Chiral chromatographic analysis of amino acids with pre-column derivatization by o-phthalaldehyde: improving the determination of enantiomers using ion-pair reagents
- 作者: Panin N.V.1, Pirogov I.V.1, Guranda D.T.1, Švedas V.K.1
-
隶属关系:
- Lomonosov Moscow State University
- 期: 卷 17, 编号 4 (2025)
- 页面: 103-109
- 栏目: Research Articles
- URL: https://journal-vniispk.ru/2075-8251/article/view/365064
- DOI: https://doi.org/10.32607/actanaturae.27703
- ID: 365064
如何引用文章
详细
The development of effective and accessible methods for the chiral analysis of amino acids is an important scientific and practical necessity. One of the most common and convenient techniques is the chromatographic determination of individual enantiomers of amino acids with preliminary conversion of enantiomers into diastereomers, which can then be separated on conventional achiral columns. We have shown that by adding ion-pair reagents to the eluent and varying their structure, one can regulate the efficiency of a chiral amino acid analysis based on the chromatographic determination and resolution of the diastereomeric isoindoles obtained upon pre-column derivatization of amino acids by o-phthalaldehyde in the presence of N-acetyl-L-cysteine. The use of ion-pair reagents allows one to achieve a better resolution of diastereomeric isoindole peaks, while the analysis time can be reduced by increasing the ionic strength. Hence, adding ion-pair reagents and optimizing the mobile phase composition are useful approaches in the engineering of an amino acid chiral analysis, along with the synthesis of new chiral SH compounds and the choice of stationary phases.
作者简介
Nikolay Panin
Lomonosov Moscow State University
Email: panin@belozersky.msu.ru
Belozersky Institute of Physicochemical Biology
俄罗斯联邦, Moscow, 119234Ivan Pirogov
Lomonosov Moscow State University
Email: ikaesovich@mail.ru
Faculty of Chemistry
俄罗斯联邦, Moscow, 119234Dorel Guranda
Lomonosov Moscow State University
Email: dorel@belozersky.msu.ru
Belozersky Institute of Physicochemical Biology
俄罗斯联邦, Moscow, 119234Vytas Švedas
Lomonosov Moscow State University
编辑信件的主要联系方式.
Email: vytas@belozersky.msu.ru
Belozersky Institute of Physicochemical Biology; Faculty of Bioengineering and Bioinformatics
俄罗斯联邦, Moscow, 119234参考
- Knol MGE, Wulfmeyer VC, Müller RU, Rinschen ММ. Amino acid metabolism in kidney health and disease. Nat Rev Nephrol. 2024;20(12):771–788. doi: 10.1038/s41581-024-00872-8
- Chen J, Cui L, Lu S, Xu S. Amino acid metabolism in tumor biology and therapy. Cell Death Dis. 2024;15(1):42. doi: 10.1038/s41419-024-06435-w
- Gałęzowska G, Ratajczyk J, Wolska L. Determination of amino acids in human biological fluids by high-performance liquid chromatography: critical review. Amino Acids. 2021;53(7):993–1009. doi: 10.1007/s00726-021-03002-x
- Ksenofontov AL, Boyko AI, Mkrtchyan GV, et al. Analysis of free amino acids in mammalian brain extracts. Biochemistry (Mosc). 2017;82(10):1183–1192. doi: 10.1134/S000629791710011X
- Pereira V, Pontes M, Câmara JS, Marques JC. Simultaneous analysis of free amino acids and biogenic amines in honey and wine samples using in loop orthophthalaldehyde derivatization procedure. J Chromatogr A. 2008;1189(1–2):435–443. doi: 10.1016/j.chroma.2007.12.014
