Method of quantifying content of recombinant troponin I by peptide fragment isotope dilution mass-spectrography

Abstract

The invention discloses a method of quantifying the content of recombinant troponin I by peptide fragment isotope dilution mass-spectrography and relates to a method of accurately quantifying the content of recombinant troponin I. The method comprises the following steps: designing a characteristic peptide fragment; determining mass spectrometry ion pairs of the characteristic peptide fragment; performing an LC-MRM condition research; performing sample analysis pre-treatment which mainly comprises the steps: recombinant protein degeneration, reductive alkylation, enzymolysis, sample purification and concentration of the sample; data analysis: concentration of a cTnI specific peptide fragment and the appearance time of the cTnI protein peptide being 4.20min; and recovery rate and precision: the recovery rate and precision meet the specification, and the mass spectrometer condition on the recombinant cTnI specific peptide fragment liquid phase is researched and modified. The established method is good in accuracy and fast in speed, and the appearance time is 4.20min.

Description

Technical field [0001] The invention relates to a method for accurately quantifying the content of recombinant troponin I, in particular to a method for quantifying the content of recombinant troponin I by peptide isotope dilution mass spectrometry. Background technique [0002] Troponin (cTnI) is one of the most sensitive biological indicators for detecting myocardial injury. However, cTnI detection still has some problems, mainly because the detection method has not been standardized, there are certain differences between different methods, and the detection results are often not comparable across time and space. [0003] The advantages of isotope liquid chromatography-tandem massspectrometry (LC / MS / MS) are high sensitivity, high accuracy, and high specificity, and the standardization of measurement results can be directly traced to the international basic unit system. Domestic scholars [1] established an amino acid isotope dilution mass spectrometry method to quantify recombina...

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Problems solved by technology

Domestic scholars [1] established amino acid isotope dilution mass spectrometry to quantitatively recombine cTnI. This method requires high-temperature hydrolysis of amino acids, which has potential safety hazards

[0004] There are few domestic research reports on the quantification of protein content by peptide isotope dilution mass spectrometry, mainly due to the complex composition of protein samples, the degree of ionization is easily affected by other impurity ions, and the experimental conditions are relatively complicated.

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