离子交换固相萃取-超高效液相色谱-串联质谱法测定减肥食品中p-辛弗林和去氧肾上腺素

Determination of p-synephrine and phenylephrine hydrochloride in diet food by ion exchange solid-phase extraction coupled with ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  p-辛弗林  去氧肾上腺素  减肥食品  混合强阳离子交换固相萃取法  超高效液相色谱-串联质谱法

英文关键词:p-synephrine  phenylephrine hydrochloride  diet food  mixed mode cation exchange solid-phase extraction  ultra performance liquid chromatography-tandem mass spectrometry

中文摘要:

      目的 建立混合强阳离子交换固相萃取法(mixed mode cation exchange solid-phase extraction, MCX)-超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定减肥食品中p-辛弗林和去氧肾上腺素两种辛弗林含量的分析方法。方法 样品以50%甲醇为溶剂超声提取, 离心后上清液经MCX固相萃取小柱净化, 5%氨水甲醇洗脱, 氮吹近干后初始流动相复溶, 以HSS T3色谱柱(100 mm×2.1 mm, 1.8 μm)分离, 以0.1% (V:V)甲酸水溶液(含10 mmol/L乙酸铵)-乙腈作为流动相, 进行梯度洗脱。在电喷雾正离子模式下, 采用多反应监测(multiple reaction monitoring, MRM)模式, 外标法定量。结果 p-辛弗林和去氧肾上腺素在1~100 ng/mL范围内与峰面积均成良好的线性, 相关系数(r)均大于0.999。p-辛弗林和去氧肾上腺素的检出限均小于0.5 μg/kg, 定量限均小于1.0 μg/kg, 在咖啡、酵素粉、减肥茶和压片糖果4种减肥食品基质中1、2、10 ng/mL 3个水平下加标回收率为85.6%~110.4%, 相对标准偏差均未超过5.0% (n=6)。结论 该方法准确、可靠, 可满足减肥食品中两种辛弗林的定量分析。

英文摘要:

      Objective To establish an analytical method for the analysis of the concentration of p-synephrine and phenylephrine hydrochloride in diet foods by mixed mode cation exchange solid-phase extraction (MCX) coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The samples were extracted by ultrasound with 50% methanol as solvent. After centrifugation, the supernatant was purified by MCX solid-phase extraction column, and eluted with 5% ammonia and methanol. After nitrogen was nearly dried, the initial mobile phase was re-dissolved, separated by HSS T3 column (100 mm×2.1 mm, 1.8 μm), and subjected to gradient elution with 0.1% (V:V) formic acid aqueous solution (containing 10 mmol/L ammonium acetate)-acetonitrile as the mobile phase. Under the positive ion mode of electrospray ionization, multiple reaction monitoring (MRM) mode was adopted and quantification was conducted by external standard method. Results p-synephrine and phenylephrine hydrochloride exhibited good linearity over the range of 1?100 ng/mL with peak area, and the correlation coefficients (r) were greater than 0.999. The limits of detection of p-synephrine and phenylephrine hydrochloride were less than 0.5 μg/kg, and the limits of quantitation were less than 1.0 μg/kg. The recoveries of spiked samples at 1, 2 and 10 ng/mL in 4 kinds of weight loss food matrices including coffee, yeast powder, weight loss tea and tablet candy were 85.6%?110.4%, and the relative standard deviations were not more than 5.0% (n=6). Conclusion This method is accurate and reliable, and can be used for quantitative analysis of 2 kinds of synephrine in diet food.