- J-STAGEトップ
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- 日本畜産学会報
- /
- 33 巻 (1962-1963) 1 号
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- 書誌
山羊の尿中ゴナドトロピン
II. 液体クロマトグラフィーによる精製
ジャーナル
フリー
1962 年 33 巻 1 号 p. 59-64
詳細
- 発行日: 1962 年 受付日: 1961/08/28 J-STAGE公開日: 2008/03/10 受理日: - 早期公開日: - 改訂日: -
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訂正情報
訂正日: 2008/03/10 訂正理由: - 訂正箇所: PDFファイル 訂正内容: -
訂正日: 2008/03/10 訂正理由: - 訂正箇所: PDFファイル 訂正内容: -
訂正日: 2008/03/10 訂正理由: - 訂正箇所: 論文抄録 訂正内容: Wrong : In the previous report, it was mentioned that crude UGS (gonodotropic preparations extracted from the urine by the kaolin method) derived from the goat had a severe toxic effect on animals used for assay, and that it was impossible to determine the activity of partially purified preparations from crude UGS derived from goat urine by biological assay.
In the present experiment, kaolin extracts from the urine of the goat were purified further by means of the modification by the authors of the liquid-chromatography method developed by Butt and Crooke. The absorbent used was composed of 10ml of 8% suspension of tricalcium phosphate and 1.5g of hyflo-super-cel. The amount of crude UGS prepared from 100ml of original urine was dissolved in 1ml of 0.85% sodium chloride solution and poured on the column. The material was absorbed by the top of the column. The elution of this material was conducted with two eluting agents. The first fraction which had been eluted from the column by 20ml of 0.002 M disodium hydrogen phosphate was designated the GA fraction and the second fraction eluted by 10ml of 0.02 M trisodium phosphate, the GB fraction.
The sugar contents of the GA and GB fractions were measured by the use of orcinal reaction. In this reaction, 1ml of the GA or GB fraction was added to 1ml of a solution of 1.6% (w⁄v) orcinol in 30% (v⁄v) sulfuric acid, and then 3ml of 80% (v⁄v) sulfuric acid was added. The combined solution was placed in a water-bath at 80°C for 30 minutes and then immediately cooled at 4°C for 15 minutes. The color density of this solution was read in a spectrophotometer at 420mμ. Glucose was used as the standard.
The gonadotropic activity of the GA plus GB fractions was assayed biologically with the increase of the mouse uterine weight as criterion. The GA plus GB fractions were mixed slowly with 10 volumes of cold ethanol. The precipitate was named (GA+GB) EtOH ppt. and stored in a desiccator until it was subjected to biological assay.
When (GA+GB) EtOH ppt. prepared from the urine of cyclic and pregnant goats was assayed biologically for gonadotropic activity, positive responses were elicited. One mouse uterine unit (MUU) of the preparation was produced from 300 to 400ml (about 8-hour equivalent) of original urine. It was considered that the value of sugar measured by chemical assay was related closely to the biological activity of the preparation.
It was concluded that the procedure consisting of kaolin extraction and tricalcium-phosphate purification was useful for detection and determination of the gonadotrodie activity in the urine of the goat.
Right : In the previous report, it was mentioned that crude UGS (gonodotropic preparations extracted from the urine by the kaolin method) derived from the goat had a severe toxic effect on animals used for assay, and that it was impossible to determine the activity of partially purified preparations from crude UGS derived from goat urine by biological assay.
In the present experiment, kaolin extracts from the urine of the goat were purified further by means of the modification by the authors of the liquid-chromatography method developed by Butt and Crooke. The absorbent used was composed of 10ml of 8% suspension of tricalcium phosphate and 1.5g of hyflo-super-cel. The amount of crude UGS prepared from 100ml of original urine was dissolved in 1ml of 0.85% sodium chloride solution and poured on the column. The material was absorbed by the top of the column. The elution of this material was conducted with two eluting agents. The first fraction which had been eluted from the column by 20ml of 0.002 M disodium hydrogen phosphate was designated the GA fraction and the second fraction eluted by 10ml of 0.02 M trisodium phosphate, the GB fraction.
The sugar contents of the GA and GB fractions were measured by the use of orcinal reaction. In this reaction, 1ml of the GA or GB fraction was added to 1ml of a solution of 1.6% (w/v) orcinol in 30% (v/v) sulfuric acid, and then 3ml of 80% (v/v) sulfuric acid was added. The combined solution was placed in a water-bath at 80°C for 30 minutes and then immediately cooled at 4°C for 15 minutes. The color density of this solution was read in a spectrophotometer at 420mμ. Glucose was used as the standard.
The gonadotropic activity of the GA plus GB fractions was assayed biologically with the increase of the mouse uterine weight as criterion. The GA plus GB fractions were mixed slowly with 10 volumes of cold ethanol. The precipitate was named (GA+GB) EtOH ppt. and stored in a desiccator until it was subjected to biological assay.
When (GA+GB) EtOH ppt. prepared from the urine of cyclic and pregnant goats was assayed biologically for gonadotropic activity, positive responses were elicited. One mouse uterine unit (MUU) of the preparation was produced from 300 to 400ml (about 8-hour equivalent) of original urine. It was considered that the value of sugar measured by chemical assay was related closely to the biological activity of the preparation.
It was concluded that the procedure consisting of kaolin extraction and tricalcium-phosphate purification was useful for detection and determination of the gonadotrodie activity in the urine of the goat.
訂正日: 2008/03/10 訂正理由: - 訂正箇所: PDFファイル 訂正内容: -
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