Institute of Agricultural Quality Standards and Testing Technology Research, Shandong Academy of Agricultural Sciences, 202 Gongyebeilu Road, Jinan 250100, Shandong Province, China.
Shandong Veterinary Drug Quality Inspection Institute, 68 Huaicun Street, Jinan, Shandong Province, China.
Molecules. 2019 Nov 19;24(22):4199. doi: 10.3390/molecules24224199.
An ultrahigh-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the determination of lactoferrin in camel milk based on the signature peptide. The camel lactoferrin was purified by heparin affinity chromatography and then used to screen tryptic signature peptides. The signature peptide was selected on the basis of sequence database search and identified from the tryptic hydrolysates of purified camel lactoferrin by ultrahigh-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry. The pretreatment procedures included the addition of isotope-labeled winged peptide and the disposal of lipids and caseins followed by an enzymatic digestion with trypsin. Analytes were separated on an Acquity UPLC BEH 300 C18 column and then detected on a triple-quadrupole mass spectrometer in 7 min. The limits of detection and quantification were 3.8 mg kg and 11 mg kg, respectively. The recoveries ranged from 74.5% to 103.6%, with relative standard deviations below 7.7%. The validated method was applied to determine the lactoferrin in ten samples collected from Xinjiang Province.
建立并验证了基于特征肽的超高效液相色谱-串联质谱法测定驼乳乳铁蛋白。采用肝素亲和层析法对骆驼乳乳铁蛋白进行纯化，然后用于筛选胰蛋白酶特征肽。根据序列数据库搜索选择特征肽，并通过超高效液相色谱和四极杆飞行时间串联质谱从纯化的骆驼乳乳铁蛋白的胰蛋白酶水解产物中鉴定特征肽。预处理程序包括添加同位素标记的 Winged 肽和处理脂质和酪蛋白，然后用胰蛋白酶进行酶解。分析物在 Acquity UPLC BEH 300 C18 柱上分离，然后在三重四极杆质谱仪中在 7 分钟内检测。检测限和定量限分别为 3.8 mg kg 和 11 mg kg。回收率在 74.5%至 103.6%之间，相对标准偏差低于 7.7%。验证后的方法用于测定来自新疆的 10 个样品中的乳铁蛋白。
