Zhang Dan, Park Jin-A, Kim Dong-Soon, Kim Na-Hyun, Kim Seong-Kwan, Cho Kyeong-Su, Jeong Dana, Shim Jae-Han, Abd El-Aty A M, Shin Ho-Chul
Department of Veterinary Pharmacology and Toxicology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of Korea.
Biotechnology Research Institute, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, Republic of Korea.
J Sep Sci. 2015 Jul;38(14):2371-80. doi: 10.1002/jssc.201500282. Epub 2015 Jun 19.
With the overarching aim to develop a simple and reliable method for the quantitative analysis of polypeptide antibiotics in various livestock products, the content of bacitracin, and polymyxin B in pork, beef, chicken, milk, and eggs was analyzed using colistin sulfate as an internal standard. The extracted samples were eluted via solid-phase extraction using 2% formic acid in acetonitrile/methanol (1:1, v/v). The two polypeptides were identified and quantified based on the intensities of mass fragments from the respective triply charged precursor ions (bacitracin: 474.97 amu and polymyxin B: 402 amu) at the defined retention time windows using liquid chromatography with electrospray ionization tandem mass spectrometry in time-scheduled multiple reaction monitoring mode. The calibration curves showed good linearity over the concentration range 50-2500 ng/mL with determination coefficients ≥ 0.991. The mean recoveries were in the range 80.3-88.8% with relative standard deviations <13% for all samples. The limits of quantitation ranged from 30-250 ng/g. The developed method was applied to market samples, but the target analytes were not detected in any of the samples. The developed method is reliable for the simultaneous detection of bacitracin and polymyxin B in pork, beef, chicken, milk, and eggs.
为了开发一种简单可靠的方法用于定量分析各种畜产品中的多肽抗生素，以硫酸黏菌素为内标，分析了猪肉、牛肉、鸡肉、牛奶和鸡蛋中杆菌肽和多黏菌素B的含量。提取的样品通过使用2%甲酸的乙腈/甲醇（1:1，v/v）进行固相萃取洗脱。使用液相色谱-电喷雾电离串联质谱，在定时多反应监测模式下，根据在定义的保留时间窗口内各自三电荷前体离子（杆菌肽：474.97 amu和多黏菌素B：402 amu）的质量碎片强度来鉴定和定量这两种多肽。校准曲线在50 - 2500 ng/mL浓度范围内显示出良好的线性，测定系数≥0.991。所有样品的平均回收率在80.3 - 88.8%范围内，相对标准偏差<13%。定量限范围为30 - 250 ng/g。所开发的方法应用于市售样品，但在任何样品中均未检测到目标分析物。所开发的方法对于同时检测猪肉、牛肉、鸡肉、牛奶和鸡蛋中的杆菌肽和多黏菌素B是可靠的。
