Cotinine is a stable metabolite of nicotine, and is the most widely used biomarker to measure tobacco use and exposure, i.e. both active and passive smoking. Serum cotinine has a half-life of 15 to 40 h and reflects tobacco exposure during the prior 3 to 5 days. The half-life of cotinine is longer than that of nicotine. Thus, the cotinine concentration is therefore rather stable throughout the day.
Method: LC-MS/MS
Assessment of smoking status and tobacco exposure.
Matrix: Serum, EDTA plasma, saliva and urine.
Volume: Minimum volume is 60 µL, but 200 µL is optimal and allows reanalysis.
Preparation and stability: Cotinine is stable.
Frozen, on dry ice. (for general instruction on transportation, click here)
The information below applies to measurement in serum or plasma.
Non-smokers < ~5 nmol/L; passive smokers 5 – 85 nmol/L; smokers > 85nmol/L; heavy smokers (>25 cigarettes/day) > 1700 nmol/L.
To convert from SI unit (nmol/L) to conventional unit (µg/L), multiply the SI value by the conversion factor = 0.1762.
Nicotine patches and snuff give very high plasma cotinine levels.
Serum cotinine cut-off distinguishing smoker from non-smoker varies between studies from 57 to 114 nmol/l, and 85 nmol/L is the value suggested in most studies. But as smoking prevalence declines, so does exposure to environmental tobacco smoke, and lower cut-off values seem to be obtained (2).
Intraclass correlation coefficient (ICC): 0.92 (3).
1. Midttun, O., Hustad, S., and Ueland, P.M. (2009). Quantitative profiling of biomarkers related to B-vitamin status, tryptophan metabolism and inflammation in human plasma by liquid chromatography/tandem mass spectrometry. Rapid Commun Mass Sp 23, 1371-79.
2. Kim, S. (2016). Overview of cotinine cutoff values for smoking status classification. Int J Environ Res Public Health 12.
3. Midttun, O., Townsend, M.K., Nygård, O., Tworoger, S.S., Brennan, P., Johansson, M., and Ueland, P.M. (2014). Most blood biomarkers related to vitamin status, one-carbon metabolism, and the kynurenine pathway show adequate preanalytical stability and within-person reproducibility to allow assessment of exposure or nutritional status in healthy women and cardiovascular patients. J Nutr 144, 784-790.
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Amino acids: Alanine, Arginine, Asparagine, Aspartic acid, Glutamic acid, Glutamine, Glycine, Histidine, Isoleucine, Kynurenine, Leucine, Lysine, Methionine, Ornithine, Phenylalanine, Proline, Sarcosine, Serine, Threonine, Total cysteine, Tryptophan, Tyrosine, Valine
Amino acid catabolites: 2-Aminoadipic acid, 2-Hydroxybutyrate, 3-Hydroxysiobutyrate, α-Hydroxyglutaric acid, β-Alanine, β-Aminoisobutyrate, β-Hydroxy B-methylbutyric acid, Phenylacetylglutamine
Acylcarnities: BB, C0, C2, C3, C3-DC, C4, C4-OH, C4-DC, iC5, C5-DC, C5:1, C6, C8, C10, C12, C14, C14-OH, C16, C16-OH, C18, C18-OH, C18:1, C18:2
TCA metabolites: α-Ketoglutarate, Citrate, Fumarate, Isocitrate, Lactate, Malate, Pyruvate
Ketone bodies: Acetoacetate, 3-Hydroxybutyrate
AGEs: Carboxyethyllysine, Carboxymethyllysine
Kynurenines: 3-hydroxykynurenine, 3-hydroxyanthranilic acid, Anthranilic acid, Kynurenine, Kynurenic acid, Nicotinic acid, Nicotinamide, N1-methylnicotinamide, Picolinic acid, Quinaldic acid, Quinolinic acid, Xanthurenic acid
Neopterin
Proteins: C-Reactive protein, Calprotectin, Serum Amyloid A
Ratio-derived: Kynurenine/tryptophan ratio, PAr index (PLP, PL, PA)
SCFAs: Acetate, Butyrate, Formate, Isobutyrate, Isovalerate, Propionate, Valerate, α-Methylbutyrate
Indoles: 3-Indoxyl sulfate, Imidazole propionate, Indole-3-acetamide, Indole-3-acetate, Indole-3-aldehyde, Indole-3-lactate, Indole-3-propionate
Choline metabolites: Choline, Betaine, DMG, TMAO
Amino acid derived: Phenylacetylglutamine
B-vitamins, functional markers, and methyl donors: 4-Pyridoxic acid, Betaine, Choline, Cobalamin, Flavin mononucleotide, Folate, Methylmalonic acid, N1-methylnicotinamide, Nicotinamide, Nicotinic acid, Pyridoxal, Pyridoxal 5-phosphate, Pyridoxine, Riboflavin, Thiamine, Thiamine monophosphate, Total homocysteine
Fat-soluble vitamins: 25-hydroxy vitamin D2, 25-hydroxy vitamin D3, α-Tocopherol (Vit. E), All-trans retinol (Vit. A), Phylloquinone (Vit. K1), y-Tocopherol (Vit. E)
Essential amino acids: Histidine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine, Threonine, Tryptophan, Valine
Meat and fish intake: 1-Methylhistidine, 3-Methylhistidine, B-Alanine, Creatine, Creatinine, TMAO
Tobacco use and coffee intake: Cotinine, Trans-3-hydroxycotinine, Trigonelline
Turnaround time
2-6 weeks, depending on number of samples and laboratory capacity.
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