Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
권호 표지

Reverse-Phase High Performance Liquid Chromatographic Determination of Amino Acids after Precolumn Derivatization with 1-Dimethyl-aminonaphthalene-5-sulfonyl chloride

Dansyl 유도체화와 역상 고성능액체크로마토그래피에 의한 아미노산의 정량

  • 나혜경 (전남대학교 식품영양학과) ;
  • 전덕영 (전남대학교 식품영양학과) ;
  • 홍윤호 (전남대학교 식품영양학과)
  • Published : 1993.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

This experiment was carried out to determine the separation condition of 1-dimethylaminonaphthalene-5-sulfony(Dansyl) derivatives of amino acids by reverse-phase high performance liquid chromatography with Nova-Pak C18 column. Determined solvent system was solvent A(200mA phosphate buffer pH 6.8 15%, acetonitrile 11%, water 74%) and solvent B(acetonitrile 65%, methanol 28%, water 7%). Linear gradient of solvent B was applied from 12% to 80% for 50min. Complete separation of 20 amino acids including asparagine and glutamine which constitute protein was achieved within 50min. As the detection limit was the range of picomole, the resolution power was excellent. Reproducibility of the retention time was less than mean $\pm$0.05min. According to the above optimum chromatographic conditions, the amino acid composition of some food and human blood was examined. The most affluent amino acid was alanine in human blood, aspartic acid and glutamic acid in soy sauce, alanine and threonine in soy milk and proline in milk and yoghurt.

Keywords

References

  1. Anal. Chem. v.59 Optical activity and ultraviolet absorbance detection of dansyl Lamino acids separated by gradient liquid chromatography Roach,M.C.;Ungar,L.W.;Zare,R.N.;Reimer,L.M.;Pompliano,D.L.;Frost,J.W.
  2. Food Technology v.6 Quantitative determination of amino acids in food by high-pressure liquid chromatography Khayat,A.;Redenz,P.K.;Gorman,L.A.
  3. Am. J. Clin. Path. v.87 no.5 HPLC qualitative amino acid analysis in the clinical laboratories Rutledge,J.C.;Rudy,J.
  4. J. Chromatogr. v.145 Determination of di- and polyamins by high performance liquid chromatographic separation of their 5-dimethylamino naphthalene-1-sulfonyl derivatives Seiler,N.;Knodgen,B.
  5. J. Chromatogr. v.145 Sensitive fluorimetric method for the determination of putrescine, spermidine and spermine by high performance liquid chromatography and it's application to human blood Saeki,Y.;Uehara,N.;Shirakawa,S.
  6. J. Chromatogr. v.286 High-performance liquid chromatography of Dns-amino acids in the purity control of peptides Levina,N.B.;Nazimov,L.V.
  7. J. Nutrition v.118 In vivo utilization of cystine containing synthetic short-chain peptides after intravenous bolus injection in the rat Stehle,P.;Albers,S.;Pollack,L.;Furst,P.
  8. J. Chromatogr. v.205 Practical considerations in the chiral separation of Dns-amino acids by reversed-phase liquid chromatography using metal chelate additives Tapuhi,Y.;Miller,N.;Karger,B.L.
  9. J. Chromatogr. Sci. v.22 Stereoselective analysis of D and L dansyl amino acids as the mixed chelate copper(Ⅱ) complexes by HPLC Lam,S.
  10. Anal. Biochem. v.115 Dansylation of amino acids high-performance liquid chromatography analysis Tapuhi,Y.;Schmidt,D.E.;Lindner,W.;Karger,B.L.
  11. Methods in enzymology v.11 Dansyl chloride procedure Gray,W.R.;Hirs,C.H.(ed.)
  12. Chemistry and biochemistry of the amino acids Liquid chromatography of amino acids and their derivaties Perrett,D.;Barrett,G.C.(ed.)
  13. Method in enzymology v.25 End-group analysis using dansyl chloride Gray,W.R.;Hirs,C.H.W.(ed.);Timasheff,S.M.(ed.)
  14. J. Chromatogr. v.380 Determination of 27 dansyl amino acid derivatives in biological fluids by reverse-phase high-performance liquid chromatography Marquez,F.J.;Quesada,A.R.;Jimenez,S.;Nunez de Castro,I.
  15. J. Chromatogr. v.295 Micro high-performance liquid chromatography of 5-dimethylaminonaphthalenesul phonyl-amino acids Takeuchi,T.;Yamazaki,M.;Ishii,D.
  16. Anal. Biochem. v.160 The use of reverse-phase high-performance liquid chromatography and precolumn derivatization with dansyl chloride for quantitation of specific amino acids in collagen and elastin Negro,A.;Garbisa,S.;Gotte,L.;Spina,M.
  17. The peptides v.182 Amino acid analysis of peptides Benson,J.R.;Louie,P.C.;Bradshaw,R.A.;Gross,E.(ed.);Mezonehofer,J.(ed.)
  18. Methods in enzymology v.182 Amino acid analysis Ozols,J.;Deutscher,M.P.(ed.)
  19. J. Chromatogr. v.240 Quantitative high-performance liquid chromatographic determination of sulfur amino acids in protein hydrolysates Mackey,L.N.;Beck,T.A.
  20. J. Biol. Chem. v.261 no.23 Amino acid sequence of Escherichia coli glutamine synthetase deduced from the DNA nucleotide sequence Colombo,G.;Villafranca,J.J.
  21. J. Biol. Chem. v.256 no.24 Purification, physical characterization, and NH₂-terminal sequence of glutamine synthetase from the Cyanobacterium anabaena 7120 Orr,J.;Keefer,L.M.;Keim,P.;Nguyen,T.D.;Wellems,T.;Heinrikson,R.L.;Haselkorn,R.
  22. J. Chromatogr. v.499 Appraisal of four pre-column derivatization methods for the high performance liquid chromatographic determination of free amino acids biological materials Furst,P.;Pollack,L.;Graser,T.A.;Godel,H.;Stehle,P.
  23. 대한가정학회지 v.24 no.4 시판 및 재래식 간장의 일반성분 및 아미노산 성분 연구 고영수;전명진
  24. 대한가정학회지 v.24 no.4 모유 및 우유의 아미노산 조성에 관한 비교 연구 고영수;장정옥
  25. J. Pediatrics v.78 no.4 Infusion of protein hydrolysates in the newborn infant plasma amino acid concentrations Stegink,L.D.;Baker,G.L.
  26. Food Chemistry v.17 Development of soy milk-A review Ang,H.G.;Kwik,W.L.
  27. 식품과학 v.14 no.1 Soybean milk의 품질관리 김오섭
  28. Yoghurt science and technology Tamime,A.Y.;Robinsion,R.K.
  29. Milk and dairy products in human nutrition Renner,E.
  30. Yoghurt Rasic,J.L.;Kurmann,J.A.