新型在线净化前处理HPLC-MS/MS测定香精香料中的麦角甾醇和麦角甾酮

建立一种新型的在线净化前处理装置结合高效液相色谱-串联质谱(high performance liquid chromatography-tandem with mass spectrometry,HPLC-MS/MS)法测定香精香料中的麦角甾醇和麦角甾酮。该前处理装置结合基质分散萃取和柱层析对样品进行萃取和净化,之后对样品进行浓缩并利用HPLC-MS/MS分析测定。HPLC-MS/MS方法以甲醇和水为流动相,0.4 mL/min流速条件下梯度洗脱,采用C_(18)色谱柱进行液相色谱分离,大气压正离子方式电离,多重反应监测模式检测,内标法进行定量分析。结果表明,该方法前处理方便且稳定性强,溶剂可回收,净化萃取浓缩为一体;目标物质麦角甾醇和麦角甾酮能在5 min内得到分离检测,在1.5~250μg/m L范围内具有较好的线性,各待测物的线性相关系数均大于0.999,检出限为0.32~0.35μg/mL,在不同添加水平条件下,平均回收率为86%~96%,相对标准偏差在3.1%~3.6%之间。本方法能较好地应用于香精香料实际样品中麦角甾醇和麦角甾酮的测定。

Novel Online Purification Pretreatment System Coupled with HPLC-MS/MS for Determination of Ergosterol and Ergotsterone in Food Flavors and Spices

A method using a new online purifying pretreatment system and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was established for the determination of ergosterol and ergotsterone in food flavors and spices. The pretreatment method in combination with matrix solid-phase dispersion and column chromatography was used for sample extraction and purification, followed by concentration and analysis by HPLC-MS/MS. The analysis was performed on a C18 column with gradient elution with a mobile phase consisting of water and methanol at a flow rate of 0.4 mL/min. The mass spectrometer with an?atmospheric pressure chemical ionization (APCI) source in the positive ion mode was operated using multiple reaction monitoring (MRM) and an internal standard method was used for quantification. The results showed that the pretreatment method was simple and stable with solvent recycle, integrating extraction, purification and concentration. The two analytes could be separated in 5 min with good linearity in the concentration range of 1.5–250 μg/mL with correlation coefficients larger than 0.999. The limits of detection (LODs) were 0.32 μg/mL for ergosterol and 0.35 μg/mL for ergotsterone. The mean recoveries of the target compounds at three different spiked levels were 86%–96% with relative deviations (RSDs) of 3.1%–3.6%. This method is suitable for the analysis of ergosterol and ergotsterone in food flavors and spices.