Mobile Phase Of Methanol Research Articles

Greenness Assessment of High‐Performance Liquid Chromatography Method for Simultaneous Estimation of Apigenin, Apocynin, and Gallic Acid With Quality by Design Approach

ABSTRACTThis study presents the development of an eco‐friendly high‐performance liquid chromatography (HPLC) method for the simultaneous determination of apigenin, apocynin, and gallic acid, integrating analytical quality by design (AQbD) and green analytical chemistry (GAC) principles. The objective was to enhance method efficiency while reducing environmental impact, energy consumption, and solvent use. An AQbD approach was employed, starting with quality risk assessment and scouting analysis, followed by optimization using Box–Behnken design (BBD) to screen three key chromatographic parameters. The optimal chromatographic conditions were determined using a C18 column (250 mm × 4.6 mm, 5 µm) with a mobile phase of methanol: water (80%:20% v/v), a flow rate of 1 mL/min, ultraviolet detection at 228 nm, and a temperature of 25°C. The method demonstrated linearity over a concentration range of 1–5 µg/mL for the target compounds, with gallic acid as an internal standard. The method's greenness profile was assessed using various evaluation tools, confirming its compliance with GAC principles. The results indicate that the proposed HPLC method is not only effective and reliable but also environmentally sustainable, making it suitable for routine use in quality control laboratories. The integration of AQbD and GAC principles highlights the method's contribution to more sustainable and efficient analytical practices.

Development of an HPLC Method for Simultaneous Quantitation of Hyperin, Quercetin and Kaempferol in the Semen cuscutae

In this study, a method for the simultaneous quantification of hyperin, quercetin, and kaempferol in samples of Semen cuscutae was developed using high-performance liquid chromatography (HPLC). The chosen chromatographic conditions were a mobile phase of methanol – 0.1% formic acid in water, running in gradient mode, with a flow rate of 0.9 ml/min, and detection wavelengths of 354, 365, and 369 nm for hyperin, kaempferol, and quercetin, respectively. The results of the quantitative analysis of 5 samples of Semen cuscutae purchased from Phu Tho province showed hyperin content ranging from 0.950 to 1.218 mg/g; quercetin from 0.077 to 0.118 mg/g; and kaempferol from 1.051 to 1.190 mg/g based on the dry weight of the medicinal material. This research may be suitable for the quality control of Semen cuscutae in the market.

Stability‐Indicating RP‐HPLC Method for Lopinavir Estimation in Pharmaceutical Co‐Crystals: A Design of Expert Approach

ABSTRACTThe primary objective of this investigation was to develop and validate a robust reversed‐phase high‐performance liquid chromatography (RP‐HPLC) method for the quantification of lopinavir in co‐crystals formed with fumaric acid and acetamide. A Box–Behnken design within the design of the experiment framework was employed to optimize chromatographic conditions. The method utilized a Phenomenex Luna C18 column and a mobile phase of methanol and 0.1% formic acid (80:20 v/v) at a flow rate of 1 mL/min, with detection at 259 nm. Validation followed ICH Q2 (R1 and R2) guidelines, confirming the method's specificity, accuracy, precision, linearity, and robustness. Co‐crystals of lopinavir were prepared using liquid‐assisted grinding and reaction crystallization techniques. The validated HPLC method successfully quantified lopinavir in these co‐crystals, with recovery rates ranging from 96.3% to 99.8%. Forced degradation studies under various stress conditions (acidic, base, oxidative, thermal, and photolytic) demonstrated the method's capability to distinguish lopinavir from its degradation products. In conclusion, the developed RP‐HPLC method is suitable for routine analysis and quality control of lopinavir in co‐crystal formulations. This method not only provides precise quantification but also offers insights into the enhanced delivery potential of lopinavir through co‐crystallization, contributing to improved therapeutic efficacy in HIV/AIDS treatment.

Method development and validation of an analytical quality by design ultrafast liquid chromatographic method for the determination of bedaquiline from pharmaceutical bulk and nanoemulsions.

