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Assignment and quantification of 2-aminopyridine derivatized oligosaccharide isomers coeluted on reversed-phase HPLC/MS by MSn spectral library.
Anal Chem. 2004 Dec 15; 76(24):7294-303.AC

Abstract

2-Aminopyridine (PA)-derivatized oligosaccharides from IgG were analyzed by using reversed-phase HPLC/mass spectrometry (RP-HPLC/MS) and a MS(n) spectral library, in particular, focusing on two pairs of isomers incompletely separated or coeluted in chromatograms. We previously reported that MS(n) spectral matching considering both major fragment ions (m/z) and intensities is useful and applicable to the structural assignment of PA-oligosaccharide isomers. In this study, MS(n) spectral matching based on the MS(n) spectral library was applied to the assignment of these PA-oligosaccharide isomers in IgG. Its usefulness was investigated by comparing it to the conventional two-dimensional mapping method based on retention time indexes. Specifically, we focus on the assignment and quantification of the isomers, which are coeluted in chromatograms. From this, we propose a new method using MS(n) spectral matching and the working curve on which are plotted the relative intensities of selected fragment ions in their MS(2) spectra versus various mixtures of the isomers. This new method demonstrated that the obtained quantities coincide very well with those estimated after separating by a combination of lectin and reversed-phase columns. This means that separation by RP-HPLC/MS is greatly simplified because complete separation of the isomers is no longer required. Application of this new method was tested by using the two other pairs of fucosylated and nonfucosylated PA-oligosaccharides from IgG. The results showed that this method works for them as well.

Authors+Show Affiliations

Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 001-0021, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15595872

Citation

Takegawa, Yasuhiro, et al. "Assignment and Quantification of 2-aminopyridine Derivatized Oligosaccharide Isomers Coeluted On Reversed-phase HPLC/MS By MSn Spectral Library." Analytical Chemistry, vol. 76, no. 24, 2004, pp. 7294-303.
Takegawa Y, Deguchi K, Ito S, et al. Assignment and quantification of 2-aminopyridine derivatized oligosaccharide isomers coeluted on reversed-phase HPLC/MS by MSn spectral library. Anal Chem. 2004;76(24):7294-303.
Takegawa, Y., Deguchi, K., Ito, S., Yoshioka, S., Sano, A., Yoshinari, K., Kobayashi, K., Nakagawa, H., Monde, K., & Nishimura, S. (2004). Assignment and quantification of 2-aminopyridine derivatized oligosaccharide isomers coeluted on reversed-phase HPLC/MS by MSn spectral library. Analytical Chemistry, 76(24), 7294-303.
Takegawa Y, et al. Assignment and Quantification of 2-aminopyridine Derivatized Oligosaccharide Isomers Coeluted On Reversed-phase HPLC/MS By MSn Spectral Library. Anal Chem. 2004 Dec 15;76(24):7294-303. PubMed PMID: 15595872.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Assignment and quantification of 2-aminopyridine derivatized oligosaccharide isomers coeluted on reversed-phase HPLC/MS by MSn spectral library. AU - Takegawa,Yasuhiro, AU - Deguchi,Kisaburo, AU - Ito,Shinya, AU - Yoshioka,Shinji, AU - Sano,Akihiro, AU - Yoshinari,Kiyomi, AU - Kobayashi,Kinya, AU - Nakagawa,Hiroaki, AU - Monde,Kenji, AU - Nishimura,Shin-Ichiro, PY - 2004/12/15/pubmed PY - 2005/2/23/medline PY - 2004/12/15/entrez SP - 7294 EP - 303 JF - Analytical chemistry JO - Anal Chem VL - 76 IS - 24 N2 - 2-Aminopyridine (PA)-derivatized oligosaccharides from IgG were analyzed by using reversed-phase HPLC/mass spectrometry (RP-HPLC/MS) and a MS(n) spectral library, in particular, focusing on two pairs of isomers incompletely separated or coeluted in chromatograms. We previously reported that MS(n) spectral matching considering both major fragment ions (m/z) and intensities is useful and applicable to the structural assignment of PA-oligosaccharide isomers. In this study, MS(n) spectral matching based on the MS(n) spectral library was applied to the assignment of these PA-oligosaccharide isomers in IgG. Its usefulness was investigated by comparing it to the conventional two-dimensional mapping method based on retention time indexes. Specifically, we focus on the assignment and quantification of the isomers, which are coeluted in chromatograms. From this, we propose a new method using MS(n) spectral matching and the working curve on which are plotted the relative intensities of selected fragment ions in their MS(2) spectra versus various mixtures of the isomers. This new method demonstrated that the obtained quantities coincide very well with those estimated after separating by a combination of lectin and reversed-phase columns. This means that separation by RP-HPLC/MS is greatly simplified because complete separation of the isomers is no longer required. Application of this new method was tested by using the two other pairs of fucosylated and nonfucosylated PA-oligosaccharides from IgG. The results showed that this method works for them as well. SN - 0003-2700 UR - https://www.unboundmedicine.com/medline/citation/15595872/Assignment_and_quantification_of_2_aminopyridine_derivatized_oligosaccharide_isomers_coeluted_on_reversed_phase_HPLC/MS_by_MSn_spectral_library_ DB - PRIME DP - Unbound Medicine ER -