クロロフィル類の標準物質の調製とその分析化学的特性

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書誌事項

タイトル別名
  • Preparation and analysis of standard samples of chlorophylls.
  • クロロフィルルイ ノ ヒョウジュン ブッシツ ノ チョウセイ ト ソノ ブンセ

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抄録

A rapid and simple purification method for chlorophylls was established and the spectroscopic properties and/or high performance liquid chromatographic properties of chlorophylls for analysis were studied in detail. Chlorophylls were extracted from confrey (Symphytum officinalle L.) and purified by cellulose column chromatography {petroleum ether : 1-propanol = 99.9 : 0.1(v/v)}. After purification, chlorophylls were identified by FAB (fast atom bombardment)-MS. The spectroscopic properties of chlorophylls in acetone were as follows; chlorophyll a (ε663=7.4 × 104 dm3 mol-1 cm -1, Ex =-430 nm, Em = 666 nm, φf=0.396), chlorophyll b (ε645 = 5.5 × 104 dm3 mol-1 cm-1, Ex = 457 nnm, Em = 650 nm, φf = 0.141), pheophytin a (ε663 = 4.2 × 104 dm3 mol-1 cm-1, Ex = 410 nm, Em = 670 nm, φf = 0.141), pheophytin b (ε650= 3.7 × 104 dm3 mol-1, Ex = 435 nm, Em 655 nm, φf= 0.077). Chlorophyll b, which has a stronger positive charge on the central part of the atom than other chlorophylls, was especially affected by oxygen molecules. Chlorophylls could be easily separated by HPLC using Zorbax-ODS (250 mm × 4.6 mm i.d.) as the stationary phase and acetonitrile : acetone = 45 : 55 (v/v) as mobile phase at a flow rate of 0.6 cm3 min-1 HPLC behavior (retention time = tR, capacity factor = k') of the chlorophylls was as follows; chlorophyll b (tR= 10'05", k' = 1.24), chlorophyll a (tR= 13'30", k' = 2.00), pheophytin b (tR=17'24", k' = 2.87), pheophytin a (tR= 21'00", k' = 3.67). Solutions of chlorophylls that had been purified were unstable, being mainly degraded by oxygen and light at room temperature. Chlorophyll a in acetone had changed to pheophytin a and other degradation products within five days. Therefore, nitrogen was bubbled through the solvent and the solution of chlorophyll a was stored in a dark box at 0°C. Using this procedure, chlorophyll a in acetone was stable for one month.

収録刊行物

  • 分析化学

    分析化学 38 (4), 182-187, 1989

    公益社団法人 日本分析化学会

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