Bedaquiline (BDQ) is a drug used to treat multidrug-resistant tuberculosis (MDR-TB). It exhibits exposure-dependent efficacy in eliminating Mycobacterium tuberculosis (Mtb). An easy, efficient and precise reverse-phase ultrafast liquid chromatography (RP-UFLC) method was developed to validate the free base of the antitubercular medication BDQ. BDQ was separated using a 10:90 v/v mobile phase of ammonium acetate buffer solution (pH = 5.4) and high-performance liquid chromatography-grade methanol, with a flow rate of 1.5mL/min and a UV detection wavelength of 226 nm. By using the Box-Behnken design (BBD) and response surface methodology (RSM), the method was optimised by varying critical analytical attributes (CAA) and critical performance attributes (CPAs) namely ammonium acetate fraction (%), flow rate (ml/min), buffer system molarity (M) and pH. BDQ was eluted at 7.5min utilising isocratic elution. The method was linear in the concentration range of 0.5-300 μg/mL with limit of detection values of 0.039 μg/mL and limit of quantification of 0.12 μg/mL. The results indicate that this validated method can be used as an alternative method for assay of BDQ.

Possibilities and limitations of computer assisted chiral HPLC method development for ozanimod on polysaccharide based chiral stationary phases.

In this study, a direct HPLC method was developed to determine the enantiomeric purity of the immunomodulatory drug, ozanimod. A systematic method development process was followed, incorporating risk assessment, identification of critical analytical procedure parameters, initial screening of stationary phases, and software-assisted optimization of method parameters. Eight different polysaccharide-based chiral columns were selected to assess chiral separation of enantiomers under polar organic elution mode. The most promising results were obtained using a methanol:2-propanol mixture on the amylose-based Chiralpak AD column. Following this, systematic modeling was conducted using DryLab software to optimize method conditions, including isocratic eluent composition, temperature, and flow rate. Baseline separation was achieved within fifteen minutes using the optimized parameters: Chiralpak AD column thermostated at 10°C, and a mobile phase of methanol:2-propanol: diethylamine, 70:30:0.1 (v/v/v %), delivered at a flow rate of 0.8 mL/min. The developed method was validated according to current guidelines and in silico robustness testing was conducted to determine tolerance limits for critical separation parameters and their impact on enantioresolution. Our findings demonstrate the utility of DryLab, typically employed for reversed-phase achiral separations, in optimizing chiral methods even in polar organic mode. Limitations of the selected approach the development of chiral separation methods are also highlighted.

Analysis of sildenafil citrate in male stamina herbal medicines from the Magelang region, Central Java using thin-layer chromatography and UV spectrophotometry

Background: Male stamina herbal medicines are widely consumed in Indonesia, often believed to enhance sexual performance. However, the illegal addition of medicinal chemicals (BKOs) such as sildenafil citrate in traditional herbal products poses health risks. Objective: To identify and quantify sildenafil citrate in selected herbal medicine samples using thin-layer chromatography (TLC) and UV spectrophotometry. Method: Three samples of male stamina herbal medicine were purchased and subjected to organoleptic tests. Ethanol (96%) was used for extraction, followed by TLC analysis using two mobile phases (methanol: chloroform at 4:1 and 1:1 ratios). UV spectrophotometry was employed to determine the maximum absorbance wavelength and quantify sildenafil citrate. Results: TLC analysis revealed that two samples (K and L) contained sildenafil citrate, with Rf values consistent with the standard. UV spectrophotometry confirmed the presence of sildenafil citrate at a maximum wavelength of 292.5 nm. Sample K contained 0.96%, while sample L had 13.766% sildenafil citrate. Conclusion: The presence of sildenafil citrate in two samples violates Indonesian regulations prohibiting medicinal chemicals in herbal products, raising significant safety concerns.

