Quantification Of Genotoxic Impurities Research Articles

A novel UHPLC-MS/MS method for trace level identification and quantification of genotoxic impurity 2-(2-chloroethoxy) ethanol in quetiapine fumarate

A selective and sensitive new reversed-phase method was developed and validated by using Waters Acquity UHPLC system coupled with Waters Micromass Quattro PremierXE (MS/MS) to identify and quantify the genotoxic impurity 2-(2-chloroethoxy) ethanol at trace level (∼2.0 ppm) in quetiapine fumarate. The method used positive ion electrospray ionization with a multiple reaction monitoring detection mode using ACE3 C18(100 mm × 4.6 mm × 3.0 µm) column. For analysis, an isocratic program was developed using 0.01 M ammonium acetate in Milli-Q water (mobile phase A) and methanol (mobile phase B) in 20:80v/v ratio. Elution of 2-(2-chloroethoxy) ethanol was monitored using triple quadrupole mass spectrometer with an injection volume of 10 μl, a column oven temperature of 40 °C, an auto-sampler temperature of 15 °C, and flow rate of 0.5 mL/min. The retention time of 2-(2-chloroethoxy) ethanol was observed at 2.38 min. The LOQ and LOD were determined at concentrations of 0.235 ppm and 0.070 ppm respectively. The correlation coefficient R 2 was 0.9983 and the percent recoveries of method range from 98.1% to 114.0%. The method was successfully validated according to International Council for Harmonization requirements (ICH Q2(R1) and ICH M7(R1)) for genotoxic impurities. The method was sensitive, selective, accurate and precise and therefore can be used for identification and quantification of genotoxic impurity 2-(2-chloroethoxy) ethanol in quetiapine fumarate.

Quantification of Genotoxic Impurities by HPLC in Active Pharmaceutical Ingredient Lorazepam

Quantification of Genotoxic Impurities by HPLC in Active Pharmaceutical Ingredient Lorazepam

Review on identification and quantification of genotoxic impurities

Genotoxic impurities can be broadly defined as those impurities that have been demonstrated to cause harmful changes in genetic material regardless of the mechanism. Globally people suffer from various health complications due to genotoxic impurities. Recent recommendations from European and United States (USA) drug regulatory bodies mandate the management of genotoxic and possibly genotoxic contaminants in pharmaceutical ingredients at per million levels. The purpose of this review is to make a critical analysis of the techniques used to comply with the prevailing rules and regulations and very strict limits on genotoxic impurities. Possible strategies to further expand the scope of currently available technologies and regulations are also to be discussed. These strategies include redesigning the synthesis of the drug substance to avoid introducing problematic impurities; modifying relevant process parameters to eliminate or reduce such impurities to negligible levels; Using process understanding to demonstrate that a particular genotoxic impurity cannot be formed or removed efficiently and by conducting toxicity studies to demonstrate that a suspected impurity is not harmful to it at low levels.

Trace level detection and quantification of genotoxic impurities 3‐amino‐4‐methylbenzoate, 3‐amino‐4‐methylbenzoic acid, and 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline in Nilotinib dihydrochloride active pharmaceutical ingredient using liquid chromatography‐tandem mass spectrometry

AbstractA selective and sensitive novel methods were developed and validated for identification and trace level quantification (0.5 ppm) of genotoxic impurities Methyl 3‐amino‐4‐methylbenzoate, 3‐amino‐4‐methylbenzoic acid, and 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline in Nilotinib dihydrochloride active pharmaceutical ingredient by using liquid chromatography‐tandem mass spectrometry. The developed and validated methods were more accurate and capable to confirm the m/z values of parent and fragment ions through mass spectrometry and tandem mass spectrometry for further fragmentation. In addition, the developed methods were validated through current regulatory guidelines with a lower detection level of 0.15 ppm and a quantification level of 0.5 ppm for all the three genotoxic impurities. The correlation coefficient was observed as 0.9997 for Methyl 3‐amino‐4‐methylbenzoate genotoxic impurity‐I, 0.9998 for 3‐amino‐4‐methylbenzoic acid, and > 0.9999 for 3‐(4‐methyl‐1H‐imidazol‐1‐yl)‐5‐(trifluoromethyl) aniline. The recovery percentage of all the three genotoxic impurities was found to be between 93% to 105%.

A SELECTIVE AND SENSITIVE LC-MS/MS METHOD FOR QUANTIFICATION OF FOUR POTENTIAL GENOTOXIC IMPURITIES IN ATAZANAVIR SULFATE DRUG SUBSTANCE IN TRACE LEVEL

Objective: The main objective of current research work is to develop and validate a rapid, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the trace analysis of four potential genotoxic impurities in Atazanavir Sulfate drug substance.
 Methods: LC-MS/MS analysis of four potential genotoxic impurities was done on Acquity UPLC CSH C18 (100 mm × 2.1 mm, 1.7 μm) column. In this method, mobile phase A (10 mM ammonium acetate) mobile phase B (methanol: acetonitrile (90:10, v/v) with gradient run with the flow rate of 0.2 ml/min. The method was developed with the short run time of 13 min. Triple quadrupole mass detector coupled with positive electrospray ionization was used for the quantification of genotoxic impurities in multiple reaction monitoring (MRM) mode.
 Results: The method was linear in the range of 0.3 ppm to 4.5 ppm for BOC Hydrazine Acid impurity, BOC Epoxide and Keto impurity with a correlation coefficient not less than 0.9994. The accuracy of the method was in the range of 99.26% to 105.71% for all four potential genotoxic impurities (PGIs). No impurities were identified in the Atazanavir Sulfate active pharmaceutical ingredient sample.
 Conclusion: The proposed method is specific, linear, precise, accurate, robust and stable for the quantification of the four genotoxic impurities at very low levels.

