RESUMEN
Carcinogenic N-nitrosamines were recently found in the sartan family of drugs and caused many drug recalls. Both of their detection and quantification are therefore important. Methods reported for N-nitrosamine quantitation rely on the use of standards and are just applicable to simple N-nitrosamines. There is an urgent need to quantify N-nitrosamines derived from drugs with a complicated structure that lack standards. To tackle the issue, this study describes a novel absolute quantitation strategy for N-nitrosamines using coulometric mass spectrometry (CMS) without standards. In our approach, N-nitrosamine is first converted into electrochemically active hydrazine via zinc reduction under acidic condition and the resulting hydrazine can then be easily quantified using CMS. To validate our method, six simple N-nitrosamines, N-nitrosodiethylamine (NDEA), N-nitroso-4-phenylpiperidine (NPhPIP), N-nitrosodiphenylamine (NDPhA), N-nitrosodibutylamine (NDBA), N-nitrosodipropylamine (NDPA), and N-nitrosopiperidine (NPIP), were chosen as test samples, and they all were quantified with excellent measurement accuracy (quantitation error ≤1.1%). Taking this one step further, as a demonstration of the method utility, a drug-like N-nitrosamine, (R)-N-(2-(6-chloro-5-methyl-1'-nitroso-2,3-dihydrospiro[indene-1,4'-piperidin]-3-yl)propan-2-yl)acetamide (VII), was also synthesized and successfully quantified using our method at 15 ppb level in a complex formulation matrix, following solvent extraction, N-nitrosamine isolation, and reductive conversion. Because of the feature of requiring no standards, CMS provides a simple and powerful approach for N-nitrosamine absolute quantitation and has great potential for analysis of other drug impurities or metabolites.
Asunto(s)
Nitrosaminas , Cromatografía de Gases y Espectrometría de Masas/métodos , Hidrazinas/análisis , Nitrosaminas/análisis , Espectrometría de Masas en TándemRESUMEN
In 1998, the aminoglycoside antibiotic gentamicin sulfate caused several cases of deaths in the United States, after the switch from twice- to once-daily application. Endotoxins were discussed as the cause for the adverse effects and sisomicin was identified as the lead impurity; batches containing sisomicin were contaminated with more impurities and were responsible for the fatalities. In 2016, anaphylactic reactions in horses, and later in humans with one fatality, were observed after application of gentamicin sulfate contaminated with histamine. To determine whether histamine was responsible for the 1990s death cases as well, histamine was quantified by means of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in 30 samples of gentamicin sulfate analyzed in previous studies. Furthermore, a relative quantification of sisomicin was performed to check for a correlation between histamine and the lead impurity. A maximum amount of 11.52 ppm histamine was detected, which is below the limit for anaphylactic reactions of 16 ppm, and no correlation of the two impurities was observed. However, the European Medicines Agency recommends a stricter limit with regard to the maximum single dose of gentamicin sulfate to reach a greater gap between the maximum histamine exposition of 4.3 µg and the quantity known to cause hypotension of 7 µg. The low amounts of histamine and the fact that there is no connection with the contamination with sisomicin showed that histamine was not the cause for the death cases in the United States in 1998, and endotoxins remain the most probable explanation.
