市售橄榄油中铜-焦脱镁素a的 HPLC-MS/MS快速测定Rapid determination of copper pyropheophytin-a in commercial olive oil by HPLC-MS/MS

Abstract

为了快速、准确检测市售橄榄油中非法添加的脂溶性色素，建立了一种测定橄榄油中铜-焦脱镁素a的高效液相色谱串联质谱法（HPLC-MS/MS）。使用SPE柱对样品进行净化前处理，随后通过HPLC-MS/MS进行定性和定量检测，色谱分析采用C8色谱柱，以甲醇-1 mol/L乙酸铵水溶液（体积比8∶ 2）和甲醇-丙酮(体积比6∶ 4)作为流动相，进行梯度洗脱；质谱分析采用大气压化学电离，负离子扫描模式，多重反应监测模式，电子倍增电压600 V，碎裂电压260 V，用外标法进行定量。经过方法学验证，该方法的检出限和定量限分别为12 μg/kg和64 μg/kg，回收率为910%~106.0%，相对标准偏差为3.9%~5.7%。该方法具有较高的灵敏度和精密度，能够准确、快速地检测市售橄榄油中的铜-焦脱镁素a。An analytical method for determining copper pyropheophytin-a in olive oil utilizing high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) was established to accurately and quickly detect illegally added liposoluble pigments in commercial olive oil. Purification of the sample was performed using a SPE column for pre-treatment, followed by qualitative and quantitative detection using HPLC-MS/MS. The Chromatographic analysis was carried out on C8 column, with methanol-1 mol/L aqueous solution of ammonium acetate (volume ratio 8∶ 2) and methanol-acetone (volume ratio 6∶ 4) as mobile phases for gradient elution. Atmospheric pressure chemical ionization at negative ion mode and multi reaction monitoring scanning were used for mass spectrometry analysis under the conditions of electronic multiplication voltage 600 V and fragmentation voltage 260 V, and the analyte was quantified by external standard method. The results of the methodological validation showed that this method exhibited a limit of detection of 12 μg/kg and a limit of quantification of 64 μg/kg, with spiked recovery rates ranging from 91.0% to 106.0%, with relative standard deviations varying from 3.9% to 5.7%. This method had advantages of high sensitivity and precision, and can detect of copper pyropheophytin-a in olive oil rapidly.

Keywords

• 高效液相色谱串联质谱法；铜-焦脱镁素a；橄榄油；掺假
• high performance liquid chromatography tandem mass spectrometry; copper pyropheophytin-a; olive oil; adulteration