Potential Degradation Impurities Research Articles

New stability indicating RP-HPLC methods for the determination of related substances and assay of trametinib acetic acid: a mass balance approach.

New stability indicating related substances and assay methods for trametinib acetic acid by RP-HPLC have been developed and validated through forced degradation and mass balance studies for the first time. In related substances (RS) method, trametinib acetic acid was successfully separated from its six related substances namely, cyclopropanamide impurity, desiodo trametinib, desacetyl trametinib, trione acetamide intermediate, trione intermediate, and trione PTSA intermediate using YMC-Triart C18 (150 × 4.6mm, 3µm) column. Orthophosphoric acid (0.15%) in water was used as buffer. Gradient elution was programmed using mobile phase-A (buffer and acetonitrile mixture in 80:20 v/v ratio) and mobile phase-B (buffer and acetonitrile mixture in 20:80 v/v ratio). Acetonitrile and methanol mixture (1:1 v/v) was used as diluent. Flow rate, injection volume, column temperature, and wavelengths were kept as 0.8mL/min, 10µL, 55°C, and 240nm, respectively. Desacetyl trametinib and cyclopropanamide impurity were identified as potential degradation impurities in acid and base degradation conditions, respectively. Assay method specific to above six related substances was also developed. Assay of forced degradation samples was also determined, and the mass balance was established by adding total impurities formed in RS method to assay of trametinib acetic acid.

A mechanistic explanation on degradation behavior of flibanserin for identification and characterization of its potential degradants using LC-DAD/ESI/APCI-Q-TOF-MS/MS

Sprout Pharmaceuticals got marketing approval for Flibanserin (FLB) from USFDA. It is a well-known fact that post-marketing research on impurity profiling significantly contributes to the improvement of quality, safety and efficacy of a drug. In this study, we performed a comprehensive forced degradation of FLB to identify and characterize its degradation impurities and establish the degradation pathways. All the potential degradation impurities were chromatographically separated and identified. Five new potential degradation products (DPs) were characterized using LC-DAD/ESI/APCI-Q-TOF-MS/MS. The plausible structure for all the DPs has been established employing their mass fragmentation pattern. Elemental compositions of the DPs were derived from accurate mass measurements and mass error in ppm. A probable mechanistic explanation has been established for formation of the DPs. Till date, no study has been reported on the degradation behavior of FLB before this. The degradation profiling of FLB reported in this study will provide a complete understanding of the entire possible degradation impurities of FLB. One or more identified degradation impurities can be recommended by the regulatory bodies as the marker for stability study of FLB. The analytical method we have developed can be used for routine quality control of this molecule to quantitate it in presence of their potential impurities. In continuation to this research, degradation kinetic study of FLB to establish the degradation behavior in different stress conditions, mass balance study to correlate the amount loss of parent drug with formation of DPs and evaluation of toxicity potential of the DPs are recommended to be conducted in future.

LC-Q-TOF-MS driven identification of potential degradation impurities of venetoclax, mechanistic explanation on degradation pathway and establishment of a quantitative analytical assay method

Venetoclax is a selective orally active Bcl-2 protein inhibitor very recently approved by USFDA to treat chronic lymphocytic leukemia and other hematological malignancies. Postmarketing surveillance of any drug depends on its acceptability based on risk to benefit ratio. When risk outweighs the benefits, withdrawal of an already marketed drug is warranted. Presence of impurity is the primary cause of increased risk in a drug substance or drug product. With the discovery of newer molecules, it is of great importance to establish advanced analytical techniques for quantification of the drugs as well as their related impurities to address the prospective regulatory queries even if it is already in the market. In this study, a quantitative analytical assay method has been developed and validated for quantification of venetoclax in presence of its degradation impurities. A stress study was performed to examine the stability of the drug in hydrolytic, oxidative, thermolytic and photolytic environments. Venetoclax was found to be prone to degradation in acidic hydrolytic and oxidative stress conditions. Three new degradation impurities have been identified and characterized with the help of LC-Q-TOF-MS with accurate mass measurement and their putative structures have been proposed. Furthermore, for the first time, a possible degradation pathway has been established with mechanistic explanation. Moreover, the analytical method developed in this study will be of immense help for routine analysis of quality control and stability study samples of venetoclax in industry and research laboratories.

