应用HPLC-MS技术测定健康受试者和肝癌患者血浆中13种胆汁酸的含量

吕磊; 赵亮; 兰红涛; 金柔男; 张国庆

doi:10.3969/j.issn.1006-0111.2014.04.008

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应用HPLC-MS技术测定健康受试者和肝癌患者血浆中13种胆汁酸的含量

吕磊, 赵亮, 兰红涛, 金柔男, 张国庆

文章导航 > 药学实践与服务 > 2014 > 32(4): 270-272,293

吕磊, 赵亮, 兰红涛, 金柔男, 张国庆. 应用HPLC-MS技术测定健康受试者和肝癌患者血浆中13种胆汁酸的含量[J]. 药学实践与服务, 2014, 32(4): 270-272,293. doi: 10.3969/j.issn.1006-0111.2014.04.008

引用本文:

LV Lei, ZHAO Liang, LAN Hongtao, JIN Rounan, ZHANG Guoqing. Determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(4): 270-272,293. doi: 10.3969/j.issn.1006-0111.2014.04.008

Citation:

LV Lei, ZHAO Liang, LAN Hongtao, JIN Rounan, ZHANG Guoqing. Determination of 13 kinds of bile acid in healthy people and patients with primary liver cancer by HPLC-MS[J]. Journal of Pharmaceutical Practice and Service, 2014, 32(4): 270-272,293. doi: 10.3969/j.issn.1006-0111.2014.04.008

应用HPLC-MS技术测定健康受试者和肝癌患者血浆中13种胆汁酸的含量

doi: 10.3969/j.issn.1006-0111.2014.04.008

第二军医大学附属东方肝胆外科医院, 上海 200438

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

摘要: 目的建立HPLC-MS方法同时检测健康人和原发性肝癌患者血浆中13种胆汁酸的含量。方法血浆样品前处理采用乙腈-蛋白沉淀法,色谱分离采用SHISEIDO MG C₁₈ (100 mm×3.0 mm,3.0 μm)柱,流动相:乙腈-水(含0.1%甲酸和0.5%氨水),梯度洗脱,流速0.4 ml/min,柱温25℃,进样量5 μl。质谱监测采用ESI源负离子模式,选择离子(SIM)分段监测。结果 13种胆汁酸在3.3~4 400 ng/ml的浓度范围内线性关系良好;日内和日间精密度均<7%;平均回收率为82.67%~109.75%。样品测定结果显示,与健康对照组相比,原发性肝癌患者血浆中胆汁酸含量升高,其中以结合型胆汁酸TUDCA、TCDCA、TCA、GCA和GCDCA的升高最为明显。结论所建HPLC-MS方法灵敏度高、选择性好,可用于原发性肝癌患者与健康人体内血清胆汁酸的含量差异比较,该研究为肝癌患者体内胆汁酸的代谢和疾病的临床诊治奠定了基础。
- 血浆 /
- 胆汁酸测定 /
- 质谱 /
- 肝癌
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.
- plasma /
- bile acid determination /
- MS /
- liver cancer

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应用HPLC-MS技术测定健康受试者和肝癌患者血浆中13种胆汁酸的含量

doi: 10.3969/j.issn.1006-0111.2014.04.008

第二军医大学附属东方肝胆外科医院, 上海 200438

收稿日期: 2013-06-09
修回日期: 2013-11-25

关键词:

血浆 /

胆汁酸测定 /

质谱 /

肝癌

摘要: 目的建立HPLC-MS方法同时检测健康人和原发性肝癌患者血浆中13种胆汁酸的含量。方法血浆样品前处理采用乙腈-蛋白沉淀法,色谱分离采用SHISEIDO MG C₁₈ (100 mm×3.0 mm,3.0 μm)柱,流动相:乙腈-水(含0.1%甲酸和0.5%氨水),梯度洗脱,流速0.4 ml/min,柱温25℃,进样量5 μl。质谱监测采用ESI源负离子模式,选择离子(SIM)分段监测。结果 13种胆汁酸在3.3~4 400 ng/ml的浓度范围内线性关系良好;日内和日间精密度均<7%;平均回收率为82.67%~109.75%。样品测定结果显示,与健康对照组相比,原发性肝癌患者血浆中胆汁酸含量升高,其中以结合型胆汁酸TUDCA、TCDCA、TCA、GCA和GCDCA的升高最为明显。结论所建HPLC-MS方法灵敏度高、选择性好,可用于原发性肝癌患者与健康人体内血清胆汁酸的含量差异比较,该研究为肝癌患者体内胆汁酸的代谢和疾病的临床诊治奠定了基础。

English Abstract

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

Keywords:

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.