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2.2. Plant extraction

The plant materials were air-dried and ground to a fine

powder. Extraction was performed by soaking samples (50 g

dry weight) in 70% EtOH (500 ml) for 24 h at 20 ◦C. After filtration

through filter paper, the residues were washed twice

with 70% EtOH, followed by drying using a rotary evaporator

at 35 ◦C. The Selaginella labordei extract was further

extracted twice with petroleum ether to remove chlorophyll.

For all experiments, the residues were dissolved in dimethyl

sulphoxide (DMSO) except in the TEAC assay, where the

residues were dissolved in water.

2.3. Inhibition of XOD and LOX

Xanthine oxidase (XOD) activity was determined spectrophotometrically

at 290 nm as described by Cos et al.

(1998). The assay mixture, containing 40mM phosphate

buffer, pH 7.5 containing 0.2mM EDTA, 5–50_M xanthine

and one unit of superoxide dismutase (SOD), was incubated

in a quartz cuvette at 37 ◦Cfor 2 min. The reactionwas started

by the addition of 10mU XOD and the increase in absorption

at 290 nm was recorded at 20 s intervals for 5 min. The procedure

was repeated with the addition of extracts at a range

of concentrations.

Lipoxygenases (LOX) activity was determined spectrophotometrically

at 234 nm as described by Chen and

Whitaker (1986). The substrate stock solution, containing

linoleic acid, Tween 20 and deionised water, was clarified by

the addition of 1Msodium hydroxide. The stock solutionwas

diluted four-fold with 0.2Mphosphate buffer, pH 7.0 to give

2.5mM substrate, and flushed with oxygen for 2 min. The

reaction was started by the addition of 200U of LOX and

the increase in absorbance at 234 nm was recorded at 20 s

intervals for 5 min at 37 ◦C. The linoleic acid concentration

rangewas between 0.25mMand 2.5 mM. The procedurewas

repeated with the addition of extracts at a range of concentrations.

1 Answer

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  • 1 decade ago
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    2.2 。植物提取物

    植物材料,空氣乾燥,地面至罰款

    粉末。萃取是由浸泡樣品( 50克

    幹重) , 70 %乙醇( 500毫升) 24 小時後,在20 ◦長過濾後

    透過濾紙,殘渣水洗兩次

    用70 %乙醇,其次是乾燥用旋轉蒸發器

    35 ◦長該卷柏labordei提取物還

    提取兩次,用石油醚,以消除葉綠素。

    所有實驗中,殘留物溶解於二甲

    亞砜(二甲基亞砜) ,除了在教學方法,而

    殘留物溶解於水。

    2.3 。抑制xod的和液氧

    黃嘌呤氧化酶( xod的)活性測定spectrophotometrically

    在290 nm處形容產地來源證等。

    ( 1998 ) 。測定混合物,含有40毫米磷酸鹽

    緩衝區, ph值7.5含0.2毫米乙二胺四乙酸, 5 - 50_m黃嘌呤

    和一個單位的超氧化物歧化酶(超氧化物歧化酶) ,孵育

    在一個石英試管,在37 ◦ cfor 2分鐘。該reactionwas開始

    通過添加10mu xod的提高和增加吸收

    在290 nm處被記錄在20 s的時間間隔為5分鐘。程序

    重複與另外的提取物,在一系列

    的濃度。

    脂氧合(液氧)活性測定spectrophotometrically

    在234 nm的形容:陳和

    惠特克( 1986 ) 。基質溶液儲存器,其中載有

    亞油酸,吐溫20和去離子水,是澄清

    加上1msodium氫氧化物。股票solutionwas

    稀釋4倍,與0.2mphosphate緩衝區, ph值7.0 ,讓

    2.5毫米基板,並衝入氧氣為2分鐘。該

    反應是,一開始便加入200u的液氧和

    增加吸光度在234 nm處記錄為20 s

    間隔時間為5分鐘,在37 ◦長亞油酸濃度

    rangewas之間0.25mmand 2.5毫米。該procedurewas

    多次與另外的提取物在不同濃度。

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