Bioanalytical Method Development Research Articles

A Review on Various Analytical Methodologies of Naproxen

Naproxen is a Nonsteroidal Anti-Inflammatory Drug (NSAID). It is mostly used to treat pain or inflammation caused by conditions such as arthritis, gout, tendinitis or menstrual cramps. Naproxen is available in isolated dose with various similar anti-inflammatory drugs, i.e.; esomeprazole, pantoprazole, paracetamol, ranitidine, sumatriptan and ibuprofen. This survey evaluates various methods for the analysis of Naproxen in bulk drugs and formulated products. Analytical procedures are critical for determining compositions, they allow as to obtain both qualitative and quantitative results utilizing by the advanced analytical tools. This include HPLC, HPTLC, UV- spectrophotometry, capillary electrophoresis, and electrochemical meth-ods. The UV-spectrophotometry method is applied for the investigation of Naproxen in biologi-cal media, bulk samples and in various dosage formulations. The HPLC technique of Naproxen alone and the combination, including parameters such as matrix, stationary phase, mobile phase, wavelength detection, etc. HPTLC method parameters such as stationary phase, mobile combi-nation phase, RF value, etc. To maintain high commercial product quality standards and to ad-here to regulatory requirements, analytical technique development is necessary. Drug concen-trations are measured using bioanalytical techniques. Development and validation of bioanalyt-ical method is important to understand the pharmacokinetics of any drug and/or its metabolites.

Bioanalytical method development and Validation for determination of efonidipine hydrochloride ethanolate in rat plasma by high-performance liquid chromatography: applications for pharmacokinetic study

Efonidipine hydrochloride ethanolate (EFE), a dihydropyridine calcium channel blocker, is used for the treatment of hypertension. To find the EFE in pre-clinical samples (rat plasma), an HPLC method was devised for the current experiment. The biological samples were prepared using the methanolic extraction procedure to eliminate solvent effects. Using a C18 column, a pH-adjusted mobile phase of methanol and water (90:10, v/v) at a flow rate of 1 mL/min, and UV detection at 252 nm were utilized for chromatographic separation. The retention time of EFE was found to be 5.2 min. The developed bioanalytical method was validated with different validation parameters. The validation parameter results are within the acceptable range of the International Council for Harmonization guidelines. The method was successfully employed to evaluate the pharmacokinetic properties in Wistar albino rats following a single oral administration of 10 mg/kg of EFE. Therefore, the simple method would make it possible to detect EFE quickly and affordably from biological material.

Development and validation of a new bioanalytical method for quantification of CDK4/6 inhibitor in Spiked Human Plasma by HPLC-UV

Abstract A sensitive and accurate high performance liquid chromatography method utilizing ultraviolet/visible light detection (HPLC-UV) for the quantification of Ribociclib in Spiked Human Plasma by HPLC-UV was developed and validated. Ribociclib (RCB) and the internal standard (IS), Trifluridine, were first extracted from plasma samples by a simple Protein Precipitation extraction using Acetonitrile. Plasma concentration of RCB and internal standard were then analyzed by applying reversed phase chromatography using Orochem orosil C18 (4.6 mm × 250 mm, 5 μ) and elution with a isocratic mobile phase consisting of 10 mM phosphate buffer – Acetonitrile (60:40, v/v) adjusted to pH 3.0 at a flow rate of 1.0 ml/min. Detection of RCB and the IS was subsequently done at a wavelength of 260 nm. The limit of quantification was 10 ng/ml. The calibration curve was linear (R2>0.998) over the concentration range of 10-1000 ng/ml. For human plasma, the accuracy and precision were within ±15% and ≤15%, respectively, for all concentrations, except for the lower limit of quantification, where they were within ≤20%.