- Cuchiaro H, Laurens LML. Total Protein Analysis in Algae via Bulk Amino Acid Detection: Optimization of Amino Acid Derivatization after Hydrolysis with O-Phthalaldehyde 3-Mercaptopropionic Acid (OPA-3MPA). J Agric Food Chem. 2019;67(19):5672–5679. doi: 10.1021/acs.jafc.9b00884
- Hall AE, Moraru CI. Comparative effects of high pressure processing and heat treatment on in vitro digestibility of pea protein and starch. NPJ Sci Food. 2022;6(1):2. doi: 10.1038/s41538-021-00116-0
- Tanwar S, Bhushan R. Enantioresolution of Amino Acids: A Decade’s Perspective, Prospects and Challenges. Chromatographia. 2015;78:1113–1134. doi: 10.1007/s10337-015-2933-8
- Ilisz I, Péter A, Lindner W. State-of-the-art enantioseparations of natural and unnatural amino acids by high-performance liquid chromatography. Trends Anal Chem. 2016;81:11–22. doi: 10.1016/j.trac.2016.01.016
- Grishin DV, Zhdanov DD, Pokrovskaya MV, Sokolov NN. D-amino acids in nature, agriculture and biomedicine. All Life. 2020;13(1):11–22. doi: 10.1080/21553769.2019.1622596
- Uifălean A, Iacobescu M, Salantă LC, Heghes SC, Moldovan RC, Iuga C-A. Biological and Analytical Perspectives on D-Amino Acids in Cancer Diagnosis and Therapy. Pharmaceuticals(Basel). 2025;18(5):705. doi: 10.3390/ph18050705
- Pollegioni L, Kustrimovic N, Piubelli L, Rosini E, Rabattoni V, Sacchi S. d-amino acids: new functional insights. FEBS J. 2025;292(17):4395–4417. doi: 10.1111/febs.70083
- Nemoto A, Horie M, Imai EI, Honda H, Hatori K, Matsuno K. Enantiomeric Excess of Amino Acids in Hydrothermal Environments. Orig Life Evol Biosph. 2005;35(2):167–174. doi: 10.1007/s11084-005-0638-y
- Jiang L, Dziedzic P, Spacil Z, et al. Abiotic synthesis of amino acids and self-crystallization under prebiotic conditions. Sci Rep. 2014;4:6769. doi: 10.1038/srep06769
- Kimura T, Hamase K, Miyoshi Y, et al. Chiral amino acid metabolomics for novel biomarker screening in the prognosis of chronic kidney disease. Sci Rep. 2016;6:26137. doi: 10.1038/srep26137
- Chang X, Zhang Y, Chen X, et al. Gut microbiome and serum amino acid metabolome alterations in autism spectrum disorder. Sci Rep. 2024;14:4037. doi: 10.1038/s41598-024-54717-2
- Lella C, Nestor L, De Bundel D, Vander Heyden Y, Van Eeckhaut A. Targeted Chiral Metabolomics of D-Amino Acids: Their Emerging Role as Potential Biomarkers in Neurological Diseases with a Focus on Their Liquid Chromatography–Mass Spectrometry Analysis upon Chiral Derivatization. Int J Mol Sci. 2024;25(22):12410. doi: 10.3390/ijms252212410
- Nimura N, Kinoshita T. o-Phthalaldehyde-N-acetyl-L-cysteine as a chiral derivatization reagent for liquid chromatographic optical resolution of amino acid enantiomers and its application to conventional amino acid analysis. J Chromatogr A. 1986;352:169–177. doi: 10.1016/S0021-9673(01)83377-X
- García Alvarez-Coque MC, Medina Hernández MJ, Villanueva Camañas RM, Mongay Fernández C. Studies on the formation and stability of isoindoles derived from amino acids, o-phthalaldehyde and N-acetyl-L-cysteine. Anal Biochem. 1989;180(1):172–176. doi: 10.1016/0003-2697(89)90107-3
- Švedas VJ, Galaev IJ, Borisov IL, Berezin IV. The interaction of amino acids with o-phthaldialdehyde: a kinetic study and spectrophotometric assay of the reaction product. Anal Biochem. 1980;101(1):188–195. doi: 10.1016/0003-2697(80)90059-7
