TY - JOUR
T1 - Highly sensitive detection of non-labeled peptides using UV excitation thermal lens microscope/liquid chromatography
AU - Hiki, Shinichiro
AU - Tokeshi, Manabu
AU - Kakuta, Masaya
AU - Mawatari, Kazuma
AU - Kikutani, Yoshikuni
AU - Sato, Kiichi
AU - Hibara, Akihide
AU - Shimura, Kiyohito
AU - Uchida, Naoyuki
AU - Kitamori, Takehiko
PY - 2007
Y1 - 2007
N2 - 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.
AB - 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.
KW - Gradient elution method
KW - Liquid chromatography
KW - Non-labeled proteins
KW - Thermal lens microscope
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U2 - 10.2116/bunsekikagaku.56.1
DO - 10.2116/bunsekikagaku.56.1
M3 - Article
AN - SCOPUS:33847657884
VL - 56
SP - 1
EP - 7
JO - Bunseki Kagaku
JF - Bunseki Kagaku
SN - 0525-1931
IS - 1
ER -