Journal of Pharmaceutical and Biomedical Analysis Open最新文献

Nowadays, breast cancer is the most affecting and globally diagnosed malignancy among women, yet Letrozole (LTZ) was considered the first-line treatment as hormonal anticancer drug. Unfortunately, LTZ develops osteoporosis as a main side effect which was overcome by using the co-administered; Zoledronic Acid (ZDA). Thus, there was a crucial need for this simultaneous quantification innovation, especially there were no any previously reported methods regarding both drugs together. In this study, an integrated framework was conducted between the experimental analytical quality-by-design (AQbD) approach and green analytical chemistry (GAC), emerging sensitive and robust RP-HPLC method. Box-Behnken Design was the developed model for optimizing an isocratic chromatographic separation on C₁₈ Equisil® ODS (4.6 × 250 mm, 5.0 μm) column at ambient temperature, using the mobile phase of 0.1 % aqueous trifluroacetic acid (pH 2.8): acetonitrile (54.5:45.5, v/v), at 1.0 mL/min flow rate with PDA detection at 254.0 nm and 210.0 nm for LTZ and ZDA, respectively. Model statistical and residual plots analysis was significant and normally distributed. Method was fully validated as per ICH guidelines, where good linearity was 0.20–10.00 µg/mL for both drugs in presence of Tadalafil (TDF) as an internal standard, obtaining adequate correlation coefficients (r) values. Calculated LOD results were 0.058 and 0.040 µg/mL while calculated LOQ results were 0.175 and 0.122 µg/mL for LTZ and ZDA, respectively. The proposed method was effectively applied on bulk, pharmaceutical dosage forms, and spiked human plasma. Statistical comparison of the anticipated results with the reported ones was performed. Greenness assessment was evaluated using Green Analytical Procedure Index (GAPI) and Analytical Greenness (AGREE) tools; where superiority results were achieved relative to other reported methods. Finally, an EVG method evaluation tool was assessed, and the attained results were represented through its radar chart.

The aim of this study is to validate a high-performance liquid chromatography (HPLC) – UV assay method for a stability assay of an oral suspension of PYR developed for newborns. Analyses were performed by HPLC with a Kinetex Coreshell C18 column, thermostated at 40°C, coupled to a diode array (λ=230 nm, 280 nm, scan from 190 to 400 nm) with an acetonitrile/methanol/KH₂PO₄ buffer gradient (pH 3, 10 mM). A forced degradation study was performed to validate the stability-indicating character. PYR was subjected to acidic (pH 2) and basic (pH 12) hydrolysis stress, thermal hydrolysis stress at native pH (60°C and 4°C), H₂O₂ oxidation stress (3 % and 15 %) and photolysis stress (UV and natural light) at day 0, 3 and 16. Linearity, specificity, precision, and accuracy of this assay method have been tested. Forced degradation tests demonstrated the stability indicator character of the method and showed that PYR was very sensitive to oxidation, sensitive to light (UV) and insensitive to hydrolysis (thermal and pH). The method of quantification was linear, specific, precise, and accurate according to ICH recommendations. A stability study on the formulation under development will be carried out with this HPLC – UV method.

In recent years, mutagenic nitrosamines, such as N-nitrosodimethylamine, have been detected in medicine. This has led to global product recalls and long-term supply suspensions by pharmaceutical companies and consequent clinical impacts. Measures to control nitrosamines in medicine, including detection methods and clarification of contamination routes, are being implemented worldwide. In this review, we focus on case reports of nitrosamine contamination of drug products in Japan, nitrosamine formation mechanisms during manufacturing and storage, as well as detection methods. We also discuss the acceptable nitrosamine intake (ng/day) in chemically synthesized drug substances in human drugs (including drug products) in the US, EU, and Japan. Overall, nitrosamine contamination of medicines is expected to remain a global public health issue. Therefore, detection methods using new technologies and detailed analysis of the formation mechanisms are necessary. However, excessive regulation may cause essential drug shortages owing to product recall; therefore, a realistic and prudent response based on regulatory science is needed.

This work introduces a novel strategy to selectively and sensitively determine Hemoglobin (Hb) using a microwave-synthesized molecularly imprinted polymer (MIP) and chemiluminescence (CL) as the detection method. Studies conducted on both MIP and non-imprinted polymer (NIP) evidenced that the MIP material exhibits a high adsorption capacity (48 µg/mg) and an imprinted factor of about 3, in addition to a selective adsorption property towards Hb in the presence of other proteins, whose structures are very similar to that of Hb. The high selectivity and sensitivity of the proposed biosensor were also assured by the CL detection, whose mechanism is based on Hb's ability to enhance luminol oxidation in an alkaline medium in the presence of hydrogen peroxide, emitting a blue light whose intensity is related to the Hb concentration. The emitted light intensities from the CL reaction, directly proportional to Hb concentration, were captured and analyzed using a smartphone and RGB Color application, avoiding the use of sophisticated equipment and facilitating the measurement process. In order to streamline and speed up the detection process, all the experimental steps including Hb adsorption on MIP_Hb, supernatant removal, CL reaction, and smartphone readout occur in a single 8-well strip. Therefore, the proposed approach offers one-pot detection for the sensitive (LOD = 0.03 µg.mL⁻¹, equivalent to 1.5 nmol.L⁻¹) and selective determination of Hb in blood samples, for biomedical or forensic applications, through a rapid, low-cost, simple, and affordable approach.

