Optimization and validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous quantification of nicotine, cotinine, trans-3′-hydroxycotinine and norcotinine in human oral fluid

Shakleya, Diaa M.; Huestis, Marilyn A.
December 2009
Analytical & Bioanalytical Chemistry;Dec2009, Vol. 395 Issue 7, p2349
Academic Journal
An analytical procedure was developed and validated for the simultaneous identification and quantification of nicotine, cotinine, trans-3′-hydroxycotinine, and norcotinine in 0.5 mL of human oral fluid collected with the Quantisal™ oral fluid collection device. Solid phase extraction and liquid chromatography-tandem mass spectrometry with multiple reaction monitoring were utilized. Endogenous and exogenous interferences were extensively evaluated. Limits of quantification were empirically identified by decreasing analyte concentrations. Linearity was from 1 to 2,000 ng/mL for nicotine and norcotinine, 0.5 to 2,000 ng/mL for trans-3′-hydroxycotinine, and 0.2 to 2,000 ng/mL for cotinine. Correlation coefficients for calibration curves were >0.99 and analytes quantified within ±13% of target at all calibrator concentrations. Suitable analytical recovery (>91%) was achieved with extraction efficiencies >56% and matrix effects <29%. This assay will be applied to the quantification of nicotine and metabolites in oral fluid in a clinical study determining the most appropriate nicotine biomarker concentrations differentiating active, passive, and environmental nicotine exposure.


Related Articles

  • Determinants of salivary cotinine levels among current smokers in Mexico. Campuzano, Julio; Hernandez-Avila, Mauricio; Jaakkola, Maritta; Ponce, Eduardo Lazcano; Morales, Pablo Kuri; Bautista, Pablo; Benowitz, Neal; Ceraso, Marion; Blackford, Amanda; Samet, Jonathan // Nicotine & Tobacco Research;Dec2004, Vol. 6 Issue 6, p997 

    The present study describes salivary cotinine levels and their relationship to cigarettes smoked per day in Mexican smokers. Using a sampling strategy based on the number of cigarettes per day, we recruited 1,222 smokers from Mexico City and the state of Morelos in Mexico during 1999. Smoking...

  • Simultaneous Determination of Nicotine and Metabolites by IC-MS Using an Acclaim Mixed-Mode HILIC-1 Column. Wang, Leo; Xiaodong Liu; Chatfield, Dale; Schnute, William // LC-GC North America;Feb2008 Applications Notebook, Vol. 26, p56 

    The article offers information about a study which demonstrates a simultaneous determination of nicotine and its metabolites through liquid chromatography-mass spectrometry. The method uses an Acclaim® Mixed-Mode hydrophilic interaction chromatography-1 (HILIC-1) column from Dionex Corp. and...

  • Chromatographic Comparison of Caries-immune and Caries-susceptible Salivas. GREEN, GORDON E.; WILSON, ROBERT M. // Journal of Dental Research;Nov1963, Vol. 42 Issue 6, p1380 

    The article compares caries-immune and caries-susceptible human salivas by analyzing chromatographic fractionations. There were no differences in the patterns of protein, amylase, anthrone-reacting carbohydrates, sialic acids, and electrophoretic mobilities. The conclusion is that the...

  • Chromatographic Separation of Ether-Extractable Components of Aerobically Incubated Human Whole Saliva. MISHIRO, YUKIHIKO; KIRIMURA, KAZUKO; KOYAMA, TOSHIE // Journal of Dental Research;Jul1969, Vol. 48 Issue 4, p601 

    The article presents a study which examined the chromatographic separation of ether-extractable components of whole human saliva following aerobic incubation. The author suggests that ultraviolet spectrophotometry may be used to estimate the content of phenol and indole in ether extracted from...

  • Chairside quantitative immunochromatographic evaluation of salivary cotinine and its correlation with chronic periodontitis. Surya, Chamarthi; Narasimha Swamy, Devulapally; Chakrapani, Swarna; Sunil Kumar, Surapaneni // Journal of Indian Society of Periodontology;Oct-Dec2012, Vol. 16 Issue 4, p508 

    Background: Cigarette smoking is an established and modifiable risk factor for periodontitis. Periodontitis appears to be dose-dependent on smoking. The purpose of this study was to assess a reliable marker of tobacco smoke exposure (salivary cotinine) chairside and to confirm the quantitative...

  • The Isolation of a Family of Cysteine-containing Phosphoproteins from Human Submandibular-Sublingual Saliva. SHOMERS, J. P.; TABAK, L. A.; LEVINE, M. J.; MANDEL, I. D.; ELLISON, S. A. // Journal of Dental Research;Aug1982, Vol. 61 Issue 8, p973 

    A family of acidic proteins has been isolated from human submandibular-sublingual saliva using Sephadex G-200 gel filtration, DE-52 cellulose chromatography, and immuno-affinity chromatography. These molecules were homogenous in size and by immunological criteria, but exhibited charge...

  • Argentine Division.  // Journal of Dental Research;Apr1985, Vol. 64 Issue 4, p638 

    The article presents abstracts on various dentistry topics which include salivary secretion results from several types of receptors, a comparison of anethesia of propolis and lidocaine, and chromatography.

  • Free Amino Acids in Human Saliva. PAPANAYOTOU, PAN H.; DOZI-VASSILIADES, JOANNA; KOVATSIS, ANAST. // Journal of Dental Research;May1973, Vol. 52 Issue 3, p418 

    By the use of thin layer chromatographic techniques and exchange column chromatography, 20 free amino acids were found in human saliva. No qualitative or semiquantitative differences in the amino acid content were observed among caries-susceptible and caries-resistant individuals 20 to 30 years old.

  • Excretion of mutagens, nicotine and its metabolites in urine of cigarette smokers. Granella, Manuela; Priante, Eugenia; Nardini, Barbara; Bono, Roberto; Clonfero, Erminio // Mutagenesis;Mar1996, Vol. 11 Issue 2, p207 

    Urine samples from 26 cigarette smokers on a restricted diet were collected in the late afternoon. Urine extracts on XAD-2 resin were tested for mutagenicity in the microsusp-ension assay using Salmonella typhimurium strain TA98 in the presence of metabolizing and deconjugating enzymes. Levels...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics