HPLC-FLによる配合飼料中ノシヘプタイド定量法の妥当性評価

摘要

配合飼料中NHのHPLC-FLによる定量法の開発および妥当性評価を行い，以下の結果を得た.1)分析試料5gを酢酸(1→20)25mLで浸潤した後，アセトン75mLを加え，かくはんした後超音波処理する抽出法を採用し，本法は現行法では十分抽出されないほ乳期子豚育成用配合飼料にも適用が可能であった.2)NH添加試料を用い，添加回収試験および試験室間共同試験を実施した結果，本法の真度，併行精度および室間再現精度はCodexの評価基準を満たし良好であった.本法は，ノシヘプタイドの成分規格の全範囲2.5〜20g(力価)/tにおいて適用可能であり，さらに，検出下限値0.5g(力価)/tの残留レベルの試験にも適用可能であった.%A determination method of nosiheptide in formula feeds by HPLC-FL was developed and validated, including an inter-laboratory study. Formula feeds were extracted with acetone after adding acetic acid. Liquid chromatographic separation was performed using a ZORBAX Eclipse XDB-C18 column, with acetonitrile and water containing acetic acid as the mobile phase. Detection of NH was carried out with a fluorescence detector. Recovery tests and an inter-laboratory study were conducted using chicken and swine formula feeds fortified with nosiheptide at 0.5-27 mg-potency/kg. Mean recoveries in recovery tests ranged from 91.4 to 103%, and the repeatability in terms of relative standard deviation was within 7.8%. Mean recoveries of the inter-laboratory study ranged from 98.4 to 108%, the repeatability and reproducibility in terms of the relative standard deviations were within 8.1% and 13% respectively, and the HorRat values ranged from 0.21 to 0.75.