RP-UFLC based Bioanalytical Method Development, Optimization, and Validation for the Estimation of Isradipine in Rabbit Serum

Introduction: The objective of this study is to provide a rapid, sensitive, consistent, and costeffective method for quantifying isradipine using ultra-fast liquid chromatography. Methods: Quality by Design principles will form the basis of this approach, grounded on response surface analysis. Shimadzu liquid chromatographic system equipped with a photodiode array detector and LC solution software was used to conduct the RP-UFLC method development and validation. An ODS C18 (250 x 4.6 mm; 5 μm) UFLC column was used to complete the analysis. The RSM methodology utilized a central composite design to perform the optimization studies. Results: The mobile phase ratio and flow rate were considered crucial method parameters, as well as the peak area, retention time, and USP plate were considered critical analytical attributes. The optimal conditions for chromatographic separation were followed using 80% acetonitrile and water (20% v/v) as mobile phase, a 1 mL/min flow rate, an injection volume of 20 μL, 40°C of column oven temperature, and maximum absorption at λmax 254 nm using graphical optimization technique. When examining concentrations between 5 and 150 ng/mL, linearity was observed with an R2 of 0.999. The method created was validated by employing stability testing per the recommendations provided by ICH Q2 (R1). The analysis of blood serum was modified so that it could be used to examine the pharmacokinetic parameters. Conclusion: As a result, high accuracy, precision, sensitivity, linearity, and robustness were established for predicting the amount of isradipine present in its freeze-dried nano-formulations.

Bioanalytical method development for quantification of trifluridine with its metabolites and tipiracil in spiked human plasma by solid-phase extraction (SPE) and HPLC with PDA detection

A novel high pressure liquid chromatography method coupled with diode array detector (HPLC-DAD) was developed and found to be efficient, sensitive, and selective for the simultaneous analysis of Trifluridine (TRI), and its metabolites 5-trifluoromethyl uracil (TFMU) and 5-carboxydeoxyuridine (CDMU) and Tipiracil (TIP) in a spiked human plasma. The HPLC-DAD technique that developed was validated in accordance with the most recent ICH M10 criteria. Sample pre-treatment was performed using a solid phase extraction with InertSep HLB reversed phase sorbent 30 mg/ml cartridge. The compounds were analyzed by liquid chromatography UV detection on a Inertsil C18 (250 mm × 4.6 mm ID, Particle size: 5 µm) with a mobile phase containing sodium acetate buffer pH (3.8; 10 mM) - acetonitrile (70:30, v/v) with flow rate of 0.8 ml/min with UV detection at 254 nm. The method was linear in the range 200–5000 ng/ml for TRI, TFMU, CDMU, and 100–1500 ng/ml for TIP. With the exception of the lower limit of quantification, where they were within ≤ 20%. The accuracy and precision for spiked human plasma were within ≤ 15% for all concentrations.

Bioanalytical Method Development and Validation of Eprosartan Mesylate and Hydrochlorthiazide using RP-HPLC in Human plasma

A precise, very rapid and cheaper RP-HPLC method that is reproducible and stable is developed aiming for estimating Eprosartan mesylate and hydrochlorothiazide simultaneously in an active pharmaceutical ingredient and tablet dosage form. This method relies on the simplicity and easy to run fashion. The diluent/solvent was selected based on the solubility of the drugs. The RP-HPLC analysis was performed using an Agilent system C18 (150mm x 4.6mm, 5m) column with a mobile phase of 0.1% orthophosphoric acid pH (2.2): Acetonitrile (60:40), Flow rate 1.0ml/min, Detector wavelength 240nm. Calibration was linear for Eprosartan (80-3200ng/ml) and Hydrochlorothiazide (8.5-340ng/ml). The value of r2 was more than 0.999 consistently. The accuracy within the same day and during the course of estimation of plasma samples were investigated, and it was discovered that the mean percent accuracy was very highwith ranges from 100.13 to 100.66% intra-day and 99.16 to 100.87% inter-day. On the basis of the analytes and plasma samples, the precision (percent CV) was computed and found to be 0.91-1.76% for intraday and 0.98-1.58% for inter-day. Eprosartan and Hydrochlorothiazide had 98.85% and 98.67% mean recovery.The percent mean stability of Eprosartan was determined to be 99.90% at 28°C and 100.16% at 80°C. The percent mean stability of Hydrochlorothiazide was determined to be 100.94 at 28°C and 101.11 at 80°C. The newly discovered approach has been effectively used to the regular examination of pharmaceutical formulations for pharmaceutical drugs.

