穀物中のエルゴステロールの分析法の簡易化について Analysis of Ergosterol in Cereals without Saponification

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Abstract

エルゴステロール(ERG)は真菌の細胞膜の構成成分の一種であり,多くの植物や動物はERGを産生しないことから,ERGは穀物中のかび加害のバイオマーカーとして考えられている.以前,私達はメタノールとアルカリ溶液を用いたけん化方法による穀物中のERGの分析法を開発し,その妥当性を確認した.しかし,けん化による分析法は時間やコスト,手間がかかるため,迅速さが求められるスクリーニングには適していない.そのため,私達は穀物中のERGの分析法の簡易化を試みた.ERGはメタノールで一時間振とう抽出し,ろ過後,ERGは液―液分配にてヘキサンに転溶し,逆相HPLCで分析した.振とう法によるERGの回収率をけん化法によるERGの回収率で比較した.3mg/kgのERGのトウモロコシサンプルにおける振とう法による回収率はけん化法による回収率の84%で,振とう法による相対標準偏差(RSD)は1.5 %であった.また,12mg/kgのERGの小麦サンプルにおける振とう法の回収率はけん化法に対して108%で,振とう法のRSD値は2.8%であった.今回開発した振とう法による穀物中のERGの分析法は,スクリーニング目的として用いるには十分適用可能であると考えられる.

  Ergosterol (ERG) is a sterol found in fungal cell membranes and is not produced by most plants and animals. Thus, ERG is a potential biomarker of fungal invasion in grain. Previously, we developed an analytical method for ERG in grains by ref uxing samples with methanol-alkali, and performed a single laboratory validation of the method. However, the method is not suitable for screening purpose, which needs to be rapid and simple. Therefore a simplified method was developed for analysis of ERG in grains. In this simplified method, ERG was first extracted with methanol by horizontal shaking for 1 hr. After f ltration, ERG was partitioned into hexane, and then analyzed with reverse phase high performance liquid chromatography (HPLC). Recoveries of ERG obtained with this shaking method were compared with recoveries obtained with the refluxing method. In maize, recovery of ERG at 3 mg/kg using this new shaking method was 84 % of that of the ref uxing method and the relative standard deviation (RSD) of the shaking method and the ref uxing method were 1.5 % and 16 %, respectively. In wheat, recovery of ERG at 12 mg/kg by the shaking method was 108 % of that of the ref uxing method, and the RSD of the shaking method and the ref uxing method were 2.8 % and 5.9 %, respectively. This study demonstrates this simplif ed protocol for ERG analysis in grains to be a suitable method for screening purposes.

Journal

  • JSM Mycotoxins

    JSM Mycotoxins 61(1), 1-9, 2011-01-31

    Japanese Society of Mycotoxicology

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Codes

  • NII Article ID (NAID)
    10030541180
  • NII NACSIS-CAT ID (NCID)
    AN00334513
  • Text Lang
    ENG
  • Article Type
    ART
  • ISSN
    02851466
  • Data Source
    CJP  IR  J-STAGE 
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