固相萃取―高效液相色谱法测定烤烟中角鲨烯含量.docVIP

固相萃取―高效液相色谱法测定烤烟中角鲨烯含量 　　摘要：通过优化提取剂、提取方法、净化剂等前处理方法和色谱条件，建立了烤烟中角鲨烯含量测定的SPE-HPLC方法。烤烟样品用三氯甲烷超声提取40 min，硅胶柱净化，高效液相色谱仪[检测波长：210 nm；流动相：甲醇/乙腈=75/25（V/V）]检测，可以准确地测定烤烟中角鲨烯含量；角鲨烯浓度在0.5～10 mg/L 范围内线性关系良好（r=0.9976）。方法的检出限（S/N=3）为 0.12 mg/L，定量限（S/N=10）为 0.39 mg/L，平均加标回收率为 88.12%。利用该方法对5个不同地区的同品种烤烟中角鲨烯含量进行测定的结果表明，不同地区的烤烟中角鲨烯含量有所差异。该方法准确度高、重现性好，可用于烤烟中角鲨烯的含量测定。 　　关键词：烤烟；角鲨烯；固相萃取；高效液相色谱仪 　　中图分类号：S572文献标识号：A文章编号：1001-4942（2017）12-0130-05 　　Abstract A SPE-HPLC method to determinate squalene in flue-cured tobacco was established by optimizing the chromatographic conditions and pretreatment method （extractant， extraction method and purification agent）. After validation， the method was used to measure the squalene in flue-cured tobacco from five different areas. Flue-cured tobacco samples were extracted by trichloromethane for 40 minutes and purified with the silica gel column. And the squalene content in samples was detected by the high performance liquid chromatograph （detection wavelength： 210 nm； mobile phase： methanol/acetonitrile = 75/25（V/V））. The squalene concentration showed a good linear relationship in the range of 0.5～10 mg/L （r=0.9976）. The limit of detection （LOD） and the limit of quantification （LOQ） of established method were 0.12 mg/L and 0.39 mg/L， respectively. The average recovery was 88.12% at three spiked levels. The results showed that the squalene content in flue-cured tobacco from different areas was different. The established SPE-HPLC method had high accuracy and good reproducibility， which could be used for the determination of squalene in flue-cured tobacco. 　　Keywords Flue-cured tobacco； Squalene；Solid phase extraction； High performance liquid chromatograph 　　角?烯是一种高度不饱和的直链三萜类化合物，具有提高血红蛋白的携氧能力，促进新陈代谢，提高体内超氧化物歧化酶（SOD）活性、增强机体免疫能力、抗肿瘤等多种生理功能，是一种具有防病治病作用的生物活性物质[1-3]。角鲨烯不仅存在动物体内，在植物中的分布也十分广泛，橄榄油和棕榈油中含有较丰富的角鲨烯，但其它植物中也有角鲨烯的存在，如鼠尾草、蕨类植物、绿色藻类植物、苔藓植物及烟叶等[4-8]。 　　卷烟烟气中含有多种有害化合物，其中自由基具有特殊的化学结构，性质较活泼，可以参与人体的多种生化反应，诱发多种疾病，对人体危害较大[9-14]。目前，清除卷烟烟气自由基的主要方法是向烟丝和滤嘴中加入