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学科主题: 分析化学
题名: Microfluidic chip-based high performance liquid chromatography coupled to mass spectrometry method for small molecule analysis
作者: Zhao CX(赵春霞) ;  Chang YW(常玉玮) ;  Wu ZM(吴泽明) ;  Zhou J(周佳) ;  Lu X(路鑫) ;  Xu GW(许国旺)
会议文集: Proceeding of HPLC 2011
会议名称: 37th International Symposium on High Performance Liquid Phase Separations and Related Techniques
会议日期: 2011-10-8
出版日期: 2011
会议地点: 大连
通讯作者: 许国旺
出版者: 待补充
出版地: 待补充
合作性质: 分会口头报告
部门归属: 1808
主办者: 中国化学会色谱专业委员会
摘要: Nanoflow liquid chromatography/mass spectrometry (nano-LC/MS), which has been developed from 1990s, generates increasing interest because it offers high detection sensitivity, reduces sample consumption and analysis time. In order to get a more stable and robust nano-LC/MS platform, a new generation of HPLC-Chip/MS system has been developed. All functional components, including enrichment column, analytical separation column and nano-electrospray emitter, were integrated onto this microfluidic device, which can significantly reduce the possibility of leaks and dead volumes, and greatly improve the ease of use during analysis. This miniaturized analytical platform results in less sample consumption, dilution and improved overall analyte ionization efficiency, and ultimately leads to the pronounced sensitivity advantage compared to conventional LC/MS. At present, most of the applications of new HPLC-Chip/MS system have been focused on proteomic analysis. However, the high sensitivity, ease of use and reusable nature of this device makes it attractive for small molecule analysis. In this study, a new UHC-Chip, including a 500 nL enrichment column and a 75 μm (ID) × 150 mm (L) separation column that were both packed with ZORBAX 80Ǻ SB-C18 5 μm stationary phase, was custom-made and used here for small molecule analysis, the HPLC-UHC-Chip/Q-TOF MS system was evaluated for qualitative and quantitative analysis of different small molecules covering a wide range of hydrophilicity. Compared with the standard chip, UHC-Chip shows higher enrichment capability, comparable chromatographic precision and separation performances. Good linearity and low LOD were also obtained by using the HPLC-UHC-Chip/Q-TOF MS. When compared with conventional LC/MS, 4-30 times increases in sensitivity were obtained for tested compounds. It was clearly demonstrated that the HPLC-UHC-Chip/Q-TOF MS system could be a valuable tool for the quantitative analysis of small molecule, satisfactory results of serum samples also proved its practicality and potential for use in drug testing and development. Moreover, a metabolic profiling method for soybean flavonoids based on HPLC-UHC- Chip/Q-TOF MS system and simple sample preparation was developed and validated. Seventy-three flavonoids were detected in a single run, including thirteen newly found flavonoids in soybean seed. To show the usefulness, the method was then applied to the profiling of different varieties of soybean sowed at the same place. A clear discrimination was observed among different cultivars, three isoflavones, accounting for nearly 80% of total flavonoid contents were found increased in the spring soybeans compared with the summer cultivars.
英文摘要: Nanoflow liquid chromatography/mass spectrometry (nano-LC/MS), which has been developed from 1990s, generates increasing interest because it offers high detection sensitivity, reduces sample consumption and analysis time. In order to get a more stable and robust nano-LC/MS platform, a new generation of HPLC-Chip/MS system has been developed. All functional components, including enrichment column, analytical separation column and nano-electrospray emitter, were integrated onto this microfluidic device, which can significantly reduce the possibility of leaks and dead volumes, and greatly improve the ease of use during analysis. This miniaturized analytical platform results in less sample consumption, dilution and improved overall analyte ionization efficiency, and ultimately leads to the pronounced sensitivity advantage compared to conventional LC/MS. At present, most of the applications of new HPLC-Chip/MS system have been focused on proteomic analysis. However, the high sensitivity, ease of use and reusable nature of this device makes it attractive for small molecule analysis. In this study, a new UHC-Chip, including a 500 nL enrichment column and a 75 μm (ID) × 150 mm (L) separation column that were both packed with ZORBAX 80Ǻ SB-C18 5 μm stationary phase, was custom-made and used here for small molecule analysis, the HPLC-UHC-Chip/Q-TOF MS system was evaluated for qualitative and quantitative analysis of different small molecules covering a wide range of hydrophilicity. Compared with the standard chip, UHC-Chip shows higher enrichment capability, comparable chromatographic precision and separation performances. Good linearity and low LOD were also obtained by using the HPLC-UHC-Chip/Q-TOF MS. When compared with conventional LC/MS, 4-30 times increases in sensitivity were obtained for tested compounds. It was clearly demonstrated that the HPLC-UHC-Chip/Q-TOF MS system could be a valuable tool for the quantitative analysis of small molecule, satisfactory results of serum samples also proved its practicality and potential for use in drug testing and development. Moreover, a metabolic profiling method for soybean flavonoids based on HPLC-UHC- Chip/Q-TOF MS system and simple sample preparation was developed and validated. Seventy-three flavonoids were detected in a single run, including thirteen newly found flavonoids in soybean seed. To show the usefulness, the method was then applied to the profiling of different varieties of soybean sowed at the same place. A clear discrimination was observed among different cultivars, three isoflavones, accounting for nearly 80% of total flavonoid contents were found increased in the spring soybeans compared with the summer cultivars.
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/116061
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Zhao CX,Chang YW,Wu ZM,et al. Microfluidic chip-based high performance liquid chromatography coupled to mass spectrometry method for small molecule analysis[C]. 见:37th International Symposium on High Performance Liquid Phase Separations and Related Techniques. 大连. 2011-10-8.
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