Phospholipid classification detection and quantification method based on stable isotope labeling

Abstract

The present invention discloses a phospholipid classification detection and quantification method based on stable isotope labeling, and belongs to the technical field of phospholipid quantification detection methods. According to the method, mainly trimethylsilyl diazomethane is used to respectively generate diazomethane and deuterium-labeled diazomethane in a methyl tert-butyl ether / methanol / 1N hydrochloric acid solution system and a methyl tert-butyl ether / deuteromethanol / 1N deuterium chloride solution system in an in-situ manner so as to further respectively carry out methyl esterification on the phosphoric acid group or carboxylic acid group in the phospholipid molecule to generate the light isotope labeled-phospholipid methyl esterification derivative and the heavy isotope labeled-phospholipid methyl esterification derivative, wherein the light isotope labeled-phospholipid methyl esterification derivative and the heavy isotope labeled-phospholipid methyl esterification derivative have the same physical and chemical properties and different molecular weights; and the relative intensity of the light isotope labeled-mass spectrum peak signal and the heavy isotope labeled-mass spectrum peak signal are compared to associate with the phospholipid molecule amount in the sample so as to achieve the relative quantification on the phospholipid. The method of the present invention has characteristics of enhanced sensitivity and completion within a shor time.

Description

technical field [0001] The invention relates to the technical field of phospholipid quantitative detection methods, in particular to a stable isotope label-based phospholipid classification detection and quantitative method. 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 between phospholipid metabo...

AI Technical Summary

Problems solved by technology

However, at present, mass spectrometric identification and quantification of phospholipid molecules in different samples are usually analyzed separately, and then referenced or quantified by internal standards, which has certain limitations

For example, the current commercial phospholipid internal standards are not only expensive and limited in variety, but also have certain restrictions on storage because phospholipid molecules contain unsaturated double bonds and are easily oxidized.

The ionization efficiency of phospholipid molecules is also easily affected by different factors such as ion spray stability at different times, the effect of fatty acid chain length and degree of unsaturation on ion efficiency, which may lead to certain errors in mass spectrometry quantification

On the other hand, when using electrospray ionization-mass spectrometry (ESI-MS) to analyze and quantify phospholipid lipid molecules, different types of phospholipid molecules have different characteristics due to their own structure (mainly polar head) and solution composition. ionization propensity, which leads to the combination of positive and negative ion modes when using ESI-MS to qualitatively and quantitatively analyze the phospholipid group in biological samples, resulting in an increase in analysis time

At the same time, in the negative ion mode, the detection sensitivity of some phospholipids is relatively low

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