临床戒烟研究中采用敏感、特异性UPLC-串联质谱检测血浆尼古丁、可替宁、反式-3-羟基甲基可丁尼和伐尼克兰含量
2011/11/21
摘要
我们开发和验证了一种敏感和特异的超高效液相色谱-串联质谱法，能同时检测人体血浆中尼古丁、尼古丁代谢产物（可替宁）、反式3-羟基甲基可丁尼和伐尼克兰。基于混合模式阳离子交换，通过固相提取0.5 ml血浆中的目标复合物和内部标准品（尼古丁-d4, 可替宁-d3, 反式3-羟基甲基可丁尼-d3和CP-533,633 [伐尼克兰的结构类似物]）。采用亲水液相色谱柱(HILIC BEH 2.1×100mm, 1.7μm)进行色谱分离。采用梯度程序，10 mM甲酸铵缓冲液（pH 3）/乙腈流动相，流速0.4 mL/min，采用三重四极杆质谱仪的正离子模式电喷雾接口检测复合物，运用多极反应监测进行定量。基质效应采用成功来评价，变异系数小于8%。该检测方法完全按照美国食品药品监督局和法国制药科学和技术协会的指南要求进行了验证。尼古丁的浓度范围为2～500 ng/mL；可替宁为1～1000 ng/mL；反式3-羟基甲基可丁尼为2～1000 ng/mL；伐尼克兰为1～500 ng/mL。该方法检测血浆，具有较好的正确度(86.2-113.6%)、可重复性(1.9-12.3%)、中等精确度(4.4-15.9%)和稳定性。该检测方法成功定量分析了临床戒烟研究中400个血浆样本的尼古丁、尼古丁代谢产物和伐尼克兰。
（陈欣 审校）
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Oct 1. [Epub ahead of print]
Quantification of nicotine, cotinine, trans-3’-hydroxycotinine and varenicline in human plasma by a sensitive and specific UPLC-tandem mass-spectrometry procedure for a clinical study on smoking cessation.
Dobrinas M, Choong E, Noetzli M, Cornuz J, Ansermot N, Eap CB.
Source
Unit of Pharmacogenetics and Clinical Psychopharmacology, Centre for Psychiatric Neuroscience, Department of Psychiatry, CHUV, University of Lausanne, Hospital of Cery, Prilly, Switzerland.
Abstract
A sensitive and specific ultra performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of nicotine, its metabolites cotinine and trans-3’-hydroxycotinine and varenicline in human plasma was developed and validated. Sample preparation was realized by solid phase extraction of the target compounds and of the internal standards (nicotine-d4, cotinine-d3, trans-3’-hydroxycotinine-d3 and CP-533,633, a structural analog of varenicline) from 0.5mL of plasma, using a mixed-mode cation exchange support. Chromatographic separations were performed on a hydrophilic interaction liquid chromatography column (HILIC BEH 2.1×100mm, 1.7μm). A gradient program was used, with a 10mM ammonium formate buffer pH 3/acetonitrile mobile phase at a flow of 0.4mL/min. The compounds were detected on a triple quadrupole mass spectrometer, operated with an electrospray interface in positive ionization mode and quantification was performed using multiple reaction monitoring. Matrix effects were quantitatively evaluated with success, with coefficients of variation inferior to 8%. The procedure was fully validated according to Food and Drug Administration guidelines and to Société Française des Sciences et Techniques Pharmaceutiques. The concentration range was 2-500ng/mL for nicotine, 1-1000ng/mL for cotinine, 2-1000ng/mL for trans-3’-hydroxycotinine and 1-500ng/mL for varenicline, according to levels usually measured in plasma. Trueness (86.2-113.6%), repeatability (1.9-12.3%) and intermediate precision (4.4-15.9%) were found to be satisfactory, as well as stability in plasma. The procedure was successfully used to quantify nicotine, its metabolites and varenicline in more than 400 plasma samples from participants in a clinical study on smoking cessation.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Oct 1. [Epub ahead of print]
