A rapid, accurate and sensitive stable isotope dilution ultra performance liquidchromatography electrospray ionization tandem mass spectrometry (ID-UPLC-ESI–MS/MS) method for the determination of glycocholic acid (GCA) in human serum was developed and validated. Serum samples were spiked with D₅-glycocholic acid and then pretreated with protein precipitation. The analysis was performed on a Waters BEH C18 column (100 mm × 2.1 mm, 1.7 μm), followed by ESI–MS/MS detection in negative ion mode under multiple reaction monitoring mode. The calibration curves covered a concentration range from 0.2 to 400 ng/mL. The limit of detection and limit of quantification was 0.01 ng/mL and 0.05 ng/mL, respectively. The method showed satisfactory precision on intra-day (2.3–6.1%) and inter-day (2.4–4.6%) analyses and achieved good recovery at three spiked levels (103.7–114.3%). Moreover, this established method was successfully applied for quantification of GCA in serum samples from healthy volunteers, patients with hepatocellular carcinoma (HCC) and patients with other cancers. We demonstrated that the level of GCA in patients with HCC was significantly higher not only than that in healthy controls, but also than that in patients with other cancer, whereas no significant difference of GCA level was observed between healthy control group and other cancers group.

建立并验证了一种快速，准确，灵敏的稳定同位素稀释超高效液相色谱电喷雾电离串联质谱法（ID-UPLC-ESI-MS / MS）。血清样品掺入D ₅-乙醇酸，然后用蛋白质沉淀进行预处理。在Waters BEH C18色谱柱（100 mm×2.1 mm，1.7μm）上进行分析，然后在多反应监测模式下以负离子模式进行ESI-MS / MS检测。校准曲线的浓度范围为0.2到400 ng / mL。检测限和定量限分别为0.01 ng / mL和0.05 ng / mL。该方法在日内（2.3–6.1％）和日间（2.4–4.6％）分析中显示出令人满意的精度，并在三个加标水平（103.7–114.3％）上实现了良好的回收率。此外，该建立的方法已成功用于健康志愿者，肝细胞癌（HCC）患者和其他癌症患者血清样品中GCA的定量分析。