A Review on Bioanalytical Method Development and Validation for Estimation of Drugs by Using Hyphenated Techniques and its Applications on Routine Analysis

Bioanalytical methods are used to create a protocol that can identify and quantify bioactive molecules and their metabolites in human and animal samples. For determining the medication and its metabolites present in human/animal body, the bioanalytical method is efficient. One of the prominent bioanalytical roles is creating new techniques, validating existing processes, and analyzing samples. Most importantly, a compound can be calculated using many methods and can also be detected using various analytical methods. In specific biological samples, the drugs can be evaluated by different extraction procedures, such as solid-phase extraction, liquid extraction, protein precipitation etc.., All the steps in methods were investigated to know how environment, sample, or techniques influence the matrix assessment up to the study's time. The analytical methods, including high-pressure (HPLC), liquid chromatography combined with dual-mass spectroscopy (LCMS-MS), and Ultra-performance extraction (UPLC), can be used to obtain the in-depth analysis of medications. All have their strengths and weaknesses. Currently, bioanalysis is usually done by HPLC-MS and GCMS-MS. Linearity, repeatability, Accuracy, precision, selectivity, recovery and consistency are essential criteria. In this review we proposed to give importance of bioanalytical system production and validation parameters to aid in drug quality, purity, safety and efficacy.

A review on bioanalytical method development and validation of anticancer drugs by using lc/ms/ms and its applications on routine analysis

A protocol is used to detect and measure biomolecules and metabolites in human and animal tissues using bimolecular methods. The biosanalinity method is effective at determining the number of drugs and metabolites in a biological system. New methods, the validation of existing procedures, and the analysis of samples are one of the prominent tasks for bioanalysis. Above all, a compound can be measured using several methods and identified by different methods of analysis. Drugs may be tested by several extraction techniques, including liquid extraction, solid-phase extraction, and protein precipitation in complex plasma and biological samples. To determine how the environment, matrix, or procedures impact the matrix estimation to the time of the analysis, all steps in the process must be investigated. The more detailed study of drug products can be performed with higher-pressure analytical techniques, such as high- extraction (HPLC), liquid chromatography coupled with double-mass spectrometry (LCMS/MS), and ultra-performance Liquid chromatography (UPLC). Both of them have flaws and strengths. At present, HPLC and GC usually perform biolysis. The parameters are linearity, repeatability, accuracy, selectivity, and continuity. We are proposing the development and validation of bioanalytical systems to assist in the quality assurance of drugs.

Bioanalytical Method Development and Validation for Determination of Rifampicin and Quercetin in Rat Plasma by UHPLC-MS/MS: Applications to Pharmacokinetic Study

UHPLC-MS/MS was utilised to quantitatively analyse Rifampicin and Quercetin-loaded Liquisolid compact in rat plasma. The UPLC Acquity C18 (1.7 μm, 2.1 × 15 mm) column and 0.5 ml/min were used to separate the analyte. In a low-pressure gradient mode, A: In water 0.1 % formic acid and B: In acetonitrile 0.1 % formic acid as the mobile phase. Sample pre-treatment was performed by protein precipitation technique with methanol and acetonitrile (1:1) from rat plasma. As an internal standard (IS), the analyte was found by tracking precursor-to-product ion transformations of 823 → 791.3 m/z for rifampicin, 303 → 257 m/z for quercetin, and 138 → 121 m/z for isoniazid in MRM mode. The proposed technique was validated for precisions (Intraday and Interday) accuracy, linearity, quantification at lower limits, and analyte recovery. The findings showed that the plasma samples inter and intra-day precision and consistency values were determined to be within the acceptable range. After orally administering Liquid solid compact and pure drug solution, which showed a substantial difference in the pace and extent of absorption, the method’s suitability for determining the pharmacokinetic profile of each drug was tested.