To investigate the stability of Azilsartan kamedoxomil (AKM) under stress conditions and to identify the degradation products, it was subjected to force degradation/stress under basic, acidic, oxidative, thermal, humidity, and photolytic conditions as per international council for harmonisation (ICH) guidelines. AKM degradation was found under basic, acidic, oxidative, thermal, photolytic, and humidity stress. Separation of the four degradation products of AKM was carried out utilizing a C-18 column, employing preparative High-performance liquid chromatography (HPLC) and gradient mode elution. The structures were elucidated with spectroscopic HRMS, 1D, and 2D NMR and the products were identified as 2-hydroxy-1-[[2′-(5-oxo-4,5-dihydro-1,2,4-oxadiazol −3-yl)[1,1′-biphenyl]-4-yl]-methyl]-1 H-benzimidazole-7-carboxylic acid (AKMDP-1), 2-ethoxy-1-[[2′-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)biphenyl-4-yl]methyl]-1 H-benzimidazole-7-carboxylic acid (AKMDP-2), (5-methyl-2-oxo-1,3-dioxol-4-yl)methyl 2-hydroxy-1-[[2′-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)[1,1′-biphenyl]-4-yl]methyl]-1 H-benzimidazole-7-carboxylate (AKMDP-3) and 2-hydroxy-3-oxobutyl 2-ethoxy-1-((2′-(5-oxo-4,5-dihydro-1,2,4-oxadiazol-3-yl)-[1,1′-biphenyl]-4-yl)methyl)-1 H-benzo[d]imidazole-7-carboxylate (AKMDP-4). Out of them, two degradants AKMDP-1 and AKMDP-3 were reported earlier, AKMDP-2 was one of the process intermediate (Azilsartan) used in the preparation of Azilsartan kamedoxomil.

This research focuses on evaluating the data quality and reliability of Size Exclusion Chromatography with Multi-Angle Light Scattering and Refractive Index (SEC-MALS/RI) chromatography in determining molecular weight-related parameters in poly(lactic-co-glycolic) acid (PLGA) polymers. The study utilizes key metrics, particularly percent relative standard deviation (%RSD), to evaluate the precision across different data processing conditions. Exploring baseline selection of the chromatograms reveals that selection of the baseline based on the intrinsic shape of the chromatogram, rather than relying on elution peaks, improves consistency. The study emphasizes the critical role of result fitting in addressing systematic errors and sample heterogeneity, recommending the minima of the elution peak selection range for realistic molecular weight distribution representation. Analysis of light-scattering (LS) detector chromatograms highlights varying effects on different PLGA polymers, emphasizing the need for careful assessment and potential exclusion for precision improvement, especially in the presence of experimental artifacts. Across diverse PLGA polymers, the utilization of different column types, along with considerations such as baseline determination and peak selection, shows minimal variations in Mn, Mw, and polydispersity index (PDI), affirming the consistency of the methodology. The %RSD analysis further supports the methodology's precision, with values well within acceptable limits. In summary, this comprehensive evaluation of methodological conditions, including baseline selection, result fitting, LS detector data management, and column types, establishes a reliable SEC-MALS/RI chromatographic approach by Absolute Method for determining molecular weight parameters in PLGA polymers.

The term ‘tumor’ describes an atypical development of cells that forms a mass inside an organ; depending on the organ's invasive nature and propensity for metastasis, the growth may be benign or malignant. Improving patient outcomes requires the early detection of malignant tumors. Despite its lower resolution and noise problems, ultrasound, which is frequently used to diagnose uterine tumors, is safer and more economical than magnetic resonance imaging (MRI) scans and biopsies. Ultrasound pictures can be processed using methods including denoising, enhancement, segmentation, and feature extraction to get around these restrictions and boost their quality. Enhanced ultrasound images can reach even higher accuracy, cementing them as plausible alternatives to MRI. A comparative analysis of copious relevant image de-speckling, image enhancement, segmentation, and feature extraction methods are carried out. Higher resolution and superior quality, strong segmented real-time ultrasound uterus tumour images are produced by using diffusion-based hybrid filters, Super Resolution Convolutional Neural Networks (SRCNN), and U-net segmentation technique. The Grey Level Co-occurrence Matrix (GLCM) and Discrete Wavelet Transform (DWT) are used to extract textural features. With the help of several machine learning approaches, such as Support Vector Machine (SVM), K-Nearest Neighbour (KNN), and Random Forest classifiers (RFC), the extracted characteristics are immediately sent to classifiers to classify uterus tumours from ultrasound images between benign and malignant. For the categorization of uterine tumors, the RFC classifier outperformed the other classifiers. The viability of cancer detection using ultrasound pictures is significantly strengthened by the suggested machine learning methodology. Multiple hospitals provided data on ultrasound pictures of uterine tumors, which were used to develop the model and obtain the prediction findings. A radiologist with 17 years of expertise in diagnostic radiology further assessed this dataset. We could produce high-quality ultrasonic real-time images of uterine tumor datasets with the suggested machine learning model at a 97.8 % accuracy rate utilizing RFC.