Multiscale simulation of liquid chromatography: Effective diffusion in macro–mesoporous beds and the B-term of the plate height equation

We performed multiscale simulations of analyte sorption and diffusion in hierarchical porosity models of monolithic silica columns for reversed-phase liquid chromatography to investigate how the mean mesopore size of the chromatographic bed and the analyte-specific interaction with the chromatographic interface influence the analyte diffusivity at various length scales. The reproduced experimental conditions comprised the retention of six analyte compounds of low to moderate solute polarity on a silica-based, endcapped, C18 stationary phase with water‒acetonitrile and water–methanol mobile phases whose elution strength was varied via the volumetric solvent ratio. Detailed information about the analyte-specific interfacial dynamics received from molecular dynamics simulations was incorporated through appropriate linker schemes into Brownian dynamics diffusion simulations in three hierarchical porosity models received from physical reconstructions of silica monoliths with a mean macropore size of 1.23 µm and mean mesopore sizes of 12.3, 21.3, or 25.7 nm. The mean mesopore size was found to have a similar influence on the effective mesopore diffusivity as the analyte polarity and the mobile-phase elution strength, which together determine the analyte residence time on a column. A smaller mesopore size attenuated the increase of the effective mesopore diffusivity with increasing mobile-phase elution strength significantly. The effective bed diffusivity was limited by the analyte residence time rather than by morphological details of the mesopore space. The stronger an analyte was retained by the chromatographic interface inside the mesopores, the slower became the mass transfer between the pore space hierarchies and the lower was the effective bed diffusivity. The B-terms of the plate height equation were finally generated with the bed diffusivities and phase-based retention factors derived from the hierarchical porosity models using additional information about the stationary-phase limit obtained from the analysis of analyte–bonded phase contacts. The B-terms highlight analyte- and mobile phase-specific behavior relevant to isocratic and gradient elution conditions in chromatographic practice. In particular, U-shaped B-term curves are observed due to the dominating contribution of the retention factor and the bed diffusivity to the B-term at low and high elution strength of the mobile phase, respectively.

Optimization of HPLC Conditions for Analysis of Paracetamol in Jamu Pegal Linu in Pasar Besar Malang City Using Response Surface Methodology-Box-Behnken Design (RSM-BBD)

Based on the Indonesian Ministry of Health Regulation No.7 in 2012, herbal medicine is disallowed to contain synthetic chemicals or isolated products with medicinal properties. However, from the findings of BPOM, there are still many herbal medicine producers who add medicinal chemicals to herbal medicine, one of which is the addition of paracetamol in jamu pegal linu. This study aimed to obtain optimum HPLC conditions for the analysis of paracetamol in jamu pegal linu using the Response Surface Methodology-Box-Behnken Design, ensure the HPLC method has met the validation parameters, and determine the presence and levels of paracetamol contained in jamu pegal linu sold in Pasar Besar, Malang City. HPLC conditions that were optimized were the percentage of methanol mobile phase in water, flow rate, and column temperature. The HPLC condition optimization results were obtained at a percentage of methanol mobile phase in water of 34.4%, mobile phase flow rate of 1 mL/min, and column temperature of 30°C. Analysis of paracetamol content of paracetamol in jamu pegal using HPLC and method validation with test parameters including selectivity (λmax 245 nm; paracetamol retention time ± 2.6 minutes; resolution ± 2.5), linearity (r2 = 0.9984), LOD (2.44 ppm), LOQ (7.40 ppm), accuracy (97-102%), and precision (0.26-0.69%). The analysis of paracetamol content in the three samples of jamu pegal linu analyzed showed that negative for paracetamol and fulfilled the regulation of the Indonesian Ministry of Health Regulation No.7 in 2012.

Advancing Bioanalytical Method Validation: A Comprehensive ICH M10 Approach for Validating LC-MS/MS to Quantify Fluoxetine in Human Plasma and Its Application in Pharmacokinetic Studies.

