An approach to on-line electrospray mass spectrometric detection of polypeptide antibiotics of enramycin for high-speed counter-current chromatographic separation

被引:22
作者
Inoue, Koichi [1 ]
Hattori, Yasuko [2 ]
Hino, Tomoaki [1 ]
Oka, Hisao [1 ,2 ]
机构
[1] Kinjo Gakuin Univ, Sch Pharm, Dept Phys & Analyt Chem, Moriyama Ku, Aichi 4638521, Japan
[2] Kinjo Gakuin Univ, Grad Sch Human Ecol, Kinjo Gakuin, Japan
关键词
Polypeptide antibiotics; Enramycin; Purification; High-speed counter-current chromatography; Electrospray mass spectrometry; NATURAL-PRODUCTS; IONIZATION; ENDURACIDIN;
D O I
10.1016/j.jpba.2009.11.010
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
In the field of pharmaceutical and biomedical analysis of peptides, a rapid on-line detection and identification for a methodology have been required for the discovery of new biological active products. In this study, a high-speed counter-current chromatography with electrospray mass spectrometry (HSCCC/ESI-MS) was developed for the on-line detection and purification of polypeptide antibiotics of enramycin-A and -B. The analytes were purified on HSCCC model CCC-1000 (multi-layer coil planet centrifuge) with a volatile solvent of two-phase system composed of n-butanol/hexane/0.05% aqueous trifluoroacetic acid solution (43/7/50, V/V/V), and detected on an LCMS-2010EV quadrupole mass spectrometer fitted with an ESI source system in positive ionization following scan mode (m/z 100-2000). The HSCCC/ESI-MS peaks indicated that enramycin-A (major m/z 786 [M+3H](3+) and minor m/z 1179 [M+2H](2+)) and enramycin-B (major m/z 791 [M+3H](3+) and minor m/z 1185 [M+2H](2+)) have the peak resolution value of 2.9 from 15 mg of loaded enramycin powder. The HSCCC collected amounts of the peak fractions were additionally 4.3 mg (enramycin-A), and 5.9 mg (enramycin-B), respectively. These purified substances were analyzed by LC/ESI-MS with scan positive mode. Based on the LC/ESI-MS chromatograms and spectra of the fractions, enramycin-A and -B were estimated to be over 95% purity. The overall results indicate that this approach of HSCCC/ESI-MS is a powerful technique for the purification and identification of bioactive peptides. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:1154 / 1160
页数:7
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