Establishment of measurement method for <i>phospholipase-D</i> activity in blood cells

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  • KAWANO Miki
    School of Health Sciences at Narita, International University of Health and Welfare
  • ISHIDA Ayano
    Former: Department of Medical Technology, Kouhoukai Takagi Hospital
  • HOKAZONO Eisaku
    Department of Health Sciences, Faculty of Medical Sciences, Kyushu University
  • OSAWA Susumu
    Department of Health Sciences at Narita, Graduate School of Health Sciences, International University of Health and Welfare

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Other Title
  • 赤血球中<i>phospholipase-D</i>活性測定法の構築
  • 赤血球中phospholipase-D活性測定法の構築
  • セッケッキュウ チュウ phospholipase-D カッセイ ソクテイホウ ノ コウチク

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<p>Background: We established a measurement method for phospholipase-D activity in red blood cells as a pre-investigation for the establishment of a measurement method for phosphatidylethanol (PEth), an alcohol-intake marker, since PLD is related to the generation of PEth. Methods: The present method we developed consists of two reactions (main reaction and removal reaction). First, endogenous choline is removed by choline oxidase (COD) and catalase. Then, the main reaction is started. PLD in blood cells hydrolyzes phosphatidyl choline included in the reagent to phosphatidic acid and choline. Afterward, COD acts on choline and produces H2O2. With this H2O2, the color reagent changed the color of the reaction mixture. We can detect the PLD activity by this color change. Results: The mean within-run imprecision of three different blood samples was 11.2–12.1% (coefficient of variation). The reaction rate showed a linear relationship with PLD activity up to 120 U/L. The recovery percentages were 94.4% (100 U/L of PLD) and 84.0% (50 U/L of PLD). The comparison of PLD activity between the healthy control group and the patient groups showed the following: the anemia group showed a lower PLD activity, and the obstructive jaundice group showed a higher PLD activity (p < 0.05). The mean PLD activity divided by the hemoglobin concentration was higher in the anemia group than in the healthy control group. This result indicates that the hemoglobin concentration affects the PLD activity. Conclusions: We established a measurement method for PLD activity in red blood cells. By this method, we can determine the PLD activity; however, the quality of precision is not good. We are planning to clarify the relationship between the PLD activity and the generating ability of PEth.</p>

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