A fast and sample cleanup approach for fluoxetine in human plasma was developed using protein precipitation coupled with LC-MS-MS. Samples were treated with methanol prior to LC-MS-MS analysis. Chromatographic separation was performed on a reverse phase column with an isocratic mobile phase of methanol and 10 mM ammonium formate pH acidified with formic acid (80:20, v/v) at a flow rate of 0.2 mL/min. The run time was 4 min. Mass parameters were optimized to monitor transitions at m/z [M + H]+ 310 > > 148 for fluoxetine and m/z [M + H]+ 315.1 > > 153 for fluoxetine-d5 as an internal standard. The lower limit of quantification and the dynamic range were 0.25 and 0.25-50 ng/mL, respectively. Linearity was good for intra-day and inter-day validations (R2 = 0.999). The matrix effect was acceptable with CV% < 15 and accuracy% < 15. The hemolytic effect was negligible. Fluoxetine was stable in human plasma for 48 h at room temperature (25 °C), for 12 months frozen at -25 °C, for 48 h in an auto-sampler at 6 °C, and for three freeze/thaw cycles. The validated method was applied in a pharmacokinetic study to determine the concentration of fluoxetine in plasma samples. The study provides a fast and simple bioanalytical method for routine analysis and may be particularly useful for bioequivalence studies. The method was successfully applied to a pharmacokinetic study of fixed-dose fluoxetine in nine healthy volunteers.

Concurrent Analysis of Amlodipine, Rosuvastatin, and Hydrochlorothiazide in Solution and Biological Fluid Using LC-MS/MS.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was created and verified in this investigation. This methodology offers a fast, easy, affordable, economical, and reliable approach to simultaneously determining amlodipine, rosuvastatin, and hydrochlorothiazide in solution and human plasma. Through the extraction process, acetonitrile was utilized as a precipitating agent for the proteins in plasma. The C18 column was injected with a volume of 10 µL, including ondansetron as an internal standard. The analytes were eluted using an isocratic mobile phase (0.1% acetic acid and 1:4 (v/v) methanol). The method was sensitive and selective for the analytes over a dynamic concentration range of 5 to 600 μg/mL in solution and 0.5 to 60.0 μg/mL in plasma. At a minimum of 0.9988 coefficient of determination, linearity was attained. Within and between runs, the precision and accuracy were 2.87 to 11.41% and 86.2 to 113%, respectively. A comparatively quick run time of less than two minutes (high throughput) was achieved. The current method can be used in many applications, such as bioequivalence investigations and therapeutic medication monitoring, to quantify the relevant analytes.

Study on the Related Substance of Lamivudine by A Validated HPTLC Method in Oral Dosage Forms

Lamivudine is an anti-retroviral drug available in the formulation of tablets and oral solution. The selected compound Salicylic acid is categorized as Lamivudine impurity C. An HPTLC method was developed and validated for the estimation of Lamivudine in bulk and oral dosage forms. The chromatograms were developed using a mobile phase of methanol: water: ethyl acetate in the ratio 4:2:4, v/v/v on pre-coated plate of silica gel 60 F254 and quantified by densitometric absorbance mode at 233nm. The Rf values were 0.76±0.03 and 0.65±0.02 for Lamivudine and Salicylic acid respectively. The linearity was found to be within the range of 500-3000 ng/band for Lamivudine and 50-300ng/band for Salicylic acid. The lower limits of detection and quantification were 21.24ng/band and 64.37ng/band for Lamivudine and 8.47ng/band and 25.68ng/band for Salicylic acid. The statistical evaluation by the %RSD calculation for accuracy and precision were below 2. The validated method was applied to detect and quantify Salicylic acid in the tablets of two dosage forms and the oral solution prepared as admixture. The quantified amount of the impurity C was found to be within the limits in the tablets and not detectable in the oral solution. This method can be adopted to determine Lamivudine and its impurity C in the API and different dosage forms.

IMIDAZOLIUM-BASED IONIC LIQUIDS FOR THE SELECTIVE SEPARATION OF TOCOTRIENOL HOMOLOGUES FROM PALM FATTY ACID DISTILLATE VIA LIQUID-LIQUID EXTRACTION

