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紫外励起熱レンズ顕微鏡/液体クロマトグラフィーを用いるペプチドの無標識高感度検出

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Title: 紫外励起熱レンズ顕微鏡/液体クロマトグラフィーを用いるペプチドの無標識高感度検出
Other Titles: Highly Sensitive Detection of Non-Labeled Peptides Using UV Excitation Thermal Lens Microscope/Liquid Chromatography
Authors: 比企, 伸一郎1 Browse this author
渡慶次, 学2 Browse this author →KAKEN DB
角田, 正也3 Browse this author
馬渡, 和真4 Browse this author
菊谷, 善国5 Browse this author
佐藤, 記一6 Browse this author
火原, 彰秀7 Browse this author
志村, 清仁8 Browse this author
内田, 直行9 Browse this author
北森, 武彦10 Browse this author
Authors(alt): Hiki, Shinichiro1
Tokeshi, Manabu2
Kakuta, Masaya3
Mawatari, Kazuma4
Kikutani, Yoshikuni5
Sato, kiichi6
Hibara, Akihide7
Shimura, Kiyohito8
Uchida, Naoyuki9
Kitamori, Takehiko10
Keywords: thermal lens microscope
liquid chromatography
non-labeled proteins
gradient elution method
Issue Date: Jan-2007
Publisher: 日本分析化学会
Journal Title: 分析化学
Volume: 56
Issue: 1
Start Page: 1
End Page: 7
Publisher DOI: 10.2116/bunsekikagaku.56.1
Abstract: A UV excitation thermal-lens microscope (UV-TLM) with an excitation beam wavelength of 266 nm and liquid chromatograph (LC) were connected to build a new system (LC/UV-TLM) for the separation and highly sensitive label-free determination of biomolecules, such as peptides. A variation in the composition of the mobile phase with time, as in gradient elution, is very commonly utilized in LC for improved separations. Thermal-lens signals are generally highly dependent on the properties of the media surrounding the photo-absorber. This effect must be carefully considered in such situations where the composition of a solution drastically changes in time. Therefore, we tested the developed LC/UV-TLM system operated in a gradient elution mode, by observing the effects of the variation in the solution composition upon TLM signal intensities. In water/acetonitrile gradient elution, no serious effects were observed as long as the acetonitrile content was less than 40% v/v. We then tested the developed system in the separation of a standard sample solution of mixed synthetic peptides; we found that the developed system was 10-times more sensitive than conventional system with the spectrophotometer used as a detector.
ペプチド等の生体由来成分の無標識高感度分離分析を目的として,励起光に波長266 nmの紫外光を有する紫外励起熱レンズ顕微鏡(UV-TLM)と液体クロマトグラフィー(LC)を組み合わせた測定システム(LC/UV-TLM)を構築した.構築したシステムの性能を評価するために,グラジエント溶出法を適応した際の熱レンズ信号に対する影響を検討した.熱レンズ信号は溶媒によって信号強度が大きく変化するため,グラジエント溶出法のような溶媒組成が時間とともに変化するような系では,その影響を検討する必要がある.水-アセトニトリル系グラジエントにおいては,アセトニトリルの体積分率が40% 未満では,熱レンズ測定に大きな影響を及ぼさないということが分かった.標準試料を用いてLC/UV-TLMの検出性能を評価したところ,通常の分光光度計を検出器としたシステムに比較して10倍以上高感度であることが分かった.
Type: article
URI: http://hdl.handle.net/2115/71686
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 渡慶次 学

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