Bioanalytical Method Development Research Articles

Bioanalytical method development and validation of docetaxel and carvacrol in mice plasma using LC-QqQ-MS/MS.

Present work describes the development of a liquid chromatography tandem mass spectrometry-based bioanalytical method for the reliable simultaneous quantification of docetaxel (DXL) and carvacrol (CVL) in the mice plasma. A rapid and sensitive bioanalytical method was developed and optimized in mice plasma using Paclitaxel as an internal standard. Validation of the bioanalytical method was performed according to the ICH M10 guideline covering the range of 9.62-1923.08 ng/mL in the mice plasma milleu at the low, mid, and high-quality control concentrations of 28.86 ng/mL, 961.54 ng/mL, and 1346.15 ng/mL, respectively for both the analytes. Validation parameters such as accuracy, precision, carryover-test, matrix effect, and reinjection reproducibility were carried out and were found in limits. Stability studies (Benchtop, autosampler, freeze-thaw, and long-term) were performed and found to be within limits. The developed bioanalytical method was found to be suitable for the simultaneous quantification of DXL and CVL in the mice plasma.

Development and Validation of a New Eco-Friendly HPLC-PDA Bioanalytical Method for Studying Pharmacokinetics of Seliciclib

Background and Objectives: Seliciclib (SEL) is the first selective, orally bioavailable potential drug containing cyclin-dependent kinase inhibitors. Preclinical studies showed antitumor activity in a broad range of human tumor xenografts, neurodegenerative diseases, renal dysfunctions, viral infections, and chronic inflammatory disorders. To support the pharmacokinetics and aid in therapeutic monitoring of SEL following its administration for therapy, an efficient analytical tool capable of quantifying the concentrations of SEL in blood plasma is needed. In the literature, there is no existing method for quantifying SEL in plasma samples. This study introduces the first HPLC method with a photodiode array (PDA) detector for the quantitation of SEL in plasma. Materials and Methods: The chromatographic resolution of SEL and linifanib as an internal standard (IS) was achieved on Zorbax Eclipse Plus C18 HPLC column (150 mm length × 4.6 mm internal diameter, 5 µm particle size), with a mobile phase composed of acetonitrile–ammonium acetate, pH 5 (50:50, v/v) at a flow rate of 1.0 mL min−1. Both SEL and IS were detected by PDA at 230 nm. The method was validated according to the ICH guidelines for bioanalytical method validation. Results: The method exhibited linearity in concentrations ranging from 50 to 1000 ng mL−1, with a limit of quantitation of 66.1 ng mL−1. All remaining validation parameters satisfied the ICH validation criteria. The environmental sustainability of the method was verified using three extensive tools. The proposed HPLC-PDA method was effectively utilized to study the pharmacokinetics of SEL in rats after a single oral administration of 25 mg/kg. Conclusions: The proposed method stands as a valuable tool for studying SELs for pharmacokinetics in humans. It aids in achieving the targeted therapeutic advantages and safety of treatment with SEL by optimizing the SEL dosage and dosing schedule.

A Liquid Chromatography‐Tandem Mass Spectrometry Bioanalytical Method Development and Validation for the Determination of Pacritinib in Plasma

ABSTRACTThe primary objective of the current work is to establish and verify an accurate and linear liquid chromatography‐electrospray ionization‐tandem mass spectrometry procedure for quantifying Pacritinib. An effective chromatographic resolution was obtained using the Hypersil Gold (50 × 4.6 mm, 2.1 µm) C18 column. The mobile phase is a mixture of methanol, 0.1% formic acid, and acetonitrile in the proportion of 20:15:65 (%v/v/v). The procedure involved closely monitoring executed ionic transitions of m/z 473.1/376.2 for Pacritinib and 584.26/101.1 for Brigatinib internal standard in multiple reaction monitoring mode. A strong correlation value (r2) of 0.9997 is associated with the linear plot regression line, which can be represented as y = 0.0001x − 0.0008. The relative standard deviation (RSD) results for the matrix effect were 3.63% and 3.57%, respectively, when the quality control (QC) level was low and high, respectively. Over the course of the study, the percentage average recoveries for Pacritinib were found to be 103.27% in high QC, 97.59% in medium QC, and 94.28% in low QC, respectively. The values that were obtained for the QC samples (0.378, 1.058, 7.56, and 11.34 µg/mL) varied from 2.00% to 4.03%. The developed method was useful for evaluating Pacritinib in biological samples for the purposes of QC, forensics, and bioavailability investigations for individuals.

