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In tobacco studies, biomarkers to assess exposure or of potential harm are both of interest. Biomarkers of exposure are tobacco smote constituents or their metabolites in a biological fluid or tissue, hair or exhaled breath. They are not measurements of how these constituents interact with the body process/functions.   Biomarkers of harm or potential harm are measurements of an effect from exposure. These biomarkers could be indicative of exposure to tobacco products or could be related to diet or environmental exposure. 

The FDA has established a list of harmful and potentially harmful constituents (HPHCs) in tobacco products and tobacco smoke. Additionally, a list of 20 HPHCs from the full list of HPHCs, for which testing methods are well established and widely available. These The HPHC list focuses on chemicals that are linked to the five most serious health effects of tobacco use (cancer, cardiovascular disease, respiratory effects, reproductive problems, and addiction.) 

The use of tobacco products results in the uptake of nicotine and a wide range of other chemicals. Biomarkers of exposure to tobacco and nicotine delivery products are limited to the chemicals taken up during product use or during exposure to product emissions. Total nicotine equivalents (TNE) is a biomarker of nicotine consumption, defined as the molar sum of the urinary concentrations of nicotine and all of its known metabolites. Typically this includes total NIC, total COT, total 3HC, and nicotine N-oxide, NNO.  Cotinine is a  a metabolite of nicotine and is  widely used biomarker of nicotine exposure. Nicotine replacement therapy and tobacco use can be distinguished by the detection of a tobacco-specific alkaloid, such as anabasine. Cotinine has 6 notable metabolites including: 3′-hydroxycotinine, cotinine glucuronide, 5′-hydroxycotinine, cotinine N-oxide, cotinine methonium ion, and norcotinine.

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