Abstract
Peptide interaction is normally monitored by liquid chromatography (LC), liquid chromatography coupled to mass spectrometry (LC-MS), mass spectrometric (MS) methods such as MALDI-TOF/MS or capillary electrophoresis (CE). These analytical techniques need to apply either high pressure or high voltages, which can cause cleavage of newly formed bondages. Therefore, near infrared reflectance spectroscopy (NIRS) is presented as a rapid alternative to monitor the interaction of glutathione and oxytocin, simulating physiological conditions. Thereby, glutathione can act as a nucleophile with oxytocin forming four new conjugates via a disulphide bondage. Liquid chromatography coupled to UV (LC-UV) and mass spectrometry via an electrospray ionisation interface (LC-ESI-MS) resulted in a 82% and a 78% degradation of oxytocin at pH 3 and a 5% and a 7% degradation at pH 6.5. Capillary electrophoresis employing UV-detection (CE-UV) showed a 44% degradation of oxytocin. LC and CE in addition to the NIRS are found to be authentic tools for quantitative analysis. Nevertheless, NIRS proved to be highly suitable for the detection of newly formed conjugates after separating them on a thin layer chromatography (TLC) plate. The recorded fingerprint in the near infrared region allows for a selective distinct qualitative identification of conjugates without the need for expensive instrumentation such as quadrupole or MALDI-TOF mass spectrometers. The performance of the established NIRS method is compared to LC and CE; its advantages are discussed in detail.
Similar content being viewed by others
References
Alexander T, Tran DC (2001) Near-infrared spectrometric determination of di- and tripeptides synthesized by a combinatorial solid-phase method. Anal Chem 73:1062–1067
Amerov AK, Chen J, Small GW, Arnold MA (2005) Scattering and absorption effects in the determination of glucose in whole blood by near-infrared spectroscopy. Anal Chem 77:4587–4594
Axon TG, Brown R, Hammond SV, Maris SJ (1998) Focusing near infrared spectroscopy on the business objectives of modern pharmaceutical production. J Near Infrared Spectrosc 6:A13–A19
Barabás B (1998) A simple testing prodecure for near infrared instruments. J Near Infrared Spectrosc 6:A163–A170
Bokobza L (1998) Near infrared spectroscopy. J Near Infrared Spectrosc 6:3–17
Burns DA, Ciurczak EW (1992) Handbook of near-infrared analysis. Marcel Dekker, New York
Camera E, Picardo M (2002) Analytical methods to investigate glutathione and related compounds in biological and pathological processes. J Chromatogr B 781:181–206
Chen QU, Zhao J, Zhang H, Wang X (2006) Feasibility study on qualitative and quantitative analysis in tea by near infrared spectroscopy with multivariate calibration. Anal Chim Acta 572:77–84
Ciurczak EW, Drennen JK (2002) Pharmaceutical and medical applications of near-infrared spectroscopy. Marcel Dekker, New York
De la Haba MJ, Garrido-Varo A, Guerrero-Ginel JE, Pérez-Marín DC (2006) Near-infrared reflectance spectroscopy for predicting amino acids content in intact processed animal proteins. J Agric Food Chem 54:7703–7709
Delpy DT, Cope M (1997) Quantification in tissue near-infrared spectroscopy. Phil Trans R Soc Lond B 352:649–659
Esbensen KH (2001) Multivariate data analysis in practice, 5th edn. Aalborg University, Esbjerg
Fernández-Ahumada E, Garrido-Varo A, Guerrero-Ginel JE, Wubbels A, Van der Sluis C, Van der Meer JM (2006) Unterstanding factors affecting NIR-analysis of potato constituents. J Near Infrared Spectrosc 14:27–35
Forina M, Casolino MC, de la Pezuela Martinez C (1998) Multivariate calibration: applications to pharmaceutical analysis. J Pharm Biomed Anal 18:21–33
Guggenbichler W, Huck CW, Kobler A, Popp M, Bonn GK (2006) Near infrared spectroscopy, cluster and multivariate analysis—contributions to wine analysis. J Food Agri Environ 4(2):98–106
