超高效液相色谱-紫外检测法和液相色谱-串联质谱法测定乳品中乳铁蛋白的比较研究

doi:10.7506/rykxyjs1671-5187-20260127-006

乳业科学与技术 ›› 2026, Vol. 49 ›› Issue (3): 33-40.DOI: 10.7506/rykxyjs1671-5187-20260127-006

• 分析检测 • 上一篇

超高效液相色谱-紫外检测法和液相色谱-串联质谱法测定乳品中乳铁蛋白的比较研究

武伦玮, 李培珍, 段国霞, 张立佳, 唐烁, 刘丽君, 吴晓莉, 宫慧丽, 陈静, 刘春霞

内蒙古伊利实业集团股份有限公司,国家市场监督管理总局重点实验室(牛羊乳肉风险防控与关键技术), 内蒙古呼和浩特 010110

发布日期:2026-07-06
基金资助:
内蒙古自治区重点研发与成果转化计划项目（2023YFHH0093）

Determination of Lactoferrin in Dairy Products: A Comparative Study of Ultra-high Performance Liquid Chromatography with Ultraviolet Detection and Liquid Chromatography-Tandem Mass Spectrometry

WU Lunwei, LI Peizhen, DUAN Guoxia, ZHANG Lijia, TANG Shuo, LIU Lijun, WU Xiaoli, GONG Huili, CHEN Jing, LIU Chunxia

Lactoferrin, a key functional protein in dairy products, serves the dual functions of nutritional supply and physiological regulation. Its accurate quantification serves as a critical link in the quality control and quality evaluation of dairy products. This study systematically compared the detection performances, operational procedures and application scopes of two mainstream analytical techniques for lactoferrin determination in dairy products, namely ultra-high performance liquid chromatography with ultraviolet detection (UPLC-UV) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The UPLC-UV method adopted a C? column for chromatographic separation, with 280 nm set as the characteristic detection wavelength. The pretreatment was accomplished by phosphate buffer extraction followed by heparin affinity column purification, and quantification was performed using the external standard calibration curve method. This method featured high instrument availability, with spiked recoveries ranging from 82.32% to 98.07%, a limit of quantification (LOQ) of 1 mg/100 g, and a correlation coefficient above 0.999, indicating good reproducibility. It was suitable for routine testing of batch samples in dairy enterprises. Nevertheless, it presented moderate anti-matrix interference capability and insufficient sensitivity for the detection of trace lactoferrin in complex matrices, and was incapable of determining denatured lactoferrin. In contrast, the LC-MS/MS method involved sample dissolution and enzymatic hydrolysis to screen for signature peptides. Quantification was performed in multiple reaction monitoring (MRM) mode using the internal standard calibration curve method. It possessed excellent anti-matrix interference capacity, with a LOQ as low as 0.2 mg/100 g, and enabled the determination of total lactoferrin. This approach was applicable to the detection of low-abundance lactoferrin in complex matrix samples such as infant formula powder and high-temperature sterilized milk. However, it was constrained by high instrument cost, insufficient supporting reference materials, cumbersome operation and relatively low detection efficiency.

Online:2026-07-06

摘要/Abstract

摘要： 乳铁蛋白是乳品中兼具营养供给与生理调节价值的关键功能活性蛋白,精准测定其含量是乳品质量管控与品质评价的重要环节?对目前乳品乳铁蛋白检测的主流技术手段超高效液相色谱-紫外检测(ultra-high performance liquid chromatography with ultraviolet detection,UPLC-UV)与液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)2 种方法的检测性能?操作流程及适用范围开展系统比较?UPLC-UV法采用 C?色谱柱分离,以280 nm作为特征检测波长,通过磷酸盐缓冲液提取结合肝素亲和柱净化完成前处理,以标准曲线外标法定量?该方法仪器普及度高,加标回收率可达82.32%~98.07%,定量限为1 mg/100 g 相关系数>0.999,重现性良好,适配乳品企业常规批量样品检测需求,但方法抗基质干扰能力一般,对复杂基质中痕量乳铁蛋白的检测灵敏度不足,不能测定变性的乳铁蛋白?LC-MS/MS法则通过样品溶解?酶解筛选特异性特征肽段,采用多反应监测模式?标准曲线内标法定量,具备极强的抗基质干扰能力,定量限可低至0.2 mg/100 g,可实现乳铁蛋白总量的测定,适配婴幼儿配方乳粉?超高温灭菌纯牛乳等复杂基质低含量乳铁蛋白样品检测,但该方法仪器成本高,标准物质配套不足,操作流程繁琐,检测效率相对偏低?

中图分类号:

TS252.1

武伦玮, 李培珍, 段国霞, 张立佳, 唐烁, 刘丽君, 吴晓莉, 宫慧丽, 陈静, 刘春霞. 超高效液相色谱-紫外检测法和液相色谱-串联质谱法测定乳品中乳铁蛋白的比较研究[J]. 乳业科学与技术, 2026, 49(3): 33-40 https://doi.org/10.7506/rykxyjs1671-5187-20260127-006

Key Laboratory of Cattle and Sheep Milk and Meat Products Risk Control and Key Technology, State Administration for Market Regulation, Inner Mongolia Yili Industrial Group Co. Ltd., Hohhot 010110, China. Determination of Lactoferrin in Dairy Products: A Comparative Study of Ultra-high Performance Liquid Chromatography with Ultraviolet Detection and Liquid Chromatography-Tandem Mass Spectrometry[J]. Journal of Dairy Science and Technology, 2026, 49(3): 33-40 https://doi.org/10.7506/rykxyjs1671-5187-20260127-006

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超高效液相色谱-紫外检测法和液相色谱-串联质谱法测定乳品中乳铁蛋白的比较研究

Determination of Lactoferrin in Dairy Products: A Comparative Study of Ultra-high Performance Liquid Chromatography with Ultraviolet Detection and Liquid Chromatography-Tandem Mass Spectrometry

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