柱后衍生阳离子交换色谱法同时测定大蒜中18种水解氨基酸
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摘要
目的:建立一种柱后衍生阳离子交换色谱法同时测定大蒜中18种氨基酸含量的方法。方法:采用线性梯度洗脱,进样量为50μL,于440nm(脯氨酸)和570nm(其余氨基酸)波长下进行柱后衍生化检测分析,根据色谱分离效果选定最佳检测方法,对比并计算大蒜中18种水解氨基酸与游离氨基酸的含量。结果:在一定的浓度范围内,各种氨基酸的线性关系良好,相关系数r均大于0.9977,其中谷氨酸的含量最高,胱氨酸的含量最低。结论:水解氨基酸色谱分离情况优于游离氨基酸,方法准确度和灵敏度高,具有良好的重复性和回收率,可选用该法对大蒜中多种氨基酸进行同时测定。
Objective: To establish a method of simultaneous determination of 18 kinds of amino acid in garlic by post-column derivatization cation exchange chromatography. Methods: Linear gradient elution was used, and detection and analysis of postcolumn derivatization was performed, injection volume was 50 μL, detection wavelength was in the 440 nm(proline) and 570 nm(the rest of the amino acids). The content of 18 hydrolytic amino acid and free amino acid in garlic samples were compared and calculated, the best determination method was selected according to the chromatographic separation effect. Result: In a certain concentration range, all kinds of amino acids were in good linear relationship. r≥0.9977, with the highest content of glutamic acid and lowest content of cystine. Conclusion: Chromatographic separation of hydrolytic amino acid is better than free amino acid, with high accuracy and sensitivity, good repeatability and recovery rate, which can be used for simultaneous determination of a variety of amino acids in garlic.
Objective: To establish a method of simultaneous determination of 18 kinds of amino acid in garlic by post-column derivatization cation exchange chromatography. Methods: Linear gradient elution was used, and detection and analysis of postcolumn derivatization was performed, injection volume was 50 μL, detection wavelength was in the 440 nm(proline) and 570 nm(the rest of the amino acids). The content of 18 hydrolytic amino acid and free amino acid in garlic samples were compared and calculated, the best determination method was selected according to the chromatographic separation effect. Result: In a certain concentration range, all kinds of amino acids were in good linear relationship. r≥0.9977, with the highest content of glutamic acid and lowest content of cystine. Conclusion: Chromatographic separation of hydrolytic amino acid is better than free amino acid, with high accuracy and sensitivity, good repeatability and recovery rate, which can be used for simultaneous determination of a variety of amino acids in garlic.
引文
[[1]马丽娜,李峰杰,陈坚,等.大蒜主要活性成分及药理作用研究进展[J].中国药理学通报,2014,(6):760-763.
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[4] Marchese A, Barbieri R. Antifungal and antibacterial activities of allicin:A review[J]. Trends in Food Science and Technology, 2016, 52:49-56.
[5] Panan R, Vorraruthai T. Fresh garlic extract inhibits Staphylococcus aureus biofilm formation under chemopreventive and chemotherapeutic conditions[J]. Songklanakarin Journal of Science and Technology, 2016, 38(4):381-389.
[6] Florinda F, Maria N 0, Autilia C. Phenolic constituents, antioxidant, antimicrobial and antiproliferative activities of different endemic Italian varieties of garlic(Allium sativum L.)[J]. Journal of Functional Foods, 2016,(21):240-248.
[7]赵东升,李新霞,张海波,等.以大蒜辣素为指标研究大蒜质量标准[J].药物分析杂志,2013, 33(9):1587-1592.
[8] Block E. The chemistry of garlic and onions[J].Scientific American, 1985,252(3):114-119.
[9]Ramirez D A, Locatelli D A. Analytical methods for bioactive sulfur compounds in Allium:An integrated review and future directions[J]. Journal of Food Composition and Analysis, 2017:4-19.
[10]赵东升,马晓丽,李新霞,等.柱前衍生-高效液相色谱法同时测定可食药用葱属植物中18种游离氨基酸含量[J].药物分析杂志,2013,(6):963-968.
