基于二维液相色谱体系分析特殊医学用途奶粉中的牛磺酸含量
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(上海市质量监督检验技术研究院 上海 200233)

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卞华(1989—),男,硕士,工程师

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上海市科学技术委员会:食品添加剂质量鉴定关键技术与质量标准研究(17DZ2201200)


Analysis on the Taurine in the Milk Powder with Special Medical Purpose Based on the Two-dimensional Liquid Chromatography System
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(Shanghai Institute of Quality Inspection and Technical Research, Shanghai 200233)

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    摘要:

    建立特殊医学用途全氨基酸水解配方奶粉中牛磺酸新的衍生方法,结合二维液相色谱体系精确定量。奶粉基质经超纯水溶解,亚铁氰化钾和乙酸锌沉淀蛋白,用高质量浓度30 mg/mL丹磺酰氯衍生,有机碱三乙胺调节衍生环境,60 ℃水浴加热1 h。衍生液在Agilent XDB-C18色谱柱上,以甲醇和0.1%三氟乙酸为一维流动相分离其余氨基酸,锁定牛磺酸的出峰时间,通过柱切换技术,经捕集和中心切割后,被切换进入Thermal AccucoreTM PFP色谱柱,以乙腈和0.1%甲酸为二维流动相进行梯度洗脱,用荧光检测器进行定量分析。研究不同的衍生液浓度,衍生液pH值,衍生时间和二维液相色谱条件对牛磺酸回收率的影响,优化其条件。所建立的方法确保牛磺酸在1.0~20.0 mg/L范围线性良好,线性相关系数(R2)≥0.9996,对企业提供的实际样品平均回收率为95.2%,相对标准偏差为1.86%。本试验方法准确性高,抗干扰能力强,克服了全氨基酸水解配方奶粉中其余氨基酸对目标物衍生的影响,并提升了衍生的效率。解决了国标无法准确定量特殊医学用途奶粉中牛磺酸的含量,被AOAC委员会认可为检测特殊奶粉基质中牛磺酸含量的推荐性方法。

    Abstract:

    A method for the determination of taurine in amino-acid-hydrolyzed formula milk powder with special medical purpose was developed using a novel derivative way, coupled with two-dimensional liquid chromatography (2D-LC) system on precise quantification. The milk powder matrixes were dissolved with ultra pure water, precipitated proteinaceous substance by potassium ferrocyanide and zinc acetate, derived by high concentration dansylchloride (30 mg/mL), adjusted by triethylamine on pH value, and heated in 60 ℃ water bath for 1 hour. The derivative was performed on the Agilent XDB-C18 column with methanol and 0.1% trifluoroacetic acid as the one-dimensional LC mobile phase, so as to separate other amino-acids and locate the target. Subsequently, taurine entered into the Thermal AccucoreTM PFP column after the capture and center-cutting by column switching technique, acetonitrile and 0.1% formic acid were utilized as the two-dimensional LC mobile phase to gradient elute the taurine, combined with fluorescence detector on quantification analysis. The derivative parameters (concentration, pH value and time) and 2D-LC condition were optimized to promote the target recovery. Taurine showed good linear ranged from 1.0-20.0 mg/L, while the correlation coefficients(R2) was higher than 0.9996. The average recovery of the actual samples was 95.2%, with RSD 1.86%. This method, with high accuracy and strong anti-interference ability, overcame the influence of other amino acids on target derivation, and improved the derivative efficiency. This method solves the problem that national standard cannot accurately quantify the content of taurine in those milk powder with special medical purpose, and has been recognized by AOAC committee as the recommended method on taurine detection of those milk powder with complicated matrix.

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卞华,翁史昱,王亮.基于二维液相色谱体系分析特殊医学用途奶粉中的牛磺酸含量[J].中国食品学报,2022,22(3):288-296

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  • 收稿日期:2021-03-19
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  • 在线发布日期: 2022-04-13
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