Akiyama Hiroshi, Isuzugawa Kazuto, Harikai Naoki, Watanabe Hiroko, Iijima Ken, Yamakawa Hirohito, Mizuguchi Yamato, Yoshikawa Reiji, Yamamoto Miho, Sato Hidetaka, Watai Masatoshi, Arakawa Fumihiro, Ogasawara Takeshi, Nishihara Rikuka, Kato Hisashi, Yamauchi Atsushi, Takahata Yoshihisa, Morimatsu Fumiki, Mamegoshi Shinichi, Muraoka Shiroo, Honjoh Tsutomu, Watanabe Takahiro, Sakata Kozue, Imamura Tomoaki, Toyoda Masatake, Matsuda Rieko, Maitani Tamio
National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
Shokuhin Eiseigaku Zasshi. 2004 Jun;45(3):120-7. doi: 10.3358/shokueishi.45.120.
Extracts of sausage, sauce, cookie, cereal and pasta sauce spiked with milk 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 three ELISA methods using a Milk Protein Casein ELISA Kit (Casein kit), a Milk Protein Beta-Lactoglobulin ELISA Kit (Beta-Lactoglobulin kit) and a FASTKIT Milk ELISA Kit (Milk ELISA kit) were mostly below 10%. Mean recoveries of the milk standard protein from the food extracts were over 40% in the three ELISA methods with a few excertions. The recoveries of milk standard protein from the sauce extract in Casein kit were improved by adjusting the extract to neutrality before the Casein kit assay. The recoveries of milk standard protein from cookie, cereal and pasta sauce were improved by the increasing the amount of antibody coated in the Milk ELISA kit. The detection limits of all the ELISA methods were 1 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of milk protein levels in extracts of sausage, sauce, cookie, cereal and pasta sauce.
将添加了浓度为5 - 20 ng/mL乳标准蛋白的香肠、酱料、饼干、谷物和意面酱提取物作为样品溶液，在10个实验室中进行重复分析。使用乳蛋白酪蛋白ELISA试剂盒（酪蛋白试剂盒）、乳蛋白β-乳球蛋白ELISA试剂盒（β-乳球蛋白试剂盒）和FASTKIT乳ELISA试剂盒（乳ELISA试剂盒）的三种ELISA方法的变异系数（CV）大多低于10%。在三种ELISA方法中，从食品提取物中回收的乳标准蛋白的平均回收率超过40%，有少数例外情况。在酪蛋白试剂盒检测前，将酱料提取物调节至中性，可提高酪蛋白试剂盒中乳标准蛋白从酱料提取物中的回收率。通过增加乳ELISA试剂盒中包被抗体的量，可提高饼干、谷物和意面酱中乳标准蛋白的回收率。所有ELISA方法在样品溶液中的检测限均为1 ng/mL。这些结果表明，所通报的ELISA方法对于检测香肠、酱料、饼干、谷物和意面酱提取物中的乳蛋白水平是可靠且可重复的。
