Determination of Nicotine in Tobacco by High Performance Liquid Chromatography

A method for the determination of nicotine in tobacco by high performance liquid chromatography (HPLC) was developed. Tobacco sample powder of 0.5 g was dissolved in 75% methanol solution containing 0.1 mol·L^-1 sodium hydroxide and made its volume up to 100 mL. After ultrasonic extraction for 30 min, HPLC was used for determination. As shown by the results, linear relationship between mass concentration of nicotine and its peak area was kept in the range of 3.00-100 mg·L^-1, with detection limit (3S/N) of 0.5 mg·L^-1. RSDs of intraday (n=6) and interday (n=9) were 0.47%-1.1% and 0.44%-0.97%, respectively. Recovery test was made on tobacco sample at 3 concentration levels by standard addition method, giving results in the range of 97.4%-100%. This method was used to analyze the actual samples and compared with gas chromatography (YC/T 246-2008) and spectrophotometry (GB/T 23225-2008), with the determined values by this method basically consistent with those of spectrophotometry, and RSDs of the determined values by the proposed method were less.