Asunto(s)
Antibacterianos/análisis , Gentamicinas/análisis , Histamina/análisis , Sisomicina/análisis , Antibacterianos/efectos adversos , Antibacterianos/química , Cromatografía Liquida , Contaminación de Medicamentos , Gentamicinas/efectos adversos , Gentamicinas/química , Espectrometría de Masas en TándemRESUMEN
The ICH M7 (R1) guideline recommends the use of complementary (Q)SAR models to assess the mutagenic potential of drug impurities as a state-of-the-art, high-throughput alternative to empirical testing. Additionally, it includes a provision for the application of expert knowledge to increase prediction confidence and resolve conflicting calls. Expert knowledge, which considers structural analogs and mechanisms of activity, has been valuable when models return an indeterminate (equivocal) result or no prediction (out-of-domain). A retrospective analysis of 1002 impurities evaluated in drug regulatory applications between April 2017 and March 2019 assessed the impact of expert review on (Q)SAR predictions. Expert knowledge overturned the default predictions for 26% of the impurities and resolved 91% of equivocal predictions and 75% of out-of-domain calls. Of the 261 overturned default predictions, 15% were upgraded to equivocal or positive and 79% were downgraded to equivocal or negative. Chemical classes with the most overturns were primary aromatic amines (46%), aldehydes (45%), Michael-reactive acceptors (37%), and non-primary alkyl halides (33%). Additionally, low confidence predictions were the most often overturned. Collectively, the results suggest that expert knowledge continues to play an important role in an ICH M7 (Q)SAR prediction workflow and triaging predictions based on chemical class and probability can improve (Q)SAR review efficiency.
Asunto(s)
Contaminación de Medicamentos , Mutágenos/química , Relación Estructura-Actividad Cuantitativa , Simulación por Computador , Pruebas de Mutagenicidad , Estudios Retrospectivos , Medición de RiesgoRESUMEN
The plastics used in drug packaging systems and medical devices are composed of homologous polymers and generally contain additives such as antioxidants, plasticizers and others, to improve their physicochemical properties. However, these additives have potential drawbacks due to possible migration or leaching towards the drug product. Leaching can cause a change in the chemical composition of the drug which, in turn, could modify its therapeutic action and, in some cases, its organoleptic properties. Leachables may also be considered a health hazard due to their inherent toxicological properties. The analytical characterization (detection, identification, typification/qualification and quantification) of leachable substances is mandatory and this information must be included in the application dossier for the drug before it can receive regulatory approval. The main aim of this paper is to collect and contextualise the reported analytical approaches for characterising and/or controlling organic leachables from plastic materials in contact with drugs. We also describe the state of the art of leachables in conjunction with a valuable, broad-based compilation of directives and guidelines. We end by presenting an updated collection of leachables both gas and liquid chromatography studies as separation techniques over the last eight years. We decided to focus our review exclusively on organic leachables as there is already a wide body of research on inorganic impurities.
Asunto(s)
Contaminación de Medicamentos , Embalaje de Medicamentos , Plásticos/química , Técnicas de Química Analítica , Contaminación de Medicamentos/prevención & control , Preparaciones FarmacéuticasRESUMEN
BACKGROUND: The presence of impurities in some drugs may compromise the safety and efficacy of the patient's treatment. Therefore, establishing of the biological safety of the impurities is essential. Diabetic patients are predisposed to tissue damage due to an increased oxidative stress process; and drug impurities may contribute to these toxic effects. In this context, the aim of this work was to study the toxicity, in 3 T3 cells, of the antidiabetic agents sitagliptin, vildagliptin, and their two main impurities of synthesis (S1 and S2; V1 and V2, respectively). METHODS: MTT reduction and neutral red uptake assays were performed in cytotoxicity tests. In addition, DNA damage (measured by comet assay), intracellular free radicals (by DCF), NO production, and mitochondrial membrane potential (ΔψM) were evaluated. RESULTS: Cytotoxicity was observed for impurity V2. Free radicals generation was found at 1000 µM of sitagliptin and 10 µM of both vildagliptin impurities (V1 and V2). A decrease in NO production was observed for all vildagliptin concentrations. No alterations were observed in ΔψM or DNA damage at the tested concentrations. CONCLUSIONS: This study demonstrated that the presence of impurities might increase the cytotoxicity and oxidative stress of the pharmaceutical formulations at the concentrations studied.
