Analysis of Glycocholic Acid in Human Plasma and Urine from Hepatocellular Carcinoma Patients

Glycocholic acid (GCA) has been identified as endogenous biomarker for hepatocellular carcinoma (HCC). To dissolve protein and liberate GCA from protein, ionic liquids (ILs) that contain chaotropic ions were used for pretreatment of liquid biological samples. Coupling with solid-phase extraction (SPE) and reversed-phase high-performance liquid chromatography (RP-HPLC), the novel sample pretreatment method was applied for quantitative determination of GCA in urine and plasma samples. Compared with the traditional organic solvents pretreatment of biological samples, the proposed method is “greener” and simpler, due to no use of volatile organic solvent, and avoiding centrifugation. Under the optimal conditions, when the GCA-free urine and plasma samples were spiked with GCA at 0.05–1.0 and 0.2–10 μmol L^?1, the recoveries of GCA ranged between 95.8–101.6 and 96.9–100.4%, respectively. These procedures only required 1 mL of urine and 3 mL of 3 mM ILs aqueous solution and 100 μL of plasma and 4 mL of 2 mM ILs aqueous solution, respectively. The proposed method has been successfully validated on a small sample size of 14 HCC patients and 14 healthy volunteers. For HCC patients, the mean concentration of GCA was 24.79 ± 6.86 and 31.98 ± 11.12 μmol L^?1 in urine and plasma samples, and was about 3 times and 24 times as much as in healthy people, respectively. The proposed method opens up a new possibility in early diagnosis of HCC.