高效液相色谱法测定羊乳中的乳铁蛋白
Determination of lactoferrin in goat milk by HPLC method
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摘要: 为开发羊乳中乳铁蛋白的高效液相色谱测定方法及报告羊乳中乳铁蛋白的分布规律。本研究采集哈尔滨地区羊乳样品(n=24)和经过巴氏杀菌的羊乳样品(n=24),采用高效液相色谱测定其乳铁蛋白质量浓度。样品脱脂后,调整pH沉淀酪蛋白,采用C8色谱柱分离,二极管阵列检测器210nm检测,以水、乙腈、三氟乙酸为流动相进行线性梯度洗脱。方法回收率94.2%,线性范围50~300μg/mL,检出限12μg/mL,定量限40μg/mL。原料羊乳中乳铁蛋白的质量浓度的平均值122.1μg/mL,巴氏杀菌后羊乳中乳铁蛋白的平均值103.9μg/mL。巴氏杀菌对羊乳中乳铁蛋白浓度无影响。Abstract: The aim of this study was the determination of lactoferrin in goat milk using HPLC method and the report of the concentration of lactoferrin in goat milk.Milk samples were collected at a goat farm in the Harbin Region, China.It were established raw milk ( n = 24) and pasteurized milk ( n = 24) that were collected during lactation.Lactoferrin contents were analyzed by reverse phase high- performance liquid chromatography ( RP- HPLC) with diode- array detector PDA 2996. Detection was carried out at the wavelength 210 nm. The average recovery, the concentration range of calibration curve, the limit of detection and the limit of quantification was 94.2%, 50~300μg /mL, 12μg /mL, 40μg /mL separately.The average concentration of lactoferrin in goat milk was 122.1μg /mL.The average concentration of lactoferrin in pasteurization ( at 72℃ for 30s) goat milk was 103.9μg /mL.The heat treatment resulted in no significant effect on the lactoferrin content.No statistically significant differences ( p < 0.05) were found between the values of raw and pasteurized goat milk.