Validated HPLC Method Research Articles

Development of bedaquiline nanoemulsions intended for paediatric multidrug-resistant tuberculosis: excipient selection and preformulation studies.

Preformulation investigations into the development of drug formulations, encompassing considerations related to the structure of the drug, excipients, composition, and physical attributes are crucial. This phase is pivotal in ensuring the ultimate success of nanoemulsion development. The objective of this study was to evaluate and define the properties of bedaquiline (BDQ) and the necessary excipients for the formulation of self-emulsifying BDQ-loaded nanoemulsions. To determine the saturation solubility of BDQ in various oils, an in-house validated HPLC method was used. Fourier transform infrared spectroscopy was utilised to identify and evaluate the compatibility between BDQ and the selected excipients. The water titration method was used to construct phase diagrams to identify the type of structure that resulted following emulsification and to characterise the behaviour of mixtures along dilution paths. The solubility studies revealed that BDQ exhibited the highest solubility in olive oil, with a solubility of 3.45 ± 0.041 mg/ml. The design space led to the formation of emulsions categorised as Winsor products. Importantly, the FTIR data indicated the absence of any potential interactions between BDQ and the chosen excipients. The preformulation studies were successful and facilitated the selection of compatible and suitable excipients for the formulation of BDQ-loaded nanoemulsions.

Disposition kinetics and tissue residues of tilmicosin following intravenous, subcutaneous, single and multiple oral dosing in geese (Anser anser domesticus).

Tilmicosin (TMC), a semi-synthetic macrolide antibiotic, is widely used in veterinary medicine due to its broad-spectrum, bacteriostatic properties. Frequently administered in various birds species, it is likely used off-label in geese as well. The study sought to investigate TMC's pharmacokinetics, tissue residues, in geese through invivo experiments. The study involved longitudinal open studies on 15 healthy adult males, with three phases separated by one-month washout periods. Geese were administered TMC through intravenous (IV, 5 mg/kg), subcutaneous (SC, 10 mg/kg), and oral (PO, 25 mg/kg for five consecutive days) routes, with blood samples drawn at specific intervals. Tissue samples were also collected for subsequent analysis at pre-assigned times. TMC in goose plasma was quantified by a fully validated HPLC method. Plasma concentrations were quantified up to 4 hr for the PO and IV routes, and up to 10 hr in the SC route. Significant variations in bioavailability were observed between SC (87%) and PO (4%) routes. The body extraction ratio was low at 0.03, suggesting minimal ability of the liver and kidneys to eliminate TMC. Multiple oral doses showed no plasma accumulation, but tissue data revealed extensive distribution and prolonged residence, up to 120 h, suggesting a sustained therapeutic effect despite the brief plasma half-life. Regarding the multiple PO doses, provisional withdrawal times of 6, 7.5, and 8 days were suggested for the liver, muscles, and kidneys, respectively, according to the MRL set for these matrices in chickens by EMA. In conclusion, while the practical oral administration is discouraged at the population level, SC administration of TMC may be appropriate for geese, albeit impractical for flock therapy.

A Comparative Study and Prediction of the Ex Vivo Permeation of Six Vaginally Administered Drugs across Five Artificial Membranes and Vaginal Tissue.

The theoretical interpretation of the vaginal permeability phenomenon, the evaluation of the suitability of five artificial membranes, and the prediction of the behaviors of vaginal drugs were the main objectives of this study. Franz vertical diffusion cells and different validated HPLC methods were used to measure the permeability of six vaginally administered drugs (econazole, miconazole, metronidazole, clindamycin, lidocaine, and nonoxynol-9). This study was performed (in vitro) on different membranes of polyvinylidene fluoride (PVDF), plain cellulose or cellulose impregnated with isopropyl myristate (IPM), and cellulose combined with PVDF or IPM. The results were compared with those obtained from cow vaginal tissue (ex vivo), where cellulose was proven to be the best simulant. According to the permeability profiles (Papp), the water solubility of the drugs was considered a necessary criterion for their transport in the membranes or in the tissue, while the size was important for their penetration. Furthermore, it was found that polar compounds show clear superiority when penetrating cellulose or tissue, while non-polar ones show superiority when penetrating the lipophilic PVDF membrane. Finally, a successful attempt was made to predict the Papp values (|Papp-predPapp| < 0.005) of the six drugs under study based on a PLS (Partial Least Squares) in silico simulation model.

