抽出浮選-吸光光度法による血清鉄の定量

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タイトル別名
  • 抽出浮選‐吸光光度法による血清鉄の定量
  • Extraction flotation-spectrophotometric determination of iron in serum
  • チュウシュツ フセン キュウコウ コウドホウ ニ ヨル ケッセイ テツ ノ テ

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抄録

Microgram quantities of iron in protein-free filtrates of serum were converted into the chelate by adding 3-(2-pyridy1)-5, 6-dipheny1-1, 2, 4-triazine (PDT), and then floated with the aid of sodium lauryl sulfate as a collector and small nitrogen bubbles. The floated PDT-iron(II) chelate was collected with about 97 %yield in a mixture of 4-methyl-2-pentanone and 1, 2- dichloroethane(4 : 1 in vol.) placed on the solution surface. The serum iron in the organic phase was determined by spectrophotometrically. The optimum pH for the extraction flotation of the chelate was in the range of 2.44.2 in the aqueous phase. The recommended procedure is as follows. Place (0.20.5) ml of serum and 0.5 ml of 1 M hydrochloric acid in a test tube. Heat the mixture in a water bath at (8095)°C for 2 min and cool to room temperature. After the addition of 0.5 ml of trichloroacetic acid(0.2 g/ml) solution, shake well the mixture and centrifuge at 1800 g for 15 min. Remove the supernatant and wash the precipitated serum proteins in the tube three times with 1 ml each of trichloroacetic acid(0.05 g/m1)- ascorbic acid(0.01 g/ml) solution. To the combined supernatant, add 1 ml of hydroxyl ammonium sulfate (0.1 g/ml) and 2 ml of 2 M acetate buffer solution(pH 4.4). Being stood for 15 min, add 2 ml of ethanol, 1 ml of 0.01 M PDT ethanol solution and 2 ml of 0.005 % sodium lauryl sulfate solution to the solution, and dilute to 20 ml with water. In a flotation cell, place the solution(pH 2.93.3) and 1.1 ml of the mixed solvent. Bubble nitrogen through the aqueous phase at a flow rate of 50 ml/min. After 5 min of bubbling, remove the organic phase, make up to 1 ml with ethanol and measure the absorbance at 555 nm against the reagent blank with 1 cm microcells. The analyzed values were in good agreement with those obtained by using the improved Matsubara's method. Moreover, hemoglobin iron present at a concentration up to 0.4 mg hemoglobin per 1 ml serum did not interfere with the determination of serum iron.

収録刊行物

  • 分析化学

    分析化学 33 (3), 134-138, 1984

    公益社団法人 日本分析化学会

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