Optimization of HPLC Method for Determination of Lactulose in Liquid Milk
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摘要: 为了对液态乳中乳果糖含量进行简便准确地定量，改进高效液相色谱法测定乳果糖含量的方法。采用化学稳定的改性酰胺基色谱柱进行分离，并以乙腈-水流动相梯度洗脱40 min，结果表明，样品进样体积为3 μL时乳糖与乳果糖的分离度为1.5，满足色谱分离检测。优化后的方法经验证，乳果糖在50~2500 mg/kg范围内线性关系良好，（R2=0.9997），乳果糖检出限和定量限分别为15和50 mg/kg，3个加标水平（n=6）乳果糖平均回收率在90.3%~105.6%之间，相对标准偏差为3.4%~3.9%。利用该法对市场中的27种实际乳样进行分析，结果均在合理范围之内。该方法经济、准确适用于液态乳中乳果糖含量的日常检测。Abstract: In order to quantify the content of lactulose in liquid milk easily and accurately,the HPLC method for determination of lactulose in liquid milk was improved. The method used stable modified amide-based chromatographic column for separation,and eluted with acetonitrile-water mobile phase for 40 minutes gradiently. The results indicated that when the injection volume was 3 μL,the resolution was 1.5,which met the chromatographic separation detection. The optimized method has been verified that lactulose has a good linear relationship in the range of 50~2500 mg/kg,(R2=0.9997),the detection limit and the quantitative limit of lactulose were 15 mg/kg and 50 mg/kg respectively. The spiked recoveries of lactulose were in the range of 90.3%~105.6%,and relative standard deviations(n=6)in the range of 3.4%~3.9%. The proposed method was used to detect 27 actual milk samples,and the results were within a reasonable range. This method was economical and accurate for the daily detection of lactulose in liquid milk.
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Keywords:
- lactulose /
- HPLC /
- modified X-amide column /
- liquid milk
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[1] Andrews G R. Distinguishing pasteurized,UHT and sterilized milks by their lactulose content[J]. International Journal of Dairy Technology,1984,37(3):92-95.
[2] 刘璐璐,张贵斌,徐春伟,等.不同加热条件对牛奶糠氨酸、乳果糖含量的影响[J].现代食品,2019,25(17):68-70. [3] Imène Felfoul,Jardin J,Gaucheron F,et al. Proteomic profiling of camel and cow milk proteins under heat treatment[J]. Food Chemistry,2017,21(6):161-169.
[4] 崔婧,胡建,刘悦,等.液态乳中复原乳检测方法研究[J]. 农产品质量与安全,2018,41(5):80-84. [5] Olano,Agustiín,Corzo N. Lactulose as a food ingredient[J]. Journal of the Science of Food & Agriculture,2010,89(12):1987-1990.
[6] ISO 11868-2007热处理牛奶.乳果糖含量的测定.高效液相色谱法[S]. [7] ISO 11285-2004牛奶.乳糖含量的测定.酶催化方法[S]. [8] NT/T939-2016巴氏杀菌乳和UHT灭菌乳中复原乳的鉴定[S]. [9] 刘洋,岳虹,赵贞,等.高效液相色谱-蒸发光散射检测法与酶法检测UHT灭菌乳中乳果糖含量的对比研究[J]. 乳业科学与技术,2017,40(1):19-22. [10] Sakkas L,Moutafi A,Moschopoulou E,et al. Assessment of heat treatment of various types of milk[J]. Food Chemistry,2014,15(9):293-301.
[11] 冯昕韡. 液质联用同时测定乳产品中的乳糖和乳果糖[C]//中国化学会质谱分析专业委员会.第三届全国质谱分析学术报告会摘要集-分会场7:环境与食品安全分析. 中国化学会质谱分析专业委员会:中国化学会,2017:28. [12] Tommaso R I Cataldi,Massimiliano Angelotti,Giuliana Bianco. Determination of mono-and disaccharides in milk and milk products by high-performance anion-exchange chromatography with pulsed amperometric detection[J]. Analytica Chimica Acta,2003,485(1):110-114.
[13] 田瑶. 牛奶中乳果糖的研究及检测方法的开发[D].长沙:湖南农业大学,2017. [14] 宋卿. 高效液相色谱新型分离材料研究[D].昆明:云南师范大学,2009. [15] Silveira M F,Pessoa Masson L M,Pereira Martins J F,et al. Simultaneous determination of lactulose and lactose in conserved milk by HPLC-RID[J]. Journal of Chemistry,2015,18(6):59-67.
[16] Regina Schuster-Wolff-Bühring,Michel R,Hinrichs J. A new liquid chromatography method for the simultaneous and sensitive quantification of lactose and lactulose in milk[J]. Dairy Science & Technology,2011,91(1):27-37.
[17] 刘萌萌,宋戈. 高效液相色谱蒸发光散射检测法测定乳制品中乳果糖[J]. 中国乳品工业,2018,46(3):49-51. [18] NataliaGonzaga,Lycio Shinji Watanabe,Juliana Mareze,et al. Green method using water for lactose and lactulose extraction and determination in milk by high-performance liquid chromatography with refractive index detection[J]. LWT,2019,113.
[19] Jiří Pazourek. Rapid HPLC method for monitoring of lactulose production with a high yield[J]. Carbohydrate Research,2019,484.
[20] Manzi P,Pizzoferrato L. HPLC determination of lactulose in heat treated milk[J]. Food and Bioprocess Technology,2013,6(3):851-857.
[21] 王桂云,苏庆,张伟,等. 氨基色谱柱测定蜂蜜中4种糖方法研究[J]. 蜜蜂杂志,2019,39(6):9-11 [22] Jacopo,Gervasoni,Aniello,et al. Validation of UPLC-MS/MS method for determination of urinary lactulose/mannitol[J]. Molecules(Basel,Switzerland),2018,23(10).
[23] Martinez-Castro I,Calvo M M,Olano A. Chromatographic determination of lactulose[J]. Chromatographia,1987,23(2):132-136.
[24] 姜金斗,周红,张丽宏,等. HPLC法蒸发光散射检测器测定UHT灭菌奶中乳果糖方法的研究[J]. 中国乳品工业,2006(3):51-53. [25] 杨朝辉. 逆流色谱分离过程理论研究及应用[D].长沙:中南大学,2011. [26] 奚海明,孙梅华.分离度对色谱定量分析的影响[J]. 污染防治技术,2006,45(6):49-50. [27] 黄晓林,宋戈,张丽宏,等.乳制品中单糖、双糖和低聚果糖的同时测定[J]. 中国乳品工业,2017,45(11):37-39. [28] 马红燕,康怀彬,李芳,等.食品增稠剂在乳制品加工中的应用[J]. 农产品加工,2016,22(2):57-61. [29] 孟悦,王英哲,田志刚,等.玉米粉制备葡萄糖的糖化工艺优化[J]. 食品研究与开发,2019,40(18):97-101.
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