动物可食性组织中克拉维酸残留HPLC-MS/MS检测方法

doi:10.3864/j.issn.0578-1752.2013.23.018

摘要/Abstract

摘要： 【目的】建立动物可食性组织中克拉维酸残留检测的高效液相色谱-串联质谱方法。【方法】1 g组织样品用3 mL 0.01 mol•L-1乙酸铵和3 mL乙腈提取后，经二氯甲烷反相提取净化，取水相定容到4 mL，正己烷除脂后上机检测。流动相为乙腈和0.1%甲酸水，梯度洗脱，经Luna 5μ C8色谱柱分离，采用电喷雾电离，多反应监测负离子模式对克拉维酸进行定量分析。【结果】采用基质匹配法对组织中克拉维酸的含量进行标准校正，克拉维酸质量浓度在5—500 μg•L-1范围内与峰面积间呈现良好的线性关系（r＞0.999）；组织中加标样品的检出限（按信噪比S/N≥3计）为10 μg•kg-1，定量限（按信噪比S/N≥10计）为20 μg•kg-1。在定量限、1/2最高残留限量、最高残留限量、2倍最高残留限量4个添加水平下，组织中克拉维酸的平均回收率为76.39%—89.26%，日间相对标准偏差为1.89%—6.20%。【结论】该方法可用于动物可食性组织中克拉维酸残留的分析检测。

关键词: 高效液相色谱-串联质谱 , 克拉维酸 , 可食性组织 , 残留

Abstract: 【Objective】A confirmatory method was developed for the determination of clavulanic acid (CLAV) residue in edible tissues of animals by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). 【Method】 One gram tissues were extracted by 3 mL 0.01 mol•L-1 ammonium acetate solution and 3 mL acetonitrile，a back-extraction of the acetonitrile with dichloromethane，aqueous layer was diluted to 4mL，and the sample was determined by HPLC-MS/MS after defatted with N-hexane. The chromatographic separation was achieved on a Luna 5μ C8 column using 0.1% formic acid in water and acetonitrile as mobile phases with gradient elution. The identification of CLAV was carried out by MS/MS equipped with electrospray ionization in negative scanning and multiple reaction monitoring (MRM) modes. 【Result】Matrix-matched calibration standard was used for the quantification. Good linearity was achieved with correlation coefficients more than 0.999 for all analytes in the concentration of 5-500 μg•L-1. The limit of detection (LOD，S/N≥3) was 10 μg•kg-1 in tissues, and the limit of quantification (LOQ，S/N≥10) was 20 μg•kg-1. The mean recoveries at the four spiked levels of LOQ, 1/2MRL (the maximum residue limit), MRL and 2MRL were in the range of 76.39%-89.26% , with the inter-day relative standard deviations (RSD) of 1.89%-6.20%. 【Conclusion】In conclusion, the established method can be applied for the determination of CLAV residues in edible tissues of animals.

Key words: high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) , clavulanic acid , edible tissues , residue

李帅鹏, 黄显会, 杨刚, 郭春娜. 动物可食性组织中克拉维酸残留HPLC-MS/MS检测方法[J]. 中国农业科学, 2013, 46(23): 5012-5019.

LI Shuai-Peng, HUANG Xian-Hui, YANG Gang, GUO Chun-Na. Determination of Clavulanic Acid Residue in Edible Tissues of Animals by High Performance Liquid Chromatography-Tandem Mass Spectrometry[J]. Scientia Agricultura Sinica, 2013, 46(23): 5012-5019.

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