近年来质谱技术的持续进步促进了靶向蛋白质组的发展,在多反应监测(Multiple reaction monitoring,MRM)靶向蛋白质组定量方法的基础上衍生出平行反应监测(Parallel reaction monitoring,PRM)技术。该技术靶向定量灵敏度和通量更高,重现性也更好,但其对于高复杂性样本在定量速度和深度上均存在一定的局限性。改善PRM定量的色谱方法包括:色谱柱的优化,增加色谱柱内径、降低柱长;色谱洗脱条件的优化,提高液相洗脱流速、缩短洗脱时间;最终建立了一种简单高效的双反相色谱串联PRM靶向蛋白质组定量平台。该平台使用150μm内径和8 cm长的短色谱柱,在800 nL/min的高流速下和35 min有效洗脱梯度内,可实现对293T全细胞裂解液蛋白样本中多达400条低丰度肽段的快速定量。该研究优化了PRM靶向定量方法,将有利于PRM技术的推广,尤其为低丰度蛋白的精准定量提供了一种技术选择。 In recent years, the continuous improvement of mass spectrometry technology has promoted the development of targeted proteomics. Based on the quantitative proteome assay of multiple reaction monitoring (MRM), parallel reaction monitoring (PRM) technology was derived. The technique targets higher quantitative sensitivity and higher throughput with better reproducibility, but it has some limitations in terms of quantitative speed and depth for high-complexity samples. The methods to improve PRM quantification include: optimizing the column, increasing the inner diameter of the column, reducing the column length; optimizing the chromatographic elution conditions, increasing the liquid elution flow rate and shortening the elution time; finally, establishing a simple and efficient double Reverse phase chromatography tandem PRM targeted proteomic quantitative platform. Using 150 μm id and 8 cm long short columns, the platform achieves up to 400 low abundances in 293T whole cell lysate protein samples at high flow rates of 800 nL / min and an effective elution gradient of 35 min Rapid quantitation of peptides. This study optimizes the PRM target-based quantification method, which will be helpful for the promotion of PRM technology, especially for the accurate quantification of low-abundance proteins.