JP5147011B2 - 血清脂質の測定方法及び測定装置 - Google Patents
血清脂質の測定方法及び測定装置 Download PDFInfo
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Description
電気泳動法による小粒子LDL粒度の推定値に近い粒度の市販の標準ラテックス(粒径の表示値:22±1.5nm)を使用して、単分散ラテックスの粒度を測定した。動的光散乱法測定部を含む測定装置として大塚電子(株)から市販されているFDLS−3000を用いた。
測定モード:Contin法
測定波長:532nm(出力100mW、固体パルスレーザー、Nd−YAGのSHG波使用)
測定散乱角度:90度
サンプリングタイム:8.0マイクロ秒
積算回数:100回
コリレーションチャネル数:128chコリーレーション法:タイムインターバル法
試料の物性値:
(血清25°C)屈折率 1.3313 粘性1.33
(水、生理食塩液 25°C)屈折率 1.3313 粘性 0.8858
(血清36.3±2°C)屈折率 1.3299 粘性1.33
(水、生理食塩液 36.3±2°C)屈折率 1.3299 粘性 0.6922-0.6968
(結果)
図1に示すとおり、上記の標準ラテックスの動的光散乱法による粒度測定の結果、散乱強度約16の単峰性ピークが得られた。この粒度分布の測定値は、21.7±2.1nmであった。これは標準サンプルの表示値である22±1.5nmにきわめて近く、本発明の動的光散乱法による測定の精度が高いことが確認された。
ヒト血清測定にあたり、多分散ラテックスを混合し、実施例1と同様の測定条件で粒度を測定した。用いた粒子は、市販の平均粒径100nm、60nm、30nm及び20nmラテックスの混合物(100,60,30,20nmラテックスの混合比=23:23:1:1)であった。この混合比は正常ヒト血清をゲルろ過して得られたCM及びLDL分画のコレステロール濃度比に基づいて、ヒト血清中の粒子の粒度を近似するように設定された。
正常ヒト血清をゲルろ過法で分画し、LDL分画とHDL分画精製後の検体の粒度分布を参考例2と同様の方法で動的散乱法で測定した。
HDLと小粒子LDLとが混在する検体のような、粒度分布の異なる複数の粒子群からなる粒度分布の解析の可否を検討した。これは、(5)式において複数粒子存在下での、粒子混合比を導く手法である。粒子の混合比の実測にあたり、市販の平均粒径21nm及び28nmのラテックスを使用し、参考例2と同様の測定方法で計測した。図6に示すとおり、散乱光の揺らぎから実測された二次自己相関関数は混合比の違いによって明確に異なっていた。
小粒子LDLの粒度の精度を検討するため、1人の患者から採取した血清を用いて繰り返し測定を行った。血清試料に2価金属イオン及びポリアニオンを含有する市販のsmall dense LDL測定用試薬(デンカ生研株式会社)を添加して、血清試料から小粒子LDLよりも大型の粒子、すなわちLDL分画のうち小粒子LDL以外の画分と、CM及びVLDLとを除去した。残った小粒子LDL画分について動的光散乱法で粒度を測定した。試料1点あたりの前処理時間は約20分で、容易に測定が可能となった。
表1に結果を示すとおり、小粒子LDLの粒度として18.9nm(標準偏差σ=2.187)という値が得られた。
Claims (4)
- 前記被検試料から小粒子LDLを含む画分を分離するステップは、ゲルろ過クロマトグラフィー法と、高速液体クロマトグラフィー(HPLC)法と、超遠心法と、2価金属イオン及びポリアニオンを含む試薬を用いる沈降法とからなるグループから選択される手法によって実施されることを特徴とする、請求項1に記載のリポ蛋白サブクラスの測定方法。
- 請求項1ないし2のいずれか1つに記載のリポ蛋白サブクラスの測定方法を行うための測定装置であって、動的光散乱法測定部を含むことを特徴とする、測定装置。
- 光源としてパルスレーザーを有することを特徴とする、請求項3に記載の測定装置。
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Cited By (3)
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US10994271B2 (en) | 2016-06-14 | 2021-05-04 | Denka Company Limited | Membrane carrier for liquid sample test kit, liquid sample test kit, and method for producing liquid sample test kit |
US11162938B2 (en) | 2017-03-28 | 2021-11-02 | Denka Company Limited | Membrane carrier, kit for testing liquid sample using same, and manufacturing method thereof |
US11385227B2 (en) | 2017-03-28 | 2022-07-12 | Denka Company Limited | Membrane carrier and kit for testing liquid sample using same |
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JP5953897B2 (ja) * | 2012-03-28 | 2016-07-20 | 株式会社明日香特殊検査研究所 | リポ蛋白質の粒子径の決定法及びリポ蛋白質の粒子マーカ |
WO2013188879A1 (en) * | 2012-06-16 | 2013-12-19 | Atherotech, Inc. | Measurement of serum lipoproteins |
JP5984074B1 (ja) * | 2016-02-18 | 2016-09-06 | メディカルフォトニクス株式会社 | 体調管理装置及びその方法 |
JP6029128B1 (ja) * | 2016-05-18 | 2016-11-24 | メディカルフォトニクス株式会社 | 血中脂質濃度計測装置及びその作動方法 |
WO2018143119A1 (ja) * | 2017-01-31 | 2018-08-09 | メディカルフォトニクス株式会社 | 脂質計測装置及びその方法 |
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JPS63259435A (ja) * | 1987-04-15 | 1988-10-26 | Shimadzu Corp | 粒度分布測定方法 |
JPS63265139A (ja) * | 1987-04-23 | 1988-11-01 | Otsuka Denshi Kk | 粒径測定装置 |
ATE505731T1 (de) * | 2002-12-06 | 2011-04-15 | Denka Seiken Kk | Verfahren zur quantifizierung von kleinen lipoproteinen mit geringer dichte |
JP3819895B2 (ja) * | 2003-10-29 | 2006-09-13 | 大塚電子株式会社 | 動脈硬化評価装置 |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
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US10994271B2 (en) | 2016-06-14 | 2021-05-04 | Denka Company Limited | Membrane carrier for liquid sample test kit, liquid sample test kit, and method for producing liquid sample test kit |
US11162938B2 (en) | 2017-03-28 | 2021-11-02 | Denka Company Limited | Membrane carrier, kit for testing liquid sample using same, and manufacturing method thereof |
US11385227B2 (en) | 2017-03-28 | 2022-07-12 | Denka Company Limited | Membrane carrier and kit for testing liquid sample using same |
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