Scaling up: Improving automated, high throughput measurement of tobacco-specific nitrosamines in urine by LC-MS/MS

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In 2005, a relatively simple and sensitive method for measurement of tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) in smokers and nonsmokers was developed with sample cleanup based on a molecularly imprinted polymer (MIP) column and atmospheric pressure ionization tandem mass spectrometry (LC-MS/MS). Since then incremental method improvements, necessary for its application to large epidemiological studies, led to the development of a lower sample volume, more sensitive LC-MS/MS method with robotic sample preparation and automatic data processing for the simultaneous determination of four tobacco-specific N-nitrosamines (TSNA), including both free and conjugated forms in urine. Improvements to these methods were applied to development of a new rapid method for measuring NNAL in smokers' urine with further decreased sample volume and sample cleanup time. LC-MS/MS analysis for NNAL, N'-nitrosonornicotine (NNN), N'-nitrosoanatabine (NAT) and N'-nitrosoanabasine (NAB) was performed using Shimadzu LC-30AD HPLC modules with post-column addition coupled with Sciex API 6500 triple quadrupole systems. Automatic data processing by Indigo Ascent was implemented, combining peak integration and quality assurance and resulting in decreased data review time and improved precision and reproducibility.

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Proceedings 64th Annual ASMS Conference on Mass Spectrometry and Allied Topics, Vol.1: San Antonio(US).June 6-9, 2016

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