Development Of HPLC Method Research Articles

Co-axial electrosprayed RAD001-loaded polycaprolactone/polyvinyl alcohol core–shell particles for treating pediatric brain tumours

Core-shell particles composed of polycaprolactone/polyvinyl alcohol (PCL/PVA) with pH sensitive properties were successfully fabricated by co-axial electrospraying in which PVA and PCL formed the shell and core layers respectively. The core–shell structure was confirmed by FTIR, DSC and SEM analysis. No chemical interaction between PVA and PCL core–shell were observed in the FTIR analysis. The RAD001 loaded core–shell particles showed a sustained and pH dependent drug release and was assayed via our previously developed HPLC method. After indirect treatment of the PF-A cells with the core–shell particles for 24 h and 5 days a decrease in cell viability was observed. Additionally, a comparison was made with our previously developed nanoparticles containing 2 %PVA-14 %SOL®-0.6 % RAD001, for the cell viability study on ependymoma. Our findings show that optimised core–shell particles exerted a significant effect for the 24 h and 5 day treatment however further studies are required to ensure toxicity of the control core–shell particles with no drug is reduced. In comparison, the 2 %PVA-14 %SOL®-0.6 %RAD001 uniaxial electrosprayed nanoparticles also exerted a toxicity effect decreasing cell viability with no toxicity observed for the control nanoparticles as well. Such pH-sensitive core–shell particles, which can degrade effectively in either acidic or neutral condition, have great potential for application in the biomedical field.

Development and Validation of a Novel HPLC Method for the Simultaneous Determination of Pregabalin and Etoricoxib in Bulk and Pharmaceutical Dosage Form

Development and Validation of a Novel HPLC Method for the Simultaneous Determination of Pregabalin and Etoricoxib in Bulk and Pharmaceutical Dosage Form

Greenness Assessment and Development of an Innovative HPLC Method for the Determination of Rifaximin and Sodium Benzoate in Oral Suspension

Greenness Assessment and Development of an Innovative HPLC Method for the Determination of Rifaximin and Sodium Benzoate in Oral Suspension

Development of HPLC method for isomeric impurities of key starting material of novel oral anticoagulant drug; Edoxaban Tosylate Monohydrate

Reverse-phase high-performance liquid chromatography method has been developed for the determination of EDO-S1 stereoisomeric impurities such as isomer 1, isomer 2, isomer 3, isomer 4, isomer 5, isomer 6 and isomer 7 with good resolution using the column, Bakerbond C18 (150 × 4.6 mm; 3 μm). The separation was achieved with mobile phase-A (10 mM dipotassium hydrogen phosphate pH-7.0 with 10% orthophosphoric acid solution in Milli-Q water) and mobile phase-B (n-Propanol: Acetonitrile ratio of 20:30 % V/V), which consisted of mobile phase mixture in the combination of moilephase-A: mobile phase-B (85:15). The total run time was 30 min at 0.8 mL/min flow rate, 20 µL injection volume and 30 ℃ column oven temperature. The column eluate was monitored at 210 nm to quantify the impurities The method showed adequate specificity, sensitivity, linearity, accuracy, precision, and robustness inline to ICH tripartite guidelines. The limit of detection and quantification limits were 0.1 and 0.3 μg mL−1, respectively, for all isomeric impurities and EDO-S1. The developed method was found to be linear over the concentration range of LOQ to 150% of specification range for isomeric impurities with a correlation coefficient >0.999. The method was precise (%RSD < 5.0), robust, and accurate (with 85%–115% recovery).

Development and Validation of Novel HPLC Methods for Quantitative Determination of Vitamin D3 in Tablet Dosage Form.

