摘要
本研究旨在建立一种准确测定饲料中大观霉素的方法。试样中大观霉素用2%三氯乙酸水溶液(含0.4%乙二胺四乙酸二钠)提取,提取液经调节pH至(4.7±0.2),由混合型阳离子交换(MCX)小柱固相萃取净化,采用亲水相互作用液相色谱法(HILIC)色谱柱分离,流动相为0.01 mol/L甲酸铵水溶液(含0.4%甲酸)和0.01 mol/L甲酸铵乙腈-水溶液(含0.4%甲酸),梯度洗脱,以电喷雾离子(ESI)正离子扫描方式,在多反应监测(MRM)模式下进行测定,基质匹配外标法定量。结果表明:本试验建立的固相萃取-超高效液相色谱-串联质谱法对饲料中大观霉素检出限为1.0 mg/kg,定量限为2.0 mg/kg。不同饲料基质中大观霉素在0.1~10.0μg/mL浓度范围内线性关系良好,相关系数(R^(2))均大于0.99。在4个浓度水平(2.0、20.0、200.0和1000.0 mg/kg)进行添加试验,大观霉素的回收率为91.3%~104.2%,相对标准偏差(RSD)为1.5%~9.4%。由此可见,本试验建立的固相萃取-超高效液相色谱-串联质谱法灵敏、可靠,可应用于饲料中大观霉素的准确测定。
The aim of this study was to establish an accurate method for the determination of spectinomycin in feed.The sample was extracted with 2%trichloroacetic acid aqueous solution[containing 0.4%ethylenediaminetetraacetic acid disodium salt(EDTA-2Na)].After its pH was adjusted to(4.7±0.2)and the sample was cleaned-up on the mixed-mode cation-exchange(MCX)solid-phase extraction(SPE).The spectinomycin was separated by a hydrophilic interaction liquid chromatography(HILIC)column with gradient elution using 0.01 mol/L ammonium formate aqueous solution(containing 0.4%formic acid)and 0.01 mol/L ammonium formate acetonitrile-aqueous solution(containing 0.4%formic acid)as mobile phases and finally confirmed and quantified in multiple reaction monitoring(MRM)mode with the electrospray ionization(ESI)positive ion scanning by matrix matching external calibration method.The results show that the limit of detection and limit of quantification of spectinomycin in feed using this solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry method was 1.0 and 2.0 mg/kg,respectively.The regression coefficient(R^(2))of linear calibration curve for spectinomycin in different feed substrates was over 0.99 within the concentration range from 0.1 to 10.0μg/mL.At the four spiked levels(2.0,20.0,200.0 and 1000.0 mg/kg),the average recovery rate of spectinomycin was from 91.3%to 104.2%and the relative standard deviation(RSD)was from 1.5%to 9.4%.As a result,the solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry method was sensitive and practical for the determination of spectinomycin in feed.
作者
王凤芹
开丽霞
程远之
汪以真
WANG Fengqin;KAI Lixia;CHENG Yuanzhi;WANG Yizhen(Key Laboratory of Animal Nutrition and Feed Science of Zhejiang Province,Key Laboratory of Animal Nutrition and Feed Science in Eastern China,Ministry of Agriculture and Rural Affairs,Key Laboratory of Molecular Animal Nutrition,Ministry of Education,College of Animal Sciences,Zhejiang University,Hangzhou 310058,China)
出处
《动物营养学报》
CAS
CSCD
北大核心
2023年第3期2004-2017,共14页
CHINESE JOURNAL OF ANIMAL NUTRITION
基金
现代农业生猪产业体系(CARS-36)。
关键词
固相萃取
超高效液相色谱-串联质谱
饲料
大观霉素
solid phase extraction
ultra performance liquid chromatography-tandem mass spectrometry
feed
spectinomycin