Asunto(s)
Composición de Medicamentos/normas , Contaminación de Medicamentos , Fibroblastos/efectos de los fármacos , Hipoglucemiantes/toxicidad , Fosfato de Sitagliptina/toxicidad , Vildagliptina/toxicidad , Células 3T3 , Animales , Supervivencia Celular/efectos de los fármacos , Daño del ADN , Fibroblastos/metabolismo , Fibroblastos/patología , Hipoglucemiantes/química , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Ratones , Óxido Nítrico/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Fosfato de Sitagliptina/química , Vildagliptina/químicaRESUMEN
Simultaneous analysis of drug compounds and their impurities of degradation and synthesis became constant in the modern pharmaceutical analysis. Likewise, analytical techniques must improve sensitivity and selectivity for the monitoring of pharmaceutical products, allowing a full assessment of impurities in drug products and, therefore, ensure safety and efficacy of pharmacological treatments. The application of Quality by Design (QbD) principles has proved to be feasible on the elaboration of analytical methods, allowing the comprehensive evaluation and measurement of different analytical parameters and their effects on critical properties of the methodology in development. QbD approach was applied to the development of a fast and selective HPLC method for the analysis of the antiplatelet aggregation drug ticagrelor and its degradation products in presence of three impurities of synthesis. Fractional factorial resolution V was the screening experimental design applied to five method parameters. Response surface methodology was carried by central composite star face design on the two critical method parameters selected. Analytical design space, established after the application of Monte-Carlo simulations, verified whether predicted results were in accordance with critical quality attributes. The developed and validated HPLC method with DAD detection at 225â¯nm was able to resolve eight related compounds in less than three minutes.
Asunto(s)
Adenosina/análogos & derivados , Cromatografía Líquida de Alta Presión/métodos , Adenosina/análisis , Contaminación de Medicamentos , Diseño de Fármacos , Estabilidad de Medicamentos , Inhibidores de Agregación Plaquetaria/análisis , Reproducibilidad de los Resultados , TicagrelorRESUMEN
A simple, fast and sensitive analytical method by high-performance liquid chromatography (HPLC) was developed and validated for the simultaneous determination of ticagrelor and two synthesis impurities. The HPLC method was established using an Agilent 1200 Series equipment coupled to photodiode array detector (PDA) at 270nm with a Zorbax Plus C8 column (150×4.6mm, 5.0µm), injection volume of 20µL, and a constant temperature of 25°C. The mobile phase consisted of acetonitrile: ammonium acetate 50mM (57:43, v/v) and pH adjusted to 8.2 with ammonium hydroxide 6M, at a flow rate of 0.7mL/min. No interference peaks from excipients and diluent system indicated the specificity of the method. The calibration curves showed determination coefficients (r2)>0.99, calculated by linear regression. The limit of quantitation (LOQ) for impurities 1 and 2 were 2.0 and 0.2µg/mL, respectively. Intra and interday relative standard deviations (RSDs) were <2% for ticagrelor and <6% for the impurities, proving the precision of the method. Besides, two mayor degradation products formed when sample solutions of ticagrelor were exposed to UVC radiation were elucidated and the mechanisms involved in the photolytic degradation of ticagrelor were proposed.
