STABILITY EVALUATION OF TARTRAZINE BY LIQUID CHROMATOGRAPHY-DIODE ARRAY DETECTOR AND HIGH-RESOLUTION ELECTRON SPRAY IONIZATION QUADRUPOLE TIME-OFFLIGHT MASS SPECTROMETRY/MASS SPECTROMETRY ANALYSIS

Objective: This study investigates the degradation behavior of tartrazine, a synthetic azo dye used in pharmaceutical products.Methods: For this purpose, standard solutions containing E102 were subjected to pH, oxidation, photolytic conditions and were analyzed by the developed high-performance liquid chromatography (HPLC) method.Results: The results show degradation in standard solution with the formation of single degradation product in oxidative visible light condition. The formed unknown degradation product was isolated by semipreparative HPLC and characterized using ultra-HPLC coupled with high resolution tandem mass spectrometer (MS).Conclusion: The isolated component was vacuum dried and subjected to high-resolution MS analysis for the plausible structural elucidation. Based on the fragmentation pattern of one degradation product (ODP) from their MSn studies, the ODP may be identified as 2,5-dihydroxy-1-phenyl-3- (phenyldiazenyl)-2,3-dihydro-1H-pyrazole-4-carboxylic acid.Â

Toxicity Profile Study of Antihypertensive Drug Prazosin in Pregnant Wistar Rats

The purpose of this research is to investigate and assess the cytotoxicity and genotoxicity of Prazosin HCL in pregnant rats. Prazosin (PZ) was administered to the animals intraperitoneally (IP) at dosages of 5, 15, and 25 mg/kg/body weight for single dose (14-day) toxicity tests. The following parameters have been examined for evaluating genotoxicity: bone marrow micronucleus assay, peripheral blood micronucleus assay, DNA damage is measured using the metaphase chromosomal analysis, DNA damage is measured using the DNA fragmentation test, and cytotoxicity is measured using a histological analysis. The results obtained clearly demonstrate that PZ induced hazardous responses at the higher dose in the hepatocytes, as evidenced by DNA damage, and increased DNA fragmentation in pregnant rats. Observing that PZ significantly increased DNA strand breakage and structural chromosomal aberrations in bone marrow cell lines is also fascinating. As a result, it is thought to be genotoxic to bone marrow and mouse hepatocyte cells. The current study showed that Prazosin had significant genotoxic effects in pregnant rats at the matching hepatotoxic dose level

Development and Validation of Stability Indicating RP HPLC method for the estimation of armodafinil and Characterization of its base Degradation Product by LC-MS/MS

The present work focuses on the development and validation of a rapid, selective and reliable stability indicating reverse phase High Performance Liquid Chromatographic method for the quantitative analysis of Armodafinil (ARM). Armodafinil is a eugeroics drug used for treatment of narcolepsy and shift work sleep disorder and also as an adjunctive treatment for obstructive sleep apnea. The separation was carried out on a Hibar Purospher C18 (250 mm × 4.6 mm; 5μ) column by using 0.01 M ammonium formate (pH 4.5, Adjusted with acetic acid) and methanol as mobile phase in the ratio of 45:55 % v/v. The flow rate was maintained at 1 mL/min and the eluents were monitored at 220 nm. Stress studies were carried out with 1 mg/mL of the drug solution, starting with mild conditions and followed by intrinsic conditions to achieve sufficient degradation. Armodafinil underwent extensive degradation under basic hydrolysis condition. The unknown degradation product formed (DP I; 6.59±0.2 min) under the basic degradation condition was separated on a semi preparative HPLC, characterized by UPLC-Q-TOF and its fragmentation pathway was proposed. The proposed structure of the degradation product was confirmed by HRMS analysis. The developed stability indicating LC method was validated with respect to accuracy, precision, specificity/selectivity and linearity. The degradation product was characterized and reported

Development of a Forced Degradation Profile of Alosetron by Single Mode Reversed-Phase HPLC, LC-MS, and its Validation

