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Determination of Acetyl-Tryptophan in Human Albumin by Reversed-Phase High Performance Liquid Chromatography
Corresponding Author(s) : Li Mao
Asian Journal of Chemistry,
Vol. 27 No. 6 (2015): Vol 27 Issue 6
Abstract
A reliable reversed-phase high performance liquid chromatography combined with perchloric acid deproteinization is presented for the determination of acetyl-tryptophan in human albumin. HPLC was achieved on an Accucore XL-C18 column by gradient elution of acetonitrile and phosphate buffered solution (pH 2.3) at a flow rate of 0.7 mL min–1 and detection was by UV adsorption at 220 nm. The standard curve of acetyl-tryptophan was linear over the range of 1-60 μg mL–1. The limits of quantitation and detection for acetyl-tryptophan were 0.167 and 0.050 μg mL–1, respectively. The recoveries of acetyl-tryptophan in human albumin were 90.5-96.8 % with the relative standard deviations (n = 3) of 1.4-4.3 %. This method was found to be superior to the current UV method and other liquid chromatographic methods.
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- Human Albumin, United States Pharmacopoeia, The National Formulary, Official Monographs, USP 32, NF27, p. 1436 (2009).
- Human Albumin Solution, European Pharmacopoeia, The Council of Europe, 67075 Strasbourg Cedex, France, edn 6, p. 2057 (2008).
- National Commission of Chinese Pharmacopoeia, Pharmacopoeia of the People’s Republic of China, China Medical Science and Technology Press, Beijing, Vol. 3, p. 408 (2010).
- M.W. Yu and J.S. Finlayson, Vox Sang., 47, 28 (1984); doi:10.1111/j.1423-0410.1984.tb01558.x.
- M.W. Yu and J.S. Finlayson, J. Pharm. Sci., 73, 82 (1984); doi:10.1002/jps.2600730122.
- L.Q. Fang, R. Parti and P.F. Hu, J. Chromatogr. A, 1218, 7316 (2011); doi:10.1016/j.chroma.2011.08.044.
- W.R. Xu and Y.F. Wei, Chinese Pharmacol. Bull., 12, 143 (1996).
- X.P. Wei, J. Wang and L.J. Fan, Chinese J. Biologicals, 23, 536 (2010).
- P.F. Rao, S.T. Liu, R.Z. Zhang, G.R. Chen, Y.Q. Zheng and B.H. Shi, Chinese J. Chromatogr., 15, 193 (1997).
- H.J.C.F. Nelis, M.F. Lefevere, E. Baert, W. D'Hoore and A.P. de Leenheer, J. Chromatogr. A, 333, 381 (1985); doi:10.1016/S0021-9673(01)87367-2.
- J. Blanchard, J. Chromatogr. B, 226, 455 (1981); doi:10.1016/S0378-4347(00)86080-6.
- Tryptophan, European Pharmacopoeia, The Council of Europe, 67075 Strasbourg Cedex, France, edn 6, p. 3142 (2008).
References
Human Albumin, United States Pharmacopoeia, The National Formulary, Official Monographs, USP 32, NF27, p. 1436 (2009).
Human Albumin Solution, European Pharmacopoeia, The Council of Europe, 67075 Strasbourg Cedex, France, edn 6, p. 2057 (2008).
National Commission of Chinese Pharmacopoeia, Pharmacopoeia of the People’s Republic of China, China Medical Science and Technology Press, Beijing, Vol. 3, p. 408 (2010).
M.W. Yu and J.S. Finlayson, Vox Sang., 47, 28 (1984); doi:10.1111/j.1423-0410.1984.tb01558.x.
M.W. Yu and J.S. Finlayson, J. Pharm. Sci., 73, 82 (1984); doi:10.1002/jps.2600730122.
L.Q. Fang, R. Parti and P.F. Hu, J. Chromatogr. A, 1218, 7316 (2011); doi:10.1016/j.chroma.2011.08.044.
W.R. Xu and Y.F. Wei, Chinese Pharmacol. Bull., 12, 143 (1996).
X.P. Wei, J. Wang and L.J. Fan, Chinese J. Biologicals, 23, 536 (2010).
P.F. Rao, S.T. Liu, R.Z. Zhang, G.R. Chen, Y.Q. Zheng and B.H. Shi, Chinese J. Chromatogr., 15, 193 (1997).
H.J.C.F. Nelis, M.F. Lefevere, E. Baert, W. D'Hoore and A.P. de Leenheer, J. Chromatogr. A, 333, 381 (1985); doi:10.1016/S0021-9673(01)87367-2.
J. Blanchard, J. Chromatogr. B, 226, 455 (1981); doi:10.1016/S0378-4347(00)86080-6.
Tryptophan, European Pharmacopoeia, The Council of Europe, 67075 Strasbourg Cedex, France, edn 6, p. 3142 (2008).