固相萃取-超高效液相色谱-多级质谱联用法测定烟叶中5种类胡萝卜素的含量
Simultaneous determination of five carotenoids in tobacco leaves by solid-phase extraction combined with ultra-high performance liquid chromatography- multistage mass spectrometry
-
摘要: 【目的】 建立同时测定烟叶中5种类胡萝卜素的定性和定量分析方法。【方法】 利用超声波提取结合固相萃取对烟叶进行提取净化,采用超高效液相色谱-多级质谱联用(UHPLC-MSn)法对类胡萝卜素进行定量分析。【结果】 适宜的提取条件为:以丙酮为超声提取溶剂,料液比1∶40、超声功率120 W和超声时间50 min;适宜的净化条件为:HF Bond Elut C18柱、淋洗液为乙腈、洗脱液为二氯甲烷;在多级质谱选择反应监测(SRM)模式下,5种类胡萝卜素的标准曲线线性关系良好,相关系数r均大于0.999 5;方法的检出限(LOD)为66.00~224.00 μg/kg,定量限(LOQ)为18.20~67.20 μg/kg,回收率为88.3%~93.3%,相对标准偏差均小于5%。【结论】 该方法简便、灵敏度高、精密度好,适合烟叶中主要类胡萝卜素的定性和定量分析,可为烟草香气前体物的代谢机制研究及烟叶风味品质定向提升提供参考。
-
关键词:
- 烟叶 /
- 固相萃取 /
- 超高效液相色谱-多级质谱联用 /
- 类胡萝卜素
Abstract: 【Objective】 To develop a qualitative and quantitative analytical method for simultaneously determining five carotenoids in tobacco leaves. 【Methods】 Tobacco leaves were extracted and purified by ultrasonic extraction combined with solid-phase extraction, and the carotenoids were quantitatively analyzed by ultra-high performance liquid chromatography-multistage mass spectrometry (UHPLC-MSn). 【Results】 The optimal extraction conditions were as follows: acetone was used as the ultrasonic extraction solvent, with a solid-to-liquid ratio of 1∶40, ultrasonic power of 120 W, and ultrasonic time of 50 min; while the optimal purification conditions were: HF Bond Elut C18 column, acetonitrile as the leaching solution, and dichloromethane as the eluent. Under the selected reaction monitoring (SRM) mode, the standard curves of the five carotenoids showed a good linear relationship with correlation coefficients (r) all greater than 0.999 5. The limits of detection (LOD) of the method were 66.00~224.00 μg/kg, the limits of quantification (LOQ) were 18.20~67.20 μg/kg, the average recoveries were 88.3%~93.3%, and the relative standard deviations (RSDs) were all less than 5%. 【Conclusion】 This method is simple, highly sensitive and precise, which is suitable for the qualitative and quantitative analysis of the main carotenoids in tobacco leaves. It provides a reference for the research on the metabolic mechanisms of tobacco aroma precursors and the targeted improvement of tobacco leaf flavor quality.-
Key words:
- tobacco leaves /
- solid-phase extraction /
- UHPLC-MSn /
- carotenoids
-
-
[1]
刘涛,曹建敏,郭存,等.不同烤烟品种化学成分及致香物质差异分析[J].中国烟草科学,2023,44(2):74-82.
LIU T,CAO J M,GUO C,et al.Difference analysis on chemical composition and aroma components of different flue-cured tobacco cultivars[J].Chinese Tobacco Science,2023,44(2):74-82. -
[2]
LI C,WANG C L,CHENG Z Y,et al.Carotenoid biosynthesis genes LcLCYB,LcLCYE,and LcBCH from wolfberry confer increased carotenoid content and improved salt tolerance in tobacco[J].Scientific Reports,2024,14:10586.
-
[3]
龙章德,王敏,薛云,等.烟叶中β-胡萝卜素高效降解菌株的筛选鉴定及发酵条件优化[J].轻工学报,2022,37(4):86-93.
LONG Z D,WANG M,XUE Y,et al.Screening and identifying of β-carotene degrading strains from tobacco leaves and its optimization of fermentation conditions[J].Journal of Light Industry,2022,37(4):86-93. -
[4]
王玉华,王德权,高政绪,等.烤烟自然醇化过程中类胡萝卜素降解产物含量变化及其与感官质量的关系[J].浙江农业科学,2020,61(2):340-344
,394. WANG Y H,WANG D Q,GAO Z X,et al.Changes of carotenoid degradation products during natural aging of flue-cured tobacco and its relationship with sensory quality[J].Journal of Zhejiang Agricultural Sciences,2020,61(2):340-344,394. -
[5]
吴巧茵,施友志,李林林,等.类胡萝卜素降解菌株的原位筛选及其在雪茄提质增香中的应用[J].生物技术通报,2023,39(9):192-201.