In the present work, an efficient isocratic HPLC method was developed for the precise and accurate estimation of vitamin D3 in tablet form. The chromatographic conditions comprised an L3 silica column (5 µm in particle size, 4.6 mm × 250 mm) with a mobile phase n-hexane/ethyl acetate (85:15 v/v) with a flow rate of 2.0 mL/min and a detection wavelength of 292 nm. The new methodology was validated for accuracy, precision, specificity, robustness, and quantification limits according to an official monograph of USP/BP and ICH guidelines. The peak areas of the six replicates of the homogeneous sample were recorded. The mean value obtained was 67,301, and the relative standard deviation (RSD) was 0.1741. The linearity and range were in the acceptable bounds, i.e., 0.999, which was calculated using regression line analysis. The results show that the method is truly acceptable as the RSD, as the flow rate was 0.81%, while for the mobile phase composition, it was 0.72%, which lies in the acceptable range. The limit of detection (LOD) and the limit of quantification (LOQ) values were 0.0539 µg/mL and 0.1633 µg/mL, respectively. The % RSD of the intra and inter-day precision of the method was deemed acceptable according to the international commission for harmonization guidelines. The developed method has potential to be used for the detection and quantification of vitamin D3 during routine analysis for tablets in dosage form.

Development and validation of simple stability indicating RP-HPLC method for piperine estimation in Piper nigrum, marketed formulations and nanoformulation

The objective of the current research is to develop simple, rapid, precise and sensitive RP-HPLC method for piperine estimation and its application to marketed Ayurvedic and herbal formulations. A reversed-phase C-18 column (5 mm, 4.6 mm, 250 mm, ZORBAX) was used to achieve the separation of piperine. Optimal chromatographic conditions were established by using mobile phase consisting of methanol and water in a 75:25 ratio with flow rate of 1 mL/min and detection wavelength of 342 nm. The developed HPLC method to estimate piperine was validated in compliance with ICH Q2 (R1) guidelines. The newly developed method demonstrated linearity in the range of 0.5-16 μg/mL, with a regression coefficient R2 > 0.999. The accuracy of the developed method was confirmed by a recovery rate ranging from 98.20% to 99.42%. Precision was also ensured, with a relative standard deviation (%RSD) less than 2%. Additionally, the method exhibited robustness. The limit of detection and limit of quantification for piperine was found to be 0.00056 μg/ml and 0.0016 μg/ml respectively. Moreover, the degradation characteristics of piperine were examined in compliance with ICH Q1A (R2) guidelines. Piperine demonstrated pronounced degradation behaviour in both oxidative and thermal investigations. The developed analytical technique was successfully employed to quantify piperine in Piper nigrum, in commercially available Ayurvedic and herbal products and in nanotransfersomes. Also, the developed method was used to estimate the percent drug entrapped within nanotransfersomes. Therefore, it can be concluded that the established approach is well-suited for performing routine quality analysis of formulations containing piperine.

Simultaneous Estimation and Stability Indicating Method of Vildagliptin And Dapagliflozin by RP-HPLC in Pharmaceutical Dosage Form

Vildagliptin and Dapagliflozin drugs are used for the management of diabetes.The primary objective of the proposed research was to develop and validate analytical methods for simultaneous quantification of vildagliptin and dapagliflozin in pharmaceutical dosage form. Both the drugs were subjected to force degradation to ensure stability of pharmaceutical products. The HPLC method development was carried out on C18 column (250mm*4.6mm,5µm particle size). The chromatographic separation was achieved by isocratic mode with a mixture of Methanol: 0.01% Trifluoro acetic acid (pH-2.78) in the ratio of 95:05 % v/v as mobile phase. Flow rate was kept at 0.8 ml/min and selected wavelength for estimation was 210 nm. Stress testing with acid, alkaline, oxidative and thermal degradation was performed. The retention time was found to be 2.282 min and 4.070 min for vildagliptin and dapagliflozin, respectively. The drugs response found to be linear with correlation coefficient value was nearly 0.999 for vildagliptin and dapagliflozin with linearity range of 10 – 60 µg/ml. The recoveries from studies were found to be 99.78% and 100.17% for vildagliptin and dapagliflozin respectively. LOD and LOQ were found to be lower, hence the method proved sensitivity. Method was validated as per ICH guideline Q2(R1). Drugs had shown more degradation in oxidative condition, 10.62% and 13.52% for vildagliptin and dapagliflozin, respectively. The proposed method would be successfully applied for the stability indicating, impurity separation, qualitative and quantitative estimation of vildagliptin and dapagliflozin in pharmaceutical dosage form and in process quality control analysis.

