毛细管区带电泳测定液态奶及奶粉中的A2 <i>β</i>-酪蛋白及总<i>β</i>-酪蛋白

引用本文: 冯慿, 丁晓静, 高铁, 杜茹芸, 陈泓序. 毛细管区带电泳测定液态奶及奶粉中的A2 β-酪蛋白及总β-酪蛋白[J]. 色谱, 2020, 38(6): 722-729. doi: 10.3724/SP.J.1123.2019.11005 shu

Citation: FENG Ping, DING Xiaojing, GAO Tie, DU Ruyun, CHEN Hongxu. Determination of A2 β-casein and total β-casein in cow milk and milk powder by capillary zone electrophoresis[J]. Chinese Journal of Chromatography, 2020, 38(6): 722-729. doi: 10.3724/SP.J.1123.2019.11005 shu

毛细管区带电泳测定液态奶及奶粉中的A2 β-酪蛋白及总β-酪蛋白

冯慿 ^2, ,
丁晓静 ^3, ,
高铁 ^1, ,
杜茹芸 ^4, ,
陈泓序 ^1,

1.
SCIEX中国, 北京 100015;
2.
惠氏营养品(中国)有限公司, 上海 200041;
3.
北京市疾病预防控制中心, 北京 100013;
4.
上海市质量监督检验技术研究院, 上海 200233

摘要: β-酪蛋白（β-CN）是牛乳中的主要酪蛋白。它包含13种变异体，其中A2是牛β-CN的主要变异体之一。定量测定A2 β-CN对A2乳制品的评价十分重要。该文建立了对液态奶和奶粉中A2 β-CN及总β-CN分离并定量的毛细管区带电泳（CZE）分析方法。使用熔融石英毛细管（50 μm×30/40 cm有效/总长度），以50 mmol/L Na₂HPO₄+140 mmol/L柠檬酸+0.2%（质量分数） HPMC+4 mol/L尿素溶液（pH为2.7）作为分离缓冲溶液，在214 nm下检测。总β-CN和A2 β-CN的校准峰面积与其质量浓度间呈良好的线性关系，相关系数在0.9968~0.9997之间，加标回收率为85.5%~106.4%。对两款巴氏奶和两款奶粉中A2 β-CN和总β-CN进行分析，A2 β-CN和总β-CN的日内精密度分别为2.4%~4.7%和2.6%~4.8%，日间精密度分别为4.0%~6.3%和3.9%~6.7%。所建立的CZE方法可分离液态奶及奶粉中的A2 β-CN，并可对A2 β-CN及总β-CN进行定量。通过计算A2 β-CN在总β-CN中的占比来评价A2乳制品的质量，为保护消费者的利益提供了方法依据。

关键词:

English

Determination of A2 β-casein and total β-casein in cow milk and milk powder by capillary zone electrophoresis

1.
SCIEX China, Beijing 100015, China;
2.
Wyeth Nutrition, Shanghai 200041, China;
3.
Beijing Center for Diseases Prevention and Control, Beijing 100013, China;
4.
Shanghai Institute of Quality Inspection and Technical Research, Shanghai 200233, China
Received Date: 05 November 2019

Abstract: β-casein (β-CN) is one of the major casein proteins in cow milk. There are 13 different variants documented for β-CN in cow milk, among which A1 and A2 are the major variants. The separation and quantitation of A2 β-CN are imperative for A2 dairy products. A new capillary zone electrophoresis (CZE) method with UV detection at 214 nm was established for the separation and quantification of the A2 variant and total β-CN content in cow milk and milk powders. The separation of β-CN variants was achieved on bare fused silica capillaries (50 μm×30/40 cm (effective/total length)). The separation buffer was a mixture of 4 mol/L urea, 0.2% (mass fraction) hydroxypropyl methylcellulose, 140 mmol/L citric acid, and 50 mmol/L disodium hydrogen phosphate buffer (pH 2.7). The corrected peak areas and the concentrations of total β-CN and the A2 variant showed good linearity, with correlation coefficients (r²) ranging from 0.9968 to 0.9997. The intra-day precisions for A2 β-CN and total β-CN determination in four samples (two pasteurized milk samples and two milk powder samples) were in the ranges of 2.4%-4.7% and 2.6%-4.8%, respectively. The inter-day precisions for A2 β-CN and total β-CN determination in four samples were in the ranges of 4.0%-6.3% and 3.9%-6.7%, respectively. The recoveries of A2 and total β-CN ranged from 85.5% to 106.4%. With the established CZE method, the A2 β-CN variant and total β-CN protein in liquid and powder bovine milk products could be separated and accurately quantified. By calculating the A2 β-CN content in the total β-CN, the quality of A2 dairy products can be evaluated, and this in turn would aid in the protection of consumer rights.

Key words:

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毛细管区带电泳测定液态奶及奶粉中的A2 β-酪蛋白及总β-酪蛋白

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Determination of A2 β-casein and total β-casein in cow milk and milk powder by capillary zone electrophoresis

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