Objective: Tocotrienols have now stepped into the limelight of vitamin E research and have proven to have some exceptional benefits that are not shared by their “older” tocopherol siblings. Unlike tocopherols, tocotrienols are able to inhibit cholesterol biosynthesis, have specific neuroprotective activities stronger antioxidant effects, antihypertensive and anti-cancer. The purpose of this study was to carry out selective separation of tocotrienols homologues from Palm Fatty Acid Distillate (PFAD) by liquid-liquid extraction, using Ionic liquids (ILs) as extractants in the presence of diluent. Methods: Four kinds of imidazolium-based ILs that used are 1-butyl-3-methylimidazolium chloride ([Bmim]Cl), 1-Butyl-3-methylimidazolium acetate ([Bmim]Ac), 1-Hexyl-3-methylimidazolium chloride ([Hmim]Cl) and 1-Ethyl-3-methylimidazolium chloride ([Emim]C]). The extraction is carried out by creating a two-phase system between ILs and PFAD in n-hexane. Quantification of extracted tocotrienols was performed using High-Performance liquid Chromatography (HPLC) with a C18 column, mobile phase methanol: water (97.5:2.5), flow rate 1 ml/min and Ultraviolet (UV) detector at 295 nm. Results: The results showed that the concentration (extraction efficiency) of tocotrienols extracted using ILs in order were [Bmim] Ac 1611.09 mg/Kg (75.41%) and [Hmim][Cl] 1603.39 mg/Kg (75.05%), [Bmim]Cl 1523.60 mg/Kg (71.32%) and [Emim]Cl 1174.24 mg/Kg (54.96%). Conclusion: [Bmim]Ac yielded the highest tocotrienols concentration and extraction efficiency.

Modern chromatographic method for estimating Loratadine and affections on healthcare

PurposeThe purpose of this study is to establish Loratadine [LRD] quantification in purified and capsule formulations using a precise and specific Reversal Phase with a very high-performance liquid Chromatographic [RP-HPLC] technique. The approach was evaluated in agreement with the principles of the International Conference on Harmonization [ICH]. Arcus EP-C18 Ion Pac column, 5 m, 4.6 mm, 250 mm, mobile phase Methanol: Acetonitrile (60:40) v/v. Dibasic potassium phosphate buffer, pH 7.2, flow rate 1.0 ml/min.Design/methodology/approachThe HPLC system used a 340 nm UV detector for testing. A 10-min run time was used for the analysis. At concentrations ranging from 2 to 10 g/ml, the technique was linear (R2 = 0.9998), exact (intra-day and inter-day relative standard deviation [RSD] values 1.0%), accurate (range recovery = 96%–102%), exclusive and strong.FindingsThe detecting and quantitation limits were 0.92 g/ml and 2.15 g/ml, respectively.Originality/valueThe findings demonstrated that the proposed method could accurately determine LRD in bulk and pill dose formats quickly and accurately.

Box-Behnken Design employed stability demonstrating RP-HPLC method development of sorafenib tosylate for rapid and sensitive quantification

Sorafenib tosylate (SFN) is a tyrosine kinase inhibitor used clinically to treat liver, kidney, and thyroid carcinoma. This research aims to accurately quantify SFN using a quality-by-design (QbD) approach with RP-HPLC. Chromatographic settings were optimized using a Box-Behnken design, measuring responses such as retention time, tailing factor, theoretical plate, and peak area. At the same time, independent variables were flow rate, mobile phase composition, and wavelength. A C18 column (4.6 × 250 mm, 5 µm) served as the stationary phase, with a mobile phase of methanol and 0.1% o-phosphoric acid in a 41:59 v/v ratio. SFN detection occurred at 267 nm in isocratic mode at a flow rate of 1.1 mL min−1. Method validation followed International Council for Harmonization guidelines, yielding a coefficient of determination (R2) of 0.9998, indicating linearity in the 5-25 µg mL−1 range. Results showed detection (LOD) and quantification (LOQ) limits of 0.04 and 0.12 µg mL−1, respectively. Additionally, the method demonstrated precision, accuracy, and robustness consistent with ICH criteria. Overall, this simple, accurate, rapid, and robust RP-HPLC method is suitable for routine SFN analysis in various formulations.

