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Hygromycin B EvoPure®

Hygromycin Impurities

Structure and Impurities of Hygromycin B

LC-MASS (liquid chromatography-mass spectroscopy) is used to identify the mass of the impurities
isolated during the preparation of high purity Hygromycin B. NMR is used to determine
the basic chemical structures of the impurities. IR spectroscopy and MASS-MASS
(tandem) spectroscopy are used to further confirm the structures. The structure
of Hygromycin B is as follows:


The above structure is Hygromycin B where R=CH3 and Impurity F where R=H.

All these impurities are Hygromycin B related and the compositions of the impurities are as following:

  Composition of impurities MW
Impurity A Destomic acid 189.2
Impurity C D-Talose+ 3-Demethyl- Hyosamine 324.5
Impurity D Hydrolysis of Impurity C (D-Talose ring opens up) 326.5
Impurity E D-Talose+Hyosamine 338.5
Impurity F Destomic acid +D-Talose+ 3-Demethyl- Hyosamine 513.5
Hygromycin B Destomic acid +D-Talose+ Hyosamine 527.5

The following structure is Impurity C where R=H and Impurity E where R=CH3.

The following structure is Impurity A.

The following structure is Impurity D.

Pardo et al. (J. Gen. Microbiol. 131:1289-1298, 1985) found that the phosphorylation of the destomic acid ring in Hygromycin B makes Hygromycin B lose its biological activity both in vivo and in vitro. Impurities C, D and E have no destomic acid ring, but still have the hyosamine to maintain bioactivity.