Determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS

LV Lei; ZHAO Liang; LAN Hongtao; JIN Rounan; ZHANG Guoqing

doi:10.3969/j.issn.1006-0111.2014.04.008

Message Board

Respected readers, authors and reviewers, you can add comments to this page on any questions about the contribution, review, editing and publication of this journal. We will give you an answer as soon as possible. Thank you for your support!

Name

E-mail

Phone

Title

Content

Verification Code

Objective To develop a method for determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS. Methods Acetonitrile was used to precipitate protein in plasma samples.The separation was performed on a SHISEIDO MG C₁₈ reverse phase column (3.0 mm×100 mm,3 μm).The mobile phase were acetonitrile and water (containing 0.1% formic acid and 0.5% ammonia), gradient elution.The flow rate was 0.4 ml/min, the temperature of column was 25℃ and the injection volume was 5 μl.Electrospray ion source (ESI) and selected ion mode (SIM) were applied for quantitative analysis under negative ion mode. Results The calibration curves of 13 kinds of bile acids were linear in the range of 3.3 - 4 400 ng/ml, intra-day and inter-day precision results were all less than 7%, the average recovery were 82.67%-109.75%.The result showed that the content of conjugated bile acid increased in the plasma of patients with primary liver cancer compared with the healthy control group, especially in TUDCA, TCDCA, TCA, GCA and GCDCA. Conclusion The LC-MS method was sensitive and selective which could be suitable for determination of bile acid in healthy people and patients with primary liver cancer.The study laid a foundation for the metabolism of bile acid in vivo and the clinical diagnosis and treatment of patients with hepatocellular carcinoma.

Determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS

Eastern Hepatobiliary Surgery Hospital affiliated to Second Military Medical University, Shanghai 200438, China

Received Date: 2013-06-09

Rev Recd Date: 2013-11-25

Key words:

Abstract: Objective To develop a method for determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS. Methods Acetonitrile was used to precipitate protein in plasma samples.The separation was performed on a SHISEIDO MG C₁₈ reverse phase column (3.0 mm×100 mm,3 μm).The mobile phase were acetonitrile and water (containing 0.1% formic acid and 0.5% ammonia), gradient elution.The flow rate was 0.4 ml/min, the temperature of column was 25℃ and the injection volume was 5 μl.Electrospray ion source (ESI) and selected ion mode (SIM) were applied for quantitative analysis under negative ion mode. Results The calibration curves of 13 kinds of bile acids were linear in the range of 3.3 - 4 400 ng/ml, intra-day and inter-day precision results were all less than 7%, the average recovery were 82.67%-109.75%.The result showed that the content of conjugated bile acid increased in the plasma of patients with primary liver cancer compared with the healthy control group, especially in TUDCA, TCDCA, TCA, GCA and GCDCA. Conclusion The LC-MS method was sensitive and selective which could be suitable for determination of bile acid in healthy people and patients with primary liver cancer.The study laid a foundation for the metabolism of bile acid in vivo and the clinical diagnosis and treatment of patients with hepatocellular carcinoma.

HTML

Reference (12)

[1]	周毅. 胆汁酸的临床诊断及应用进展[J].放射免疫学杂志, 2012, 25(3): 267-268.
[2]	李淑华, 阮桂芝. 胆汁酸在肝胆疾病诊断中的价值[J].中国医药指南, 2011, 9(13): 253-254.
[3]	段凤梅. 血清胆汁酸测定在肝病诊断中临床应用[J].中国医药指南, 2011, 9(28): 18-19.
[4]	翟庆云. 总胆汁酸及肝功指标检测在肝病诊断中的意义[J].中国社区医师, 2008, 10(9): 64-65.
[5]	王孔明, 林红.不同肝胆疾病患者血清总胆汁酸测定的临床意义[J].河北医科大学学报, 2011, 32(4): 475-476.
[6]	王文娟, 邓文平, 王猛, 等. 高效液相色谱法同时测定胆汁中9种结合型胆汁酸[J].重庆医科大学学报, 2009, 34(1): 59-62.
[7]	Batta AK, Salen G. Gas chromatography of bile acid[J].J Chromatogr B, 1999, 723(1-2): 1-5.
[8]	Rodriguez VG, Lucangioli SE, Fernandez GC, et al.Determination of bile acid in pharmaceutical formulations using micellar electrokinetic chromatography[J].J Pharm Biomed Anal, 2000, 23(2-3): 375-380.
[9]	李水军, 刘罡一, 印其友, 等. 液相色谱-串联质谱法测定血清中15种胆汁酸[J].分析测试学报, 2007, 26(3): 315-322.
[10]	Tagliacozzi D, Mozzi AF, Casetya B, et al.Quantitative analysis of bile acid in human plasma by liquid chromatography-electrospray tandem mass spectrometry: a simple and rapid one-step method[J].Clin Chem Lab Med, 2003, 41(12): 1633-1640.
[11]	邓文平, 王文娟, 邵勇, 等. 高效液相色谱-串联质谱法测定妊娠肝内胆汁淤积患者血清牛磺结合型胆汁酸[J]. 重庆医科大学学报, 2009, 34(6): 732-734.
[12]	丁敏, 叶炼, 王猛, 等. 高效液相色谱-串联质谱法同时测定血清中游离和结合型胆汁酸[J].分析化学, 2007, 35(10):1506-1508.

Message Board

Determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS

doi: 10.3969/j.issn.1006-0111.2014.04.008

Abstract

References

Proportional views

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Related

Proportional views