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    Captiva EMR-Lipid固相萃取柱净化-高效液相色谱-串联质谱法测定猪肉中37种兽药的残留量

    Determination of Residues of 37 Veterinary Drugs in Pork by High Performance Liquid Chromatography-Tandem Mass Spectrometry after Purification with Captiva EMR-Lipid Solid Phase Extraction Column

    • 摘要: 取均质后的猪肉2.00 g,加入1.0 mg·L−1混合内标溶液20 μL,混匀并静置10 min。加入陶瓷均质子,充分混合,再加入6.0 mL含0.1%(体积分数,下同)甲酸的乙腈溶液,涡旋混匀,超声5 min。加入由4.0 g 硫酸钠和1.0 g 氯化钠组成的萃取盐包,振荡3 min,于4 ℃离心10 min,收集上清液。在残渣中加入4.0 mL含5%(体积分数)甲酸的乙腈溶液,同上重复提取1次。合并2次提取液,混匀后于4 ℃离心10 min,上清液全部过Captiva EMR-Lipid固相萃取柱。控制常压下以2~3滴·s−1速率流出洗脱液,在洗脱终点施加真空排空固相萃取柱。收集全部洗脱液,在40 ℃下氮吹至近干,加入体积比1∶9的乙腈-0.1%甲酸溶液的混合溶液1.0 mL,涡旋1 min,过0.22 μm滤头,滤液按照高效液相色谱-串联质谱法测定。目标物在Agilent Poroshell 120 EC-C18色谱柱上采用0.1%甲酸溶液-含0.1%甲酸的乙腈溶液体系(正离子模式)或水-乙腈体系(负离子模式)分离后,以电喷雾离子源正、负离子模式及多反应监测(MRM)模式检测,内标法或外标法定量。结果显示:37种兽药的质量浓度在2~100 μg·L−1内与定量离子峰面积或其与内标定量离子峰面积比值呈线性关系,检出限为0.05~2.0 μg·kg−1;按照标准加入法进行回收试验,回收率为63.2%~106%,测定值的相对标准偏差(n=6)为2.7%~7.9%。

       

      Abstract: The 2.00 g of homogenized pork was taken, and 20 μL of 1.0 mg·L−1 mixed internal standard solution was added. After mixing well, the mixture was settled for 10 min. A ceramic homogenizer was added, the mixture was mixed thoroughly, and 6.0 mL of acetonitrile solution containing 0.1% (volume fraction, the same below) formic acid was added. The mixture was mixed by vortex, and sonicated for 5 min. An extraction salt package consisting of 4.0 g of sodium sulfate and 1.0 g of sodium chloride was added, and the mixture was shaken for 3 min, and centrifuged at 4 ℃ for 10 min. Then, the supernatant was collected, 4.0 mL of acetonitrile solution containing 5% (volume fraction) formic acid was added into the residue, and the extraction was repeated once following the same procedure. The two extracts were combined, mixed well, and centrifuged at 4 ℃ for 10 min. The entire supernatant was passed through a Captiva EMR-Lipid solid phase extraction column. The eluate was controlled to flow out at the rate of 2–3 drops·s−1 under atmospheric pressure, and the solid phase extraction column was emptied by applying vacuum at the end of the elution. All the eluate was collected, and evaporated to near dryness with nitrogen at 40 ℃. The mixed solution (1.0 mL) of acetonitrile and 0.1% formic acid solution at volume ratio of 1∶9 was added, the mixed solution was vortexed for 1 min, and passed through a 0.22 μm filter. The analytes in the filtrate were determined according to the high performance liquid chromatography-tandem mass spectrometry. The target compounds were separated on the Agilent Poroshell 120 EC-C18 chromatographic column using the mobile phase system of 0.1% formic acid solution and acetonitrile solution containing 0.1% formic acid (positive ion mode) or the water-acetonitrile system (negative ion mode), detected using the electrospray ion source in positive and negative ion modes with multiple reaction monitoring mode, and quantified by the internal/external standard method. It was shown that linear relationships between the mass concentrations of the 37 veterinary drugs and peak areas of the quantitative ion or the peak area ratios of quantitative ions of target compounds to internal standards were kept in the range of 2‒100 μg·L−1, with detection limits in the range of 0.05–2.0 μg·kg−1. Test for recovery was performed using the standard addition method, giving recoveries in the range of 63.2%–106%, and RSDs (n=6) of the determined values ranged from 2.7% to 7.9%.

       

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