Bio-Analytical Method Development and Validation of Asciminib and its Application to Pharmacokinetic Studies in Rat Plasma by Using RP-HPLC

The aim of the proposed study was to validate a rapid, uncomplicated, accurate, robust and sensitive bio-analytical method for quantifying asciminib using ivosidenib as an internal standard in rat plasma. This work provides a summary of recent advances in bio-analytical HPLC methodologies employing a Luna phenyl hexyl column (150 × 4.6 mm, 3.5) with an organic mobile phase of potassium dihydrogen phosphate (adjusted to pH 3.0 with orthophosphoric acid) and acetonitrile in the ratio 40:60. With a correlation coefficient of 0.999, the asciminib calibration curve was linear over the range of 100-2000 ng/mL. Using liquid-liquid extraction, the percent of recovered ASB ranged between 97.8 and 99.9%. The plasma stability of the bench-top and auto sampler remained stable for 24 h in the auto sampler. The T1/2 is 5 h and Tmax is 3 h, as calculated from the pharmacokinetic parameters. The application indicates that all parameters of system suitability, specificity, linearity and accuracy are in good agreement with the prescribed limits; consequently, the current method is highly stable, rapid and capable of analyzing plasma samples. This method is ideally adapted for determining asciminib in pure or dose form due to its novel technology.

Bioanalytical method development and validation of folic acid from rat plasma using reverse phase high performance liquid chromatography

Bioanalytical method development and validation of folic acid from rat plasma using reverse phase high performance liquid chromatography

Bioanalytical method development and validation of a novel antiseizure agent Cenobamate using LC-MS/MS

Bioanalytical method development and validation of a novel antiseizure agent Cenobamate using LC-MS/MS

Bio-Analytical Method Development and Validation for Ramipril and Its Metabolite by Liquid Chromatography-Tandem Mass Spectrometry

Bio-Analytical Method Development and Validation for Ramipril and Its Metabolite by Liquid Chromatography-Tandem Mass Spectrometry

Bioanalytical method development and validation for the quantitation of larotrectinib in human plasma: Application to pharmacokinetics in healthy rabbits

Bioanalytical method development and validation for the quantitation of larotrectinib in human plasma: Application to pharmacokinetics in healthy rabbits

Development and Validation of Bioanalytical Method for the Quantification of Febuxostat in Human K2 EDTA Plasma by LC-MS/MS: Pharmacokinetic Studies in Wister Rats

A specific, linear and accurate LC-MS/MS method was developed for the determination of febuxostat in human K2 EDTA plasma and it was successfully applied for the pharmacokinetic study in wister rats. Chromatographic isolation of febuxostat and febuxostat D9 were attained on Purosphur C18, 100 × 4.6 mm, 5 μ column with 1.0 mL/min flowing rate. The technique was linear over the standard concentrations ranging from 24.995-7001.401 ng/mL and the regression coefficient was perceived to be ≥ 0.9997. All LLOQ samples % RSD of back computed concentrations varied from 1.00 to 4.01. The analysis of plasma from healthy rats was successfully conducted using the validated LC-MS/MS procedure to quantify the presence of febuxostat. From the pharmacokinetic studies, Tmax, Cmax, T1/2 and AUC0-∞ of the febuxostat tablets were 2.0 ± 0.03 h, 8.85 ± 1.87 ng/mL, 6.34 ± 0.53 h, 95.58 ± 6.37 ng h/mL, respectively.

Development and Validation of a Novel Bioanalytical Method for Estimating Epigallocatechin 3 Gallate in Wistar Rat Plasma by RP-HPLC Employing Gradient Elution Techniques

Development and Validation of a Novel Bioanalytical Method for Estimating Epigallocatechin 3 Gallate in Wistar Rat Plasma by RP-HPLC Employing Gradient Elution Techniques

Bioanalysis of Endogenous Isotretinoin in Altered and Unaltered Human Plasma by Liquid Chromatography-Tandem Mass Spectrometry Coupled with Electrospray Ionization.

Bioanalytical method development and validation of endogenous Isotretinoin with Isotretinoin D5 as internal standard was done as per current regulatory guidelines. The method is simple, rugged and sensitive enough to estimate endogenous Isotretinoin using the chromatography-tandem mass spectrometry technique. An alternative approach has been adopted for quantitative analysis of endogenous Isotretinoin in human plasma. Isotretinoin free matrix (surrogate matrix) was prepared and further used for the development and validation of Isotretinoin. The method was validated in altered and unaltered plasma. The chromatographic optimization was done with column (ACE C18, 100 × 4.6mm I.D. 5μm particle size), using a mobile phase containing 1mM ammonium acetate, pH3.0 as a solvent A and solvent B (1mM ammonium acetate (pH3.0) with acetonitrile in a ratio of 10:90). A flow rate was set at 0.75mL/min in a binary gradient mode. The analyte was recovered by liquid-liquid extraction method with diethyl ether as an extraction solvent. Multi-reaction monitoring mode in negative polarity was implemented for the quantification of endogenous Isotretinoin in plasma. The calibration curve of Isotretinoin was linear (r2 > 0.9992) over the concentration range of 0.5-1000ng/mL. The intra-day precision was found in a range of 2.0-3.9% CV for altered samples and 0.9-3.7% CV for unaltered samples. The inter-day precision was found 2.6-6.1% CV for altered samples and 1.3-3.8% CV for unaltered samples. The average recovery of the extraction procedure was found 64.6% for altered samples and 62.2% for unaltered samples.

