畜禽粪污中7种磺胺类和8种氟喹诺酮类兽药残留UPLC-MS/MS检测方法研究

Study on UPLC-MS/MS detection method of 7 sulfonamides and 8 fluoroquinolones veterinary drug residues in livestock and poultry manure

关键词:  超高效液相色谱-串联质谱法  磺胺类  氟喹诺酮类  畜禽粪污

Key words:  Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)  Sulfonamide antibiotics  Fluoroquinolone antibiotics  Livestock and poultry manure

摘要:

对猪、鸡、牛、羊粪污中同时测定7种磺胺类和8种氟喹诺酮类兽药残留的超高效液相色谱-串联质谱方法进行了研究。样品采用2%氨化乙腈+0.1 mol/L 氢氧化钠溶液（50:50，V/V）提取，提取液浓缩后用磷酸盐缓冲液溶解，HLB 固相萃取小柱净化，浓缩富集后用乙腈：0.1%甲酸水溶液（20:80，V/V）复溶，过0.22 μm微孔滤膜，上机，基质加标液外标法定量。结果显示，猪、鸡、牛、羊粪污中15种兽药浓度在1~100 μg/mL范围内线性关系良好（R2>0.994）。方法的检出限为20.0 μg/kg，定量限为50.0 μg/kg。在50、100、500 μg/kg三个添加浓度下，15种兽药平均回收率为60.5 %~115.5 %，批内RSD为1.2 %~17.6 %，批间RSD为0.2 %~17.6 %。该方法具有较好的准确度、精密度，能快速、高效、方便、准确地测定猪、鸡、牛、羊粪污中15种兽药残留，为进一步调查掌握我国畜禽养殖粪污中的残留兽药种类及残留量，提供强有力的技术支撑。

Abstract:

An ultra-high performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of 7 sulfonamides and 8 fluoroquinolones veterinary drug residues in pig, chicken, cattle and sheep manure was studied. The sample was extracted with 2% ammoniated acetonitrile + 0.1 mol/L sodium hydroxide solution (50:50, V/V), then dissolved in phosphate buffer, purified by HLB solid phase extraction cartridge, concentrated and enriched with acetonitrile: 0.1% formic acid aqueous solution (20:80, V/V) was reconstituted, passed through a 0.22 μm microporous membrane, and tested on the machine. Quantification was performed using the matrix-spiked external standard method. The results showed that the concentration of 15 veterinary drugs in pig, chicken, cattle and sheep manure had a good linear relationship in the range of 1~100 μg/mL (R2>0.994). The detection limit of the method was 20.0 μg/kg, and the quantification limit was 50.0 μg/kg. At three additive concentrations of 50, 100, and 500 μg/kg, the average recoveries of 15 veterinary drugs were 60.5%~115.5%, the average intra-batch RSD was 1.2%~17.6%, and the inter-batch RSD was 0.2%~17.6%. The method has good accuracy and precision, and can quickly, efficiently, conveniently and accurately determine 15 kinds of veterinary drug residues in pig, chicken, cattle and sheep manure. It will provide strong technical support for further investigation of the kinds and residues of veterinary drugs in livestock and poultry breeding manure in China.