Quantification of Combined Amount of Isopentenyl Diphosphate and Dimethylallyl Diphosphate by LC/MS/MS Using Selective Solid-phase Extraction

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  • SATO Tomomi
    Department of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
  • KUWABARA Naoko
    Department of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
  • NAKAGAWA Saori
    Department of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences
  • YAMATO Susumu
    Department of Bio-analytical Chemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences

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Other Title
  • 選択的固相抽出法を用いるイソペンテニルピロリン酸とジメチルアリルピロリン酸との合量のLC/MS/MS定量
  • センタクテキコソウ チュウシュツホウ オ モチイル イソペンテニルピロリンサン ト ジメチルアリルピロリンサン ト ノ ゴウリョウ ノ LC/MS/MS テイリョウ

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Abstract

Isoprenoids are intermediates of cholesterol biosynthesis, and are also related to Ras prenylation associated with cell proliferations. Recently, an inhibitory effect of bisphosphonate on the biosynthesis of isoprenoids has been noted as a target of an anti-cancer drug. There were small amounts of isporenoids in the biological fluid; therefore, a highly sensitive determination method of isoprenoids by LC/MS/MS is required. On the other hand, LC/MS/MS requires one to remove any suppressed ionization substance, e.g., phospholipids, from the matrix before analysis. We tried to separate isoprenoids from phospholipids using Hybrid SPETM-Phospholipid (HybridSPE) solid-phase extraction to determine isoprenoids, and then applied to determine them in a HepG2 cell culture medium. We could effectively separate isopentenylpyrophosphate (IPP) and dimethylallylpyrophosphate (DMAPP) from phospholipids. Also, this method was successfully applied to determine the total amount of IPP and DMAPP in a cell culture medium. Bisphoshonates, inhibitors of farnesyl-pyrophosphate (FPP) synthetase, increase IPP and DMAPP in a cell culuture medium with a dose dependence. Functional evaluation of bisphosphonates could be performed by determining IPP and DMAPP.

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 62 (8), 731-735, 2013

    The Japan Society for Analytical Chemistry

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