UFLC Method Development and Validation for Anti-retroviral Drugs, Investigation of Greenness Assessment using Complex GAPI, AGREE and AMGS SPREADSHEET

Introduction: The study’s proposal was to establish a method of development and validation by green chromatography technique for antiviral drugs, as well as to assess the proposed method’s greenness using various tools, which is one of the emerging developments in the analytical field. The chromatographic separation is achieved by using Eclipse plus C18 (250 x 4.6, 5 μm) column by applying isocratic elution using the mobile phase containing methanol and isopropyl acetate in the ratio of (60:40% v/v) with 1-mL/min flow rate. The separation of drugs is achieved by using a greener mobile phase. Results: The retention time of ritonavir and ombitasvir was found to be 1.854 and 8.09 minutes, respectively. The regression coefficient (R2) is 0.997 for both drugs. Accuracy and precision is evaluated for the method and found be within the limit and the results were reproducible. Assessment of the method was carried out using the three different tools. The developed method is anticipated to be eco-friendly, an alternative to the developed method HPLC method in regard to safe solvent, less toxic and less run time. The developed method was found suitable for the simultaneous estimation in their combined dosage form.

Solubility of Poorly Soluble Drugs in Phosphatidylcholine-Based Drug Delivery Systems: Comparison of the Loading Capacity in the Bulk Formulation and Its Dispersed State.

The aim of this study was to determine the drug loading capacity of phosphatidylcholine-based formulations for four poorly water-soluble drug substances (clofazimine, fenofibrate, artemether, cannabidiol). Two self-dispersing lipid formulations were investigated, which consisted of soybean phospholipids, medium-chain triglycerides and ethanol with a different phospholipid-oil ratio. The direct loading of the bulk formulation was conducted with dual centrifugation, which proved to be a suitable method for screening experiments with the highly viscous formulations. To estimate possible precipitation after dispersion in the gastrointestinal fluids, the solubility of the drugs was investigated in the dispersed formulations. For this purpose, nanodispersions were prepared from the bulk formulations via high pressure homogenization and subsequently subjected to passive loading. A newly developed HPLC method with Charged Aerosol Detection allowed a simultaneous evaluation of the content of soybean lecithin and medium-chain triglycerides in the nanodispersions. When comparing the two phosphatidylcholine-based formulations, a high content of oil was advantageous with regard to a high loading capacity. Drug substances with melting points below 150 °C exhibited a high solubility in the phospholipid-based formulations. A surprisingly high solubility was observed for artemether and cannabidiol with up to 13.0% and 33.3% drug loaded to the formulations, respectively. In the dispersions, a similar solubility as in the bulk formulations was obtained for fenofibrate and cannabidiol. Clofazimine yielded a higher loading result in the nanodispersions than in the bulk formulation.

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

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.

Simultaneous HPLC method development for azelastine HCl and fluticasone propionate nasal spray formulation

Abstract: Azelastine HCl and Fluticasone Propionate nasal spray drug product is commonly used to treat moderate to severe allergic rhinitis and rhino-conjunctivitis. To date, a simultaneous HPLC method for active sub-stances and impurities of this product has not been reported. Acetonitrile and buffer (Octane-1-Sulfonic Acid Sodium Salt and Potassium Dihydrogen Phosphate) were used as a mobile phase. The separation was achieved using Phenomenex Phenyl-hexyl (150 cm × 4.6 mm, 5 µm) column at 1 mL/min flow rate in gradient elution mode. The chromatograms were monitored at 240 nm. The validation parameters such as linearity, limit of quantification, limit of detection, accuracy, precision, robustness and specificity were investigated for the nasal spray. Overall, this study provides valuable insights into the development and validation of an efficient analytical method for assessing the quality and stability of a combination of Azelastine HCl and Fluticasone Propionate nasal spray.

High Performance Liquid Chromatography Method for Analysis of Alpha Arbutin and Niacinamide in Facial Whitening Creams

The combination of alpha arbutin and niacinamide is often used in facial whitening creams at relatively small levels so required valid analysis method. This study aimed to develop HPLC method for analyzing alpha arbutin and niacinamide in three brands of whitening creams. The HPLC system developed was a reverse phase type with a C₁₈ column and the mobile phase was a mixture of metanol: aquabidest (20:80, v/v) with a flow rate of 1.0 ml/min and a detection wavelength of 274 nm. The results showed that all parameters met the requirements. The linearity test yielded a correlation value was r= 0.999. The LOD value for alpha arbutin was 0.405 µg/mL and the LOD for niacinamide was 0.398 µg/mL. The LOQ value of alpha arbutin was 1.351 µg/mL, the LOQ for niacinamide was 1.327 µg/mL and the selectivity was good. Precision test results with %RSD ≤ 2% and accuracy tests yielded values in the 99.04-100.97% range. The content of alpha arbutin in brand A was 0.651%, brand B was 0.551%, and brand C was 0.878%. The content of niacinamide in brand A was 0.526%; brand B was 0.680% and brand C was 1.053%. The results of the analysis meet the requirements.

Chromatographic analysis of triple cough therapy; bromhexine, guaiafenesin and salbutamol and pharmaceutical impurity: in-silico toxicity profile of drug impurity

Bromhexine (BR), guaiafenesin (GUF) and salbutamol (SAL) are formulated as Ventocough syrup® (with and without sugar), labeled to contain propyl paraben and sodium benzoate as inactive ingredients. They are used to make coughing more productive and easier. A crucial element and a major issue in the pharmaceutical industry is the control of organic related impurities to obtain safe and effective treatment. Guaiacol (GUL) is reported to be GUF related impurity that was proved to be extremely toxic (toxic rating class 5), and its use should be banned. In this work, In-Silico study and ADMET estimation were conducted to predict GUL pharmacokinetic properties and its toxicity profile. Additionally, two chromatographic methods were conducted to analyze the studied components along with GUF impurity in the presence of the labeled dosage form excipients. The In-Silico study assured that GUL has oral rat acute toxicity and it is considered to be skin sensitizer. On the other hand, the developed TLC- densitometeric method depended on using a mobile phase mixture of hexane: methylene chloride: triethylamine (5.0:6.0:0.3, by volume) as a developing system. UV-Scanning was performed immediately at 275 nm for SAL, GUF and GUL, while scanning at 310 nm was used for scanning BR. Linearity was established in the ranges of 0.25–4.0, 0.25–4.0, 0.5–8.0 and 0.1–1.6 µg/band for BR, SAL, GUF and GUL, respectively. In the developed HPLC method, separation was performed on X-Bridge® C18 column (250 × 4.6 mm, 5 μm) using a solvent mixture of 0.05M disodium hydrogen phosphate pH 3 with aqueous phosphoric acid: methanol (containing 0.3%, v/v triethylamine) (40:60, v/v). Detection was done at 225 nm and separation was achieved within 10 min. Linearity was proved in the range of 2–50 µg/mL for the proposed drugs. Validation of the developed methods was done and all the calculated parameters were within the acceptable limits recommended by ICH guidelines. After that, methods were used to examine the potency of the selected marketed dosage forms and concentrations of all drugs were within the acceptable limits. Additionally, complete separation between the studied drugs and the additives were observed. The developed methods can be used during routine quality control analysis of the proposed drugs when the required issues concern on sensitivity, selectivity and analysis time.Graphical

Quantification of mirtazapine in tablets via DNA binding mechanism; development of a new HPLC method

