Chromatographia最新文献

Regulatory agencies prioritize the safety and efficacy of pharmaceuticals. In this context, our research aims to develop a simple, sensitive, and eco-friendly LCMS-compatible high-performance liquid chromatography stability indicating analytical method (SIAM) technique to identify impurities in Acalabrutinib (ACB) under various stress conditions. This method is designed in compliance with ICH QIA (R2) and Q3, incorporating Green Chemistry principles and Quality by Design (QbD). To optimize the method, an ideal split-plot design was used to evaluate critical method parameters (CMPs), followed by a central composite design (CCD) to refine the conditions. Statistical analysis revealed p values < 0.001 for the model and 0.05 for lack of fit, indicating the most suitable statistical model for the evaluated responses (peak resolutions R1–R4). The optimized method attributes include an ACN:ammonium acetate buffer (pH 5) ratio of 50:50 (v/v), a flow rate of 0.8 mL/min, and a column oven temperature of 35 °C, derived from CCD. An isocratic elution method using the Waters e2998 HPLC Kromasil 100-5-C18 (250 × 4.6 mm; 5 µm) analytical column enabled superior separation of components, with a run time of 35 min and a wavelength of 254 nm. Stress studies revealed that ACB is sensitive to hydrolysis and photolytic conditions, with ACN identified as the most suitable diluent. Method validation was conducted according to ICH Q2 guidelines, and showed a correlation coefficient (r²) exceeding 0.992, with RSD values (n = 6) ranging from 0.76 to 1.93% across the LOQ-150% range. Specificity studies confirmed no interference between impurities and active analytes. Through stress testing, 28 degradation products were identified, and the method was assessed for eco-friendliness using seven different tools, confirming its sustainable nature for pharmaceutical applications.

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A novel HPLC system utilizing a phase-separation multiphase flow as the eluent has been developed, referred to as phase-separation mode. This research explores the influence of the porous structure in an octadecyl-modified silica (ODS) column (with a pore diameter of 12 nm) on chromatographic outcomes under the phase-separation mode in HPLC. The chromatograms obtained from the porous ODS column were compared to those generated with a non-porous ODS column. In preliminary experiments, twenty-four mixed solutions, comprising combinations of water/acetonitrile/ethyl acetate and water/acetonitrile, were introduced as eluents at a column temperature of 20 °C. A model mixture of 2,6-naphthalenedisulfonic acid (2,6-NDS) and 1-naphthol (1-NA) was injected into the system, with separation achieved in most solutions except for some highly organic solvent-rich solutions where 2,6-NDS eluted faster than 1-NA, indicating reverse-phase mode operation. Subsequently, the separation of the model mixture was assessed at 0 °C, and four specific ternary mixtures were analyzed in detail at both 20 °C and 0 °C. These ternary mixtures, defined by their volume ratios, exhibited a two-phase separation, establishing a phase-separation multiphase flow. Consequently, the solution flow was homogeneous at 20 °C and heterogeneous at 0 °C. For instance, solutions with water/acetonitrile/ethyl acetate ratios of 20:60:20 (organic solvent-rich) and 70:23:7 (water-rich) were introduced as eluents at both 20 °C and 0 °C. At 0 °C in the organic solvent-rich eluent, 1-NA eluted faster than 2,6-NDS, characteristic of the phase-separation mode. In contrast, the water-rich eluent resulted in faster elution of 2,6-NDS at both temperatures. The porous ODS column displayed improved separation efficiency at 0 °C compared to the non-porous column, which can be attributed to the porous effect under phase-separation conditions.

Combination nasal spray products containing fluticasone furoate and oxymetazoline hydrochloride are commonly used to treat asthma, chronic pulmonary diseases, and perennial allergic rhinitis. However, testing for impurities in pharmaceutical products is a challenging task due to the similarity in properties between the impurities and the drugs. It is important to strictly maintain the product safety and quality to ensure the regulatory compliance. An attempt has been made to develop and validate a new simple, single-run RP-HPLC method for the simultaneous estimation of seven known and several unknown impurities from fluticasone furoate and oxymetazoline hydrochloride nasal spray products. The separation of multiple impurity peaks was achieved by using a 5 µm, Inertsil ODS 3 V, 250 × 4.6 mm column. Dual-wavelength UV detector (220 nm and 240 nm) was used for the detection of oxymetazoline hydrochloride, fluticasone furoate and their impurities. The relative response factor was calculated for all the known impurities to make the method accurate, simple and economical. The developed method was validated following the guidelines outlined in the ICH Q2(R2). The forced degradation study results confirmed the stability-indicating nature of the method. The developed impurity profiling methodology will greatly help the quality control labs in maintaining the safety and efficacy of finished products and to comply with the regulatory requirements of various countries. Overall, the proposed method offers unparalleled precision, accuracy, sensitivity, and cost-effectiveness making it a valuable addition to the pharmaceutical industry.

