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

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Abstract

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.

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Authors and Affiliations

  1. Pathology, Xingtai Medical College, 618 Gangtiebei Road, Qiaoxi District, Xingtai, 054000, China

    Hui Li, Hongxia Zhao & Zhanhua Li

  2. Statistics, Xingtai Technician College, Xingtai, 054000, China

    Qian Li

  3. Pathology, The Second Affiliated Hospital of Xingtai Medical College, Xingtai, 054000, China

    Dongyue Meng

Authors
  1. Hui Li
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  2. Hongxia Zhao
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  3. Qian Li
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  4. Dongyue Meng
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  5. Zhanhua Li
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Correspondence to Zhanhua Li.

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The authors have declared no conflict of interest.

This study was approved by the Ethical Committee of Xingtai Health Bureau. Written informed consent was obtained from each patient for the use of his/her urine and plasma samples.

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Li, H., Zhao, H., Li, Q. et al. Analysis of Glycocholic Acid in Human Plasma and Urine from Hepatocellular Carcinoma Patients. Chromatographia 80, 209–215 (2017). https://doi.org/10.1007/s10337-016-3237-3

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  • DOI: https://doi.org/10.1007/s10337-016-3237-3

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