Highly sensitive detection of non-labeled peptides using UV excitation thermal lens microscope/liquid chromatography

Shinichiro Hiki, Manabu Tokeshi, Masaya Kakuta, Kazuma Mawatari, Yoshikuni Kikutani, Kiichi Sato, Akihide Hibara, Kiyohito Shimura, Naoyuki Uchida, Takehiko Kitamori

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

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 a conventional system with the spectrophotometer used as a detector.

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalBUNSEKI KAGAKU
Volume56
Issue number1
DOIs
Publication statusPublished - 2007

Keywords

  • Gradient elution method
  • Liquid chromatography
  • Non-labeled proteins
  • Thermal lens microscope

ASJC Scopus subject areas

  • Analytical Chemistry

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