Investigation of vancomycin and related substances by liquid chromatography/ion trap mass spectrometry

Abstract

Liquid chromatography (LC) methods compatible with mass spectrometry (MS) that are suitable for impurity profiling of vancomycin mixtures have not been described in the literature. The mobile phases of the existing methods contain non-volatile additives and/or solvents that give problems in combination with MS. In this paper, a reversed-phase LC/tandem mass spectrometry method is described for the investigation of vancomycin and related substances. The LC method uses a Zorbax Extend C18 column (250 x 4.6 mm i.d.), 5 microm, and a mobile phase consisting of methanol, water and ammonium acetate solution (pH 9.0). This method allows us to separate vancomycin and its impurities. Mass spectral data are acquired on an LCQ ion trap mass spectrometer equipped with an electrospray interface operated in the positive and negative ion modes. The LCQ is ideally suited for identification of impurities and related substances because it provides on-line LC/MSn capability, which allows efficient identification without time-consuming isolation and purification procedures. Using this method, the fragmentation of vancomycin and known derivatives was studied and the structures of six substances occurring in commercial samples were elucidated.

Product Quality of Parenteral Vancomycin Products in the United States

“…Many of these impurities have now been characterized, and the mechanisms for formation have been elucidated (9,11,13). The main related-substance impurities of vancomycin are monodechlorovancomycin, demethylvancomycin, desamidovancomycin B (CDP-1M and CDP-1m), aglucovancomycin, and desvancosaminylvancomycin.…”

Quality Assessment of U.S. Marketplace Vancomycin for Injection Products Using High-Resolution Liquid Chromatography-Mass Spectrometry and Potency Assays

“…An ultrahigh-pressure liquid chromatographic (UHPLC) method, based on a modification of the work of Diana et al, was used to evaluate vancomycin products (4). Using a Phenomenex Kinetex C 18 column (100 by 2.1 mm, 1.7 m) operating at 40°C, vancomycin B and any impurities were separated using a binary gradient where mobile phase A was 95:5 (vol/vol) H 2 O-0.2 M ammonium acetate, pH 9, and mobile phase B was 65:30:5 (vol/vol/vol) H 2 O-methanol-0.2 M ammonium acetate, pH 9.…”

“…Many methods for the determination and characterization of vancomycin in various matrices exist in the literature (2,4,5,7,16,17). Significantly, both the United States Pharmacopeia and British Pharmacopoeia publish methods for the determination of vancomycin purity and potency (3,19).…”

Ultra high performance liquid chromatography-tandem mass spectrometry vs. commercial immunoassay for determination of vancomycin plasma concentration in children. Possible implications for everyday clinical practice

“…The main disadvantage of these methods is the lack of specificity related to cross-reacting substances, and variable accuracy and precision. 4,5 In addition, the large volumes of plasma needed for automated measurement can be critical in paediatrics, especially in low birth weight neonates.…”