Quantification of Genotoxic Impurities in Key Starting Material of Sertraline Hydrochloride by Simple and Sensitive Liquid Chromatography Technique

Background:Schiff base is an advanced key starting material of Sertraline hydrochloride. Schiff base is synthesized using two raw materials 1-Naphthol and 1,2-Dichlorobenzene which are potentially genotoxic impurities.Objective:Genotoxic impurities need to be controlled in key starting material to avoid carry forward in the active pharmaceutical ingredient. For trace level quantification of impurities a sensitive, accurate and cost effective method is developed by simultaneous estimation of both impurities.Methods:Reverse phase high performance liquid chromatography (HPLC) method was developed and validated for determination of both impurities in Schiff base. HPLC column Cosmosil MS-II C18, 100 mm X 4.6 mm, 3 μm particle size with ultra-violet detector (UV) was used.Results:The calibration curve of 1-Naphthol and 1,2-Dichlorobenzene showed good linearity over the concentration range of 0.25 µg/g to 7.5 µg/g and 1.5 µg/g to 7.5 µg/g and the regression coefficient was 0.999 and 0.998 respectively. Method had very low limit of detection (LOD) and limit of quantification (LOQ) of both analytes which proves that the method is sensitive and suitable for quantification of compounds at trace level.Conclusion:The proposed method is specific, linear, accurate, rugged and precise. Genotoxic impurities 1-Naphthol and 1,2-Dichlorobenzene are quantified and controlled in the key starting material of Sertraline hydrochloride. The validated method can be used in quality control unit in pharmaceutical industry.

Quantification of Genotoxic Impurities in Tadalafil by Simple and Sensitive Gas Chromatography Technique

Chloroacetyl chloride and thionyl chloride are used in the manufacturing of tadalafil. On basis of structure-activity relationship (SAR) both compounds are potentially genotoxic. A sensitive gas chro- matography (GC) method was developed and validated for the determination of both impurities in the active pharmaceutical ingredient tadalafil. GC column Agilent DB-1 of length 15 m, internal diameter 0.32 mm and film thickness 3 μm with flame ionization detector (FID) was used. The proposed method is specific, linear, accurate, rugged, robust and precise. The calibration curve of chloroacetyl chloride and thionyl chloride showed linear response over the concentration range of 7.5 μg/g to 56.4 μg/g with respect to test solution concentra- tion and the regression coefficient was 0.999 for both analytes. Method had very low limit of detection (LOD) and limit of quantification (LOQ) of both analytes which proves that the method is sensitive and suitable for quantification of both genotoxic impurities at very low level.

Confirmation and Quantification of Genotoxic Impurity 2- Dimethylaminoethyl chloride hydrochloride (DMC HCl) by GCMS in Chlorpheniramine/Chlorphenamine Maleate

Confirmation and Quantification of Genotoxic Impurity 2- Dimethylaminoethyl chloride hydrochloride (DMC HCl) by GCMS in Chlorpheniramine/Chlorphenamine Maleate

Trace Level Determination of Three Genotoxic Impurities in Drug Samples of Rizatriptan Benzoate by Liquid Chromatography-Tandem Mass Spectrometry

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was developed and validated for the trace analysis of three potential genotoxic impurities in rizatriptan benzoate drug sample. LC-MS/MS analysis of these impurities was done on symmetry C-18 (100 mm x 4.6 mm, 3.5 μm) column. In this method, 0.2% formic acid was used as mobile phase A and mixture of methanol and acetonitrile (5:95, v/v) used as mobile phase B in gradient mode with the flow rate of 0.8 mL/min. The method was developed with the short run time of 12 minutes. Triple quadrupole mass detector coupled with positive electrospray ionization was used for the quantification of genotoxic impurities in multiple reaction monitoring (MRM) mode. The method was linear in the range of 2.97 ppm to 74.16 ppm for N-(3-chloropropyl)-N,N- dimethylamine (CDA), 3.02 ppm to 75.44 ppm for l-(4-hydrazinophenyl) methyl-l,2,4-triazole (HMT) and 3.02 ppm to 75.52 ppm for l-(4-nitrophenyl) methyl-1,2,4-triazole (NMT) with a correlation coefficient not less than 0.9995. The accuracy of the method was in the range of 92.75% to 102.49% for all three potential genotoxic impurities (PGIs). No impurities were identified in the rizatriptan benzoate active pharmaceutical ingredient sample. This method is able to quantify the impurities at very low levels.

