Analytical and Bioanalytical Chemistry

, Volume 400, Issue 7, pp 2109–2117 | Cite as

Application of molecularly imprinted polymer solid-phase extraction for salivary cotinine

  • Ricardo Vilela VitorEmail author
  • Matheus Coutinho Gonçalves Martins
  • Eduardo Costa Figueiredo
  • Isarita Martins
Original Paper


A method constituted by molecularly imprinted solid-phase extraction (MISPE) with high-performance liquid chromatography coupled to diode array detector (HPLC-DAD) was developed for cotinine analysis in saliva samples. For this purpose, the separation was carried out with a C18 reversed-phase column at 20 °C. The mobile phase which was composed of a mixture of 09:91 (v/v) acetonitrile/phosphate buffer, pH 6.3, was delivered with isocratic flow rate at 1.4 mL min−1. Employing MISPE, the best conditions were achieved with 1.5 mL of saliva plus 1.5 mL of 0.1 mol L−1 of acetate buffer, pH 5.5, which were then passed through a cartridge previously conditioned with 2 mL acetonitrile, 2 mL methanol, and 2 mL of 0.1 mol L−1 sodium acetate buffer, pH 5.5. The washing was carried out with 1 mL deionized water, 1 mL of 0.1 mol L−1 sodium hydroxide, and 1 mL hexane; finally; the cotinine elution was carried out with 3 mL methanol/water (97.5: 2.5, v/v). Linearity ranged from 30 to 500 ng mL−1 with r > 0.99. Intra-assay, interassay precision, and accuracy ranged from 3.1% to 10.1%, 5.2% to 15.9%, and 99.22% to 111.17%, respectively. The detection and quantification limits were 10 and 30 ng mL−1, respectively. This investigation has provided a reliable method for routine cotinine determination in saliva, and it is an important tool for monitoring cigarette smoke exposure in smokers. The method was applied in five smokers’ samples who consumed around five to 20 cigarettes per day and the values of cotinine in saliva were from 66.7 to 316.16 ng mL−1.


Molecularly imprinted polymer Solid-phase extraction Cotinine Saliva Liquid chromatography 



This research was supported by the National Council for Scientific and Technological Development (CNPq)/Brazil and by “Fundação de Amparo à Pesquisa do Estado de Minas Gerais” (FAPEMIG)/Brazil (processes number CDS-APQ-4487-4.04/07 and CDS-PPM-00055-09).


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Ricardo Vilela Vitor
    • 1
    Email author
  • Matheus Coutinho Gonçalves Martins
    • 1
  • Eduardo Costa Figueiredo
    • 1
  • Isarita Martins
    • 1
  1. 1.Laboratory of Analysis of Toxicants and DrugsFaculty of Pharmaceutical Sciences, Federal University of AlfenasAlfenasBrazil

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