Separation and characterization of unknown impurities in cefonicid sodium by trap-free two-dimensional liquid chromatography combined with ion trap time-of-flight mass spectrometry.

RATIONALE

Seven unknown antibiotic impurities in cefonicid sodium were separated and characterized by a trap-free two-dimensional liquid chromatography coupled to high-resolution ion-trap time-of-flight mass spectrometry (2D-LC/IT-TOF MS) using both positive and negative modes of electrospray ionization. Trap-free 2D-LC and an online demineralization technique made it possible to characterize cefonicid sodium under the conditions of the official standard, and the TIC chromatogram obtained by LC/MS was in conformity with the LC chromatogram obtained by the official analytical method in the peak sequence of impurities.

METHODS

In the first dimension, the column was a GRACE Alltima C (250 mm × 4.6 mm, 5 μm), and the gradient elution used 0.02 mol·L ammonium dihydrogen phosphate solution and methanol as mobile phase. In the second dimension, the analytical column was a Shimadzu Shim-pack GISS C (50 mm × 2.1 mm, 1.9 μm) with 10 mmol·L ammonium formate solution and methanol as mobile phase. Full scan LC/MS was first executed to obtain the exact m/z values of the molecules. Then LC/MS and LC-MS experiments were performed on the compounds of interest.

RESULTS

The structures of seven unknown degradation products in cefonicid sodium were deduced based on the high-resolution MS data using both positive and negative mode.

CONCLUSIONS

The problem of incompatibility between the non-volatile salt mobile phase and mass spectrometry was solved completely by multidimensional heart-cutting approaches and an online demineralization technique, which is worthy of widespread use and application for the advantages of stability and repeatability.