Abstract: To addresses technical bottlenecks such as the complexity of the camel milk matrix and the interference of endogenous proteins with target detection, this study established an analytical method to detect exogenous bovine insulin in camel milk using dispersive liquid-liquid microextraction (DLLME) coupled with high performance liquid chromatography (HPLC). The extraction conditions and the instrumental parameters were optimized. The results showed that efficient enrichment and accurate quantification of target analytes in complex milk matrices were achieved by HPLC combined with DLLME using 0.01 mol/L HCl as the extractant, volume fraction 60% aqueous acetone as the extraction solvent, and n-hexane as the separation phase. The method exhibited good linearity for bovine insulin over the concentration ranges of 5–500 μg/mL (R2 = 0.999). Recoveries for blank sample of Bactrian camel milk from Dabancheng District, ürümqi ranged from 64.1% to 86.9% at spiked levels of 5, 50 and 500 μg/mL, with relative standard deviations (RSDs) of 4.9%–12.4%. The limit of detection and limit of quantification were 0.5 and 5 μg/mL, respectively. The DLLME-HPLC method is characterized by simple operation, short pretreatment time, and high analytical efficiency, providing technical support for rapid screening of exogenous bovine insulin in camel milk.

Key words: camel milk; dispersive liquid-liquid microextraction; bovine insulin; high performance liquid chromatography

摘要： 针对驼乳基质复杂、内源性蛋白易对目标物检测产生干扰等技术瓶颈，基于分散液-液微萃取（dispersive liquid-liquid microextraction，DLLME）与高效液相色谱（high performance liquid chromatography，HPLC）构建驼乳中外源牛胰岛素快速检测方法，并对DLLME体系与色谱条件进行优化。结果表明，以0.01 mol/L HCl为DLLME萃取剂、体积分数60%丙酮为提取剂、正己烷为分离相，结合HPLC可实现复杂乳基质中目标物的高效富集与准确定量，在5～500 μg/mL范围内，牛胰岛素线性关系良好（R2＝0.999），乌鲁木齐市达坂城区双峰驼乳空白样品在5、50、500 μg/mL加标量下的回收率为64.1%～86.9%，相对标准偏差为4.9%～12.4%，检出限与定量限分别为0.5 μg/mL与5 μg/mL。综上，本研究建立的DLLME-HPLC法操作简便、前处理时间短、分析效率高，可为驼乳中外源牛胰岛素快速筛查提供技术支撑。

关键词: 驼乳；分散液-液微萃取；牛胰岛素；高效液相色谱