Heigl N, Huck CW, Rainer M, Ul-Haq N, Bonn GK (2006) Near infrared spectroscopy, cluster and multivariate analysis hyphenated to thin layer chromatography for the analysis of amino acids. Amino Acids 31:45–53
Herschel W (1800) Investigation of the powder of the prismatic colours to heat and illuminate objects. Phil Trans 255–283
Huck CW, Maurer R, Popp M, Basener N, Bonn GK (1999) Quantitative Fourier transform near infrared reflectance spectroscopy (NIRS) compared to high performance liquid chromatography (HPLC) of a flavone in Primulae veris Flos extracts. Pharm Pharmacol Lett 9(1):26–29
Huck CW, Guggenbichler W, Bonn GK (2005) Analysis of caffeine, theobromine and theophylline in coffee by near infrared spectroscopy (NIRS) compared to high-performance liquid chromatography (HPLC) coupled to mass spectrometry. Anal Chim Acta 538(1–2):195–203
Huck CW, Pezzei V, Schmitz T, Bonn GK, Bernkop-Schnürch A (2006) Oral peptide delivery: are there remarkable effects on drugs through sulfhydryl conjugation? J Drug Target 14(3):117–25
Lauridsen RK, Everland H, Nielsen FL, Engelsen SB, Norgaard L (2003) Exploratory multivariate spectroscopic study on human skin. J Chromatogr 9:137–146
McIntosh LM, Summers R, Jackson M, Mantsc HH, Mansfield JR, Howlett M, Crowson AN, Toole JWP (2001) Towards non-invasive screening of skin lesions by near-infrared spectroscopy. J Invest Dermatol 116:175–181
Miyazawa M, Sonoyama M (1998) Second derivative near infrared studies on the structural characterization of proteins. J Near Infrared Spectrosc 6:A253–A257
Morgenstern I, Kirch W (2003) Das glutathionsystem im menschlichen gastrointestinaltrakt. Dtsch Med Wochenschr 128:507–511
Pezzaniti JL, Jeng TW, McDowell L, Oosta GM (2001) Preliminary investigation of near-infrared spectroscopic measurements of urea, creatinine glucose, protein, and ketone in urine. Clin Biochem 34:239–246
Reich G (2003) Near infrared spectroscopy and imaging: basic principles and pharmaceutical applications. Adv Drug Del Rev 57:1109–1143
Schmitz T, Huck CW, Bernkop-Schnürch A (2006) Characterisation of the thiol-disulphide chemistry of desmopressin by LC, μ-LC, LC-ESI-MS and Maldi-Tof. Amino Acids 30(1):35–42
Siesler HW, Ozaki Y, Kawata S, Heise HM (2002) Near-infrared spectroscopy principles, instruments, applications. Wiley–VCH, Germany
Sissons M, Osborne B, Suissons S (2006) Application of near infrared reflectance spectroscopy to a durum wheat breeding programme. J Near Infrared Spectrosc 14:17–25
Shaw RA, Kotowich S, Mantsch HH, Leroux M (1996) Quantitation of protein, creatinine, and urea in urine by near-infrared spectroscopy. Clin Biochem 29(1):11–19
Shaw RA, Mansfield JR, Kupriyanov VV, Mantsch HH (2000) In vivo optical/near-infrared spectroscopy and imaging of metalloproteins. J Inorg Biochem 79:285–293
Sowa MG, Kohlenberg E, Payettw JR, Leonardi L, Levasseur MA, Riley CB (2006) Detecting intestinal ischemia using near infrared spectroscopy. J Near Infrared Spectrosc 14:1–7
Tran DC, Kong X (2000) Near-infrared spectrophotometric determiantion of tri- and tetrapetides. Anal Biochem 286:67–74
Acknowledgments
The Austrian Nano-Initiative co-financed this work as part of the Nano-Health project (no. 0200), the sub-project NANO-N-0204 being financed by the Austrian FWF (Fonds zur Förderung der Wissenschaftlichen Forschung) (Project no. N-0204-NAN).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Petter, C.H., Heigl, N., Bachmann, S. et al. Near infrared spectroscopy compared to liquid chromatography coupled to mass spectrometry and capillary electrophoresis as a detection tool for peptide reaction monitoring. Amino Acids 34, 605–616 (2008). https://doi.org/10.1007/s00726-007-0014-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-007-0014-5