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Chemical derivatization-based phosphatide classification detection and quantification method

A technology for classification and detection of phospholipids, applied in measurement devices, scientific instruments, material separation, etc., can solve the problems of unclear detection and quantification of phospholipid molecules, and achieve the effect of fast detection and high sensitivity

Active Publication Date: 2015-11-04
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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Problems solved by technology

Third, how methylated-derived phospholipid molecules can be detected and quantified by mass spectrometry is unclear

Method used

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  • Chemical derivatization-based phosphatide classification detection and quantification method
  • Chemical derivatization-based phosphatide classification detection and quantification method
  • Chemical derivatization-based phosphatide classification detection and quantification method

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Embodiment

[0108] Sample and Reagent Preparation

[0109] ① Reagents: All chemical reagents of analytical grade (unless otherwise specified) were purchased from Sigma-Aldrich Company (St. ) produced Milli-Q water; trimethylsilyl diazomethane (IUPAC name=(diazomethyl)-trimethylsilane, TMS-diazomethane) (2M dissolved in n-hexane) was purchased from Acros Organics company; chromatographic grade purity methyl tert Butyl ether (methyl tert-butyl ether, MTBE), methanol (methanol), chloroform (Choloform), acetic acid (Acetic acid) are products of J.T.Baker Company (Phillipsburg, PA, USA).

[0110] ②Internal standard: You can choose isotope-labeled phospholipid molecules that can be detected separately or select phospholipids that do not exist or have a very low content in the sample and add them to the sample as an internal standard. This embodiment selects C17:0 / C14:1-PtdCho, C17:0 / C14:1-PtdEtn, C17:0 / C14:1-PtdGro, C17:0 / C14:1-PtdSer, C17:0 / C14: 1-PtdH, C17:0 / C14:1-PtdIns, C17:1-lysoPtdCho, ...

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Abstract

The present invention discloses a chemical derivatization-based phosphatide classification detection and quantification method, and belongs to the quantitative detection method technical field. According to the method, trimethylsilyldiazomethane is used for methylation derivation of a phosphate group in a phosphatide molecule in a to-be-tested sample; a plurality of ions to be tested by mass spectrometer are produced by the methylation derivatived phosphatide molecule in the ionized sample; in the selection of corresponding ions to be tested by the mass spectrometer, the ions added with H <+> or NH4 <+> are selected by different phosphatide kinds; different neutral loss or precursor ion scanning models can be set according to mass spectrometer fragmentation rules of different kinds of ions, and amounts of mass spectrometer ion models of different kinds of ions can be obtained; and finally, by comparison with the internal standard and calculation, the amounts of different types of phosphatide molecules in the sample can be obtained. The method is fast in detecting speed and high in sensitivity.

Description

technical field [0001] The invention relates to the technical field of quantitative detection methods for phospholipids, in particular to a method for the classification, detection and quantification of phospholipids based on chemical derivation. Background technique [0002] Phospholipids are not only an important component of cell membranes, but also widely involved in the regulation of various life processes, including energy conversion, material transport, information recognition and transmission, cell development and differentiation, and cell apoptosis. Abnormal metabolism of phospholipids is also closely related to arteriosclerosis, diabetes, obesity, Alzheimer's disease and tumorigenesis. Therefore, from the perspective of "omics", systematic and large-scale research on the dynamic changes in the composition and relative quantity of phospholipid molecules under different physiological or pathological conditions will be of great help in revealing the relationship betwe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 蔡潭溪杨福全舒清博郭晓静
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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