Urinary sarcosine was considered to be a potential biomarker for prostate cancer (Pca). In this work, an integrated strategy of multiplex tags chemical isotope labeling (MTCIL) combined with magnetic dispersive solid phase extraction (MDSPE), was proposed for specific extraction and high-throughput determination of sarcosine by ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). In the past three months, we have developed 8-plex MTCIL reagents with excellent qualitative and quantitative performance. In this work, the multiplexing capacity of MTCIL reagents (MTCIL360/361/362/363/364/365/366/375/376/378/379/381) was increased 1.5-fold from 8-plex to 12-plex. MTCIL359 was prepared and used to label sarcosine standard as internal standard (IS). The structural analogue derivative (MTCIL373-sarcosine) of all targeted MTCIL-sarcosine derivatives was synthesized and used as a novel dummy template to prepare dummy magnetic molecularly imprinted polymers (DMMIPs). The integration of MTCIL and DMMIPs procedures were extremely favorable to excellent chromatographic separation and efficient mass spectrometric detection. The labeling efficiency, chromatographic retention and mass spectrometry responses of MTCIL reagents were consistent for sarcosine. In a single UHPLC-MS/MS run (2.0 min), this method can simultaneously quantify sarcosine in 12-plex urine samples and achieve unbiased concentrations comparison between different urine samples. Analytical parameters including linearity (R² 0.989–0.997), detection limits (0.02 nM), precision (2.6–11.5%), accuracy (96.1–107.4%), matrix effect, labeling and extraction efficiency were carefully validated. The proposed method was successfully applied for urinary sarcosine determination of healthy male individuals and Pca patients. It was found that the sarcosine concentrations in these two groups were statistically extremely significantly different (P < 0.001). The developed method was a powerful analytical tool to substantially promote the analysis throughput and large-scale experiments about the potential biomarker research.

尿肌氨酸被认为是前列腺癌（Pca）的潜在生物标志物。在这项工作中，提出了一种结合标签化学同位素标记（MTCIL）和磁分散固相萃取（MDSPE）的集成策略，用于超高效液相色谱串联质谱法（UHPLC）的肌氨酸特异性提取和高通量测定。 -MS / MS）。在过去的三个月中，我们开发了具有出色定性和定量性能的8重MTCIL试剂。在这项工作中，MTCIL试剂（MTCIL360 / 361/362/363/364/365/366/375/376/378/379/381）的多路复用能力从8重提高到12重。制备了MTCIL359，并用于将肌氨酸标准品标记为内标（IS）。合成了所有目标MTCIL-肌氨酸衍生物的结构类似物衍生物（MTCIL373-肌氨酸），并将其用作新型的虚拟模板，以制备虚拟磁性分子印迹聚合物（DMMIP）。MTCIL和DMMIPs程序的集成非常有利于出色的色谱分离和有效的质谱检测。MTCIL试剂的标记效率，色谱保留率和质谱响应与肌氨酸一致。在一次UHPLC-MS / MS运行中（2.0分钟），该方法可以同时定量12样尿样中的肌氨酸，并实现不同尿样之间的无偏浓度比较。分析参数包括线性（R MTCIL和DMMIPs程序的集成非常有利于出色的色谱分离和有效的质谱检测。MTCIL试剂的标记效率，色谱保留率和质谱响应与肌氨酸一致。在一次UHPLC-MS / MS运行中（2.0分钟），该方法可以同时定量12样尿样中的肌氨酸，并实现不同尿样之间的无偏浓度比较。分析参数包括线性（R MTCIL和DMMIPs程序的集成非常有利于出色的色谱分离和有效的质谱检测。MTCIL试剂的标记效率，色谱保留率和质谱响应与肌氨酸一致。在一次UHPLC-MS / MS运行中（2.0分钟），该方法可以同时定量12样尿样中的肌氨酸，并实现不同尿样之间的无偏浓度比较。分析参数包括线性（R 该方法可同时定量12样尿样中的肌氨酸，并在不同尿样之间实现无偏浓度比较。分析参数包括线性（R 该方法可同时定量12样尿样中的肌氨酸，并在不同尿样之间实现无偏浓度比较。分析参数包括线性（R² 0.989–0.997），检出限（0.02 nM），精密度（2.6–11.5％），准确度（96.1–107.4％），基质效应，标记和提取效率得到了验证。该方法已成功用于健康男性和Pca患者的尿肌氨酸测定。结果发现，两组的肌氨酸浓度在统计学上有显着差异（P  <0.001）。所开发的方法是功能强大的分析工具，可大大提高分析通量和有关潜在生物标记物研究的大规模实验。