DSpace コレクション: 2005-12-10
http://hdl.handle.net/11173/585
2005-12-102024-03-29T13:08:51Z小麦Lipid Transfer Proteinに対する酵素免疫測定法の確立
http://hdl.handle.net/11173/1422
タイトル: 小麦Lipid Transfer Proteinに対する酵素免疫測定法の確立
著者: 山口(村上), 友貴絵; Murakami-Yamaguchi, Yukie; 枝, 裕子; Eda, Yuko; 本庄, 勉; Honjoh, Tsutomu; 成田, 宏史; Narita, Hiroshi
抄録: Human serum showing high IgE contents to wheat reacted with about 9kDa protein in crude extract of wheat flour on Western analysis. The 9kDa protein was purified from the extract by ammonium sulfate precipitation and anion-exchange, cation-exchange, and hydrophobic chromatography successively. The Nterminal amino acid sequence of this protein was determined to be homologous to non-specific Lipid Transfer Protein (LTP). Sandwich enzyme-linked immunosorbent assay (ELISA) was constructed with rabbit polyclonal IgG raised against this protein. The detection limit of this ELISA was 5 ng/ml for wheat LTP and the cross-reactivity to barley LTP was negligible. Since 2002 in Japan, commercial ELISAs for gliadin were noticed ministerialy to use for the determination of the content of wheat proteins in foods. It is difficult for the noticed ELISA to be applied to fermented foods because of proteolysis of gliadin. However, LTP could be determined in soy sause and wheat-supplemented beer by our ELISA for LTP. It was shown that LTP is an appropriate target protein for the determination of ingredient contents even in fermented foods because of its proteolysis-resistant nature.2005-12-09T15:00:00Z尿中の遊離γ-カルボキシグルタミン酸定量のためのHPLCによる改良法
http://hdl.handle.net/11173/1419
タイトル: 尿中の遊離γ-カルボキシグルタミン酸定量のためのHPLCによる改良法
著者: 桒原, 晶子; Kuwabara, Akiko; 木戸, 詔子; Kido, Shoko
抄録: A rapid and sensitive high performance liquid chromatographic (HPLC) method for the determination of free γ-carboxyglutamic acid (Gla) in urine using precolumn fluorescent derivatization with OPA/ET reagent was previously developed. In the present study, we improved the method for quantitative analysis of urinary free Gla. 1) Fluorescent strength of OPA/ET reagent reached a maximum level after 3 days from the preparation of the reagent. Then the fluorescent strength was maintained at least after 3 weeks by the addition of ethanthiol every 3 days. 2) An urine sample was diluted by 20 to 60-folds and a 2.5μl aliquot of the diluted urine sample containing about 1 to 2 pmol of Gla was injected into the ChemcoPak Liquid Chromatography Columns packed with Nucleosil 5SB. The mobile phase consisted of 0.12 M sodium citrate buffer (pH 5.28) and acetonitrile in the ratio 60:40. Under these conditions, Gla peak appeared in a retention time of about 7 to 10 minutes and was completely resolved from the other amino acids. Since the peak disappeared after the sample was subjected to decarboxylation treatment, the peak was confirmed to contain only Gla. 3) This method gave a linear standard curve in a range of 0.10-150pmol and allowed quantitative analysis of Gla in an amount as low as 0.1pmol. We used the standard curve in a range of 0.10-4.0pmol for urinary Gla analysis. This is a sensitive and simple assay of free Gla in urine which was subjected to only dilution without further treatment.2005-12-09T15:00:00Zモノクローナル抗体を用いた殺菌剤クロロタロニルに対する酵素免疫測定法の開発
http://hdl.handle.net/11173/1420
タイトル: モノクローナル抗体を用いた殺菌剤クロロタロニルに対する酵素免疫測定法の開発
著者: 山口(村上), 友貴絵; Murakami-Yamaguchi, Yukie; 小川, 加那子; Ogawa, Kanako; 伊東, 茂寿; Ito, Shigehisa; 成田, 宏史; Narita, Hiroshi
抄録: Chlorothalonil (tetrachloroisophthalonitrile) is a compound which has been widely used as germicide. A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the detection of residual chlorothalonil in crops. Three monoclonal antibodies, mAbs 12E, 9A, and 11D were generated against bovine serum albumin-comjugated pentachlorophenol as a homologue of chlorothalonil. By using these mAbs, a direct competitive ELISA has been done. Detectable range of chlorothalonil in the ELISA was 1-3ng/ml for mAb 12E and 0.3-3ng/ml for both mAbs 11D and 9A. MAb 9A reacted with chlorothalonil 3 times stronger than another structurally related germicide fthalide, while mAb 11D reacted with fthalide 5 times stronger than chlorothalonil. The proposed ELISA with mAb 9A would be useful for convenient monitoring of residual chlorothalonil in crops.2005-12-09T15:00:00Zアミノカラムを用いたガラクツロン酸と中性糖類のHPLCによる同時分析
http://hdl.handle.net/11173/1421
タイトル: アミノカラムを用いたガラクツロン酸と中性糖類のHPLCによる同時分析
著者: 吉野, 世美子; Yoshino, Yomiko
抄録: 本研究ではアミノカラムを用いたHPLC分析によりペクチン質を構成するガラクツロン酸と中性糖類を同時に分離する方法を検討した。その結果,ショウデックスアサヒパックNH2P-50 4Eカラムの温度を40℃とし,1.75%燐酸を含む95%アセトニトリル溶液で溶出し,示差屈折計で検出することによりガラクツロン酸と中性糖類であるグルコース,ガラクトース,アラビノース,マンノース,ラムノース,キシロース,フコース,リボース,および内部標準物質のグリセロールを完全に分離,検出することができた。; The method for simultaneous analysis of galacturonic acid and neutral sugars which constitute pectic substances was examined by HPLC analysis using the amino column (Shodex Asahipak NH2P-50). The temperature of column was 40℃ and the refractive index detector was used. This method using 95% acetonitrile solution including 1.75% phosphoric acid allowed to clearly separate galacturonic acid, glucose, galactose, arabinose, mannose, rhamnose, xylose, fucose and ribose with glycerol as an internal standard.2005-12-09T15:00:00Z