人血清catecholamine高精度迅速定量法について

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Various methods^<1-4> using trihydroxyindol reaction for the measurement of catecholamines in human plasma have been designed by workers hitherto. However, in these methods^<1,3,4>, isolation of the catecholamines with an adsorbent presents considerable difficulties. Thus, in spite of the availability of these methods^<1,3,4> reproduction by other laboratories are difficult. The main objective of the present work was to improve the method for simultaneous determination of epinephrine (Ep.) and norepinephrine (NE.) in human plasma. A preliminary experiment was set up in order to run a comparison against these methods with respect to the recovery, accuracy and rapidity. Of these methods PRICE's method^3 showed satisfactory recovery and rapidity. Thus several improvements of PRICE's methods^3 were made: 1) Purification of acid-washed aluminum oxide (Merck acid washed #71695) was made. 2) A large amount of EDTA disodium was used in several steps. 3) The timing procedure becomes essential due to the fact that fluorophor of both Ep. and to a smaller degree NE. decays at a considerable rate. Thus, a means with sufficiently high accuracy and rapidity was obtained. Material and Method A preparation of acid-washed aluminum oxide (for chromatographic use). a) 300-500 g of alumina (Merck-71695 acid washed) was boiled using a reflux condenser with frequent shaking in 1 L 2N HCl for 1 hour. The slurry was allowed to cool to room temperature. This was transferred to a 1 L chromato-graphic tube with a fritter disc (pyrex #38450). and was cycled with deionized water until the pH of the supernatant rose to 5.0 after which a stirring of the column was followed by cleaning of the supernatant for 2 minutes. This rapid cleaning indicates that most of the fine grain alumina is removed. Next collect the alumina in a suction funnel with filter paper, allow to dry overnight on large sheet of filter paper (Whatman No.3) at room temperature. Then heat in an oven at 200℃ for two hours, place in desiccator. b) All solutions of mineral acids, bases, and buffers were prepared from reagent grade chemicals with water previously distilled and passed through a commercial deionizer. Water purified in this manner is used throughout the procedure.
社団法人日本循環器学会の論文
1969-09-20

人血清catecholamine高精度迅速定量法について

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