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Five hundred microliter of the methylene chloride layer containing the nicotine was then vortexed with 500 l of 50 mM HCl solution and the nicotine of the HCl solution was estimated by HPLC analysis at 254 nm

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They completed the Contemplation Ladder [40] to evaluate their readiness to quit smoking and the Cigarette Withdrawal Scale (CWS-21) [41] to assess symptoms of withdrawal on six dimensions: craving, insomnia, depressionanxiety, appetiteweight gain, irritabilityimpatience, and difficulty concentrating

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