Dalbergia sissoo Roxb., rich in isoflavones and isoflavone glycosides, is widely used to treat osteogenic effects, pain, and other diseases. This study employed simple procedures based on preparative reversed-phase high-pressure liquid chromatography (prep-RP-HPLC) for large-scale isolation of isoflavones and isoflavone glycosides present in Dalbergia sissoo leaves (DSL). The seven compounds were isolated from the ethanolic extract by the preparative HPLC–PDA method, and their purity was > 95%. The authors carried out an in silico analysis to find possible anti-aging compounds. Isolated compounds biochanin-A-7-O-[β-Dapiofuranosyl-(1–5)-β-D-apiofuranosyl-(1–6)-β-D-glucopyranoside (DS2) and biochanin-A-7-O [β-D-apiofuranosyl-(1–6)-β-D-glycopyranoside (DS3) demonstrated docking scores of −10.466 and −9.430 with docking energies of -81.864 and -77.531 kcal/mol, respectively, to the Hyaluronidase enzyme. For the Collagenase enzyme, they showed docking scores of −10.734 and −10.777 with docking energies of −107.102 and −98.317 kcal/mol, respectively. Against the Tyrosinase enzyme, Compounds DS2 and DS3 had docking scores of −7.333 and −7.993 with docking energies of −59.933 and −84.825 kcal/mol, respectively. For the elastase enzyme, they exhibited docking scores of −9.840 and −7.993 with docking energies of −68.490 and −84.815 kcal/mol, respectively. This study found that the isolation and purification method is simple and produces neat chromatograms with good selectivity and reproducibility. At the same time, docking results revealed that the compounds exhibited anti-aging properties.

Graphical abstract

The purpose of this study was to develop and validate a robust, precise, and selective high-performance liquid chromatography (HPLC) method for the separation and determination of related impurities in fosaprepitant dimeglumine API. The chromatographic separation was performed on a Supersil ODS-2 column (250 × 4.6 mm, 5 µm) at a wavelength of 215 nm using a mixture of phosphate buffer (pH 2.15) and acetonitrile as the mobile phase in gradient elution mode. The validation results demonstrate that the method exhibits acceptable specificity, linearity, accuracy, precision, and robustness. The detection limits and quantitation limits ranged from 1.5 to 12.5 ng mL⁻¹ and from 3.0 to 37.5 ng mL⁻¹, respectively. A linear relationship was observed between the peak area and concentration of fosaprepitant and its eight related impurities with a correlation coefficient value of r² ≥ 0.999. The analysis of commercial fosaprepitant dimeglumine products revealed a significantly higher purity than expected, with all known impurities falling below specification limits. The new HPLC method has been successfully applied to analyze commercial bulk drug samples and is suitable for quality-control laboratories for both qualitative and quantitative assessment of eight related substances in the fosaprepitant dimeglumine API.

The objective of the present numerical investigation is to propose a microseparator specifically engineered for the separation of platelets from white blood cells (WBCs). This microfluidic device, which incorporates circular electrodes operating at a minimal voltage of 1.4 V, and a frequency of 100 kHz, is recommended for the targeted separation of platelets and WBCs utilizing dielectrophoresis (DEP) force. The utilization of a low-voltage level ensures the preservation of the viability of biological cells, a fundamental consideration in medical applications. Simulation results are presented to illustrate the electric potential, electric field, velocity, pressure, and DEP force profiles concerning two distinct particles. Through a comparative analysis employing the finite element method, we delineate the implications of modulating the inlet velocity field on the pressure exerted upon the particles. Subsequently, the impact of varying the fluid conductivity and input voltage on electrodes within the microchannel on particle separation was examined. It is anticipated that this comprehensive design is exceptionally conducive to the realization of DEP-based practical biochips for cell separation.