Formulation development and optimization of taste-masked azithromycin oral suspension with ion exchange resins: Bioanalytical method development and validation, in vivo bioequivalence study, and in-silico PBPK modeling for the paediatric population

The study is based on preparing taste-masked oral suspension of azithromycin using ion exchange resins Kyron T135 and Doshion-P542 AB. The complexation process was optimized through Design of Experiment (DOE), and the resins-complexes were characterized by bitterness score, differential scanning calorimetry (DSC), fourier transform infra-red spectrophotometry (FTIR) and powder x-ray diffraction (PXRD). The complexes were further formulated and optimized through Central Composite Design (CCRD) into oral suspension using xanthan gum, hydroxypropyl cellulose, tri-basic sodium phosphate, and sucrose. To obtain the optimized product, the trial batches were tested for sedimentation ratio and viscosity in comparison to the reference product followed by assay and studies of drug-release kinetics. To perform bioequivalence of the optimized formulations against the reference product, an LC-MS/MS based bio-analytical method was developed and validated followed by a three-period cross over study. The pharmacokinetic parameters Cmax, Tmax, AUC0-t and AUC 0-∞ were obtained through non-compartmental analysis (NCA) using log-transformed data. Moreover, physiologically based pharmacokinetic (PBPK) modelling of the in-vitro data was carried out to predict the pharmacokinetics of the optimized formulations in the intended paediatric population. Kyron T-135 yielded the best loading efficiency at a drug resin ratio of 1:1.35 and Doshion-P542 AB at 1:3.25, while stirring and swelling time of 30 min for Kyron T-135 yielded optimum drug loading. In case of Doshion P-542 AB the optimum time was 22.5 and 30 min for stirring and swelling respectively. The bitterness score proved the tastelessness of the drug resin complex. The optimized formulation with Kyron (formulation code K4) and Doshion (formulation code D6) were selected as optimum on the basis of similar quality attributes to the reference product. The drug dissolution kinetics revealed first order drug release. The developed bioanalytical method was found to be sensitive and linear in the range of 2–500 ng/mL showing accuracy within 101.32–106.68%. Both optimized formulations (K4 and D6) were found to be bioequivalent with geometric mean ratio of Cmax, Tmax, AUC0-t and AUC 0-∞ within 90% C.I. in silico PBPK based predictive pharmacokinetics in the paediatric population was successfully estimated for the developed formulations.

Review on bioanalytical and analytical method development of Antiepileptic racetams

Review on bioanalytical and analytical method development of Antiepileptic racetams

Bioanalytical methods for pharmacokinetic studies of antileishmanial drugs.

Bioanalytical method development and validation for the quantification of antileishmanial drugs are pivotal to support clinical trials and provide the data necessary to conduct pharmacokinetic (PK) analysis. This review provides a comprehensive overview of published validated bioanalytical assays for the quantification of antileishmanial drugs amphotericin B, miltefosine, paromomycin, pentamidine, and pentavalent antimonials in human matrices. The applicability of the assays for leishmaniasis clinical trials as well as their relevance to PK studies with emphasis on the choice of matrix, calibration range, sample volume, sample preparation, choice of internal standards, separation, and detection was discussed for each antileishmanial drug. Given that no published bioanalytical methods included multiple antileishmanial drugs in a single assay although antileishmanial shortened combination regimens currently were under investigation, it was recommended to combine various drugs in a single bioanalytical method. Furthermore, bioanalytical method development regarding target site matrix as well as applying microsampling strategies was recommended to optimize future clinical PK studies in leishmaniasis.