Determination of alosetron in the presence of its degradation products was studied and validated by a novel HPLC method. The separation of the drug and its degradation products was achieved with the Jones Chromatography C18 analytical column (150 mm x 4.6 mm; 3 μm) with a stationary phase in isocratic elution mode. The mobile phase used was 0.01 M ammonium acetate, pH-adjusted to 3.5 with glacial acetic acid and acetonitrile in the ratio of 75:25 (V/V) at a flow rate of 1 ml/min and UV detection was carried out at 217 nm. Further, the drug was subjected to stress studies for acidic, basic, neutral, oxidative, and thermal degradations as per ICH guidelines and the drug was found to be labile in base hydrolysis and oxidation, while stable in acid, neutral, thermal, and photolytic degradation conditions. An MS study has been performed on the major degradation products to predict the degradation pathway of alosetron. The method provided linear responses over the concentration range of 100–1500 ng/ml and regression analysis showed a correlation coefficient value (r2) of 0.994. The LOD and LOQ were found to be 1 ng/ml and 3 ng/ml, respectively. The developed LC method was validated as per ICH guidelines with respect to accuracy, selectivity, precision, linearity, and robustness

Characterization of Oxidative Degradation Product of Darunavir by LC-MS/MS

A rapid, selective, and reliable LC-MSn method has been developed and validated for the isolation and structural characterization of the degradation product of darunavir (DRV). DRV, an HIV-1 protease inhibitor, was subjected to intrinsic oxidative stress conditions using 30% hydrogen peroxide and the degradation profile was studied. The oxidative degradation of DRV resulted in one degradation product. The unknown degradation product was separated on a Hibar Purospher C18 (250 mm × 4.6 mm; 5 µm) column by using 0.01 M ammonium formate (pH 3.0) and acetonitrile as mobile phase in the ratio of 50:50, v/v. The eluents were monitored at 263 nm using a UV detector. The isolated degradation product was characterized by UPLC-Q-TOF and its fragmentation pathway was proposed. The proposed structure of the degradation product was confirmed by HRMS analysis. The developed stability-indicating LC method was validated with respect to accuracy, precision, specificity/selectivity, and linearity. No prior reports were found in the literature about the oxidative degradation behavior of DRV

Evaluation of antiparkinson activity of PTUPB by measuring dopamine and its metabolites in Drosophila melanogaster: LC-MS/MS method development.

Soluble epoxide hydrolase (sEH) inhibition is reported to elevate endogenous epoxyeicosatrienoic acids (EET's), which are known to play an important role in neuroprotection by inhibiting oxidative stress and neuroinflammation. In the present study, PTUPB, a dual inhibitor of sEH and COX-2, has been tested for its antiparkinson activity against rotenone (ROT) induced neurodegeneration in Drosophila model of Parkinson's disease (PD). To determine the efficacy and brain bioavailability of PTUPB a simple, rapid and sensitive LC-MS/MS method was developed and validated for the estimation of PTUPB (Method-I), dopamine (DA) and its metabolites (Method-II) in fly head. Mass spectrometric acquisitions of analytes signals were performed in positive and negative electron spray ionization MRM mode by monitoring the daughter ions. The isocratic elution using formic acid (0.1% v/v) and acetonitrile (20:80v/v) (for method I), and acetic acid (0.1% v/v) and methanol (for method II) on Jones C18 was carried out to achieve the separation. The results of brain PTUPB, DA and its metabolites estimation shows a dose dependent increase in PTUPB concentration and a dose dependent prevention of ROT induced changes in DA and its metabolites levels (p<0.05), indicating a significant neuroprotection activity of PTUPB. In the present study, we have successfully developed and validated LC-MS/MS methods to identify and quantify PTUPB, DA and its metabolites using a UFLC-ESI-QqQ mass spectrometer for the screening of neuroprotective agents in Drosophila Melanogaster