Development and Validation of a Stability-Indicating RP-HPLC Method for the Determination of Erythromycin Related Impurities in Topical Dosage Form

The aim of this research work was to develop and validate a stability-indicating, single reversed-phase HPLC method for the separation of five impurities, including enantiomers, diastereomers, and degradation products in sacubitril-valsartan tablets.The method was developed using a Chiralcel OJ-RH column (150 Â 4.6 mm, 5 μm) at 45 C with a gradient program of (T/%B) 0.01/25, 10.0/25, 25/38, 37.0/45, 39.0/25, and 45.0/25 at a flow rate of 0.8 ml/min. Mobile phase A consisted of 1 ml of trifluoroacetic acid in 1000 ml of Milli-Q water. Mobile phase B consisted of 1 ml of trifluoroacetic acid in a mixture of acetonitrile and methanol in the ratio of 950:50 (v/v). Sacubitril, valsartan, and their five impurities were monitored at 254 nm. Degradation was not observed when sacubitril-valsartan was subjected to heat, light, hydrolytic, and oxidation conditions. In acid degradation study (1 N HCl/60 C/2 h) impurity 1 (m/z 383.44) was formed, and in base degradation study (0.1 N NaOH/40 C/1 h) impurities 1 and 5 (m/z 265.35) were formed; both impurities were confirmed using LC-MS. The degradation products, enantiomers, and diastereomers were well separated from sacubitril and valsartan, proving the stability-indicating power of the method. The developed method was validated per the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. The inter-and intra-day percentage relative standard deviation for sacubitril, valsartan, and their five impurities was less than 5.2%, recovery of the five impurities was between 93 and 105%, and linearity was ≥0.999. The limit of detection was 0.030-0.048 μg/ml, and the limit of quantification was 0.100-0.160 μg/ml.

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development and validation of a stability‐indicating, single HPLC method for sacubitril–valsartan and their stereoisomers and identification of forced degradation products using LC–MS/MS

Vijaykumar

Yalavarti²,

Aparna

2022

“…The forced degradation (stress testing) gives information on the degradation products, degradation pathways and intrinsic stability of drug moiety. The forced degradation studies of ALM tablets were employed with reference to the ICH guidelines (Blessy et al, 2014; Ettaboina et al, 2022; ICH, 1996, 2003; Narasimha & Chandrasekar, 2019) using a concentration of 0.5 mg/ml of ALM. With the intention of developing a stability‐indicating method for the quantitative estimation of ALM assay and its impurity profiling, the stress testing studies were conducted using hydrolysis (acid, alkali, neutral and humidity conditions), oxidation, photolysis (UV and fluorescence lights) and thermal stress testing.…”

Section: Methodsmentioning

confidence: 99%

Section: Stress Testingmentioning

confidence: 99%

Degradation pathways and impurity profiling of the anticancer drug apalutamide by HPLC and LC–MS/MS and separation of impurities using Design of Experiments

Lakka

Kuppan

Vadagam

et al. 2022

Apalutamide, an androgen receptor inhibitor, is used to treat prostate cancer. A stability-indicating high-performance liquid chromatography method was developed for the estimation of assay and organic impurities of apalutamide in drug substance and in tablet dosages using Design of Experiments. The chromatographic separation was achieved within 30 min using Atlantis dC 18 , 100 Â 4.6 mm, 3.0 μm and the binary gradient program (10 mM KH 2 PO 4 , pH 3.5; acetonitrile). The detection wavelength, flow rate, column temperature and injection volume used were 270 nm, 1.0 ml/min, 45 C and 10 μl, respectively. The interaction of independent variables (pH, column temperature and flow rate) and their influences on HPLC parameters were studied using a central composite design, and then the peak separation and elution behaviors between apalutamide and its seven impurities were determined. The method validation was performed for linearity, detection limit, quantitation limit, accuracy, precision and robustness as per the International Conference on Harmonization. A high-quality recovery with good precision (91.7-106.0%) and correlations (r 2 > 0.997) within a linear range of 0.12-2.24 μg/ml (0.05-0.3%, w/w) were achieved consistently for assay and organic impurities of apalutamide. The stabilityindicating characteristics of the proposed method were assessed through forced degradation and mass balance studies. An effort was made to figure out the chemical structures of newly formed degradation products (DP1-DP5) using LC-MS/MS.

“…Rivaroxaban is used as an antithrombotic agent and exhibits potent anticoagulant effects (Perzborn et al, 2005; Roehrig et al, 2005). In the literature, a number of analytical methods have been described for the estimation of rivaroxaban and its impurities employing various techniques such as spectrophotometric methods (Celebier et al, 2014; Satyanarayana & Madhavi, 2012a; Sekaran et al, 2013), HPLC (Celebier et al, 2013; Dongala et al, 2021; Ettaboina et al, 2021, 2022; Jebaliya et al, 2015; Katakam, Dongala, & Ettaboina, 2021; Pinaz kasad, 2013; Pinaz kasad & Krishna, 2013; Prajapati & Patel, 2015; Rao et al, 2015; Rao & Sekar, 2013; Satyanarayana & Madhavi, 2012a, 2012b; Sekaran et al, 2013; Sekhar Chandra et al, 2012; Sunitha et al, 2014; Yenda et al, 2022), chiral HPLC (Ettaboina et al, 2021; Katakam et al, 2020; Katakam, Dongala, & Ettaboina, 2021; Prabhune et al, 2015), and LC–MS (Abdallah et al, 2015; Katakam, Dongala, & Ettaboina, 2021; Rao Nageswara & Krishna Rama, 2014). The main aim of this present research work is to develop a single method for the separation of enantiomers and its process‐related known and unknown impurities in drug substances containing rivaroxaban.…”

Section: Introductionmentioning

confidence: 99%

Stability‐indicating HPLC method development and validation of rivaroxaban impurities and identification of forced degradation products using LC–MS/MS

Vijaykumar

Y.²,

Aparna

et al. 2022

The current aim of this study is to develop a simple, sensitive, and robust analytical method that can separate the Rivaroxaban and its related impurities by using HPLC.This new method can separate enantiomers and all the process-related impurities of rivaroxaban. This method can also be used to determine the assay of rivaroxaban in drug substances and drug products. This new method was developed using a Chiralpak IC (250 Â 4.6 mm, 5 μm) column at 27 C with a gradient program of 25.0 min at a flow rate of 0.8 mL/min. Mobile phase A consisted of acetonitrile, ethanol, and n-butyl amine in the ratio of 95:5:0.5 (v/v/v), respectively. Mobile phase B consisted of a mixture of Milli-Q water, methanol, and n-butyl amine in the ratio of 50:50:0.5 (v/v/v), respectively. The detector wavelength was 254 nm. Rivaroxaban was subjected to the forced degradation conditions (stress conditions) of hydrolysis, base, acid, thermal, photolysis, and oxidation. The degradation products were well separated from rivaroxaban peak and enantiomer peak, and its potential impurities prove the stability-indicating power of the method. The proposed new stability-indicating method was validated with respect to specificity, precision, accuracy, limit of quantification, limit of detection, linearity robustness, and roundness per International Conference on Harmonization guidelines. The %RSD (relative standard deviation) for six sample preparations for rivaroxaban and impurities less than 10% and the recovery is between 90 and 110%. The current method can be used in quality control labs for analysis of commercial products.