基于低共熔溶剂的分散液液微萃取/超高效液相色谱-串联质谱法测定马肉中非甾体抗炎药

Determination of Non-steroidal Anti-inflammatory Drugs in Horse Meat by Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry with Deep Eutectic Solvent Based on Dispersive Liquid-Liquid Microextraction

采用氯化胆碱和苯酚合成低共熔溶剂(DES),以其为萃取剂建立了分散液液微萃取(DLLME)结合超高效液相色谱-串联质谱(UPLC-MS/MS)测定马肉中10种非甾体抗炎药的方法。优化了提取溶剂、萃取剂、分散剂和盐的加入量等条件,最佳萃取条件为:以氯化胆碱和苯酚(比例为1∶2)合成的低共熔溶剂为萃取剂,马肉样品经5 mL含10 mmol/L抗坏血酸的20 mmol/L醋酸铵缓冲液(乙酸调至pH 3.5)提取,加入1.0 g氯化钠、0.8 mL萃取剂和0.8 mL分散剂,旋涡混合1 min,离心后取上清液经超高效液相色谱-串联质谱测定,外标法定量。结果显示,10种非甾体抗炎药在各自质量浓度范围内呈良好线性关系,相关系数(r 2)均大于0.996,检出限为0.05~2.0μg/kg,定量下限为0.10~5.0μg/kg,平均回收率为73.5%~94.6%,相对标准偏差(RSD)为1.1%~8.1%。该方法操作简单、准确高效、绿色环保,可用于马肉中非甾体抗炎药的测定。

Non-steroidal anti-inflammatory drugs,are one of the most widely used pharmaceutically active compounds in veterinary medicine.Dispersive liquid-liquid microextraction(DLLME)is a mini-extraction technique with merits of low cost,environment friendliness and high extraction efficiency.Moreover,this method is easy to operate and requires only a short operation time.In this paper,a deep eutectic solvent(DES)based on choline chloride was used for the first time as the liquid-liquid microextraction solvent for efficient and safe extraction of non-steroidal anti-inflammatory drugs in horse meat.DES was simply synthesized via just mixing choline chloride and phenol at room temperature.The analytes were extracted into acetate buffer,and then back-extracted to the DES phase simply by oscillation.Meanwhile,acetonitrile was used as a dispersive solvent and the DES phase was collected by centrifugation.An ultra high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)with liquid-liquid microextraction based on the DES was established for the determination of ten non-steroidal anti-inflammatory drugs in horse meat.A series of analytical factors such as extraction agent,dispersing agent and dosage of sodium chloride were optimized.The optimum conditions were as follows:DES was prepared with choline chloride and phenol at a 1∶2 molar ratio,while the horse meat sample was extracted with 5 mL 20 mmol/L acetate buffer(adjusting to pH 3.5 with acetic acid)containing 10 mmol/L ascorbic acid,and subsequently added 1.0 g sodium chloride,0.8 mL extractant and 0.8 mL dispersant,then vortex extracted for 1 min and followed by a centrifugation,finally detected by UPLC-MS/MS.Results showed that there existed good linear relationships for ten non-steroidal anti-inflammatory drugs in the respective concentration ranges with their correlation coefficients(r 2)larger than 0.996.The limits of detection(LOD)and the limits of quantitation(LOQ)were in the ranges of 0.05-2.0μg/kg and 0.10-5.0μg/kg,respectively.Recoveries for ten non-steroidal anti-inflammatory drugs at three spiked levels ranged from 73.5%to 94.6%with relative standard deviations of 1.1%-8.1%.Results indicated that the DES could be used as an alternate of traditional solvents for dispersive liquid-liquid microextraction.The method was simple,rapid,accurate and reliable,and could be used for the analysis of non-steroidal anti-inflammatory drugs in horse meat.