中国食品添加剂

本研究旨在建立一种基于离心超滤前处理技术结合高效液相色谱-荧光检测法（HPLC-FLD）的乳与乳制品中溶菌酶定量分析方法，以解决现有检测方法在灵敏度、基质适用性及操作效率方面的局限性。样品经氯化钠溶液提取，以强酸变性沉淀法进行样品初步净化，再用超滤离心管过滤后，在反相色谱柱（C4-T,250 mm×4.6 mm,300Å，5μm）上分离，用0.1%的三氟乙酸水溶液和0.1%的乙腈溶液作为流动相进行梯度洗脱，之后使用荧光检测器检测。溶菌酶在0.5～20.0 mg/kg浓度范围内线性良好，加标试验结果显示回收率在91.0%～99.5%，相对标准偏差（RSD）在0.7%～2.0%之间，牛奶基质检出限为3 mg/kg，定量限为10 mg/kg；奶酪基质检出限为7 mg/kg，定量限为15 mg/kg。该方法灵敏度高、操作简单快速、定量准确可靠，适用于乳与乳制品中溶菌酶的定量检测。本研究不仅有效弥补了现有检测方法的不足，还可为其他食品基质中溶菌酶的检测提供技术参考。

To address the limitations of existing detection methods in terms of sensitivity,matrix applicability,and operational efficiency,a robust quantitative method was established for determining lysozyme in milk and dairy products using centrifugal ultrafiltration pretreatment combined with high-performance liquid chromatographyfluorescence detection(HPLC-FLD). Samples were extracted with a sodium chloride solution,initially purified using strong acid denaturation and precipitation,and subsequently filtered through ultrafiltration centrifuge tubes. Chromatographic separation was achieved on a reversed-phase column(C4-T,250 mm×4.6 mm,300 Å,5 μm) using a gradient elution of 0.1% trifluoroacetic acid and acetonitrile,with a fluorescence detector. Lysozyme exhibited excellent linearity within the concentration range of 0.5 to 20.0 mg/kg. Matrix-spiked experiments yielded recoveries between 91.0% and 99.5%,with relative standard deviations(RSDs) ranging from 0.7% to 2.0%. The LODs were 3 mg/kg for milk matrix and 7 mg/kg for cheese matrices,with corresponding LOQs of 10 and 15 mg/kg,respectively. This rapid,highly sensitive,and reliable method is well-suited for routine quantitative control of lysozyme in dairy products and provides a valuable technical reference for application in other complex food matrices.