Quantification of Arbutin and its degradation products, hydroquinone and p-Benzoquinone, in hyperpigmentation topical formulation: Effect of extraction procedure and interference assessment

The utilization of Hydroquinone (HQ) in over-the-counter skincare items is subject to restrictions. Consequently, Arbutin (AR) serves as a reliable alternative for addressing hyperpigmentation in non-prescription topical formulations. Nevertheless, AR undergoes decomposition into HQ and p-Benzoquinone (BZ) when exposed to temperature stress, ultraviolet light, or dilution in an acidic environment, all of which can induce skin toxicity. The intention of this paper is to investigate the effect of extraction procedure on the conversion of AR to HQ and or BZ and to evaluate kinetics of AR hydrolysis to HQ. Meanwhile this study aims to evaluate AR and BZ interference with the United States Pharmacopoeia (USP) identification and assessment method for HQ Hydrolytic stress during extraction conditions underwent optimization through systematic screening tests. Subsequent assessment of the residual drug and its degradation products were achieved by HPLC method. The resulting data were meticulously fitted to various kinetic models. To analyze the potential interference of AR in HQ measurement using USP method, the standard concentrations of AR and HQ were analyzed through UV–VIS spectrophotometry. For enhanced certainty, a validated HPLC method analysis was also conducted. Notably, the acid hydrolysis of AR exhibited independence from its initial concentration. So, the hydrolytic degradation of AR exhibited a Zero-order kinetic profile. Furthermore, the proven interference of AR in the UV–VIS spectrophotometry method was identified within the context of the USP method. This study successfully utilized an adopted HPLC method for the concurrent quantification of AR, HQ, and BZ. The potential interference of AR in the UV–VIS spectrophotometric assay for HQ may lead to false results especially for regulatory purposes.

Pharmacokinetics of tildipirosin in estuarine (Crocodylus porosus) and freshwater (Crocodylus siamensis) crocodiles

Tildipirosin is a macrolide antimicrobial. It is authorised for the treatment and prevention of respiratory disease in cattle and pigs. There are no data on its administration in crocodiles. Therefore, this study evaluated the disposition kinetics of tildipirosin after intravenous (dose: 2 mg/kg) and intramuscular (doses: 2 and 4 mg/kg) administration in two crocodilian species (estuarine and freshwater; n = 5). Tildipirosin plasma concentrations were quantified by a validated HPLC method. Plasma concentrations obtained at each extraction time were analysed by non-compartmental methods. In the estuarine and freshwater crocodiles, the apparent volumes of distribution of tildipirosin after intravenous administration were 0.36 ± 0.10 and 1.48 ± 0.26 L/kg, respectively. These values, suggesting poorer tissue distribution, were much lower than those obtained in mammals. There was complete bioavailability of tildipirosin after intramuscular route at a dose of 2 mg/kg; however, at a dose of 4 mg/kg the bioavailability decreased by about 20–25 %. Furthermore, the pharmacokinetics of tildipirosin were markedly different in the two crocodilian species. Considering a MIC of 0.5 µg/mL, the surrogate marker AUC0–24/MIC indicates that tildipirosin would greatly exceed the value of 65 h for both crocodile species and dose levels tested. This suggests that both doses (2 and 4 mg/kg) may provide a bactericidal effect. Therefore, based on the absence of adverse reactions following the administration of tildipirosin in both crocodilian species, and considering its favourable pharmacokinetic properties, tildipirosin may be useful in treating infections in these reptiles.

Quantitative Analysis of Swertiamarin Content from Fagraea fragrans Leaf Extract using HPLC Technique and its Correlation to Antibacterial Activity

The rise in drug resistance poses escalating challenges for antibacterial medications, leading to an urgent demand for the exploration and innovation of new antibacterial drugs. Fagraea fragrans Roxb., belonging to the Gentianaceae family, is one of the common herbal medicines which can be found abundantly in Southeast Asia. The secoiridoid glucoside swertiamarin, one of the major compounds in F. fragrans leaf, exhibits antimicrobial effects. To guarantee the medicinal effectiveness of F. fragrans leaves, it is essential to identify a standardized analytical method for quantifying the active compound. In this study, the optimized HPLC method following ICH guideline was validated for the quantitative analysis of swertiamarin content in F. fragrans leaf in terms of linearity (y = 5733.5x - 369.1; R2 = 0.9999), accuracy (93.57-96.39% recovery), precision (0.91% RSD for repeatability precision; 1.19% RSD for intermediate precision), limit of detection (0.73 µg/mL), limit of quantitation (2.23 µg/mL), specificity (peak purity index = 0.999995) and robustness (% RSD &lt;1).The maximum wavelength of swertiamarin was found to be at 238 nm. The amount of swertiamarin content in F. fragrans leaf extract conducted from the validated HPLC method was found to be 0.0259 ± 0.0005 g/100 g crude drug. The leaf extract exhibited antimicrobial activity against clinical isolates of Enterobacter cloacae, Klebsiella pneumonia, and Escherichia coli at 0.125 mg/mL, and Pseudomonas aeruginosa at 0.5, showing minimum inhibitory concentration (MIC) values. Whereas swertiamarin exhibited even lower MIC values. The developed HPLC analysis effectively determines swertiamarin content as a chemical marker to ensure the antimicrobial potential of F. fragrans leaves.

Does sildenafil citrate affect the pharmacokinetics of metformin in rats? Screening of mechanism through analytical and molecular docking approach.

In the present study, the effect of sildenafil on the pharmacokinetics of metformin was studied in experimental rats, and we also postulated the molecular mechanism by performing molecular docking studies. Analysis of metformin and sildenafil (SIL) from rat plasma was done by high performance liquid chromatography. Optimum chromatographic separation and quantification of MET, SIL and Cetirizine was achieved on Phenomenex EVO C18 column with triethyl amine (0.3%): Methanol: Acetonitrile (70:05:25 v/v) as mobile phase maintaining flow rate of 1 ml/min, the detector was tuned at 224 nm. The extraction of MET and sildenafil from rat plasma was achieved by solid-phase extraction using Strata-X cartridges. The method was validated as per the ICH guidelines. For docking studies, the crystal structure of organic cation transporter 1 (OCT1) protein and multidrug and toxin extrusion (MATE) protein (5XJJ) were downloaded from the PubChem database. The docking study was performed by PyRx virtual screening software, and the results were analyzed by BIOVIA Discovery Studio. The validation of HPLC method was done, intraday and interday precision study of HPLC method demonstrated %RSD values less than 5%, the extraction recovery for MET and SIL were near to 80 % for low, medium and high QC samples. The plasma stability of MET and SIL showed % RSD values <10% for low, medium, and high QC samples. A sensitivity study for MET and SIL in rat plasma suggested a lower limit of quantification values of 8 and 10 ng/mL, respectively. The pharmacokinetic parameters were recorded, Cmax of experimental and control rats was 611.2 and 913.2 ng/mL; t1/2 1.66 and 1.98, AUC (0-t) 1637.5 and 2727.24, AUC (0-∞) 1832.38 and 2995.24 for MET. The results suggested that the Cmax of MET in experimental rats (MET + SIL) was 33.07% lower than the control (MET only) and also the t1/2 was 0.32 h shorter. Docking analysis suggested a higher binding affinity of sildenafil with MATE protein (5XJJ) compared to OCT1, suggesting possible involvement of MATE family proteins for pharmacokinetic alterations of MET. The HPLC and solid-phase extraction method were developed and applied successfully for the pharmacokinetics of MET and SIL. Intake of SIL altered the pharmacokinetics of MET in rats. Molecular docking studies suggested the involvement of MATE family proteins for alterations of MET pharmacokinetics.

Effect of chemical penetration enhancers on the transdermal delivery of olanzapine in human skin in vitro

This research study involves the development of an olanzapine (OLZ) formulation using various chemical penetration enhancers (CPEs) for transdermal delivery. The aim of this study was to obtain the initial data needed about the effects of various CPEs on the skin permeation of OLZ. The effects of the selected CPEs were examined by studying the permeation profiles of OLZ from formulations applied to human cadaver skin samples. A control formulation of OLZ in propylene glycol (PG) was prepared and compared against formulations containing chemical penetration enhancers. Five different CPEs (oleic acid (OA), cineole (Cin), isopropyl alcohol (IPA), Tween 80 (T80), and N-methyl pyrrolidone (NMP)) at 5% w/w were individually added to the formulation containing OLZ in PG. The in vitro permeation study was carried out using vertical Franz diffusion cells mounted with human cadaver skin. Samples from the receptor compartment of the cell were collected at 2 h, 4 h, 8 h, 12 h, and 24 h at room temperature. The amount (µg/cm2) of permeated drug (OLZ) was measured using a validated HPLC method, and the percentage (%) of OLZ permeated was calculated. Based on the data obtained, different CPEs were found to have a significant impact on OLZ permeability compared to the control formulation. The most effective chemical penetration enhancer was shown to be 5% w/w OA with a 3.3-fold increase in enhancement ratio (ER). The rank of order for the highest concentration of OLZ permeated from each of CPE containing formulation was as follows: OA > Cin > IPA > T80 > NMP. The most effective chemical penetration enhancer was OA but the cytotoxic study using human fibroblast cells suggests that OA may not be safe due to its cytotoxic effects.

Determination of Genotoxic Impurities in Nilotinib using LC-MS Method

A validated HPLC method was developed for the determination of Nilotinib (Nob) in pharmaceutical formulation. Isocratic elution at a flow rate of 1.0ml/min was employed on Zorbax SB C18 5µm × 4.6mm, 150mm, or similar is used for this chromatography analysis and the column temperature is maintained as ambient. Mobile phase was a mixture of 2.00ml of acetic acid in 2000ml of water as mobile phase A and Acetonitrile 100% as mobile phase B was used. Rate of flow is identified at 1.3ml/min. a 50.0µl sample was injected. The run time is 20 minutes to Sample, blank, system suitability, and sensitivity solution. For the diluted standard solution, it is 60 minutes. The retention time was 1.30minutes. The% R.S.D Nob is identified. The mean percentage recovery for Nob is found within the specification limit. Due to this reason this proposed LC-MS method is successfully adopted to routine quality control analysis in formulations.

A new Turkish pharmacokinetics software program for therapeutic drug monitoring of theophylline

In this study, it was aimed to develop a Turkish software with pharmacokinetic (PK) data for therapeutic drug monitoring and IV dose adjustment of narrow therapeutic index theophylline. The study involved three groups, each comprising two rabbits The dose required for the target concentration (15µg/mL) was calculated with the developed program according to the weight of the rabbits in the first group. Blood samples taken at certain times were analyzed by validated HPLC method to calculate the elimination rate constant (ke) after IV bolus administration. The r2 values for ke were found to be 0.86 and 0.95. The second dose calculated according to revised PK findings was administered and blood samples were taken. When the analyzed results and theoretical results were compared, the deviation was found to be 5.53% and 8.795%. The findings were taken as the population PK for other applications. IV multiple dose bolus and IV fast-slow combined infusion were administered to the second group and the third group, respectively. The results obtained from the analysis of blood samples taken at the times determined according to the application were compared with the theoretical results. As a result, although there is a high difference between theory and practice at low concentrations, there is very little variation at high concentrations. By using this program, it has been achieved to keep theophylline at the desired level without reaching the minimum toxic concentration and without falling below the minimum effective concentration. It is thought that deviations will be reduced with larger samples.

Therapeutic drug monitoring of vancomycin in surgical patients using a validated HPLC method.

Vancomycin is being used for the treatment of a variety of infections caused by methicillin resistant Staphylococcus aureus and methicillin susceptible Staphylococcus aureus. Therapeutic drug monitoring (TDM) is highly recommended for ensuring the safe and effective therapy with vancomycin. A reliable and cost-effective bioanalytical method is required for TDM as well as pharmacokinetic studies of vancomycin. A selective, sensitive, and cost effective HPLC method was developed and validated for quantification of vancomycin concentrations in human plasma. The mobile phase was a mixture of buffer (50 mM ammonium dihydrogen phosphate, pH 2.4) and acetonitrile 88 : 12 v/v. The separation was carried on C18 column (125 × 4.6 mm, particle size 5 µm) with isocratic flow rate of 0.370 mL/min at room temperature with UV detection at 215 nm. The method was validated for sensitivity, accuracy, and precision as well as stability of vancomycin in human plasma by following European Medicine Agency (EMA) guideline. Therapeutic drug monitoring of vancomycin was performed by quantifying the trough concentrations of vancomycin in 65 human plasma samples after administration of therapeutically relevant dose. The developed method was sensitive enough to quantify vancomycin concentrations as low as 0.25 mg/L in human plasma. Moreover, the method was proved accurate and precise in terms of quantifying the unknown concentration of vancomycin. The evaluation of short-term, long-term, and freeze-thaw stability proved the stability of vancomycin in human plasma. The TDM of vancomycin by using this method showed that 39 (60%) samples were within the target trough concentration range (TTCR), i.e. 10 - 20 mg/L, while 23 samples (35.4%) were below the TTCR, and 3 samples (4.6%) were above this range. The developed method is sensitive and cost effective for quantification of vancomycin in human plasma. The results of sample analysis shows that the developed method can be used reliably for TDM of vancomycin.

Novel HPLC Method Validation for Quantification of Dapagliflozin in Bulk Drug and Formulation

A new, steady, easy-to-use, and straightforward high-performance liquid chromatography (HPCL) technique was also tested to determine dapagliflozin concentration in medicinal and API forms. For the chromatographic extraction study, a Phenomenex Gemini-NX C18 Column was used. This column was filled with a mobile phase that was 70:30 v/v methanol to sodium 1-octanesulphonate. This experiment used a 20-liter syringe amount and a flow rate of 1-mL per minute. A bunch of factors were used to make sure the new method worked. The drug dapagliflozin was released at a wavelength of 203 nm after 6.5 ± 0.5 minutes. A correlation coefficient (r2) of 0.9994 between 15 to 55 μg/mL shows that the planned process is linear. The theoretical plates and peak tailing were found to be 4129 and 1.03, respectively. In terms of healing, the percent accuracy that was reached was within the acceptable range of 98 to 100. New suggestions say that the regular testing of dapagliflozin in large amounts must be done using a proven method that is unique and easy to repeat. Forced breakdown tests with drugs were used to show that the proposed method is stable. The product was tested according to the steps laid out in the ICH Q2R1 Guidelines. The test looked at many chemical processes, such as oxidation, photolysis, heat stress, and acids and alkaline hydrolysis.

Development and Validation of Stability-indicating HPLC Method for Estimation of Azilsartan in Pharmaceutical and Solid Lipid Nanoparticles

Development and Validation of Stability-indicating HPLC Method for Estimation of Azilsartan in Pharmaceutical and Solid Lipid Nanoparticles

Validation of HPLC Method for the Assessment of Occurrence of Ochratoxin A in Wheat Grains Sold in Major Markets in Lagos

The presence of Ochratoxin A (OTA) in wheat grain is a serious threat to safety of wheat based foodstuff. In the present study, sixty wheat grain samples were collected from six major markets in Lagos for determination of OTA levels using HPLC-FLD utilizing acetonitrile- water-acetic acid (51:47:2 V/V/V) mixture as mobile phase. The limit of Quantitation (LOQ) was 0.47 μg/kg and the average recovery values of OTA for reference material and spiked wheat grain sample ranged from 87.68-107.09%. Twenty-four (40%) wheat grain samples were contaminated with detectable quantity of OTA ranging from 0.40-41.58μg/kg. OTA levels in five samples (20.8%) exceeded the NAFDAC and European Union (EU) permissible limit of 5.0μg/kg. The expanded uncertainty of measurement calculated signifies that the true value is within ±11.00% of the reported results at a confidence level of 95%. Considering that OTA levels in 20.8% of wheat grain samples analyzed are above the permissible limit of 5.0μg/kg, it could be seen as a serious public health threat. Therefore, continuous monitoring and tighter regulation of OTA in cereals is recommended.

Analysis of Tolfenamic Acid using a Simple, Rapid, and Stability-indicating Validated HPLC Method.

Tolfenamic acid (TA) belongs to the fenamates class of nonsteroidal anti-inflammatory drugs. Insufficient information is available regarding the availability of a reliable and validated stability-indicating method for the assay of TA. A relatively simple, rapid, accurate, precise, economical, robust, and stabilityindicating RP-HPLC method has been developed to determine TA in pure and tablet dosage forms. The method was validated according to the ICH guideline, and parameters like linearity, range, selectivity, accuracy, precision, robustness, specificity, and solution stability were determined. TLC and FTIR spectrometry were used to ascertain the purity of TA. The specificity was determined with known impurities and after performing forced degradation, while the robustness was established by Plackett-Burman's experimental design. The mobile phase used for the analysis was acetonitrile and water (90:10, v/v) at pH 2.5. The detection of the active drug was made at 280 nm using a C18 column (tR = 4.3 min.). The method's applicability was also checked for the yellow polymorphic form of TA. The results indicated that the method is highly accurate (99.39-100.80%), precise (<1.5% RSD), robust (<2% RSD), and statistically comparable to the British Pharmacopoeia method with better sensitivity and specificity. It was observed that the stress degradation studies do not affect the method's accuracy and specificity. Hence the proposed method can be used to assay TA and its tablet dosage form.

HPLC profiling for the simultaneous estimation of antidiabetic compounds from Tradescantia pallida

Diabetes is a long-term metabolic disease epitomized by postprandial hyperglycemia. The prolonged use of synthetic drugs renders distinct side effects, necessitating the development of safe and cost-effective substitutes. The aim of the current study is to isolate, evaluate the antidiabetic potential and HPLC method development for simultaneous estimation of antidiabetic compounds from the leaves of Tradescantia pallida. The leaves were extracted, fractionated and subjected to column chromatography. The isolated compounds' antidiabetic potential was evaluated using α-amylase and glycosylation of hemoglobin assays. The study employed molecular docking to scrutinize interactions between antidiabetic compounds and human α-amylase and hemoglobin protein. Prime MM-GBSA calculations determined binding energies of ligand–protein complexes. Further analysis of morin and catechin involved exploring dynamic and thermodynamic constraints through molecular dynamics simulations under specific biological conditions. A rapid HPLC method was developed and validated for the simultaneous estimation of isolated compounds. The column chromatography culminated in the isolation of four antidiabetic compounds (syringic acid, catechin, p-coumaric acid and morin). The in vitro analyses revealed that morin and catechin exhibited 72.67 % and 78 % α-amylase inhibition and 67 % and 71.66 % inhibition of hemoglobin glycosylation, respectively. In silico studies substantiated the in vitro assay, confirming the stability of catechin and morin complexes via root mean square deviation analysis. Interactions, encompassing hydrophilic, hydrophobic, water bridges, and ionic interactions, identified key residues involved in these processes. The validated HPLC method exhibited excellent correlation coefficients ranged from 0.9909 to 0.9997. The antidiabetic compounds were quantified from the extract in the range of 0.072 – 0.160 µg/mL. The study concluded that the isolated compounds from Tradescantia pallida have remarkable antidiabetic activity, and the developed method can be successfully used for the identification and quantification of phenolic compounds in Tradescantia pallida and other plant-derived matrix.

A Validated HPLC Method using C18 Analytical Column (Agilent) for the Estimation of Cyclobenzaprine Hydrochloride by Quality by Design Approach in Bulk and Its Tablet Dosage Form

Cyclobenzaprine hydrochloride is used to treat muscle spasms brought on by acute, uncomfortable musculoskeletal diseases. The primary objective of this work is to develop accurate, quick and precise HPLC method for Cyclobenzaprine hydrochloride. The method used is a Central Composite Design to get exact data for the procedure. Various parameters were used to validate and to develop method. The optimized model for the analysis of Cyclobenzaprine Hydrochloride was found to have an area 8.3239. This optimization was done on the C18 analytical column(Agilent) with mobile phase as methanol: 0.01 % Orthophosphoric acid (61:39 v/v) and a flow rate maintain was 0.9 ml/min with a detection wavelength of 224 nm. With the use of a correlation coefficient (r2=0.999), the linearity of Cyclobenzaprine hydrochloride in the 5–25 ug/ml range was determined. The accuracy values were discovered to fall between 99.86 and 100.71%. While the robustness was shown to be less than 0.06 for flow rate and for wavelength was 0.09 % RSD, also the intraday and interday precision were determined to be under 0.28 and 0.29 % RSD. The most effective approach for analysing Cyclobenzaprine hydrochloride is the one that has been presented. The development and validation of the HPLC technique for Cyclobenzaprine hydrochloride were assessed.

