超高效液相色谱-串联质谱法同时测定保健食品中14种功效成分

Simultaneous Determination of 14 Functional Components in Health Foods by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

摘要:

功效成分是保健食品常见的一项检测项目，是控制保健食品质量的重要指标。本文建立了超高效液相色谱-串联质谱同时测定保健食品中14种功效成分的快速检测方法。样品前期采用甲醇超声提取，采用 Venusil MP C18 （100 mm×2.1 mm，3 μm）色谱柱，0.1%甲酸水-乙腈为流动相，梯度分离后使用串联四级杆质谱进行检测，外标法峰面积进行定量。结果表明，超高效液相色谱-串联质谱的方法能在4 min内完成所有样品的出峰。芦荟苷、腺苷在0.05~10 ng/mL，芦丁、淫羊藿苷、苦杏仁苷、芍药苷在0.5~100 ng/mL，其余8种功效成分在5~1000 ng/mL浓度范围内线性关系良好（r>0.998）。方法精密度相对标准偏差<5%。绿原酸、没食子酸、丹参素钠、白藜芦醇、红景天苷、黄氏甲苷、甘草酸、大蒜素检出限为75 μg/kg，芦荟苷、腺苷检出限为0.75 μg/kg，芦丁、淫羊藿苷、苦杏仁苷、芍药苷检出限为7.5 μg/kg。通过三个浓度的加标回收实验表明，方法回收率在83.42%~108.44%。该方法灵敏度高、线性关系和重现性好，可快速地对保健食品中多种植物功效成分进行同时测定,可为保健食品的质量监控提供一定的技术参考。

Abstract:

The functional component and composition are commonly analyzed for health foods, and are important quality indices of these products. An ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for simultaneous determination of 14 kinds of plant-based functional components present in health foods. Ultrasonic extraction with methanol was applied to the samples prior to analysis by UPLC-MS/MS using a Venusil MP C18 column (100 mm×2.1 mm, 3 μm) and a gradient mobile phase composed of acetonitrile and water containing 0.1% formic acid. The chromatographic peak areas were quantified using external standards. The results showed that the chromatographic peak of all the test samples were eluted within 4 min by the present UPLC-MS/MS method. Good linearity was observed in the range of 0.05~10 ng/mL for aloin and adenosine，0.5~100 ng/mL for rutin, icariin, amygdalin and paeoniflorin, and 5~1000 ng/mL for the other 8 kinds of functional components, with the correlation coef?cients greater than 0.998. The method precision, the relative standard deviation, was less than 5%. The detection limit was 75 μg/kg for chlorogenic acid, gallic acid, sodium danshensu, resveratrol, salidroside, astragaloside, glycyrrhizin and allicin, 0.75 μg/kg for aloin and adenosine, and 7.5 μg/kg for rutin, icariin, amygdalin and paeoniflorin. The recovery rate was 83.42%~108.44% based on the recoveries of the spike-and-recovery experiments at three standard concentrations. Thus, this current method had a high sensitivity, good linear relationship and reproducibility, and can be used for simultaneous determination of various functional components in a health food, which provides a technical support for the quality control of health foods.