Preparation and Characterization of Four Major Novel Degradation Products of Pralatrexate Injection and Validation of HPLC-UV Method

Background: Four major degradation products (1-4) of pralatrexate injection were formed under hydrolytic and light stress conditions. The impurities 1 and 2 were the potential photo degradation products and the impurities 3 and 4 were the potential hydrolytic degradation products. Objective: To prepare and characterize the novel degradation impurities 1, 2, 3 and 4 of pralatrexate injection using NMR, HR MS and IR techniques; and to develop and validate stability indicating analytical reverse phase HPLC-UV method for quantitative simultaneous determination of potential degradation impurities, related substances of pralatrexate and pralatrexate active in pralatrexate liquid formulation. Methods: Gradient HPLC-UV method was developed for the quantification of degradation impurities, related substances and pralatrexate in pralatrexate injection. The separation was achieved on C18 column (250 mm X 4.6 mm, 5µm) using a mobile phase composed of sodium dihydrogen phosphate monohydrate in water (pH 3.0; 0.01M) and methanol. The components were monitored by the UVvisible detector at 242 nm with a flow rate of 1.0 mL/min. Results: The method validation parameters such as accuracy, selectivity, linearity, LOD, LOQ, precision, ruggedness and robustness were demonstrated successfully for pralatrexate and its degradation impurities. The stability-indicating capability of the developed HPLC method was demonstrated by adequate separation of all potential pralatrexate related substances from pralatrexate stressed drug product samples. Conclusion: The developed stability indicating HPLC method was found to be suitable for the simultaneous quantitative determination of potential degradation impurities and related substances of pralatrexate and pralatrexate active in pralatrexate liquid formulation.

RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF MACITENTAN WITH ITS KNOWN AND UNKNOWN DEGRADATION IMPURITIES IN ITS TABLET DOSAGE FORM

Objective: To develop and validate macitentan with its known and unknown degradation impurities in its tablet dosage form.Methods: The RP-HPLC method for macitentan and its impurities was developed and three potential degradation impurities MCA-02, MCA-01 and degradation impurity and N-propyl derivative and N-N dimethyl derivative process impurities were separated. Chromatographic separation was achieved within 70 min on Inertsil C8 (250*4.6 mm, 5 µm) column, Using mobile phase A [Ammonium acetate (ph 4.5 adjusted with glacial acetic acid)] and mobile phase B acetonitrile in gradient elution. Other hplc parameter which was optimized flow rate 1.5 ml/min, detection wavelength 266 nm, column oven temperature 30 ° C and injection volume 20μl. macitentan was subjected to forced degradation also known as stress testing. It was validated as per ICH guidelines.Results: The drug showed extensive degradation in acidic and basic conditions, a slight degradation in oxidative condition. The developed method was statistically validated for linearity (0.45-2.25 ppm). The result of precision (%RSD<5), robustness, LOD(0.15 ppm) and LOQ(0.45 ppm) are well within limits.% Recovery at LOQ, 50%, 100% and 150% was found to be within limit 80-120 %.Conclusion: RP-HPLC method was successfully developed with satisfactory separation of macitentan and its impurities. The proposed method was found to be specific, accurate, precise and robust can be used for estimation of macitentan and its impurities and can be successfully employed in the routine analysis of macitentan.

The Assessment of Impurities for Genotoxic Potential and Subsequent Control in Drug Substance and Drug Product

The strategies implemented at Eli Lilly and Company to address European Medicines Agency and US Food and Drug Administration requirements governing the control of genotoxic impurities (GTIs) are presented. These strategies were developed to provide understanding with regard to the risk and potential liabilities that could be associated with developmental and marketed compounds. The strategies systematize the assessment of impurities for genotoxic potential, addressing both actual and potential impurities. Timing of activities is designed to minimize impact to development timelines while building a data package sufficient to either discharge the risk of potential GTI formation or support the implementation of a specification necessary for long-term control. This article presents the background associated with GTI control, the types of impurities that should be assessed, and the actions to be taken when an impurity is found to be genotoxic. A systematic approach to define potential degradation products derived from stress-testing studies is outlined with a proposal to perform a genotoxic risk assessment on these impurities. Finally, an Arrhenius-based strategy is proposed for a rapid assessment of the likelihood of potential degradation impurities to form in the commercial drug product formulation. Importantly, this article makes a proposal for discharging the risk of a potential GTI with supporting data.