Development and Validation RP-HPLC Method for Estimation of Antidiabetic Drugs in Pharmaceutical Dosage Form

This study details the development and validation of an RP-HPLC method for estimating Canagliflozin and Metformin in pharmaceutical dosage forms. Canagliflozin, an SGLT2 inhibitor, and Metformin, an antihyperglycemic agent, were obtained from Arti Drug Pvt. Ltd. Marketed formulation Invokamet was sourced locally. Chromatographic separation was achieved on a C18 column (250 x 4.6 mm, 5 µm) using a mobile phase of Methanol: Water (80:20) with 0.1% ortho phosphoric acid at a 1.0 ml/min flow rate and detection at 254 nm. The retention times for Canagliflozin and Metformin were 8.183 min and 3.633 min, respectively. Method validation parameters included system suitability, accuracy, precision, and robustness. System suitability tests confirmed adequate resolution and repeatability. Accuracy was evaluated through recovery studies at 80%, 100%, and 120% concentration levels, yielding satisfactory results. Precision was demonstrated with intra-day and inter-day % RSD values below 2. Robustness was assessed by varying mobile phase composition and flow rate, showing consistent retention times and tailing factors. The method proved selective, accurate, precise, and robust, with a short run time, facilitating rapid quantification of samples. It is suitable for routine quality control analysis of Canagliflozin and Metformin in pharmaceutical formulations

Determination of Mesalamine in Bulk and Suppository Dosage Forms through the Development and Validation of Stability- indicating RP-HPLC Method

Background: In the current study, a simple and cost-effective stability-indicating RP-HPLC method was developed and validated to estimate the mesalamine from both bulk and pharmaceutical dosage forms. Methods: An isocratic HPLC method using a reverse phase HiQSilC18 column (250 x 4.6 mm, 5μm) and a mobile phase methanol: ammonium acetate buffer (90:10 v/v) were employed as the mobile phase with a flow rate of 1 mL/min at 25°C. Detection was carried out at 305 nm, and the injection volume was 20μl. The developed method was validated as per ICH Q2 guidelines. Mesalamine has been subjected to various stress testing conditions, such as hydrolysis of acid and base, thermal degradation, oxidation, and photolysis. Also, methods have been validated with regard to linearity, accuracy, precision, and robustness. Results: The RT of mesalamine was determined to be 3.550 min ± 0.024 minutes, providing a reliable marker for its identification. The method was found to be linear between 5-30 μg/mL concentration with (R²) of 0.994. This demonstrated the method's ability to measure varying concentrations of mesalamine accurately. Additionally, the percentage recovery of mesalamine was approximately 100%, confirming the accuracy of the developed method. The parameters for system suitability have also been found to be within acceptable limits. Force degradation studies reinforced the method's selectivity and sensitivity in detecting mesalamine under various degradation scenarios. Notably, mesalamine significantly degraded in an acidic environment. Conclusion: In conclusion, our proposed RP-HPLC method provides a sensitive, accurate, and precise means of analyzing mesalamine in both bulk and pharmaceutical dosage forms.

Simultaneous Quantification of Dutasteride and Silodosin Using a Stability-Indicating RP-HPLC Approach

An alternative stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) approach remained established to quantify silodosin (SLD) and dutasteride (DTS) in the mixture, validated per International Council of Harmonization (ICH) recommendations. A Shimadzu Model LC-2030 PLUS (IND) HPLC system, 231 nm detection wavelength, and 10 minutes run duration were used to create the method. A 75:25 mobile phase of methanol and water with 0.05% OPA was used at 1-mL/min. The concentrations of SLD (8–40 μg/mL) and DTS (0.5–2.5 μg/mL) were shown to be linearly related, with R2 values of 0.9998 for SLD and 0.9993 for DTS. Both of the drugs had accuracy and precision within 2% RSD. SLD and DTS mean recoveries were 97.5 to 102%. According to ICH criteria, degradation trials with acid, alkali, oxidizing agent, and heat assessed the approach’s stability. The established approach is fast, accurate, sensitive, and precise for routine SLD and DTS analysis from bulk as well as pharmaceutical dosage forms.

