高效液相色谱-质谱检测赣南脐橙幼果中酚酸

Determination of phenolic acids in Gannan navel orange young fruit by high performance liquid chromatography-mass spectrometry

通过优化色谱分离条件和质谱条件,建立了基于高效液相色谱-串联质谱测定赣南脐橙幼果中酚酸的测定方法。以咖啡酸、对香豆酸、阿魏酸、芥子酸、没食子酸和对羟基苯甲酸6种酚酸化合物标准品,对赣南脐橙幼果6种酚酸进行定量分析。色谱柱为Agilent-C_(18) column(2.1mm×100 mm,1.8μm),流动相为0.1%甲酸水溶液(A)/乙腈(B)进行梯度洗脱,柱温35℃,流速0.3 m L/min;质谱以负离子模式进行多反应监测模式(MRM)扫描。结果表明:利用该方法可同时对赣南脐橙幼果中的咖啡酸、对香豆酸、阿魏酸、芥子酸、没食子酸和对羟基苯甲酸6种酚酸进行定量分析;采用外标法建立了6种酚酸的定量方法,6种酚酸的标准物质在各自质量浓度范围内具有良好线性关系,相关系数为0.991~0.999,检出限为(0.04~8.92)μg/g,平均回收率为85.5%~103.9%,相对标准偏差(RSD)小于2.8%。该方法简便、快速、准确,不仅适合脐橙中酚酸化合物的含量测定,而且对于其他柑橘属植物中酚酸化合物含量的测定也具有一定参考价值。

A high performance liquid chromatography-mass spectrometry method for the determination of phenolic acids in Gannan navel orange young fruit was established by optimizing chromatographic conditions and mass spectrometry conditions. Standard compounds of the target phenolic acids, including caffeic acid, p-coumaric acid, ferulic acid, sinapic acid, gallic acid and p-hydroxybenzoic acid, were carried on the quantitative analysis of Gannan navel orange fruit of six phenolic acids. The liquid-chromatographic separation was performed on Agilent-C18 column（2.1 mm×100 mm, 1.8 μm）, using 0.1% formic acid aqueous solution（A）/acetonitrile（B） as the mobile phase with aflow rate of 0.3 m L/min through gradient elution; column kept at 30 ℃; mass spectrometry was performed in multiple reaction monitoring mode（MRM） with negative ion mode scanning. Results showed that six phenolic acids such as caffeic acid, p-coumaric acid, ferulic acid, sinapic acid, gallic acid and p-hydroxybenzoic acid could be quantitative analysis by this method in Gannan navel orange young fruit. Quantification was performed by an external standard method. A good linear relationship was observed for six phenol acid standards with correlation coefficients of 0.991 to 0.999. The detection limits were in the range of 0.04 μg/g to 8.92 μg/g. The average recovery rates of six phenolic acids in Gannan navel orange young fruit were 85.5% to 103.9% with relative standard deviations less than 2.8%. It was proved that the proposed method is simple, rapid, accurate, and should be used for the determination of phenolic acids in navel orange as well as other citrus plant.