我们开发和验证了一种敏感和特异的超高效液相色谱-串联质谱法，能同时检测人体血浆中尼古丁、尼古丁代谢产物（可替宁）、反式3-羟基甲基可丁尼和伐尼克兰。基于混合模式阳离子交换，通过固相提取0.5 ml血浆中的目标复合物和内部标准品（尼古丁-d4, 可替宁-d3, 反式3-羟基甲基可丁尼-d3和CP-533,633 [伐尼克兰的结构类似物]）。采用亲水液相色谱柱(HILIC BEH 2.1×100mm, 1.7μm)进行色谱分离。采用梯度程序，10 mM甲酸铵缓冲液（pH 3）/乙腈流动相，流速0.4 mL/min，采用三重四极杆质谱仪的正离子模式电喷雾接口检测复合物，运用多极反应监测进行定量。基质效应采用成功来评价，变异系数小于8%。该检测方法完全按照美国食品药品监督局和法国制药科学和技术协会的指南要求进行了验证。尼古丁的浓度范围为2～500 ng/mL；可替宁为1～1000 ng/mL；反式3-羟基甲基可丁尼为2～1000 ng/mL；伐尼克兰为1～500 ng/mL。该方法检测血浆，具有较好的正确度(86.2-113.6%)、可重复性(1.9-12.3%)、中等精确度(4.4-15.9%)和稳定性。该检测方法成功定量分析了临床戒烟研究中400个血浆样本的尼古丁、尼古丁代谢产物和伐尼克兰。
（陈欣 审校）
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Oct 1. [Epub ahead of print]
Quantification of nicotine, cotinine, trans-3’-hydroxycotinine and varenicline in human plasma by a sensitive and specific UPLC-tandem mass-spectrometry procedure for a clinical study on smoking cessation.
Dobrinas M, Choong E, Noetzli M, Cornuz J, Ansermot N, Eap CB.
Source
Unit of Pharmacogenetics and Clinical Psychopharmacology, Centre for Psychiatric Neuroscience, Department of Psychiatry, CHUV, University of Lausanne, Hospital of Cery, Prilly, Switzerland.
Abstract
A sensitive and specific ultra performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of nicotine, its metabolites cotinine and trans-3’-hydroxycotinine and varenicline in human plasma was developed and validated. Sample preparation was realized by solid phase extraction of the target compounds and of the internal standards (nicotine-d4, cotinine-d3, trans-3’-hydroxycotinine-d3 and CP-533,633, a structural analog of varenicline) from 0.5mL of plasma, using a mixed-mode cation exchange support. Chromatographic separations were performed on a hydrophilic interaction liquid chromatography column (HILIC BEH 2.1×100mm, 1.7μm). A gradient program was used, with a 10mM ammonium formate buffer pH 3/acetonitrile mobile phase at a flow of 0.4mL/min. The compounds were detected on a triple quadrupole mass spectrometer, operated with an electrospray interface in positive ionization mode and quantification was performed using multiple reaction monitoring. Matrix effects were quantitatively evaluated with success, with coefficients of variation inferior to 8%. The procedure was fully validated according to Food and Drug Administration guidelines and to Société Française des Sciences et Techniques Pharmaceutiques. The concentration range was 2-500ng/mL for nicotine, 1-1000ng/mL for cotinine, 2-1000ng/mL for trans-3’-hydroxycotinine and 1-500ng/mL for varenicline, according to levels usually measured in plasma. Trueness (86.2-113.6%), repeatability (1.9-12.3%) and intermediate precision (4.4-15.9%) were found to be satisfactory, as well as stability in plasma. The procedure was successfully used to quantify nicotine, its metabolites and varenicline in more than 400 plasma samples from participants in a clinical study on smoking cessation.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Oct 1. [Epub ahead of print]
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为什么采用两种戒烟药物疗效好于一种药物：渴望抑制的作用
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