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1 2023

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Simple HPLC method for simultaneous quantification of nicotine and cotinine levels in rat plasma after exposure to two different tobacco products

Alhusban Ala A., Hammad Alaa M., Alzaghari Lujain F.

Acta Chromatographica

2023

Vol. 35, no. 1

106--114

Purpose: Development and validation of a selective analytical method to accurately and precisely quantify nicotine and cotinine levels in rat's plasma after exposure to tobacco cigarettes and tobacco water-pipe. Methods: An easy HPLC-Photodiode-Array Detection (PDA) method was developed and validated for simultaneous determination of nicotine and cotinine levels in plasma of 15 rats (10 rats after tobacco products exposure and 5 control rats). Nicotine and cotinine were extracted in one step from plasma using acetonitrile and concentrated to lowest volume using nitrogen stream. Results: The developed method offered a rapid analysis time of 14 min with single step of analytes extraction from rat's plasma with recovery percentage range between 93 and 95% and excellent linearity with correlation factor more than 0.994 with analytical range between 50 and 1000 ng mL⁻¹ and LOD of 25 ng mL⁻¹ and 23 ng mL⁻¹ for nicotine and cotinine, respectively. The analysis of rat's plasma after 28 days of exposure to tobacco cigarettes and tobacco water-pipe revealed that the average concentrations of 376 ng mL⁻¹ for cotinine and 223 ng mL⁻¹ for nicotine were obtained after tobacco cigarettes exposure, and 220 ng mL⁻¹ for cotinine and 192 ng mL⁻¹ for nicotine after tobacco water-pipe exposure. Conclusion: Higher nicotine and cotinine levels were found in plasma after tobacco cigarettes exposure than water-pipe exposure which may have potential undesirable effects on passive smokers in both cases.