在线净化二维液相色谱快速检测食品中维生素A、D、E
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摘要
目的 利用三泵两阀二维液相色谱系统,建立食品中维生素A、D、E在线二维液相色谱快速检测方法。方法 采用中心切割方法搭建在线二维液相色谱分离系统,以高聚合物为填料的具有耐强碱溶液能力的PLRP-S色谱柱作为净化样品皂化液的固相萃叐(solid phase extraction,SPE)柱,以C_18为一维色谱柱分离维生素A和E,以多环芳烃(polycyclic aromatic hydrocarbon, PAH)色谱柱为二维色谱柱分离维生素D2和D3。一维色谱确立维生素D的切割时间,由C_18小柱捕获维生素D,建立在线二维液相色谱分析方法。利用样品和标品液对建立的方法进行方法学验证,同时比较该方法与GB 5009.82-2016对实际样品的测试结果。结果 维生素A、D、E在试验标准曲线范围内线性系数r>0.999,定量限分别为30μg/100g, 2μg/100 g和120μg/100 g,与国标一致;该方法 3水平加标回收率为90.4%～101.7%,可以满足目前食品中维生素A、D、E的检测需求。结论本文建立的测试方法操作简单高效、自动化程度高、准确度高、重复性好,适合于大批量样品的测试。
Objective To establish an on-line two-dimensional liquid chromatographic method for the determination of vitamins A, D and E in food by using a three-pump and two-valve two-dimensional liquid chromatographic system. Methods An on-line two-dimensional liquid chromatography separation system was set up by central cutting method. The PLRP-S column with high polymer as filler which had strong alkali resistance was used as the solid phase extraction(SPE) column for purifying saponification solution of samples. Vitamin A and E were separated by C_18 as one-dimensional column, and vitamin D2 and D3 were separated by polycyclic aromatic hydrocarbon(PAH) as two-dimensional column. One-dimensional chromatography confirmed the cutting time of vitamin D, VD was captured by C_18 column, and then established on-line two-dimensional liquid chromatography analysis method. Samples and standard solutions were used to conduct the validation of the establishe d method, and the test results of actual samples were compared by using established method and GB 5009.82-2016. Results The linear coefficients of vitamin A, D and E were all more than 0.999 in the range of standard curves. The limits of quantitation were 30 μ g/100 g, 2 μ g/100 g and 120 μ g/100 g, which were consistent with the national standard GB5009.82-2016. The recoveries of vitamin A, D and E at 3 levels of this method were all between 90.4% and 101.7%,which could meet the detection requirements of vitamin A, D and E in food. Conclusion The method is simple,efficient, highly automated, accurate and reproducible, which is suitable for the determination of large quantities of samples.
Objective To establish an on-line two-dimensional liquid chromatographic method for the determination of vitamins A, D and E in food by using a three-pump and two-valve two-dimensional liquid chromatographic system. Methods An on-line two-dimensional liquid chromatography separation system was set up by central cutting method. The PLRP-S column with high polymer as filler which had strong alkali resistance was used as the solid phase extraction(SPE) column for purifying saponification solution of samples. Vitamin A and E were separated by C_18 as one-dimensional column, and vitamin D2 and D3 were separated by polycyclic aromatic hydrocarbon(PAH) as two-dimensional column. One-dimensional chromatography confirmed the cutting time of vitamin D, VD was captured by C_18 column, and then established on-line two-dimensional liquid chromatography analysis method. Samples and standard solutions were used to conduct the validation of the establishe d method, and the test results of actual samples were compared by using established method and GB 5009.82-2016. Results The linear coefficients of vitamin A, D and E were all more than 0.999 in the range of standard curves. The limits of quantitation were 30 μ g/100 g, 2 μ g/100 g and 120 μ g/100 g, which were consistent with the national standard GB5009.82-2016. The recoveries of vitamin A, D and E at 3 levels of this method were all between 90.4% and 101.7%,which could meet the detection requirements of vitamin A, D and E in food. Conclusion The method is simple,efficient, highly automated, accurate and reproducible, which is suitable for the determination of large quantities of samples.
引文
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[14]林玉宙,吴宏,郭荣烁,等.二维液相色谱双检测器法同时测定婴幼儿配方奶粉,米粉中维生素A,D3和E的含量[J].食品工业科技,2015,37(20):68-77.Lin YZ,Wu H,Guo RS,et al.Simultaneous determination of vitamin A,D3 and E in infant formula milk powder and rice flour by two-dimensional liquid chromatography/double deterctor[J].Sci Technol Food Ind,2015,37(20):68-77.
[15]Davankov V,Tsyurupa M.Chapter 13-hypercrosslinked polystyrene as column packing material in HPLC[J].Compreh Anal Chem,2011,(56):503-521.
[16]Dzhabieva SA,Kurbatova SV.Sorbent effect on retention of benzotriazoles in RP HPLC[J].Russ J Phys Chem A,2015,89(10):1907-1913.
[17]Sychow CS,Ilyin MM,Davankov VA,et al.Elucidation of retention mechanisms on hypercrosslinked polystyrene used as column packing material for high-performance liquid chromatography[J].J Chromatogr A,2004,1030(1-2):17-24.