Method Development and Validation using RP-HPLC for estimation of Genotoxic impurity i.e. Melamine mainly present as contaminant in diversified categories of API

A simple and precise gradient Reversed Phase High Performance Liquid Chromatography (RP-HPLC) common method was developed for determination and quantification of genotoxic impurity i.e. Melamine. The gradient mobile phase consisted of A = 0.1%v/v Tetra butyl Ammonium Hydroxide 0.1N solution in Isopropyl Alcohol in water and B = 100% Acetonitrile, the flow rate was kept 1 mL/min using Unisphere Extend C18 column from Agela Technologies (250 mm x 4.6 mm x 5μm). Detection was performed at 220 nm using PDA detector. The common method was validated for LOD, LOQ, Linearity and Accuracy as per ICH guidelines. This method is suitable for detecting and quantifying melamine in various categories of API's such as for cardiovascular problems, Anti psychotic, API's for stomach and esophageal problems, Anti-diabetic, Antibiotic, Anti fungal, Anti gout, Antimalarial, Anti-Alzheimer and Antiadrenergic API's. Precision and accuracy was performed for all the above mentioned categories of API's on individual basis at three concentrations in three replicates i.e. at 80%, 100% and 150% of the limit concentration. Also LOQ level was shown to be precise and accurate by spiking the individual API's at LOQ level.

METHOD DEVELOPMENT AND VALIDATION STUDY FOR QUANTITATIVE DETERMINATION OF GENOTOXIC IMPURITY AND ITS PRECURSOR IN FLUCONAZOLE SAMPLE BY LIQUID CHROMATOGRAPHY–TANDEM MASS SPECTROMETRY

<p><strong>Objective: </strong>The objective of this work is method development and validation study for quantitative determination of 1-[2-(2,4-difluorophenyl)-2,3-epoxypropyl]-1H-1,2,4-triazole, a genotoxic impurity and its precursor in a fluconazole drug sample by liquid chromatography–tandem mass spectrometry.</p><p><strong>Methods: </strong>LC-MS/MS analysis of these impurities was performed on Hypersil BDS C18 (100 mm x 4.0 mm, 3 µm) column. 5 mmol ammonium acetate and acetonitrile in the ratio of 65:35 (v/v) was used as the mobile phase with a flow rate of 0.4 ml/min. The developed method was accomplished with a short run time of 10 min. Triple quadrupole mass detector coupled with positive electrospray ionization was used for the quantification of genotoxic impurities in multiple reaction monitoring (MRM).</p><p><strong>Results: </strong>The method was validated as per International Conference on Harmonization (ICH) guidelines. The method was linear in the range of 0.30 µg/g to 11.37 µg/g for impurity A and 0.30 µg/g to 11.34 µg/g for impurity B with a correlation coefficient of 0.999. The accuracy of the method was in the range of 98.25 % to 100.53 % for both impurities.</p><p><strong>Conclusion: </strong>A specific, selective, highly sensitive and more accurate analytical method using LC-MS/MS coupled with positive electrospray ionization has been developed for the quantification of genotoxic impurity (1-[2-(2,4-difluorophenyl)-2,3-epoxypropyl]-1H-1,2,4-triazole) and its precursor (1-(2,4-difluorophenyl)-2-[1,2,4]triazol-1-yl-ethanone) at 0.3 µg/g with respect to the 5.0 mg/ml of fluconazole.</p>

Characterization and quantification of genotoxic impurities complying with ICH M7: A pharma perspective

Characterization and quantification of genotoxic impurities complying with ICH M7: A pharma perspective

Quantification of Genotoxic Impurity 4-Chloro-1-Hydroxy Butane Sulfonic Acid Sodium Salt by LCMS/MS in Sumatriptan Succinate

The objective of present research work is to develop a suitable LCMS/MS method for the quantitative determination of genotoxic impurity 4-Chloro-1-Hydroxy Butane Sulfonic Acid Sodium Salt at ppm level present in Sumariptan dug substance. The LCMS/MS method was developed on Zorbax SB-C8 column using the mobile phase consists a mixture of 0.05% (v/v) Formic acid in water and Acetonirile using a isocratic composition of 90:10 (v/v) at a flow rate of 0.8 mL/min. Ion source is electospray ionization (ESI), source temperature is 325°C, gas flow is 8 L/min, Nebuliser pressure is 40 psi, capillary voltage is 4000 V. Under these conditions impurity was quantified by selecting most stable MRM pair (187/81). The limit of detection and the limit of quantitation for the impurity were established. Validation of the developed LCMS/MS method was carried out as per ICH requirements and the data shows that the proposed method is specific, linear, accurate, precise and robust. This method has been tested in a number of Sumatriptan samples and used successfully for quantification of the impurity at ppm level. The developed LCMS/MS method was found to be suitable to quantify the genotoxic impurity 4-Chloro-1-Hydroxy Butane Sulfonic Acid Sodium Salt at ppm level present in Sumatriptan Succinate.