色谱

色谱 ›› 2013, Vol. 31 ›› Issue (5): 441-446.DOI: 10.3724/SP.J.1123.2012.12049

• 研究论文 • 上一篇    下一篇

高效液相色谱法同时检测乳制品中3种β-内酰胺酶抑制剂的残留量

林钦*   

  1. 福建省产品质量检验研究院, 福建 福州 350002
  • 收稿日期:2012-12-27 修回日期:2013-01-30 出版日期:2013-05-28 发布日期:2013-05-16
  • 通讯作者: 林钦,Tel:(0591)83762015,E-mail:phlqfcii@126.com.
  • 基金资助:

    国家质量监督检验检疫总局科技计划项目(2010QK021).

Simultaneous determination of three β-lactamase inhibitor residues in dairy products by high performance liquid chromatography

LIN Qin*   

  1. Fujian Inspection and Research Institute for Product Quality, Fuzhou 350002, China
  • Received:2012-12-27 Revised:2013-01-30 Online:2013-05-28 Published:2013-05-16

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摘要：

建立了乳制品中3种β-内酰胺酶抑制剂克拉维酸钾、他唑巴坦和舒巴坦的提取和固相萃取净化方法。样品经水溶后,用丙酮沉淀蛋白质。上清液在弱酸性条件下用PWAX固相萃取小柱(60 mg/3 mL)富集、净化,0.05%(如无特殊说明均为体积分数)氨水-甲醇溶液洗脱。然后采用高效液相色谱-二极管阵列检测器(HPLC-PAD)分离检测。分析柱为资生堂CAPCELL PAK MG-C18(150 mm×4.6 mm, 5 μm);流动相为0.03%磷酸-乙腈。该方法对克拉维酸钾、他唑巴坦和舒巴坦的最低检测质量浓度均为0.03 mg/L;纯牛奶、酸奶和奶粉中的回收率在84.6%～101.7%之间,RSD在2.2%～7.4%之间(n=6);在0.05～5 mg/L范围内均呈良好的线性关系,线性回归系数r>0.999。该方法的前处理净化效果好、检测灵敏度高、回收率和重现性良好,适用于乳制品中β-内酰胺酶抑制剂的测定。

关键词: β-内酰胺酶抑制剂, 残留量, 高效液相色谱, 克拉维酸钾, 乳制品, 舒巴坦, 他唑巴坦

Abstract:

An improved method has been developed for the determination of clavulanate potassium, tazobactam and sulbactam in dairy products. High performance liquid chromatography with photodiode array detector (HPLC-PAD) was used in this method. The separation was performed on a CAPCELL PAK MG-C18 column (150 mm×4.6 mm, 5 μm). The mobile phase was 0.03% (volume fraction unless otherwise specified) phosphate-acetonitrile. This method offered effective sample preparation procedures including dissolution with water, protein isolation with acetone, clean-up and enrichment by solid-phase extraction (SPE) with PWAX cartridges (60 mg/3 mL) under weak acidic condition and further elution with 0.05% ammonia-methanol solution. The analytical method was well validated and good results were obtained with the RSDs of 2.2%-7.4% and the recoveries of 84.6%-101.7%. The detection limits of clavulanate potassium, tazobactam and sulbactam were 0.03 mg/L. All of the target compounds exhibited good linearities (r>0.999) over the mass concentration range of 0.05-5 mg/L. With the advantages of good purification effect, high sensitivity, good recovery and repeatability, the method is suitable for the detection of β-lactamase inhibitors in dairy products.

Key words: β-lactamase inhibitors, clavulanate potassium, dairy products, high performance liquid chromatography (HPLC), residue, sulbactam, tazobactam

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