Separation and characterization of two series of unknown degradation impurities caused by light irradiation and autoclaving in xinfujunsu injection using liquid chromatography tandem mass spectrometry.

RATIONALE: A series of photodegradation impurities and a series of degradation impurities produced in autoclaving in xinfujunsu injection were discovered, and these unknown impurities were separated and characterized thoroughly using liquid chromatography tandem quadrupole time-of-flight mass spectrometry. METHODS: The column was a Platisil 5 µm ODS (4.6 × 250 mm, 5 µm). For the analysis of degradation impurities caused by light irradiation and autoclaving, the mobile phase was composed of 0.01 M ammonium formate aqueous solution and acetonitrile/isopropanol (90:10, V/V). Full scan LC-MS and LC-MS2 was carried out to obtain as much structural information as possible. The fragmentation behavior of actinomycin D, actinomycin S3 , and its impurities was studied and used to obtain information about the structures of these impurities. RESULTS: Based on MS2 spectral data and exact mass measurements, the chemical structures of two series of degradation impurities were characterized, among which five unknown impurities were photodegradation impurities and seven unknown impurities were degradation impurities produced in autoclaving of xinfujunsu injection. CONCLUSIONS: Based on characterization of impurities, this study also revealed the cause of impurity production and provided guidance for enterprises to improve the process and drug packaging material to reduce impurity content. Furthermore, this study also provided scientific basis for further improvement of official monographs in pharmacopoeias.

Separation and structural elucidation of cefsulodin and its impurities in both positive and negative ion mode in cefsulodin sodium bulk material using liquid chromatography/tandem mass spectrometry.

RATIONALE: The structural identification of impurities in cephalosporins has been reported. However, to the best of our knowledge, there was no report on the impurities of cefsulodin sodium, which is necessary for the quality control. Thus, the aim of this study was to separate and characterize the impurities in cefsulodin sodium raw material using liquid chromatography/tandem mass spectrometry (LC/MS/MS). METHODS: The analytes were separated on a Kromasil 100-5C18 column (4.6 mm × 250 mm, 5 µm) using a gradient elution with a mobile phase consisting of 1% ammonium sulphate aqueous solution and acetonitrile in the first dimension. The separation in the second dimension was carried on a Shimadzu Shim-pack GISS C18 column (50 mm × 2.1 mm, 1.9 µm) with a mobile phase consisting of 10 mM ammonium formate solution and methanol. RESULTS: The fragmentation behaviors of cefsulodin and its impurities were studied and the structures of the impurities were deduced based on the MSn data. The structures of ten unknown impurities were proposed based on the work carried out in this study. The degradation behaviors of cefsulodin sodium were also studied. This revealed that cefsulodin sodium should be stored in a dry, cool and dark closed container. CONCLUSIONS: Based on the characterization of impurities, this study not only revealed the mechanism by which impurities were produced, thus providing guidance to pharmaceutical companies for manufacturing process improvement and impurity control, but also provided a scientific basis for further improvement of official monographs in pharmacopoeias.

Highly efficient prevention of radiation dermatitis using a PEGylated superoxide dismutase dissolving microneedle patch.

PEGylated superoxide dismutase (PEG-SOD) is commonly used as a cytoprotective agent in radiotherapy. However, its effectiveness in preventing radiation dermatitis is limited owing to its poor skin permeability. To address this issue, a PEG-SOD-loaded dissolving microneedle (PSMN) patch was developed to effectively prevent radiation dermatitis. Initially, PSMN patches were fabricated using a template mold method with polyvinylpyrrolidone K90 as the matrix material. PSMNs exhibited a conical shape with adequate mechanical strength to penetrate the stratum corneum. More than 90 % of PEG-SOD was released from the PSMN patches within 30 min. Notably, the PSMN patches showed a significantly higher drug skin permeation than the PEG-SOD solutions, with a 500-fold increase. In silico simulations and experiments on skin pharmacokinetics confirmed that PSMN patches enhanced drug permeation and skin absorption, in contrast to PEG-SOD solutions. More importantly, PSMN patches efficiently mitigated ionizing radiation-induced skin damage, accelerated the healing process of radiation-affected skin tissues, and exhibited highly effective radioprotective activity for DNA in the skin tissue. Therefore, PSMN patches are promising topical remedy for the prevention of radiation dermatitis.