Development and Validation of a New HPLC Bio Analytical Internal Standard Method for the Analysis of Two Immunology Drugs (Lamivudine and Dolutegravir) in Human Plasma

Bioanalytical methods are useful for the quantitative analysis of drugs and their metabolites in biological fluids. Bio-analytical internal standards methods plays vital role in areas of human clinical pharmacology. The aim of this work is to development and validation of bio-analytical internal standard method for determination of two immunology drugs Lamivudine (LMVD) and Dolutegravir (DTGR) in plasma with Abacavir(ABVR) drug as internal standard (IS). Liquid-liquid extraction with Diethyl ether and methanol in the ratio of 50:50 (v/v) was used for the extraction of drugs from the biological matrix. The optimized chromatography conditions consist of Water, Acetonitrile and Methanol in the ratio of 40:25:35 (v/v) as a mobile phase with Ascentis, C18 Column (250 X 4.6 mm, 5μ) as stationery phase. Isocratic elution with 1.2 ml flow separates theLMVD at 3.5 min, DTGR at 9.8 min and ABVR at 6.8 min. The method was validated as per ICH guidelines. The relative standard deviation (%RSD) were found to be <5%for precision studies. Hence the method was found to be suitable fort the analysis of LMVD and DTGR in spiked human plasma.

Stability Indicating Bioanalytical Method Development and Validation for Estimation of Remogliflozin Etabonate by RP-HPLC in Human Plasma

Stability Indicating Bioanalytical Method Development and Validation for Estimation of Remogliflozin Etabonate by RP-HPLC in Human Plasma

Bioanalytical Method Development and Validation of MEM-PLGANanoparticles via Nasal Route Administration.

The aim of this effort is to increase extended pharmaceutical release by systematizing PN elaboration for ME, a novel treatment for Alzheimer’s disease. Bioanalysis is an essential part in drug discovery and development. Bioanalysis is related to the analysis of analytes (drugs, metabolites, biomarkers) in biological samples and it involves several steps from sample collection to sample analysis and data reporting. A simple, sensitive, and quick LC-MS technique for quantifying MEM in rat plasma was developed and validated. Chromatography was carried out on a C18 column (4.6 × 75mm, 3.5μm). ME and its internal standard, NLB, were extracted by direct protein precipitation and were chosen as the sample treatment method rather than liquid-liquid extraction techniques and evaluated with LC-MS/MS in multiple-reaction monitoring (MRM) mode. This approach demonstrated intra- and interday precision in the ranges of 2.1-3.7 and 1.4-7.8%. Furthermore, intra- and interday accuracy ranged from 95.6 to 99.8, and 95.7-99.1%, respectively. MEM and NLB showed mean recovery rates of 86.07 ± 6.87 and 80.31 ± 5.70%, respectively. The stated method was effectively applied in the bioequivalence study of MEM and evaluated its potential for treating Alzheimer’s disease via the nasal route. MEM administered via the nasal route will demonstrate improved bioavailability and efficacy in the treatment of Alzheimer’s disease compared to current therapies.

Review on Green Bioanalytical Chemistry with Sustainable Approaches in Method Development: Unveiling the Crucial Role of Sustainable Bioanalysis in the Future of Chemistry

Green bioanalytical chemistry, with its focus on incorporating sustainable practices into developing bioanalytical methods, has the potential to significantly reduce the environmental impact of laboratory activities. This approach, while maintaining the high standards required for analytical precision and accuracy, includes key strategies such as reducing hazardous chemicals, optimizing energy use, and implementing waste-reduction practices. This review highlights recent advancements in green bioanalytical chemistry, including green solvent systems, miniaturized techniques, and eco-friendly analytical platforms. We discuss the benefits, such as reduced environmental pollution and improved resource efficiency, challenges, and future perspectives of integrating green principles into bioanalytical method development, emphasizing the balance between sustainability and analytical performance.

Bioanalytical Method Development and Validation for the Estimation of Hydroxyproline in Urine Samples of Osteoarthritic Patients Using LC–MS/MS Technique

Bioanalytical Method Development and Validation for the Estimation of Hydroxyproline in Urine Samples of Osteoarthritic Patients Using LC–MS/MS Technique

Bioanalytical Method Development, Validation and Oral Pharmacokinetic Profile of Chebulinic Acid

Terminalia chebula Retz, sometimes referred to as "Haritaki/Myrobalan," has long been used in traditional medicine. It has been widely used in many traditional medical systems, including Unani, Tibb, Ayurveda, and Siddha, to treat a wide range of conditions in people, including bleeding, diarrhoea, carminatives, liver tonics, digestive problems, antidiarrheal, analgesics, anthelmintics, antibacterials, and skin disorders. Using a variety of computerized databases, research on the pharmacological effects of T.

