同位素稀释-液相色谱-串联质谱法测定蜂蜜中的葫芦巴碱

Determination of Trigonelline in Honey by Isotopic Dilution-Liquid Chromatography-Tandem Mass Spectrometry

为建立蜂蜜中葫芦巴碱测定的高效液相色谱-串联质谱法。本研究将蜂蜜样品中葫芦巴碱用0.1%甲酸水溶液(V/V)提取后,经混合型阳离子交换固相萃取柱(MCX)净化,高效液相色谱法(HILIC)色谱柱分离,以乙腈-5 mmol·L^(-1)乙酸铵水溶液为流动相进行梯度洗脱,液相色谱-串联质谱仪测定,同位素内标法定量。结果表明,目标物在10~500 ng·mL^(-1)的浓度范围内线性良好,相关系数(R^(2))为0.9999。方法检出限为0.075 mg·kg^(-1),定量限为0.25 mg·kg^(-1)。在空白样品中进行0.25、0.5和2.5 mg·kg^(-1)3个浓度水平的添加测试,本方法的平均回收率为97.8%~101.8%,相对标准偏差(RSD)范围为0.5%~4.3%(n=6)。综上,该方法具有良好的准确度和精密度,适用于蜂蜜中葫芦巴碱含量的测定,可以为蜂蜜的品种鉴别与品质评价提供技术支撑。

A high-performance liquid chromatography-tandem mass spectrometry method was developed a method to determine trigonelline in honey.Trigonelline was extracted from the honey sample using a 0.1%formic acid water(V/V)solution,purified on a mixed-mode cation exchange(MCX)column,separated by a hydrophilic interaction chromatography(HILIC)chromatography column,gradient eluted with acetonitrile-5 mmol·L^(-1) ammonium acetate aqueous solution as the mobile phase,determined by tandem mass spectrometry,and quantified by isotope internal standard method.The linearity of the target substance is good in the concentration range of 10-500 ng·mL^(-1),with a R^(2) of 0.9999.The detection limit of the method is 0.075 mg·kg^(-1),and the quantitative limit is 0.25 mg·kg^(-1).Three concentration levels(0.25,0.5 and 2.5 mg·kg^(-1))were added to the blank sample for testing,resulting in an average recovery of 97.8%-101.8%and relative standard deviations(RSDs)ranged from 0.5%-4.3%(n=6).The results show that the method has good accuracy and precision,and is suitable for the determination of trigonelline content in honey.It can be a valuable tool for identifying honey varieties and assessing their quality.