Analysis of polymerized impurities in mezlocillin sodium and sulbenicillin sodium using two chromatographic separation mechanisms coupled to tandem mass spectrometry.

To effectively control the polymerized impurities in mezlocillin sodium and sulbenicillin sodium, a HPSEC method with TSK-gel G2000SWxl column and a RP-HPLC method with C18 analytical column were established to replace the classical gel filtration chromatography with Sephadex G-10 gel as stationary phase. By studying the chromatographic behavior of polymerized impurities in both methods with different chromatographic separation mechanisms, the polymerized impurities in mezlocillin sodium and sulbenicillin sodium were separated and detected effectively. The column switching two-dimension liquid chromatography ion trap/time-of-flight mass spectrometry was applied to characterize the structures of polymerized impurities eluted from the HPSEC method and RP-HPLC method for both drugs. The structures of the polymerized impurities in mezlocillin sodium and sulbenicillin sodium were deduced based on the MSn data. The results showed that the polymerized impurities detected by HPSEC method and RP-HPLC method were completely different. Therefore, two methods should be used meanwhile to control the polymerized impurities in mezlocillin sodium and sulbenicillin sodium.

Study of the polymerized impurities in cefotaxime sodium and cefepime by applying various chromatographic modes coupled with ion trap/time-of-flight mass spectrometry.

Polymerized impurities in ß-lactam antibiotics can induce allergic reaction, which seriously threaten the health of patients. In order to study the polymerized impurities in cefotaxime sodium and cefepime, a HPSEC method with TSK-gel G2000SWxl column and a RP-HPLC method with C18 column were established to replace the classical gel filtration chromatography with Sephadex G-10 gel as stationary phase. Two-dimensional (2D) liquid chromatography was employed to further investigate the HPSEC and RP-HPLC method and ion trap/time-of-flight mass spectrometry was applied to characterize the structures of polymerized impurities eluted from the two methods. The structures of the polymerized impurities in cefotaxime sodium and cefepime were deduced based on the MSn data, six of which were previously unknown. The corresponding relationship of impurities between the two methods was established, and the specificity of the two methods was compared. The results showed that the polymerized impurities in cefotaxime sodium and cefepime were co-eluted with other small molecular weight impurities with high polarity in HPSEC, leading to a poor specificity. The newly established RP-HPLC methods could effectively separate and detect polymerized impurities and were suitable for the quality control in daily analysis.

Characterization of eight unknown impurities and analysis of their source in xinfujunsu and injection by liquid chromatography coupled with ion trap/time-of-flight mass spectrometry.

Eight unknown impurities in xinfujunsu and its injection were characterized by liquid chromatography coupled with ion trap/time-of-flight mass spectrometry (LC-IT-TOF MS). In order to determine the m/z values of the molecular ions and predict the formulas of all detected impurities, full scan LC-MS in positive ion mode was firstly executed to obtain the m/z value of the molecules. Then LC-MS2, LC-MS3 and LC-MS4 were carried out on target compounds to obtain as much structural information as possible. Based on MSn spectral data and exact mass measurements, the chemical structures of eight unknown impurities were characterized, among which three impurities were degradation impurities and five impurities were process impurities. In addition, the source of impurities and the correlation between process and impurities were also studied. The production of degraded impurities was caused in the high pressure sterilization process of xinfujunsu injection. Based on characterization of impurities, this study revealed the cause of impurity production and provided guidance for enterprises to improve the process to reduce impurity content. Furthermore, this study also provided scientific basis for the further improvement of official monographs in pharmacopoeias.