通过液相色谱-串联质谱法（LC-MS/MS）选择用于检测婴儿配方奶粉中牛乳铁蛋白的潜在特征肽段。
Selection of possible signature peptides for the detection of bovine lactoferrin in infant formulas by LC-MS/MS.
作者信息
Yuan Mingmei, Feng Cong, Wang Shouyun, Zhang Weiwei, Chen Mo, Jiang Hong, Feng Xuesong
机构信息
Department of Pharmaceutical Analysis, School of pharmacy, China Medical University, Shenyang, PR China.
Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang, PR China.
出版信息
PLoS One. 2017 Sep 19;12(9):e0184152. doi: 10.1371/journal.pone.0184152. eCollection 2017.
An LC-MS/MS assay based on a signature peptide was developed and fully validated for the quantitation of bovine lactoferrin in infant formulas. Three unreported signature peptides were derived and identified from the tryptic peptides of bovine lactoferrin. The peptide ETTVFENLPEK was used for quantification based on assay performance. The blank matrix camel milk powder and bovine lactoferrin protein standards were mixed and spiked with stable isotope-labeled internal standard to establish a calibration curve. The established method was extensively validated by determining the linearity (R2 > 0.999), sensitivity (limit of quantitation, 0.16 mg/100 g), recovery (83.1-91.6%), precision (RSD < 5.4%) and repeatability (RSD < 7.7%). To validate the applicability of the method, four different brands of infant formulas in China were analysed. The acquired contents of bovine lactoferrin were 52.60-150.56 mg/100 g.
建立了一种基于特征肽的液相色谱-串联质谱（LC-MS/MS）分析方法，并对其进行了全面验证，用于定量婴儿配方奶粉中的牛乳铁蛋白。从牛乳铁蛋白的胰蛋白酶肽段中推导并鉴定出三种未报道的特征肽。基于分析性能，选择肽ETTVFENLPEK进行定量。将空白基质驼奶粉和牛乳铁蛋白蛋白标准品混合，并加入稳定同位素标记的内标物，建立校准曲线。通过测定线性（R2>0.999）、灵敏度（定量限，0.16 mg/100 g）、回收率（83.1-91.6%）、精密度（相对标准偏差<5.4%）和重复性（相对标准偏差<7.7%），对所建立的方法进行了广泛验证。为验证该方法的适用性，分析了中国市场上四个不同品牌的婴儿配方奶粉。测得的牛乳铁蛋白含量为52.60-150.56 mg/100 g。
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