Identification of stress degradation products of iloperidone using liquid chromatography coupled with an Orbitrap mass spectrometer.

RATIONALE: Iloperidone (ILOP) is an atypical antipsychotic drug used for the treatment of schizophrenia and related psychotic disorders. Comprehensive stress testing of the ILOP drug was carried out as per ICH guidelines to understand its degradation profile. The presence of degradation products in a drug affects not only the quality, but also the safety and efficacy of drug formulation. Thus, it is essential to develop an efficient analytical method which can be useful for the separation, identification and characterization of all possible degradation products of ILOP. METHODS: ILOP was subjected to various stress conditions such as acidic, basic, neutral hydrolysis, oxidation, photolysis and thermal conditions; and the resulting degradation products were investigated using LC-PDA-HRMS and MS/MS. An efficient and simple ultra-high-performance liquid chromatography (UHPLC) method has been developed on Acquity UPLC® BEH C18 column (2.1 × 100mm, 1.7 µm) using a gradient elution of heptafluorobutyric acid (0.1% HFBA) and acetonitrile as mobile phase. RESULTS: ILOP was found to degrade under acidic and basic hydrolysis and oxidative stress conditions, whereas it was stable under neutral hydrolysis, thermal and photolytic conditions. A total of seven degradation products (DP1 to DP7) were identified and characterized by LC/MS/MS in positive ion mode with accurate mass measurements. The hydrolytic degradation under acidic and basic conditions produced two DPs (DP1 and DP2) and four DPs (DP4 to DP7), respectively, whereas DP3 was formed under oxidative conditions. In silico toxicity predictions showed higher probability values for DP4, DP6 and DP7, which indicates these DPs have the potential to mutate DNA. CONCLUSIONS: ILOP was found to be labile under hydrolytic and oxidative conditions. The structures of the degradation products were rationalized by appropriate mechanisms. The proposed method can be effectively used for the determination and detection of ILOP and its degradation products.

Assessment of human health risks from heavy metals in outdoor dust samples in a coal mining area.

Jharia (India) a coal mining town has been affected by the consequences of mining and associated activities. Samples of outdoor fallen dust were collected at different locations of Jharia covering four different zones: commercial, petrol pump, high traffic, and residential areas. The dust samples were analysed for different trace elements (As, Cd, Co, Cr, Cu, Mn, Ni, Pb, Se, and Zn). The highest concentration of the elements in the dust samples are Mn (658 mg/kg), Zn (163.6 mg/kg), Cr (75.4 mg/kg), Pb (67.8 mg/kg), Ni (66 mg/kg), Cu (56.8 mg/kg), Co (16.9 mg/kg), As (4.1 mg/kg), and Cd (0.78 mg/kg). The concentration of selenium was below detection limit. Except Cd, contents of all the other elements in the dust samples were significantly lower in the residential area. High amount of Ni (145 mg/kg) and Pb (102 mg/kg) was observed in the high traffic and petrol pump areas, respectively. The exposure risk assessment strategies are helpful in predicting the potential health risk of the trace elements in the street dust. Selected receptors for risk assessment were infants, toddlers, children, teens, and adults. The calculated hazard quotient (HQ) for lifetime exposure was <1.0 for all the elements studied, indicating no risks from these elements for adults Among the receptors, toddlers were found to be more vulnerable, with HQ for Co, Cr, and Pb > 0.1. The finding predicts potential health risk to toddlers and children.

Some proteins of M. tuberculosis that localise to the nucleus of THP-1-derived macrophages.

Host-pathogen dialectics in tuberculosis (TB) via DNA-protein interactions are emerging. We investigated whether proteins produced by Mycobacterium tuberculosis (Mtb) could translocate to the host nucleus. Using lysates of nuclei purified from Mtb-infected THP-1-derived macrophages, we identified at least 15 proteins of Mtb-origin by electrophoretic and chromatographic separation and mass spectrometry. Western blotting confirmed time-dependent accumulation of Mtb EF-Tu, GroEL, GroES and MtrA in the host nucleus. MtrA could pull down at least 16 host proteins. Mtb proteins may have moonlighting functions that affect host gene expression.