Abstract
A new, simple, sensitive, selective, precise, and robust high-performance thin-layer chromatography (HPTLC) method was developed and validated for the determination of nicotine in tobacco extracts. The analysis of nicotine was performed on thin-layer chromatography (TLC) glass plates precoated with silica gel 60 F254 as stationary phase. Development was carried out in a horizontal chamber under sandwich configuration with the mobile phase ethyl acetate-methanol-ammonia 2.8% (v/v) (20:10:4, v/v/v) at room temperature (25 ± 2°C). A TLC scanner was used for spectrodensitometric scanning and analysis in the absorbance mode at 263 nm. The system was found to give compact spots for nicotine (RF value of 0.20 ± 0.02). The calibration plots showed good linear relationships with r2 = 0.9998 ± 0.0001 in the concentration range 90-900 ng/spot with respect to the peak area. According to the International Conference of Harmonization (ICH) guidelines, the TLC method was validated for precision, recovery, robustness, an...
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