亲水液相色谱-串联质谱法测定羊肌肉组织中安乃近代谢物的残留量

Simultaneous Determination of Residues of Four Dipyrone Metabolites in Goat Muscle by Hydrophilic Interaction Liquid Chromatography-Mass Spectrometry

建立了羊肌肉组织中安乃近4种主要代谢物——4-甲氨基安替比林、4-氨基安替比林、4-甲酰氨基安替比林和4-乙酰氨基安替比林的亲水液相色谱-串联质谱分析方法。肌肉样品均质后,用体积分数为5%的氨水乙腈提取,提取液用Cleanert PSA初步净化,60℃氮气吹干,乙腈复溶后过MAX（SPE）柱净化,采用亲水液相色谱-串联质谱测定,4种物质均采用外标法定量。结果显示：4-甲氨基安替比林、4-甲酰氨基安替比林和4-乙酰氨基安替比林在添加量为4-200μg/kg范围内线性良好;4-氨基安替比林在添加量为40-2000μg/kg范围内线性良好。在5、50、100、1000μg/kg4个添加量,回收率在75.02%-116.2%范围内。日内变异系数为1.63%-15.12%（n=5）,日间变异系数均小于11%（n=15）。4-甲氨基安替比林、4-甲酰氨基安替比林和4-乙酰氨基安替比林的检测限均为0.5μg/kg,4-氨基安替比林的检测限为5μg/kg。该方法灵敏度高、重复性好,适用于羊肌肉组织中安乃近4种主要代谢物的大批量样品检测。

A hydrophilic interaction liquid chromatography-mass spectrometry （HILIC-MS） method was developed for the determination of four dipyrone metabolites, namely 4-methylaminoantipyrine （MAA）, 4-aminoantipyrine （AA）, 4-formylaminoantipyrine （FAA）, and 4-acetylaminoantipyrine （AAA） in goat muscle. These metabolites were extracted with acetonitrile mixed with 5% ammonia. The extracts were cleaned up with Cleaner PSA, and then blow-dried by nitrogen gas, re-dissolved in acetonitrile and re-cleaned up using MAX （SPE） cartridge. After chromatographic separation, the analytes were quantified by the external standard method. The calibration curves were linear between 4 and 200 μg/kg matrix equivalent concentrations for MAA, FAA, and AAA, and between 40 and 2 000 μg/kg for AA. The mean recoveries in blank samples at four spike levels （5, 50, 100, and 1 000 μg/kg） ranged between 75.02% and 116.2%. The intra-day coefficient of variation （CV） was 1.63% to 15.12% （n = 5）, and the inter-day CV was less than 11% （n = 15）. The detection limits were 0.5μg/kg for MAA, FAA and AAA and 5μg/kg for AA. The proposed method is sensitive and repeatable, allowing analysis of a large number of samples in a working day.