WU Q Y,SHI Y Z,LI L L,et al.In situ screening of carotenoid degrading strains and the application in improving quality and aroma of cigar[J].Biotechnology Bulletin,2023,39(9):192-201. -
[6]
FU W T,ZHENG X Y,CHEN X C,et al.The potential roles of carotenoids in enhancing phytoremediation of bisphenol a contaminated soil by promoting plant physiology and modulating rhizobacterial community of tobacco[J].Chemosphere,2023,316:137807.
-
[7]
王静,王杰,阿娜尔,等.枸杞类胡萝卜素指纹图谱的建立及其抗氧化活性谱效关系研究[J].中国药房,2022,33(5):575-578.
WANG J,WANG J,A N E,et al.Establishment of carotenoid fingerprint in Lycium barbarum and study on its antioxidant activity spectrum-effect relationship[J].China Pharmacy,2022,33(5):575-578. -
[8]
SANDMANN G.HPLC analysis of carotenoids from bacteria[J].Methods in Enzymology,2022,670:139-153.
-
[9]
PAVELKOVÁ P,KRMELA A,SCHULZOVÁ V.Determination of carotenoids in flowers and food supplements by HPLC-DAD[J].Acta Chimica Slovaca,2020,13(1):6-12.
-
[10]
王楠,周宝成,葛志伟,等.高效液相色谱法测定番茄果肉中4种天然类胡萝卜素含量[J].分析科学学报,2024,40(5):592-596.
WANG N,ZHOU B C,GE Z W,et al.Determination of four natural carotenoids in tomato flesh by high performance liquid chromatography[J].Journal of Analytical Science,2024,40(5):592-596. -
[11]
DAOOD H G,RÁTH S,PALOTÁS G,et al.Efficient HPLC separation on a core-C30 column with MS2 characterization of isomers,derivatives and unusual carotenoids from tomato products[J].Journal of Chromatographic Science,2022,60(4):336-347.
-
[12]
GRUJIĆ V J,TODOROVIĆ B,KRANVOGL R,et al.Diversity and content of carotenoids and other pigments in the transition from the green to the red stage of Haematococcus pluvialis microalgae identified by HPLC-DAD and LC-QTOF-MS[J].Plants,2022,11(8):1026.
-
[13]
LONDOÑO-GIRALDO L M,BUENO M,CORPAS-IGUARÁN E,et al.HPLC-DAD-APCI-MS as a tool for carotenoid assessment of wild and cultivated cherry tomatoes[J].Horticulturae,2021,7(9):272.
-
[14]
赖长鸿,王寒雪,李培杰,等.琯溪蜜柚果肉中类胡萝卜素组分的特征解析[J].中山大学学报(自然科学版)(中英文),2024,63(4):71-77.
LAI C H,WANG H X,LI P J,et al.Characteristics of carotenoid components in juice sacs of pomelo(Citrus grandis cv ‘Guanximiyou’)[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2024,63(4):71-77. -
[15]
FENG X Y,YU Q Q,LI B,et al.Comparative analysis of carotenoids and metabolite characteristics in discolored red pepper and normal red pepper based on non-targeted metabolomics[J].LWT-Food Science and Technology,2022,153:112398.
-
[16]
欧阳璐斯,陈森林,王铭,等.低氧贮存片烟解封过程中化学成分变化研究[J].云南大学学报(自然科学版),2025,47(1):154-162.
OUYANG L S,CHEN S L,WANG M,et al.Study on the change of chemical composition during unsealling of the tobacco in hypoxic storage[J].Journal of Yunnan University (Natural Sciences Edition),2025,47(1):154-162. -
[17]
FAN X,ZI W H,AO J C,et al.Analysis and application evaluation of the flavour-precursor and volatile-aroma-component differences between waste tobacco stems[J].Heliyon,2022,8(9):e10658.
-
[18]
余晶晶,史莉莉,刘雨欣,等.COFs材料磁固相萃取-HPLC-MS/MS法测定烘焙食品中两种主要晚期糖基化终末产物[J].轻工学报,2023,38(6):52-61.
YU J J,SHI L L,LIU Y X,et al.Determination of two major advanced glycation end products in baked food using HPLC-MS/MS based on COFs material magnetic solid phase extraction[J].Journal of Light Industry,2023,38(6):52-61. -
[19]
HAO J W,LIU X Q,CHEN N D,et al.Solid-phase extraction followed by direct TOF-MS-MS and HPLC analysis of ptaquiloside in Pteridium aquilinum from different places of China[J].Journal of Food Composition and Analysis,2021,98:103845.
-
[1]
-
点击查看大图
计量
- PDF下载量: 0
- 文章访问数: 52
- 引证文献数: 0
下载:
