超高效液相色谱法同时测定甘薯中13种类胡萝卜素的含量

Simultaneous determination of 13 carotenoids in sweetpotato by Ultra-Performance Liquid Chromatography

类胡萝卜素作为评价甘薯食用和加工品质的重要指标,对于推进甘薯育种至关重要,但由于缺乏甘薯类胡萝卜素组分及含量的快速检测方法,限制了甘薯食用品质分析和育种进展。本研究通过优化色谱条件和样品前处理过程,建立了同时测定甘薯中13种类胡萝卜素含量的超高效液相色谱分析方法。结果表明,待测样品前处理过程中乙醇∶丙酮∶正己烷(1∶1∶1,v/v/v)作为提取液,甲基叔丁基醚为复溶液类胡萝卜素得率最高。超高效液相色谱分析条件:甲醇∶乙腈(1∶3,v/v)含0.01%2,6-二叔丁基对甲酚(Butylated hydroxytoluene,BHT)和0.1%甲酸作为流动相A相,甲基叔丁基醚含0.01%BHT作为流动相B相梯度洗脱,色谱柱YMC Carotenoid C30(2 mm×100 mm,3μm),流速为0.8 mL min^(–1),柱温23℃,进样量1μL,检测波长450 nm,分析时间8 min。该方法中13种类胡萝卜素线性关系良好,相关系数大于0.997;精密度、重复性、稳定性及回收率结果均符合要求。本研究建立的超高效液相色谱方法能快速、准确的同时检测甘薯中13种类胡萝卜素的含量,可为大批量甘薯种质资源和育种材料的品质分析奠定基础。

Carotenoids as important indexes to evaluate the edible and process quality of sweetpotato,are crucial to promoting sweetpotato breeding.However,the lack of rapid detection methods for the compositions and contents of carotenoids in sweetpotato has limited the progress of edible quality analysis and breeding of sweetpotato.In this study,an Ultra-Performance Liquid Chromatography(UPLC)method was established for simultaneous determination of 13 carotenoids in sweetpotato by optimizing the chromatographic conditions and sample pretreatment process.The results showed that ethanol:acetone:hexane(1:1:1,v/v/v)was used as the extraction solution,and methyl tert-butyl ether was used as the dissolution solution to obtain the highest yield of carotenoids.Analysis conditions of UPLC:methanol:acetonitrile(1:3,v/v)containing 0.01%2,6-butylated hydroxytoluene(BHT)and 0.1%formic acid as mobile phase A,methyl tert-butyl ether containing 0.01%BHT as mobile phase B gradient elution,YMC Carotenoid C30(2 mm×100 mm,3μm),the flow rate was 0.8 mL min-1,injection volume was 1μL,the detection wavelength was 450 nm,and the analysis time was 8 min.The linear relationships of 13 carotenoids in this method were good,the correlation coefficients were all greater than 0.997.The test results of precision,repeatability,stability and recovery were all in line with the requirements.The UPLC method in this study can quickly,accurately and simultaneously detect the contents of 13 carotenoids in sweetpotato,which can lay a foundation for quality analysis of germplasm resources and breeding materials in large quantities of sweetpotato.