Analysis of Peptides in Plasma by High Performance Liquid Chromatography Coupled with Electrospray Ionization Mass Spectrometry

Abstract:

In this study, a high performance liquid chromatography coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) method for the determination of eight endogenous peptides, which included angiotensin-II, neurotensin, 5-met-enkephalin, 5-leu-enkephalin, somatostatin, bradykinin, dynorphin A and oxidized glutathione, was developed. MS parameters were optimized in order to obtain the best signal intensity. The optimum MS parameters were: fragmentor: 100 V; nebulizer gas (N²) pressure: 40 psi; drying gas (N²) flow rate: 10 L/min; drying gas temperature: 350 °C; capillary voltage: 2000 V. Validation of the method showed positive results. The correlation coefficients (R²) between the peak areas and the corresponding concentrations of peptides were greater than 0.9903. The limits of detection (LODs) (S/N = 3) of the eight peptides were 3.88 ng/mL ~ 17.5 ng/mL. The developed method had been successfully applied to analyzing the human plasma fortified with the eight peptides, which indicated the potential of the methodology for the analysis of biological sample.

• Periodical: Advanced Materials Research (Volume 340)
• Pages: 273-279
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.340.273
• Online since: September 2011
• Authors: Hai Yan Fang, Yin Liu, Shu Fang Wang
• Keywords: Endogenous Peptides, LC-ESI-MS, Plasma

References

[1] P. Schulz-Knappe, H.D. Zucht, G. Heine, M. Jurgens, R. Hess, and M. Schrader, Peptidomics: the comprehensive analysis of peptides in complex biological mixtures, Comb. Chem. High. Throughput. Screen, vol. 4, no. 2, pp.207-217, (2001).

DOI: 10.2174/1386207013331246

[2] R.S. Tirumalai, et al, Characterization of the low molecular weight human serum proteome, Mol. Cell Proteomics, vol. 2, no. 10, pp.1096-1103, (2003).

[3] M. Soloviev, and P. Finch, Peptidomics: bridging the gap between proteome and metabolome, Proteomics, vol. 6, no. 3, pp.744-747, (2006).

DOI: 10.1002/pmic.200500878

[4] L.P. Aristoteli, M.P. Molloy, and M.S. Baker, Evaluation of endogenous plasma peptide extraction methods for mass spectrometric biomarker discovery, J. Proteome. Res, vol. 6, no. 2, pp.571-581, (2007).

DOI: 10.1021/pr0602996

[5] G.B. Martin, et al, CE-MS method development for peptides analysis, especially hepcidin, an iron metabolism marker, Electrophoresis, vol. 30, no. 15, pp.2624-2631, (2009).

DOI: 10.1002/elps.200800794

[6] I.S. Zagon, W.J. Hurst, and P.J. McLaughlin, Identification of [Met5]-enkephalin in developing, adult, and renewing tissues by reversed-phase high performance liquid chromatography and radioimmunoassay, Life Sci, vol. 61, no. 4, pp.363-370, (1997).

DOI: 10.1016/s0024-3205(97)00393-7

[7] H. Harbach, et al, Beta-endorphin (1-31) in the plasma of male volunteers undergoing physical exercise, Psychoneuroendocrinology, vol. 25, no. 6, pp.551-562, (2000).

DOI: 10.1016/s0306-4530(00)00009-3

[8] E. Hernandez, F. Benavente, V. Sanz-Nebot, and J. Barbosa, Analysis of opioid peptides by on-line SPE-CE-ESI-MS, Electrophoresis, vol. 28, no. 21, pp.3957-3965, (2007).

DOI: 10.1002/elps.200700845

[9] A. Elhamili, M. Wetterhall, M. Jodie, R. Sebastiano, and J. Bergquist, Analysis of peptides using N-methylpolyvinylpyridium as silica surface modifier for CE-ESI-MS, Electrophoresis, vol. 31, no. pp.1-6, (2010).

DOI: 10.1002/elps.200900536

[10] M. Lu, L. Zhang, Q. Lu, Y. Chi, and G. Chen, Rapid analysis of peptides and amino acids by CE-ESI-MS using chemically modified fused-silica capillaries, Electrophoresis, vol. 30, no. 13, pp.2273-2279, (2009).

DOI: 10.1002/elps.200800683

[11] J.Z. Yang, K.C. Bastian, R.D. Moore, J.F. Stobaugh, and R.T. Borchardt, Quantitative analysis of a model opioid peptide and its cyclic prodrugs in rat plasma using high-performance liquid chromatography with fluorescence and tandem mass spectrometric detection, J. Chromatogr. B, vol. 780, no. 2, pp.269-281, (2002).

DOI: 10.1016/s1570-0232(02)00536-6

[12] C.T. Mant, et al, HPLC analysis and purification of peptides, Methods Mol Biol, vol. 386, no. pp.3-55, (2007).

[13] J.M. Saz, and M.L. Marina, Application of micro- and nano-HPLC to the determination and characterization of bioactive and biomarker peptides, J Sep Sci, vol. 31, no. 3, pp.446-458, (2008).

DOI: 10.1002/jssc.200700589

[14] R. Rodriguez-Gonzalez, O. Hurtado, T. Sobrino, and J. Castillo, Neuroplasticity and cellular therapy in cerebral infarction, Cerebrovasc Dis, vol. 24 Suppl 1, no. pp.167-180, (2007).

DOI: 10.1159/000107393

[15] M. Kobari, N. Ishihara, and K. Yunoki, CSF beta-endorphin and leu-enkephalin levels in the acute and chronic stages of cerebral infarction, J Neurol, vol. 234, no. 5, pp.289-291, (1987).

DOI: 10.1007/bf00314282

[16] H. Cramer, and K.M. Heitzelmann, Somatostatin in cerebrospinal fluid after generalised convulsions or cerebral infarction in humans, J Neurol Neurosurg Psychiatry, vol. 53, no. 11, pp.1015-1016, (1990).

DOI: 10.1136/jnnp.53.11.1015-a

[17] W. Zhou, and B.R. Yu, Serial Observation of Plasma Endothelin, RAS and Calcitonin Gene-related Peptide with Cerebral Infarction, Tong Ji Yi Ke Dab Xue Xue Bao, vol. 26, no. 3, pp.195-198, (1997).

[18] H.M. Zhang, D.Y. Zhang, and E.Q. Lang, The change of plasma β EP and DynA 1-13 in patients with cerebral infarction, Zhong Feng Yu Shen Jing Ji Bing Za Zhi, vol. 14, no. 3, pp.145-147, (1997).

[19] Y.H. Guo, et al, The change of red blood cells GSH and MDA in different periods with Acute cerebral infarction, Shang Hai Yi Xue Jian Yan Za Zhi, vol. 14, no. 2, pp.72-73, (1999).

[20] S.Y. Lv, M.Z. Zhu, Y.M. Zang, and S.Y. Chen, Peripheral endogenous opioid peptides on the regulation of cardiovascular function, Xin Gong Neng Za Zhi, vol. 11, no. 2, pp.106-109, (1999).

[21] Q. Chen, S.W. Xue, N.P. Luo, X.W. Zhang, and J. Xu, Levels of detection and clinical research of plasma and platelet neuropeptide and neurotensin in elderly patients with cerebral infarction, He Nan Yi Yao Xin Xi, vol. 8, no. 11, pp.19-20.