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Ionization method and apparatus using electrospray, and analyzing method and apparatus

外国特許コード F130007405
整理番号 P09-039US
掲載日 2013年6月18日
出願国 アメリカ合衆国
出願番号 201213491331
公報番号 20120248303
公報番号 9111739
出願日 平成24年6月7日(2012.6.7)
公報発行日 平成24年10月4日(2012.10.4)
公報発行日 平成27年8月18日(2015.8.18)
優先権データ
  • 特願2009-278458 (2009.12.8) JP
  • 2010JP072511 (2010.12.8) WO
発明の名称 (英語) Ionization method and apparatus using electrospray, and analyzing method and apparatus
発明の概要(英語) It is arranged so that living tissue that has not been pretreated can be adopted as a sample of interest.
A sample is introduced into at least a tip portion of a hollow insulated sample holder 11 having a small hole 11a in the tip portion.
A slender metal wire 12, which has been inserted into the sample holder 11 from the rear, is projected outwardly from the sample holder 11 through the hole 11a while being brought into contact with the sample.
A high voltage is applied to the slender metal wire 12 at least in a portion of a time period in which the tip of the slender metal wire 12 is being projected outwardly from the hole 11a, thereby ionizing, by electrospray, the sample adhering to the tip of the slender metal wire 12.
The ions are introduced into an analyzing apparatus and are analyzed.
従来技術、競合技術の概要(英語) BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to an ionization method and apparatus using electrospray, and to an analyzing method and apparatus.
2. Description of the Related Art
A typical method of imaging mass spectrometry for dealing with biological samples and industrial products and the like is mass spectrometry using matrix-assisted laser desorption ionization (MALDI).
With this method, a pretreatment, namely the preparation of a MALDI sample, is required.
The molecular analysis of live single cells has been gaining momentum in recent years, and nano-electrospray (ESI) mass spectrometry has been proposed as an effective analytical method for this purpose.
See Mizuno, Tsuyama, Harada and Masujima "Live single-cell video-mass spectrometry for cellular and subcellular molecular detection and cell classification", J. Mass Spectrom. 2008; 43: 1692-1700.
According to this method, a cell of interest or cellular solution thereof is drawn into the end of a glass ESI tip (a capillary), the end having an opening diameter (inner diameter) on the micron order, an ionization solvent (for example, acetonitrile containing 0.5% formic acid for the positive-ion mode, or 0.5% ammonia water for the negative-ion mode) is applied to the interior of the ESI tip to achieve dilution by 106 times or more, then the diluted cellular sample is ionized by electrospray and the result introduced to an analyzing apparatus.
With this method, the solvent also undergoes mass spectrometry as a matter of course, and an ion signal ascribable to molecules resulting from the solvent shows up in the mass spectrometry spectra.

特許請求の範囲(英語) [claim1]
1. An ionization method using electrospray comprising: introducing a sample into at least a tip portion of a hollow insulated sample holder having a small hole in the tip portion;
supporting an electrically conductive linear body, which has been inserted inside said sample holder, such that a tip thereof is projectable outwardly from or
retractable into said hole;
projecting the tip of said linear body outwardly from the sample holder through said hole while being brought into contact with said sample inside said sample holder; and
applying a high voltage for electrospray of several kV to 1 kV to said linear body after the tip of said electrically conductive linear body is projected outwardly from said hole of said sample holder, thereby ionizing, by electrospray, the sample adhering to the tip of said linear body,
wherein the linear body is a solid wire.
[claim2]
2. An ionization method according to claim 1, wherein the projecting and retracting of the tip of said electrically conductive linear body and the electro spraying of the sample are repeated multiple times with regard to a single sample.
[claim3]
3. An ionization method according to claim 1, wherein application of said high voltage is halted when the sample at the tip of the electrically conductive linear body has been consumed by electrospray.
[claim4]
4. An ionization method according to claim 1, wherein a sample is taken directly at the tip of said sample holder.
[claim5]
5. An ionization method according to claim 1, wherein a liquid sample is supplied to said sample holder from liquid chromatography.
[claim6]
6. An ionization method according to claim 1, wherein ionization is carried out under atmospheric pressure.
[claim7]
7. An ionization method according to claim 1, further comprising using an electrically conductive linear body at least the tip of which has been subjected to a hydrophobic or hydrophilic surface treatment.
[claim8]
8. An ionization analyzing method of analyzing molecules that have been ionized by the ionization method set forth in claim 1.
[claim9]
9. An ionization method according to claim 2, wherein a sample is taken directly at the tip of said sample holder.
[claim10]
10. An ionization method according to claim 3, wherein a sample is taken directly at the tip of said sample holder.
[claim11]
11. An ionization method according to claim 2, wherein a liquid sample is supplied to said sample holder from liquid chromatography.
[claim12]
12. An ionization method according to claim 3, wherein a liquid sample is supplied to said sample holder from liquid chromatography.
[claim13]
13. An ionization method according to claim 2, wherein ionization is carried out under atmospheric pressure.
[claim14]
14. An ionization method according to claim 3, wherein ionization is carried out under atmospheric pressure.
[claim15]
15. An ionization method according to claim 2, further comprising using an electrically conductive linear body at least the tip of which has been subjected to a hydrophobic or hydrophilic surface treatment.
[claim16]
16. An ionization method according to claim 3, further comprising using an electrically conductive linear body at least the tip of which has been subjected to a hydrophobic or hydrophilic surface treatment.
[claim17]
17. An ionization method according to claim 6, further comprising using an electrically conductive linear body at least the tip of which has been subjected to a hydrophobic or hydrophilic surface treatment.
[claim18]
18. An ionization analyzing method of analyzing molecules that have been ionized by the ionization method set forth in claim 2.
  • 発明者/出願人(英語)
  • HIRAOKA KENZO
  • UNIVERSITY OF YAMANASHI
国際特許分類(IPC)
米国特許分類/主・副
  • 2016/01/01
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