[18]Davankov VA,Tsyurupa MP.Structure and properties of porous hypercrosslinked polystyrene sorbents styrosorb[J].Pure Appl Chem,1989,61(11):1881-1888.
[19]Saifutdinov B,Davankov V,Petukhova G,et al.Thermodynamics of liquid-phase adsorption on hypercrosslinked polystyrene networks with ultimate degrees of crosslinking[J].Dokl Phys Chem,2015,462(2):135-139.
[20]Wu D,Nedev GK,Lucy CA.Retention mechanism of hypercrosslinked polystyrene silica hybrid phase in normal phase chromatography[J].JChromatogr A,2014,(1370):50-55.
[2]Finley JW,Demand JM.Principles of food chemistry[Z].
[3]Debelo H,Novotny JA,Ferruzzi MG.Vitamin A[J].Adv Nutr,2017,8(6):992-994.
[4]Fujita M,Rendon M.Nutrition in Infancy:Volume 1[M].New Jersey:Humana Press,2013.
[5]中国营养学会.中国居民膳食营养素参考摄入量[M].北京:北京科学出版社,2013.Chinese Society of Nutrition.Reference intake of dietary nutrients for Chinese residents[M].Beijing:Beijing Science Publishing Press,2013.
[6]Elizabeth DFD,Mesquita-Martiniano LV,Lopes-Lima LL,et al.Acute kidney injury due to excessive and prolonged intramuscular injection of veterinary supplements containing vitamins A,D and E:A series of 16cases[J].Nefrología,2017,37(1):61-67.
[7]Niki E.Role of vitamin E as a lipid-soluble peroxyl radical scavenger:In vitro and in vivo evidence[J].Free Rad Biol Med,2014,66(8):3-12.
[8]Fanali C,D'Orazio G,Fanali S,et al.Advanced analytical techniques for fat-soluble vitamin analysis[J].Trac-Trend Anal Chem,2017,(87):82-97.
[9]郑熠斌.食品中维生素A,D,E的检测方法研究[D].杭州:浙江工业大学,2016.Zheng YB.Study on the analysis methods of vitamin A,Vitamin D and Vitamin E in food[D].Hangzhou:Zhejiang University of Technology,2016.
[10]GB 5009.82-2016食品安全国家标准食品中维生素A,D,E的测定[S].GB 5009.82-2016 National food safety standard-Determination of Vitamin A,D,E in food[S].
[11]Woollard DC,Bensch A,Indyk H,et al.Determination of vitamin A and vitamin E esters in infant formulae and fortified milk powders by HPLC:Use of internal standardization[J].Food Chem,2016,197(A):457-465.
[12]张艳海,张大伟,曹莹,等.在线二维反相色谱法快速测定维生素AD制剂中维生素A、D的含量[J].分析测试学报,2016,35(1):28-34.Zhang YH,Zhang DW,Cao Y,et al.Simultaneous determination of Vitamins A,D in Vitamin A and D pharmaceutical by online two-dimensional reverse-phase liquid chromatography[J].J Instrum Anal,2016,35(1):28-34.
[13]Plozza T,Trenerry VC,Caridi D.The simultaneous determination of vitamins A,E andβ-carotene in bovine milk by high performance liquid chromatography-ion trap mass spectrometry(HPLC-MSn)[J].Food Chem,2012,134(1):559-563.
[14]林玉宙,吴宏,郭荣烁,等.二维液相色谱双检测器法同时测定婴幼儿配方奶粉,米粉中维生素A,D3和E的含量[J].食品工业科技,2015,37(20):68-77.Lin YZ,Wu H,Guo RS,et al.Simultaneous determination of vitamin A,D3 and E in infant formula milk powder and rice flour by two-dimensional liquid chromatography/double deterctor[J].Sci Technol Food Ind,2015,37(20):68-77.
[15]Davankov V,Tsyurupa M.Chapter 13-hypercrosslinked polystyrene as column packing material in HPLC[J].Compreh Anal Chem,2011,(56):503-521.
[16]Dzhabieva SA,Kurbatova SV.Sorbent effect on retention of benzotriazoles in RP HPLC[J].Russ J Phys Chem A,2015,89(10):1907-1913.
[17]Sychow CS,Ilyin MM,Davankov VA,et al.Elucidation of retention mechanisms on hypercrosslinked polystyrene used as column packing material for high-performance liquid chromatography[J].J Chromatogr A,2004,1030(1-2):17-24.
[18]Davankov VA,Tsyurupa MP.Structure and properties of porous hypercrosslinked polystyrene sorbents styrosorb[J].Pure Appl Chem,1989,61(11):1881-1888.
[19]Saifutdinov B,Davankov V,Petukhova G,et al.Thermodynamics of liquid-phase adsorption on hypercrosslinked polystyrene networks with ultimate degrees of crosslinking[J].Dokl Phys Chem,2015,462(2):135-139.
[20]Wu D,Nedev GK,Lucy CA.Retention mechanism of hypercrosslinked polystyrene silica hybrid phase in normal phase chromatography[J].JChromatogr A,2014,(1370):50-55.