Design of trio-colored validated HPLC method for synchronized multianalyte quantitation of four top selling antihyperlipidemic drugs in different fixed-dose combined tablets

The ongoing attempts to attain the sustainable development goals, to save the environment, to embrace the novel paradigm of white analytical chemistry and to focus on the practicality of the technique are non-stoppable. In this consensus, we designed a rapid trio-colored (green, white and blue) analytical methodology for the simultaneous estimation of Ezetimibe (EZT), Atorvastatin (ATV), Rosuvastatin (RSV) and Simvastatin (SMV). The suggested method was HPLC-DAD employing Microsorb MV-C18 column (4.6 × 250 mm, 5 μm particle size). The mobile phase composed of 0.1 % ortho-phosphoric acid and acetonitrile eluted in a gradient mode. Diode array detector was adjusted at 237 and 243 nm. Separation was fulfilled in less than 9 min and the retention times were 3.48, 4.32, 4.73 and 8.74 min for RSV, EZT, ATV and SMV respectively. The 4 drugs were quantitatively measured in the concentration range 1–50 μg/mL. In addition to validation according to the ICH guides, the greenness assessment using the novel AGREE calculator and the holistic overall evaluation applying the RGB-12 model in addition to the practicality check using the Blue Applicability Grade Index (BAGI) were performed. The results proved the efficiency and whiteness of the suggested technique to be routinely involved in quality control laboratories for assay of these drugs in their binary mixtures combination pills.

Development of a novel quality by design-enabled stability-indicating HPLC method and its validation for the quantification of nirmatrelvir in bulk and pharmaceutical dosage forms.

A systematic and novel quality by design-enabled, rapid, simple, and economic stability-indicating HPLC method for quantifying nirmatrelvir (NMT) was successfully developed and validated. An analytical target profile (ATP) was established, and critical analytical attributes (CAAs) were allocated to meet the ATP requirements. The method used chromatographic separation using a Purosphere column with a 4.6 mm inner diameter × 250 mm (2.5 μm). The analysis occurred at 50°C with a flow rate of 1.2 mL/min and detection at 220 nm. A 10 μL sample was injected, and the mobile phase consisted of two components: mobile phase A, containing 0.1% formic acid in water (20%), and mobile phase B, containing 0.1% formic acid in acetonitrile (80%). The diluent was prepared by mixing acetonitrile and water at a 90:10 v/v ratio. The retention time for the analyte was determined to be 2.78 min. Accuracy exceeded 99%, and the correlation coefficient was greater than 0.999. The validated HPLC method was characterized as precise, accurate, and robust. Significantly, NMT was found to be susceptible to alkaline, acidic, and peroxide conditions during forced degradation testing. The stability-indicating method developed effectively separated the degradation products formed during stress testing, underlining its effectiveness in stability testing and offering accuracy, reliability, and sensitivity in determining NMT.

Stability Studies of Cefuroxime in Peritoneal Dialysis Solution by Using the HPLC Method

Peritoneal dialysis is a technique for blood purification and is used in chronic kidney disease. Peritonitis is a major problem linked to peritoneal dialysis which limits its utilization. Cephalosporins are extensively used for the treatment of peritoneal dialysis-related peritonitis. Cefuroxime is the second-generation cephalosporin that is used for the prevention and cure of peritonitis. The stability of cefuroxime in peritoneal dialysis solution was investigated after storage at different temperatures. Cefuroxime in different concentrations (250 mg, 750 mg and 1.5 g) was injected into the bags of peritoneal dialysis solution which were stored at 4 oC, 25 oC, and 40 oC. Samples were taken at regular intervals and evaluated by a validated HPLC method. Cefuroxime remained stable for fourteen days at 4 and#176;C and the pattern of stability was the same for all concentrations of cefuroxime. At 25 and#176;C, cefuroxime remained stable for 24 hours in isotonic and hypertonic PD solutions and maintained more than 90% of its initial concentrations. At 40 and#176;C, cefuroxime remained stable for 6.7 and 6.0 hours in isotonic and hypertonic PD solutions respectively. The degradation of cefuroxime was higher in hypertonic solutions as compared to isotonic solutions