A rapid and efficient reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous determination of six antiviral drugs: emtricitabine (EMT), entecavir (ENT), favipiravir (FAV), ganciclovir (GAN), valaciclovir (VAL), and valganciclovir (VLG). Chromatographic separation was achieved on an ExsilTM Mono 100 C18 column using a mobile phase of 50 mM phosphate buffer (pH 4): acetonitrile (90:10, v/v) at a flow rate of 0.5 mL/min. UV detection was performed at 215 nm. The method was validated according to ICH Q2(R2) guidelines, demonstrating excellent linearity (R² > 0.9938) across the concentration ranges of 1.0–150 μg/mL for EMT, ENT and VLG, and 5.0–200 μg/mL for FAV, GAN and VAL. The LOD and LOQ values ranged from 0.02 to 0.48 μg/mL and 0.05 to 1.45 μg/mL, respectively. Intra-day and inter-day precision, expressed as relative standard deviation (RSD), were less than 1.82%, while accuracy, expressed as relative error (RE), was within ± 1.34%. Recovery studies yielded values ranging from 98.12% to 101.60%, further confirming method accuracy. The method proved to be selective, robust, and suitable for the quantitative analysis of these antiviral drugs in pharmaceutical formulations, with results obtained for commercially available tablets demonstrating excellent agreement with labeled contents. Notably, stability studies revealed the susceptibility of EMT, ENT, and FAV to degradation under refrigerated storage, highlighting the need for timely analysis of these compounds. In addition, the greenness, blueness, and whiteness of the method were evaluated, and the developed method performs strongly across environmental (AGREE score: 0.79), practical (BAGI index: 85), and sustainability (whiteness value: 74.7) criteria, reinforcing its viability as a sustainable approach for antiviral analysis. This validated RP-HPLC method provides a reliable and efficient tool for the quality control of these essential antiviral medications.

The detection and quantification of genotoxic impurities, specifically nitrosamines, in pharmaceuticals have garnered high regulatory attention due to their potential carcinogenic effects. This study presents a comprehensive validation of a liquid chromatography with Atmospheric Pressure Chemical Ionization (APCI) Coupled with Triple Quardrapole Analyser (LC-MS/MS) method tailored for the determination of multiple nitrosamine impurities including N-nitrosodimethylamine (NDMA), N-nitroso methyl butyl amine (NMBA), N-nitrosodiethylamine (NDEA), N-nitroso diethyl isopropyl amine (NEIPA), N-nitrosodiisopropylamine (NDIPA), N-nitroso methyl ethyl amine (NMPA), N-nitroso propyl amine (NDPA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitroso butyl amine (NDBA) in telmisartan tablet formulations. The method was meticulously validated according to the International Council for Harmonisation (ICH) guidelines, focusing on critical parameters such as specificity, accuracy, precision, limit of detection (LOD), limit of quantification (LOQ), and filter compatibility. The specificity of the method was rigorously established, demonstrating unequivocal differentiation between nitrosamine impurities and potential interferences from excipients and active pharmaceutical ingredients (APIs). Recovery studies validated the method’s accuracy, yielding recoveries within the acceptable range of 70–120% across various concentration levels, confirming its reliability for routine analysis. Precision was assessed through the relative standard deviation (RSD) of multiple replicate analyses, with RSD values consistently below the ICH threshold of < 15%, underscoring the method’s reproducibility. Sensitivity assessments revealed exceptional LODs as low as 10.76 ppb and LOQs around 33.00 ppb for several nitrosamines, significantly exceeding current regulatory limits for genotoxic impurities. Evaluations of filter compatibility indicated that both 0.22 µm PVDF and 0.2 µm nylon filters are effective for sample preparation, ensuring the integrity of the analytes during the filtration process. Upon application of the validated method to commercial 40 mg telmisartan tablets, all targeted nitrosamines were undetected, affirming the method’s applicability in routine quality control settings. This work culminates in the establishment of a highly sensitive, specific, and robust LC–MS/MS methodology for the detection of nitrosamine impurities in telmisartan, addressing regulatory concerns while ensuring patient safety. The low detection thresholds provided by this method position it as an invaluable tool for pharmaceutical manufacturers and regulatory bodies, facilitating stringent monitoring and compliance with safety standards, ultimately enhancing the safety profile of medicinal products in the market.