An isocratic separation of underivatized gentamicin components, 1H NMR assignment and protonation pattern

Wojciech Lesniak; John Mc Laren; Wesley R. Harris; Vincent L. Pecoraro; Jochen Schacht

doi:10.1016/j.carres.2003.08.005

An isocratic separation of underivatized gentamicin components, ¹H NMR assignment and protonation pattern

Wojciech Lesniak, John Mc Laren, Wesley R. Harris, Vincent L. Pecoraro, Jochen Schacht

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

Abstract

A simple method for the separation of the major components of commercial gentamicin sulfate (C-1, C-1a, C-2, C-2a) by high-performance liquid chromatography (HPLC) on an analytical and a semipreparative scale was developed. The method utilized ion-pair reversed-phase chromatography, isocratic elution with an aqueous solution containing 9% trifluroacetic acid and 2.5% acetonitrile as the mobile phase at a flow rate of 2 and 9 mL/min for analytical and semipreparative columns, respectively. Detection was carried out at 213 nm without derivatization. The protonation pattern of the separated gentamicins was determined by potentiometry and ¹⁵N and ¹H NMR. The full proton NMR assignment for gentamicin C-1 was obtained through the use of ¹H 1D and 2D ¹H-¹H COSY measurements.

Original language	English (US)
Pages (from-to)	2853-2862
Number of pages	10
Journal	Carbohydrate Research
Volume	338
Issue number	24
DOIs	https://doi.org/10.1016/j.carres.2003.08.005
State	Published - Nov 21 2003
Externally published	Yes

Keywords

Aminoglycosides
HPLC
NMR spectroscopy
Potentiometery

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Organic Chemistry

Access to Document

10.1016/j.carres.2003.08.005

Cite this

@article{fa2c689dfbfc486799531b91fdd081e5,

title = "An isocratic separation of underivatized gentamicin components, 1H NMR assignment and protonation pattern",

abstract = "A simple method for the separation of the major components of commercial gentamicin sulfate (C-1, C-1a, C-2, C-2a) by high-performance liquid chromatography (HPLC) on an analytical and a semipreparative scale was developed. The method utilized ion-pair reversed-phase chromatography, isocratic elution with an aqueous solution containing 9% trifluroacetic acid and 2.5% acetonitrile as the mobile phase at a flow rate of 2 and 9 mL/min for analytical and semipreparative columns, respectively. Detection was carried out at 213 nm without derivatization. The protonation pattern of the separated gentamicins was determined by potentiometry and 15N and 1H NMR. The full proton NMR assignment for gentamicin C-1 was obtained through the use of 1H 1D and 2D 1H-1H COSY measurements.",

keywords = "Aminoglycosides, HPLC, NMR spectroscopy, Potentiometery",

author = "Wojciech Lesniak and {Mc Laren}, John and Harris, {Wesley R.} and Pecoraro, {Vincent L.} and Jochen Schacht",

note = "Funding Information: This work was supported by grant DC-03685 from the National Institute on Deafness and Other Communicational Disorders, National Institutes of Health. ",

year = "2003",

month = nov,

day = "21",

doi = "10.1016/j.carres.2003.08.005",

language = "English (US)",

volume = "338",

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journal = "Carbohydrate Research",

issn = "0008-6215",

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AU - Lesniak, Wojciech

AU - Mc Laren, John

AU - Harris, Wesley R.

AU - Pecoraro, Vincent L.

AU - Schacht, Jochen

N1 - Funding Information: This work was supported by grant DC-03685 from the National Institute on Deafness and Other Communicational Disorders, National Institutes of Health.

PY - 2003/11/21

Y1 - 2003/11/21

N2 - A simple method for the separation of the major components of commercial gentamicin sulfate (C-1, C-1a, C-2, C-2a) by high-performance liquid chromatography (HPLC) on an analytical and a semipreparative scale was developed. The method utilized ion-pair reversed-phase chromatography, isocratic elution with an aqueous solution containing 9% trifluroacetic acid and 2.5% acetonitrile as the mobile phase at a flow rate of 2 and 9 mL/min for analytical and semipreparative columns, respectively. Detection was carried out at 213 nm without derivatization. The protonation pattern of the separated gentamicins was determined by potentiometry and 15N and 1H NMR. The full proton NMR assignment for gentamicin C-1 was obtained through the use of 1H 1D and 2D 1H-1H COSY measurements.

AB - A simple method for the separation of the major components of commercial gentamicin sulfate (C-1, C-1a, C-2, C-2a) by high-performance liquid chromatography (HPLC) on an analytical and a semipreparative scale was developed. The method utilized ion-pair reversed-phase chromatography, isocratic elution with an aqueous solution containing 9% trifluroacetic acid and 2.5% acetonitrile as the mobile phase at a flow rate of 2 and 9 mL/min for analytical and semipreparative columns, respectively. Detection was carried out at 213 nm without derivatization. The protonation pattern of the separated gentamicins was determined by potentiometry and 15N and 1H NMR. The full proton NMR assignment for gentamicin C-1 was obtained through the use of 1H 1D and 2D 1H-1H COSY measurements.

KW - Aminoglycosides

KW - HPLC

KW - NMR spectroscopy

KW - Potentiometery

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