高效液相色谱同时检测不同类型烟叶中的多种生物碱含量  

Simultaneous Determination of Alkaloids in Different Types of Tobacco by HPLC

作　　者：林雨晟 金洪石 张皓楠 金江华[2] 郭伟[2] 李玉娥[2] 赵铭钦 刘鹏飞 LIN Yucheng;JIN Hongshi;ZHANG Haonan;JIN Jianghua;GUO Wei;LI Yu’e;ZHAO Mingqin;LIU Pengfei(College of Tobacco Science,Henan Agricultural University,National Tobacco Cultivation&Physiology&Biochemistry Research Center,Zhengzhou 450002,China;Jilin Tobacco Industrial Co.,Ltd.,Yanji,Jilin 133001,China)

机构地区：[1]河南农业大学烟草学院,国家烟草栽培生理生化研究基地,郑州450002 [2]吉林烟草工业有限责任公司,吉林延吉133001

出　　处：《中国烟草科学》2021年第3期83-89,共7页Chinese Tobacco Science

基　　金：吉林烟草工业有限责任公司科技项目(JSZX-2019-03)。

摘　　要：为快捷准确地测定烟叶中生物碱的含量,本研究通过优化提取条件,建立了测定烟草中烟碱、新烟草碱、假木贼碱和降烟碱的高效液相色谱检测方法。优化后的检测步骤为:干烟叶和5%甲醇的料液比为100 mg/20 mL,在25℃下振荡提取30 min,色谱条件:C18柱(250 mm×4.6 mm,5μm),流动相A(90%水+10%甲醇,pH 6.8,0.15%醋酸钠),流动相B(90%甲醇+10%水,pH 3.8)。烟碱在0.1~50μg/mL、其余3种在0.005~2μg/mL的范围内,4种生物碱均具有较好的线性关系(R^(2)>0.99);烟碱检出限为0.00065μg/mL,平均回收率100.79%,RSD 3.11%;新烟草碱检出限0.0884μg/mL,平均回收率98.53%,RSD 3.75%;假木贼碱检出限0.0348μg/mL,平均回收率98.70%,RSD 3.63%;降烟碱检出限0.1509μg/mL,平均回收率101.33%,RSD 4.61%。对4种不同类型烟叶共11个不同样品进行检测验证,检测结果可靠。该方法前处理简单快捷,灵敏度高,重复性较好,可以满足烟草样品中4种生物碱的检测要求。In order to quickly and accurately determine the content of alkaloids in tobacco leaves,the extraction conditions were optimized,and a method for the determination of nicotine,anatabine,anabasine and nornicotine contents in tobacco by HPLC was established.The optimized detection steps are as follows:100 mg/20 mL solid-liquid ratio,5% methanol was extracted by oscillation at 25℃for 30 min,the chromatographic conditions were C18 column(250 mm×4.6 mm,5μm),mobile phase A(90% water+10% methanol,pH 6.8,0.15% sodium acetate),mobile phase B(90% methanol+10% water,pH 3.8).Nicotine was in the range of 0.1-50μg/mL and the other three alkaloids were in the range of 0.005-2μg/mL,with the standard curves of the four alkaloids all havig good linear relationship(R^(2)>0.99).The limits of detection(LOD)of nicotine were 0.00065μg/mL,and the average recoveries were 100.79% ,RSD 3.11%;the LOD of anatabine were 0.0884μg/mL,and the average recoveries were 98.53% ,RSD 3.75%;the LOD of anabasine were 0.0348μg/mL,and the average recoveries were 98.70% ,RSD3.63%;the LOD of nornicotine were 0.1509μg/mL,and the average recoveries were 101.33% ,RSD 4.61% .A total of 11 different samples of 4 different types of tobacco leaves were tested,and the results were reliable.The method is simple and fast,sensitive and repeatable,and can meet the requirements of detecting four alkaloids in tobacco samples.

关 键 词：烟草 烟碱 新烟草碱 假木贼碱 降烟碱 液相色谱法 

分 类 号：O657.72[理学—分析化学] TS411[理学—化学]