血合筋肉の化学的研究-10-ミオグロビンの新定量法

概要

論文の詳細を見る
A new method for the determination of fish myoglobin (Mb) was offered, which is based on a difference in heat coagulability between Mb and hemoglobin (Hb)15) (see Table 3). The procedure is as follows: Muscle is ground with dry-ice, extracted with distilled water, and both pigments are converted to cyanmet form with a trace of ferricyanide and KCN. The solution is then mixed with 1/3 volume of 10% basic lead acetate solution and precipitates are centrifuged off. To the supernatant a phosphate buffer (pH 6.7) is added to obtain a final concentration of 0.2M and precipitates are centrifuged off. After an addition of 3mg. KCN per 5ml. of the solution, the solution is heated for 5min. at a given temperature (see Table 3) under the condition described previously15). Coagulated Hb is centrifuged off, and Mb in the supernatant is measured spectrophotometrically (Method 2•2). Met form was found not to be suitable because of a partial denaturation caused by basic lead acetate, especially in the presence of excess ferricyanide (cf. Figs. 2 and 3). Total amount of the two pigments in muscle was estimated as cyanmet derivatives, using small portion of the extract. Results obtained on some kinds of fish are shown in Table 2, together with recovery of pure preparations added. As seen in Tables 1 and 2, most of the water-soluble pigments are Mb both in red muscle and in ordinary muscle, contrary to the previous quantitative results1) on Mb obtained by DRABKIN's method3), which was found to give lower values in the case of fish Mb owing to its characteristic properties such as solubility and behavior to basic lead acetate. In the cases of Mb-rich muscles (e. g., red muscle of tuna), extracts can be diluted to a farther extent and so the deproteinization process can be omitted (Method 2•1). Results obtained by this convenient method are shown in Table 1. A spectrophotometric method for simultaneous determination of Mb and Hb (Methods 1•1 and 1•2, refer to Fig. 1) was found not to be usable in the case of fish extract, because of the too slight spectral differences between these pigments.
社団法人日本水産学会の論文

血合筋肉の化学的研究-10-ミオグロビンの新定量法

スポンサーリンク

概要

著者

関連論文

スポンサーリンク