基于UPLC-MS/MS的豆瓣菜酚酸类组分分析

Analysis of phenolic acids in Nasturtium officinale R.Br.base on UPLC-MS/MS

为深入了解和挖掘豆瓣菜的功能性营养物质,本研究采用超高效液相色谱-串联质谱(UPLC-MS/MS)法对18个豆瓣菜品种的酚酸类成分进行定性和定量分析。结果表明,运用HSS T3色谱柱,流动相0.1%甲酸水溶液-乙腈梯度洗脱,流速0.3 mL/min,柱温30℃,选择电喷雾离子源在负离子模式下扫描,共鉴定出豆瓣菜的17个酚酸类物质,各化合物分离效果好,标准曲线在检测范围内具有良好的线性(R≥0.9992),检出限在0.8401~4.040μg/mL,定量限在2.001~20.50μg/mL,平均加样回收率在94.63%~102.4%,相对标准偏差≤4.4%(n=6)。该方法测定的18份豆瓣菜中17个酚酸均可定量,不同品种豆瓣菜的总酚酸含量以及同一品种的各酚酸含量均存在明显差异,总酚酸含量范围为106.0~230.0 mg/g DW,其中羟基苯甲酸、鞣花酸、绿原酸和3,5二羟基苯甲酸是主要的酚酸类物质。综上所述,基于UPLC-MS/MS方法可快速精准分析豆瓣菜中酚酸类化合物的组成和分布,为后续富含酚酸的豆瓣菜优异种质资源的挖掘建立了技术平台,有助于加快豆瓣菜品种改良。

To understand and explore the functional nutrients in Nasturtium officinale R.Br.,an ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method was established for the qualitative and quantitative analysis of 17 phenolic acids in 18 N.officinale cultivars.The optimal detection method obtained by optimizing the parameters of UPLC-MS/MS condition was as follows:the samples were separated using an HSS T3 column and eluted with 0.1%formic acid-acetonitrile as mobile phase,the flow rate was set to 0.3 mL/min and the column temperature was set to 30℃.A total of 17 phenolic acids in N.officinale were detected by negative ion mode scanning.The separation effect of compounds is good.The linear ranges of 17 phenolic acids showed good linearity(R≥0.9992),the limit of detection ranged from 0.8401μg/mL to 4.040μg/mL,the limit of quantitation ranged from 2.001μg/mL to 20.50μg/mL,and the average recovery ranged from 94.63%to 102.4%,relative standard deviation≤4.4%(n=6).All 17 phenolic acids in 18 N.officinale detected by our method were quantitatively measurable.The total phenolic acid content in different cultivars and the accumulation of 17 kinds of phenolic acids in the same N.officinale cultivar were significantly different.The total content of phenolic acids ranged from 106.0 mg/g DW to 230.0 mg/g DW.Among them,p-hydroxybenzoic acid,ellagic acid,chlorogenic acid and 3,5-dihydroxybenzoic acid were the main phenolic acids in N.officinale.In summary,the UPLC-MS/MS method can quickly and accurately analyze the composition and distribution of phenolic acid compounds in N.officinale,and establish a technical platform for the subsequent mining of excellent phenolic acid-rich germplasm resources of N.officinale,which is helpful to accelerate the improvement of N.officinale cultivars.