Identity Profiling of Complex Mixtures of Peptide Products by Structural and Mass Mobility Orthogonal Analysis

Abstract

Identity is a critical quality attribute that must be determined before releasing batches of medicinal and dietary products. However, the identities of peptide-derived products composed of a large number of diverse molecules is challenging since most analytical techniques cannot analyze multiple molecules simultaneously. Here, we proposed the determination of the weight-average molecular weight (M w) and polydispersity index (PDI) by mass spectrometry for control quality for the batch release of complex products, namely, glatiramer acetate (Copaxone), collagen hydrolysate (Colagenart), and a human dialyzable leucocyte extract (Transferon). The M w and PDI values were orthogonally determined by PFG-STE-H2O­(presaturation)-DOSY-NMR analysis. To the best of our knowledge, this is the first time that MS and NMR spectra have been combined to determine the PDI of complex products derived from protein hydrolysis that are not monodisperse. The performance of each method was evaluated by comparing the obtained results to those reported for glatiramer acetate using MALLS, the technique commonly employed to determine PDI. This combined approach demonstrates the ability of these techniques to separate peptide populations from complex mixtures to establish their identity through their mass distribution profiles.

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“…Recently, the polydispersity index was determined at 1.11 orthogonally using mass spectrometry and nuclear magnetic resonance. This proves the applicability of high-technology techniques for a characterization that will allows identifying the sequence of the active peptide components in this complex drug (Vázquez-Leyva et al, 2019).…”

“…Estos estudios han mostrado que los péptidos de Transferon ® tienen un peso molecular menor a 10 kDa, son principalmente hidrofílicos y que el aminoácido más abundante es la glicina; estas propiedades son reproducibles entre lotes (Medina-Rivero et al, 2016). Recientemente, se logró determinar el índice de polidispersión en 1.11 de forma ortogonal empleando espectrometría de masas y resonancia magnética nuclear, lo que demuestra la aplicabilidad de técnicas de última generación para una caracterización que permita identificar la secuencia de los componentes peptídicos activos de este fármaco complejo (Vázquez-Leyva et al, 2019).…”

Comparative NMR Metabolomics Profiling between Mexican Ancestral & Artisanal Mezcals and Industrialized Wines to Discriminate Geographical Origins, Agave Species or Grape Varieties and Manufacturing Processes as a Function of Their Quality Attributes

“…(a) Standard direct-excitation one-dimensional proton nuclear magnetic resonance spectra needed to prepare water-to-ethanol off-resonance multipresaturations [37][38][39] were carried out by recording a total of 64 transients, which were collected in 28,844 complex data, with a spectral width of 20 ppm (12,019 Hz), an optimized recovery delay of 6 s and acquisition times of 1.2 s, produced experimental times of 6 min per spectrum. No apodization function was applied during the Fourier Transform.…”

Polyphenol-based nuclear magnetic resonance non-targeted metabolomics of temperature- and time-controlled blue and red maize sprouting

“…1, signal to noise ratio of metabolites' proton resonances in standard 1D-1 H-NMR are severely penalized due to the presence of intense water and methanol signals. For that, a solvent multi-suppression scheme is needed for obtaining a 16-fold signal-tonoise ratio gain of maize sprouts' extracts [18][19][20], needed to prepare the output data matrix for multivariate statistical analysis. Figure 2 resumes the workflow to prepare the NMR output data matrix needed to PCA and OPLS-DA analysis: i) spectra processing (ppm calibrations, base-line corrections and frequency alignments) and ii) reduction of data dimensionality by frequency binning.…”