Development and application of a validated stability-indicating Ultra-Performance Liquid Chromatography (UPLC) method for the determination of dantrolene and its related impurities

A rapid, sensitive, and specific ultra-performance liquid chromatographic (UPLC) method was developed for the simultaneous determination of dantrolene and its potential degradation impurities. Chro...

Identification, Isolation, and Characterization of Five Potential Degradation Impurities in Candesartan Cilexetil Tablets

Five impurities were observed when candesartan cilexetil tablets were subjected to stability and forced degradation studies. These impurities were successfully isolated and characterized as desethyl candesartan cilexetil, 1N-ethyl candesartan cilexetil, 2N-ethyl candesartan cilexetil, 1N-ethyl oxo candesartan cilexetil, and 2N-ethyl oxo candesartan cilexetil. A gradient reverse phase liquid chromatography (LC) and an isocratic preparative LC method were used to detect and isolate all five degradation products impurities simultaneously. Mass spectrometry, 1H/13C, DEPT and 2D NMR experiments were extensively utilized to characterize these impurities. Even though desethyl candesartan cilexetil, 1N-ethyl candesartan cilexetil were 2N-ethyl candesartan cilexetil were documented in the literature as known impurities, the regioisomers 1N-ethyl oxo candesartan cilexetil and 2N-ethyl oxo candesartan cilexetil were never noticed. Single-crystal diffraction data has been used to confirm their structure unambiguously and synthetic preparations of all known and unknown impurities were also presented.

Gas turbine installations with divided expansion : Cårdu, M. and Baica, M. Energy Conversion and Management, 2002, 43, (13), 1747–1756

The strategies implemented at Eli Lilly and Company to address European Medicines Agency and US Food and Drug Administration requirements governing the control of genotoxic impurities (GTIs) are presented. These strategies were developed to provide understanding with regard to the risk and potential liabilities that could be associated with developmental and marketed compounds. The strategies systematize the assessment of impurities for genotoxic potential, addressing both actual and potential impurities. Timing of activities is designed to minimize impact to development timelines while building a data package sufficient to either discharge the risk of potential GTI formation or support the implementation of a specification necessary for long-term control. This article presents the background associated with GTI control, the types of impurities that should be assessed, and the actions to be taken when an impurity is found to be genotoxic. A systematic approach to define potential degradation products derived from stress-testing studies is outlined with a proposal to perform a genotoxic risk assessment on these impurities. Finally, an Arrhenius-based strategy is proposed for a rapid assessment of the likelihood of potential degradation impurities to form in the commercial drug product formulation. Importantly, this article makes a proposal for discharging the risk of a potential GTI with supporting data.

SIMULTANEOUS DETERMINATION OF LIGNOCAINE HYDROCHLORIDE, CHLORHEXIDINE GLUCONATE, AND TRIAMCINOLONE ACETONIDE IN SUSPENSION BY REVERSED-PHASE HPLC

The validation of an isocratic high performance liquid chromatographic (HPLC) procedure for the simultaneous determination of lignocaine hydrochloride, chlorhexidine gluconate and triamcinolone acetonide in suspension is reported. Reverse phase chromatography was conducted using 250 × 4.6 mm I.D. 5μm C18 column and monitored on a UV detector at 240nm. Two mobile phases were used: mobile phase A comprised a mixture of methanol/water/triethylamine (58:42:0.4) and the pH adjusted to 3 with phosphoric acid for analysing active ingredients, and mobile phase B was a mixture of methanol/0.01 N ammonium acetate (70:30) for limit testing related substances. Linear response (r > 0.999) was observed over the range of 20–240% of its label claim. The intra-day precision (RSD) of label claim amongst five independent sample preparations, was not more than 0.64% for peak area, and there was no significant difference (P < 0.05) between intra- and inter-day studies. Intermediate precision, as determined from twenty sample preparations, generated by two analysts on different HPLC systems over 7 days, indicated that the assay possessed high degrees of ruggedness by ANOVA test. Recovery studies showed good results for all solutes (99.20%–100.52%) and coefficients of variation ranging from 0.28% to 1.19%. The method showed that the excipients of the commercial suspension and potential degradation impurities caused no interferent effect in the determination of the active ingredients. Related substance limit test indicated that the amount of degradation impurities could be monitored which was important for evaluating the impact of manufacturing process or source changing on product quality. The stability studies at room temperature demonstrated that the drug is stable for at least 1 year. The method is simple, rapid, specific, and reliable, and can be successfully used for the quality control on commercial bulk product.