Thermal behavior of sildenafil citrate (SC), a vasodilator used in the treatment of both erectile dysfunction (ED) and pulmonary arterial hypertension (PAH) treatment was investigated by thermogravimetry-differential thermal analysis (TGA-DTA), differential scanning calorimetry (DSC), powder X-ray diffraction (PXRD) and evolved gas analysis by thermogravimetry coupled to infrared spectroscopy (TGA-FTIR). Under N₂ and air atmospheres SC remained thermally stable until 180.0 °C, and then decomposed in three steps under N₂ and four steps under air. DSC curves showed two endothermic events at first heating, one related to the dehydration around 63.5 °C and the second event is a concomitant melting-decomposition process (T_peak = 199.6 °C). Cold recrystallization was observed during the second heating, with peaks at 119.8 °C and 129.7 °C and melting at 179.7 °C. XRD results for powder collected at 138 °C suggested that the phase formed is the sildenafil free base. TGA-FTIR revealed that the thermal decomposition of SC initiates with release of water, carbon dioxide, and itaconic anhydride, resulting from the decomposition of the citrate counter-ion. Above 303 °C, CO, SO₂, benzene, ethanol, 1-methylpiperazine, isocyanic acid, and methylamine were identified in the gas phase. Based on these results, a thermal behavior mechanism for sildenafil citrate was proposed.

Since the discovery of potentially mutagenic nitrosamine impurities in pharmaceuticals in 2018, liquid chromatography-mass spectrometry (LC-MS) methods have been widely applied for the trace level quantification of nitrosamines. As more nitrosamines and drug samples are analyzed, method accuracy is increasingly recognized as a major analytical challenge. Matrix effects can have a significant adverse impact on method accuracy but have not attracted much attention in nitrosamine quantification literature. In this study, we observed severe matrix effects when adapting LC-MS method conditions from a published and validated method for the analysis of nitrosamines in tuberculosis drugs. Several approaches were explored to mitigate these matrix effects, including using stable isotope-labeled internal standards, modifying LC conditions and sample concentration, and quantifying with standard addition instead of an external calibration method. The results underscore the importance of monitoring matrix effects and validating nitrosamine quantification procedures for specific sample matrices. Finally, two LC-MS/MS methods are presented with validation data for the quantification of 1-methyl-4-nitrosopiperazine (MNP) in rifampin and 1-cyclopentyl-4-nitrosopiperazine (CPNP) in rifapentine and revealed high nitrosamine levels in the tested samples that far exceeded recommended acceptable intake limits.

Glucose electro-oxidation in an alkaline media was investigated using an electrode composed of carbon paste and sugarcane bagasse activated carbon (SBAC). The SBAC was prepared using an easy, affordable, and environmental friendly process. The porous activated carbon was produced using sugarcane bagasse through a pre-treatment process that involved carbonization at 650 °C and K₂CO₃. X-ray diffraction (XRD) was used to observe the formation of carbon crystallites during activation at higher temperatures. The Fourier-transform infrared (FTIR) spectrum shows functional groups that are present in the carbon material. The carbon sample's morphology was evaluated using scanning electron microscope (SEM). The surface roughness was studied using atomic force microscopy (AFM). Porous activated carbon derived from sugarcane bagasse was carefully mixed into electrode components such as graphite powder and mineral oil for glucose (GLS) detection. For precise GLS detection, the SBACs were employed as non-enzymatic electrochemical sensor probes in their as-prepared condition. According to the electrochemical experiments, the constructed sensor shows exceptional electrochemical performance towards glucose oxidation, including excellent selectivity, a low detection limit, good sensitivity, and a wide linear range. Investigation has been conducted on the oxidation of GLS in alkaline solutions with varying concentrations of halide ions such as iodide and chloride. The GLS oxidation peak current increased the performance of a SBAC modified electrode was compared to that of a bare carbon paste electrode (CPE). The role of sensitivity in minimizing halide poisoning is significant. The risk of poisoning is increased if iodide > chloride. As the modified electrode's voltage changes in a chloride solution containing glucose, the peak current gradually reduces. The redesigned electrode that is currently in use as a consequence not only promotes glucose oxidation but also shows strong resistance to electrode poisoning caused by halides. The sensitive and targeted SBAC/CPE has also improved its reliability while evaluating samples of human serum, urine, and breast milk.