Asunto(s)
Adenosina/análogos & derivados , Técnicas de Química Analítica/métodos , Contaminación de Medicamentos , Fotólisis , Adenosina/análisis , Adenosina/química , Técnicas de Química Analítica/normas , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Líquida de Alta Presión/normas , TicagrelorRESUMEN
While not acutely toxic, chronic hepatic effect of certain gadolinium chelates (GC), used as contrast agent for magnetic resonance imaging, might represent a risk in renally-impaired patients due to free gadolinium accumulation in the liver. To answer this question, this study investigated the consequences of the presence of small amounts of either a soluble gadolinium salt ("free" Gd) or low-stability chelating impurity in the pharmaceutical solution of gadoteric acid, a macrocyclic GC with high thermodynamic and kinetic stabilities, were investigated in renally-impaired rats. Renal failure was induced by adding 0.75% adenine in the diet for three weeks. The pharmaceutical and commercial solution of gadoteric acid was administered (5 daily intravenous injections of 2.5 mmol Gd/kg) either alone or after being spiked with either "free" gadolinium (i.e., 0.04% w/v) or low-stability impurity (i.e., 0.06 w/v). Another GC, gadodiamide (low thermodynamic and kinetic stabilities) was given as its commercial solution at a similar dose. Non-chelated gadolinium was tested at two doses (0.005 and 0.01 mmol Gd/kg) as acetate salt. Gadodiamide induced systemic toxicity (mortality, severe epidermal and dermal lesions) and substantial tissue Gd retention. The addition of very low amounts of "free", non-chelated gadolinium or low thermodynamic stability impurity to the pharmaceutical solution of the thermodynamically stable GC gadoteric acid resulted in substantial capture of metal by the liver, similar to what was observed in "free" gadolinium salt-treated rats. Relaxometry studies strongly suggested the presence of free and soluble gadolinium in the liver. Electron microscopy examinations revealed the presence of free and insoluble gadolinium deposits in hepatocytes and Kupffer cells of rats treated with gadoteric acid solution spiked with low-stability impurity, free gadolinium and gadodiamide, but not in rats treated with the pharmaceutical solution of gadoteric acid. The presence of impurities in the GC pharmaceutical solution may have long-term biological consequences.
Asunto(s)
Quelantes/farmacocinética , Medios de Contraste/farmacocinética , Gadolinio/farmacocinética , Compuestos Heterocíclicos/farmacocinética , Compuestos Organometálicos/farmacocinética , Insuficiencia Renal/metabolismo , Animales , Química Farmacéutica , Fémur/metabolismo , Gadolinio/sangre , Compuestos Heterocíclicos/sangre , Riñón/metabolismo , Hígado/metabolismo , Masculino , Miocardio/metabolismo , Compuestos Organometálicos/sangre , Ratas Wistar , Piel/efectos de los fármacos , Piel/metabolismoRESUMEN
(Quantitative) structure activity relationship [(Q)SAR] modeling is the primary tool used to evaluate the mutagenic potential associated with drug impurities. General recommendations regarding the use of (Q)SAR in regulatory decision making have recently been provided in the ICH M7 guideline. Although (Q)SAR alone is capable of achieving reasonable sensitivity and specificity, reliance on a simple positive or negative prediction can be problematic. The key to improving (Q)SAR performance is to integrate supporting information, also referred to as expert knowledge, into the final conclusion. In the regulatory context, expert knowledge is intended to (1) maximize confidence in a (Q)SAR prediction, (2) provide rationale to supersede a positive or negative (Q)SAR prediction, or (3) provide a basis for assessing mutagenicity in absence of a (Q)SAR prediction. Expert knowledge is subjective and is associated with great variability in regards to content and quality. However, it is still a critical component of impurity evaluations and its utility is acknowledged in the ICH M7 guideline. The current paper discusses the use of expert knowledge to support regulatory decision making, describes case studies, and provides recommendations for reporting data from (Q)SAR evaluations.
Asunto(s)
Bases de Datos Factuales/legislación & jurisprudencia , Contaminación de Medicamentos/legislación & jurisprudencia , Sistemas Especialistas , Mutágenos , Relación Estructura-Actividad Cuantitativa , Bases de Datos Factuales/normas , Contaminación de Medicamentos/prevención & control , Humanos , Pruebas de Mutagenicidad/normasRESUMEN
The review presents a survey of recent applications of high-performance capillary electromigration methods-capillary zone electrophoresis, nonaqueous capillary electrophoresis, capillary isotachophoresis, micellar electrokinetic chromatography, microemulsion electrokinetic chromatography and capillary electrochromatography-for the determination of impurities of pharmaceuticals, including chiral impurities, for the period 2007-2013. In addition, due to the missing evaluation of the determination of counterions of pharmaceuticals by capillary electromigration methods in the last 20 years, the publications dealing with this topic since 1995 are included in this review. General aspects of both these types of applications of capillary electromigration methods in pharmaceutical analysis are discussed, and detailed experimental conditions used for determination of various chemical impurities and counterions of many particular drugs are described.