[11]杨春霞.柱后衍生高效阳离子交换色谱法测定宁夏枸杞及枸杞花粉中氨基酸含量[J].食品科学,2011, 32(18):239-242.
[12] Zheng G H, Jin W W, Fan P. A novel methodfor detecting amino acids derivatized with phenyl isothiocyanate by high-performance liquid chromatography-electrospray ionization mass spectrometry[J]. International Journal of Mass Spectrometry,2015, 392:1-6.
[13]杨卫,鲜殊,李大祥,等.邻苯二甲醛柱前衍生反相高效液相色谱法测定茶叶中17种游离氨基酸[J].茶叶科学,2011, 31(3):211-217.
[14]艾则孜·莫合买提,沙丽娜,巴哈尔古丽·黄尔汗,等.柱后衍生阳离子交换色谱法测定桦菌芝中氨基酸的含量[J].药物分析杂志,2012, 32(11):1972-1975.
[15]张婕,葛永辉,向章敏,等.茚三酮柱后衍生氨基酸分析仪测定卷烟主流烟气中的氨[J].中国烟草,2013,(6):113-116.
[2] Keshab R P. Pharmacological effects of traditional herbal plant garlic(Allium sativum):A review[J].Journal of Kathmandu Medical College. 2015, 3(4):158-161.
[3] Borlinghaus J, Albrecht F, Gruhlke MC, et al.Allicin:Chemistry and Biological Properties[J].Molecules. 2014, 19(8):12591-12618.
[4] Marchese A, Barbieri R. Antifungal and antibacterial activities of allicin:A review[J]. Trends in Food Science and Technology, 2016, 52:49-56.
[5] Panan R, Vorraruthai T. Fresh garlic extract inhibits Staphylococcus aureus biofilm formation under chemopreventive and chemotherapeutic conditions[J]. Songklanakarin Journal of Science and Technology, 2016, 38(4):381-389.
[6] Florinda F, Maria N 0, Autilia C. Phenolic constituents, antioxidant, antimicrobial and antiproliferative activities of different endemic Italian varieties of garlic(Allium sativum L.)[J]. Journal of Functional Foods, 2016,(21):240-248.
[7]赵东升,李新霞,张海波,等.以大蒜辣素为指标研究大蒜质量标准[J].药物分析杂志,2013, 33(9):1587-1592.
[8] Block E. The chemistry of garlic and onions[J].Scientific American, 1985,252(3):114-119.
[9]Ramirez D A, Locatelli D A. Analytical methods for bioactive sulfur compounds in Allium:An integrated review and future directions[J]. Journal of Food Composition and Analysis, 2017:4-19.
[10]赵东升,马晓丽,李新霞,等.柱前衍生-高效液相色谱法同时测定可食药用葱属植物中18种游离氨基酸含量[J].药物分析杂志,2013,(6):963-968.
[11]杨春霞.柱后衍生高效阳离子交换色谱法测定宁夏枸杞及枸杞花粉中氨基酸含量[J].食品科学,2011, 32(18):239-242.
[12] Zheng G H, Jin W W, Fan P. A novel methodfor detecting amino acids derivatized with phenyl isothiocyanate by high-performance liquid chromatography-electrospray ionization mass spectrometry[J]. International Journal of Mass Spectrometry,2015, 392:1-6.
[13]杨卫,鲜殊,李大祥,等.邻苯二甲醛柱前衍生反相高效液相色谱法测定茶叶中17种游离氨基酸[J].茶叶科学,2011, 31(3):211-217.
[14]艾则孜·莫合买提,沙丽娜,巴哈尔古丽·黄尔汗,等.柱后衍生阳离子交换色谱法测定桦菌芝中氨基酸的含量[J].药物分析杂志,2012, 32(11):1972-1975.
[15]张婕,葛永辉,向章敏,等.茚三酮柱后衍生氨基酸分析仪测定卷烟主流烟气中的氨[J].中国烟草,2013,(6):113-116.