Atypical antidepressant mirtazapine (MIR) is mostly prescribed for the management of major depressive disorder. The identification of MIR in pharmaceutical dosage forms was made possible by developing a novel, quick, sensitive high-performance liquid chromatography (HPLC) approach that was verified in accordance with ICH recommendations. In the first part of this study, HPLC investigations were optimized with regard to variables including pH, working column, mobile phase, temperature, and flow rate. The limit of detection (LOD) was 0.013 ppm, the limit of quantification (LOQ) was 0.044 ppm, and the linear range was computed as 0.5–15 ppm (R2 = 0.9998). The recovery investigation assessed the method's accuracy, which was shown to range between 98.82 and 100.97 %. In the second part, by using UV–vis spectroscopy, HPLC, thermal denaturation, and viscosity measurements, the mechanism of binding interaction of MIR with double-stranded fish sperm deoxyribonucleic acid (dsDNA) has been thoroughly studied. The DNA binding constants (Kb) were determined using UV–Vis absorption and HPLC methods. To investigate the interactions of MIR with dsDNA, molecular docking calculations and additionally, molecular dynamics simulations were performed. Results showed that MIR is located in the minor groove of dsDNA, and in addition to hydrogen bonding, electrostatic interaction is also formed between the aromatic ring of MIR and phosphate oxygen of dsDNA. Finally, a binding characterization study using MIR tablets was also conducted in order to assess the interaction mechanism of the DNA with the drug using the validated analytical procedure developed for the MIR molecule.

Development and Validation of a Robust HPLC Method for Simultaneous Quantitative Analysis of Quercetin and β-sitosterol in Plant Extract

Development and Validation of a Robust HPLC Method for Simultaneous Quantitative Analysis of Quercetin and β-sitosterol in Plant Extract

Effect of microwave-assisted drying and extraction of 6-Shogaol from Zingiber officinale Roscoe

Ginger (Zingiber officinale Roscoe, Zingeberaceae) is a medicinal plant widely used as food, spice, or flavoring agent worldwide. 6-Shogaol is a compound of prime interest in exhibiting anti-inflammatory, antioxidant and chemopreventive effects. The objective of the study is to investigate the effect of microwave-assisted drying (MAD) followed by microwave-assisted extraction (MAE) so as to produce 6-Shogaol enriched Ginger with improved therapeutic benefits. Various drying techniques viz. shade drying, tray drying, microwave-assisted drying and osmotic dehydration as a pretreatment were used for drying Ginger rhizomes. The dried rhizomes were extracted by conventional solvent extraction and microwave-assisted extraction techniques and tested for content of 6-Shogaol using the newly developed HPLC method whereas total flavonoid and polyphenol content were determined using the UV spectrophotometric method. Subjecting the microwave dried Ginger to microwave-assisted extraction for 45 min at constant power level of 284 W resulted in a significant rise in the extractability of 6-Shogaol (1.660 ± 0.018), total polyphenols (855.46 ± 5.33) and flavonoids (617.97 ± 6.40) compared to the conventional method of extraction. The proposed Ginger processing method of microwave drying followed by microwave extraction outperforms traditional methods in terms of speed, convenience, and performance thus can be scaled up to industrial levels.

Development and Validation of HPLC Method for Estimation of Ibandronic Acid in Tablet Dosage Form

Development and Validation of HPLC Method for Estimation of Ibandronic Acid in Tablet Dosage Form

Development and validation of HPLC method for simultaneous determination of rimantadine marketed product

Development and validation of HPLC method for simultaneous determination of rimantadine marketed product

Determination of compounds with laxative action in aloe latex and aloe vera‐based products—Chemometric classification of aloe vera plants (Aloe barbadensis Miller) under different conditions

AbstractBackgroundThe bioactive compounds of latex in aloe vera are associated with several of its medicinal properties. Aloe emodin, aloins A and B, which are present in aloe latex, are known for their laxative action.ResultsThe developed HPLC method for the determination of compounds with purgative effect was performed with a C18 column, and the total analysis time was 22 min. Each leaf from the plants was studied separately for its content in aloins. It was observed that, in contrast with field cultivation plants, the concentrations of aloins in potted plants follow a specific pattern based on the position of the leaves. It was also observed that climatic and environmental conditions have a significant impact on the concentrations of aloins. Principal Component Analysis was performed to evaluate correlations among measurements. The model was able to separate the aloe plants based on their seasonal variation and growing conditions. Finally, it was observed that, in only two out of thirteen products, aloins were present, with the total content exceeding the maximum levels set by EU legislation.ConclusionThe present work provides useful information for the aloins variation among leaves in different positions and plants under various conditions.