- Meyer MW, Meyer VR, Ramseyer S. The kinetics of diastereomeric amino acids with o-phthaldialdehyde. Chirality. 1991;3(6):471–475. doi: 10.1002/chir.530030611
- Brückner H, Westhauser T, Godel H. Liquid chromatographic determination of d- and l-amino acids by derivatization with o-phthaldialdehyde and N-isobutyryl-l-cysteine applications with reference to the analysis of peptidic antibiotics, toxins, drugs and pharmaceutically used amino acids. J Chromatogr A. 1995;711(1):201–215. doi: 10.1016/0021-9673(95)00158-j
- Nimura N, Fujiwara T, Watanabe A, et al. A novel chiral thiol reagent for automated precolumn derivatization and high-performance liquid chromatographic enantioseparation of amino acids and its application to the aspartate racemase assay. Anal Biochem. 2003;315(2):262–269. doi: 10.1016/S0003-2697(02)00705-4
- Buck RH, Krummen K. High-performance liquid chromatographic determination of enantiomeric amino acids and amino alcohols after derivatization with o-phthaldialdehyde and various chiral mercaptans. J Chromatogr. 1987;387:255–265. doi: 10.1016/S0021-9673(01)94529-7
- Guranda DT, Shapovalova IV, Švedas VK. A new N-acyl derivative of (S)-cysteine for quantitative determination of enantiomers of amino compounds by HPLC with a precolumn modification with o-phthalaldehyde. Bioorg Khim. 2004;30(5):451–457. doi: 10.1023/b:rubi.0000043781.72807.f5
- Guranda DT, Kudryavtsev PA, Khimiuk AY, Švedas VK. Efficient enantiomeric analysis of primary amines and amino alcohols by high-performance liquid chromatography with precolumn derivatization using novel chiral SH-reagent N-(R)-mandelyl-(S)-cysteine. J Chromatogr A. 2005;1095(1–2):89–93. doi: 10.1016/j.chroma.2005.07.125
- Chernobrovkin MG, Shapovalova EN, Guranda DT, Kudryavtsev PA, Švedas VK, Shpigun OA. Chiral high-performance liquid chromatography analysis of α-amino acid mixtures using a novel SH reagent-N-R-mandelyl-l-cysteine and traditional enantiomeric thiols for precolumn derivatization. J Chromatogr A. 2007;1175(1):89–95. doi: 10.1016/j.chroma.2007.10.034
- Lkhagva A, Tai HC. Dimethylcysteine (DiCys)/o-Phthalaldehyde Derivatization for Chiral Metabolite Analyses: Cross-Comparison of Six Chiral Thiols. Molecules. 2021;26(24):7416. doi: 10.3390/molecules26247416
- Naghashian-Haghig A, Hemmateenejad B, Shamsipur M. Determination of enantiomeric excess of some amino acids by second-order calibration of kinetic-fluorescence data. Anal Biochem. 2018;550:15–26. doi: 10.1016/j.ab.2018.04.004
- Sychev KS. Practical Guide to Liquid Chromatography. Moscow: Tekhnosfera. 2010:192–197.
- Cecchi T. Ion Pairing Chromatography. Crit Rev Anal Chem. 2008;38(3):161−213. doi: 10.1080/10408340802038882
- Sagi-Kiss V, Li Y, Carey MR, et al. Ion-Pairing Chromatography and Amine Derivatization Provide Complementary Approaches for the Targeted LC-MS Analysis of the Polar Metabolome. J Proteome Res. 2022;21(6):1428−1437. doi: 10.1021/acs.jproteome.2c00030
- Saprykin LV, Saprykina LV. Some aspects of the practical application of dynamic modification in HPLC on silica gel sorbents. Sorption and Chromatographic Processes. 2006;6(2):284–301.
补充文件