Quality by Design Approach for the Stability Indicating Method Development and Validation of Selpercatinib Drug Formulation by using RP-HPLC

In method development quality design approach is useful for method goal identification evaluation, method selection and risk assessment. Selpercatinib is a drug used in the treatment of certain types of cancer. The method development includes selecting appropriate chromatographic conditions such as the mobile phase composition, column, and selection wave length. The aim is to achieve optimum separation and quantification of selpercatinib in the dosage form. Optimizing chromatographic conditions by using design expert software. The mobile phase methanol and 0.1% Acetic acid are used. The mobile phase range is 80:20 v/v. The range of this mobile phase is 80:20 .0.7 mL/min found flow rate. The retention time of selpercatinib is 3.20 minutes. The assay was found to be 99.01% for selpercatinib. Optimization was performed. Factorial design performed, i.e., flow rate and percentages of methanol. Analysis of variance confirmed that method parameters. UV detection at 220 nm. The International Council of Harmonization (ICH) guidelines ensure the accurate, precise and reliable results to validate the parameters is an acceptable range. Stress studies reveal that drugs were degraded acidic conditions, alkaline conditions, neutral conditions, oxidative conditions, and photolytic conditions. Hence the proposed method was stability, indicating using quality by design (QbD) approach all the method parameters were better understood. This systematic approach ensures a thorough understanding of the assay, facilitates process optimization, and enables effective control strategies to reduce variability and enhance product quality.

Development and Validation of HPTLC for the Determination of Gefitinib HCl in Tablet Dosage Form

A simple, high-performance thin-layer chromatography (HPTLC) method of gefitinib HCl in pharmaceutical formulation was developed. Separation was done on TLC aluminum plates precoated by silica gel 60GF254 (20 × 20 cm) with 200 μm layer depth. With UV detection set to 254nm, chromatography was conceded out by the mobile phase of dichloromethane and methanol (9:1, v/v). Linearity was found to range of 50 to 300 ng/spot and, the correlation coefficient was found to be 0.99 and the Rf value was found to be 0.46. The system’s suitability was considered to ascertain the performance of ascertaining quality performance of the developed chromatographic method. The method’s accuracy, precision, and specificity were all verified against the International Council of Harmonization (ICH) standards. Application of the approach to the determination of gefitinib in tablet form was successful.

Development and validation of a rapid, simple, and reliable UPLC-MS/MS method for the quantification of vancomycin in human plasma

Vancomycin is a critical antibiotic frequently utilized in clinical settings, with therapeutic drug monitoring (TDM) strongly advised to optimize treatment efficacy and mitigate the risk of adverse effects. However, current methods for measuring vancomycin levels in human plasma are hindered by long analysis times and complicated sample preparations. Thus, this study developed and validated a novel UPLC-MS/MS method for a rapid (with a running time of 3.5 min) and simple analysis of plasma vancomycin. To quantify vancomycin concentration in human plasma, we have developed and validated the UPLC-MS/MS method with high sensitivity, specificity, and accuracy, meeting the strict criteria according to the Food and Drugs Administration (FDA) guidelines for validation biological analysis methods. Vancomycin and atenolol (internal standard) underwent positive electrospray ionization (ESI+) and detection in multi-reaction monitoring (MRM) mode. The selected MRM transitions were m/z 725.66→144.16 for vancomycin and m/z 267.29→189.96 for atenolol. Plasma samples were precipitated using a simple mixture containing acetonitrile, methanol, and formic acid as a pH adjuster. The separation was performed using the Poroshell 120 Phenyl Hexyl Column (4.6 × 150 mm, 2.7 μm) maintained at 25 °C for 3.5 min. Isocratic elution with a mobile phase (methanol and 0.1% formic acid in a 40:60 v/v ratio) at a flow rate of 0.5 mL/min was employed. The method showed linearity (0.1–75 μg/mL) with a coefficient of determination above 0.9994 and a lower limit of quantification at 0.1 μg/mL. Precision, both intraday and interday, was below 10%, and accuracy ranged from 91.70% to 111.57%. System suitability, selectivity, stability, carryover, dilution, recovery, and matrix effect validation results all met acceptable criteria. The established UPLC-MS/MS method is expected to be a rapid, simple, and reliable tool for drug monitoring and pharmacokinetic studies, enhancing patient care during vancomycin administration.