Citation: CHEN Rouhan, GU Shuqing, ZHAO Chaomin, HUO Yihui, NIU Bing, DENG Xiaojun. Determination of bovine lactoferrin in milk and dairy products by ultra performance liquid chromatography-tandem mass spectrometry[J]. Chinese Journal of Chromatography, ;2020, 38(6): 663-671. doi: 10.3724/SP.J.1123.2019.10025 shu

Determination of bovine lactoferrin in milk and dairy products by ultra performance liquid chromatography-tandem mass spectrometry

1.
Department of Food Engineering, School of Life Science, Shanghai University, Shanghai 200436, China;
2.
Technical Center for Animal Plant and Food Inspection and Quarantine, Shanghai Customs, Shanghai 200135, China

Corresponding author: GU Shuqing, gushuqing@sina.cn
Received Date: 24 October 2019

Fund Project: National Key Research and Development Plan Project (Nos. 2017YFF0211303, 2018YFC1603503)

An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was established for the quantitative detection of bovine lactoferrin in dairy products. The samples were treated by degreasing and tryptic hydrolysis. Proteins of bovine lactoferrin and peptides were identified using ultra performance liquid chromatography-quadrupole/electrostatic orbitrap-high resolution mass spectrometry (UPLC-Q/Exactive-HRMS) and analyzed by Protein Pilot software. Eight species-specific marker peptides of bovine lactoferrin were identified by comparison of the basic local alignment search tool (BLAST) with the Uniprot database. Three markers with high response strength and stability were chosen for further quantitative research by UPLC-triple quadrupole mass spectrometry (QqQ-MS). The method showed a good linear relationship within its own range. The limits of detection and limits of quantification were 0.023-0.041 and 0.077-0.137 mg/kg, respectively. The observed recoveries were in the range of 93.8%-103.9%. The intra-day and inter-day RSDs were lower than 8.8% and 9.5%, respectively. This method presents various advantages such as strong anti-interference, high sensitivity and reproducibility, and it is suitable for the quantitative analysis of bovine lactoferrin in dairy products.
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