 2001. A relative abundance of coliphage, 1000-10,000 PFU/ml determined with three different Escherichia coli strains, was detected in the influent. But, no remarkable seasonal change in the phage concentration in the influent was observed during the ten-month test period. Almost ten times higher coliphage concentration was detected by the F+ E. coli strain than by the other two F- strains. The RNA phage was more stable than the DNA phage against aerobic treatment using activated sludge. Most of the phages in the influent and primary settling tank were detected as suspended forms. Anaerobic-aerobic treatment enhanced adsorption of the phage by the solid particles. Almost no phage was detected in the effluent. Aerobic treatment using activated sludge and/or the addition of flocculants such as PAC was effective for the removal of coliphage, an index of enteric viral pollution.     

Determination of vitamin K in aojiru (green juice) products by HPLC

A survey of vitamin K contents was carried out on 41 aojiru products and 10 vegetable juice products that were purchased from local markets. Aojiru is a health food made from green vegetables such as kale, ashitaba, mulberry leaf, barley grass and the like. The products are usually provided in various forms, such as frozen, powder and tablet. Vitamin K in samples was extracted with n-hexane, and separated on a C18 column with methanol-ethanol (95 : 5). After separation, vitamin K was converted to the hydroquinone form on a reduction column and determined with a fluorescence detector at lambdaex 240 nm and lambdaem 430 nm. The contents of vitamin K1 (phylloquinone) in frozen samples (n = 8), powder samples (n = 26) and tablet samples (n=7) were 90-190, 410-3,300, and 640-3,100 microg/100 g, respectively, and that in vegetable juice (n= 10) was 1-12 microg/100 g. Vitamin K2 (menaquinone) was not detected. The daily intake of vitamin K from aojiru products was estimated to be 99-380, 20-250 and 27-210 microg/day for frozen, powder and tablet types, respectively. These results suggest that patients prescribed warfarin should take care about their intake of vitamin K from aojiru products.