Fast and Sensitive Bioanalytical Method Development and Validation for Determination of Capmatinib in Human Plasma Using HPLC-MS/MS

The term “anticancer drugs” evokes memories of a time when the development of medications in that category was still necessary. Capmatinib, for example, is a kinase inhibitor that targets the C-Met receptor tyrosine kinase in the treatment of non-small cell lung cancer, which involves tissue repair and organ regeneration.1 Thus, utilizing liquid chromatography-tandem mass spectroscopy (LC-MS/MS) in human plasma in accordance with United States Food and Drug Administration (USFDA) bioanalytical technique validation criteria, a quick, simple, specific, dependable, and sensitive approach was created using deucravacitinib as an internal standard. Capmatinib and the internal standard were separated using liquid-liquid extraction. The extracted sample run through a chromatographic system with an ACE-C18 column (4.6 × 100 mm, 5 μm); the mobile phase consisted of methanol and 2 mM ammonium formate in an 80:20 ratio at a flow rate of 1.00 mL/min. The system operates for three minutes in multiple reaction monitoring mode at the ABSCIEX API 4000 mass spectrometer using electron spray ionization. For capmatinib, the ion transitions are 413.10 to 382.10 and 426.30 to 358.20 for deucravacitinib, with a concentration range of 5.00 to 4000 ng/mL, the accuracy, precision, linearity, selectivity, and selectivity were validated and found to be within the acceptability limits.

Application of immunocapture and nanoflow LC-MS/MS to overcome the barriers to the quantitation of oxytocin in plasma of women in third stage labour.

Postpartum haemorrhage (PPH) is the leading cause of maternal mortality worldwide. To prevent PPH, the WHO recommends administration of oxytocin (OT) immediately after birth, i.e. during the third stage of labour (TSL). Previous studies demonstrate that methods to quantify OT in biological matrices, e.g. enzyme-linked immunosorbent assays (ELISA), radioimmunoassays (RIA) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) lack the specificity and/or sensitivity to accurately quantify OT in plasma from women administered OT during TSL. This is due to increased metabolic clearance of OT in late-stage pregnancy and at the time of childbirth, resulting in extremely low OT plasma concentrations. This study describes the development of an ultra-sensitive bioanalytical method that overcomes the issues previously reported and enables accurate pharmacokinetic analyses of exogenously administered OT in TSL. A selective and sensitive assay to quantify OT in TSL plasma was developed. Immunoprecipitation (IP) was applied to selectively extract OT from the TSL plasma, thereby generating clean extracts compatible with nanoflow LC (nLC). nLC-MS/MS was chosen for its high sensitivity and ability to differentiate between OT and potentially co-captured OT-like immunoreactive products. The presented methodology is accurate and precise, with a good linear fit between 100-10 000 fg mL-1 OT. TSL plasma samples from a clinical phase 1 study (NCT02999100) were analysed successfully, enabling OT quantification down to 100 fg mL-1. The presented IP-nLC-MS/MS method succeeded in overcoming the sensitivity challenge related to the assay of OT in TSL plasma and thereby revealing the PK profiles of OT in TSL plasma clinical study samples.

A Review on Sample Preparation for Bio analytical Method Development by HPLC

This article compares the benefits and drawbacks of each approach while reviewing recent advancements in bioanalysis sample preparation methods. It also provides an update on fundamental concepts, theories, applications, and automation opportunities. Because every biological matrix has its own special difficulties and complexity, sample preparation is thought to be the bottleneck stage in bioanalysis. In the last ten years, new sample preparation methods for bioanalysis have developed quickly. In the preclinical and clinical phases of drug development, it is crucial to create reliable bioanalytical method(s). Consequently, it is widely acknowledged that sample preparation A number of excellent review papers have recently been published in the literature that address various scientific and technical facets of bioanalysis. The use of bioanalysis in the pharmacokinetic/pharmacodynamic characterization of novel chemical entities, beginning with their finding and continuing through their market authorization, is now generally acknowledged. Physico-chemical and biological technologies used in bioanalysis, including mass spectrometry, immunoassay, and chromatography. Liquid chromatography-mass spectrometry is a method used in labs for the qualitative and quantitative study of drug substances and metabolites. It makes use of liquid chromatography/RP HPLC. The current review concentrated on different extraction methods, including protein precipitation, solid phase extraction, and liquid-liquid extraction, all of which are crucial for sample preparation and RP HPLC sample detection.

Bioanalytical Method Development and Validation and forced degradation of Sitagliptin and determination of Pharmacokinetic application study in Human Plasma by RP-HPLC method

In the present investigation, an attempt was made for the development, validation, forced degradation and pharmacokinetic application of sitagliptin in the human plasma spiking studies by UV - HPLC method. The experimentation was developed based on the extensive literature survey and ascertained by the statistical parameters of the sampling. A simplified, accurate method was created by the liquid chromatographic system from Shimadazu LC 20AD consisting of manual injection. The optimized chromatogram was obtained with acetonitrile in the isocratic mobile phase method at a 1.0 mL/min flow rate. A thermo C-8 column (4.6X250mm,5μm) was used as a stationary phase, and 265.0nm was selected as the detection wavelength with the aid of a UV-Vis detector. The proposed method was validated as per ICH guidelines. The technique was linear in the range of 10-50μg/mL with correlation coefficient R2 = 0.9746, respectively. Recovery studies postulated that % RSD 19.14, 3 & 9.95 respectively. Injection repeatability values were found to be % RSD 17 & 10.63 for intraday and interday, respectively. Stress degradation studies revealed that sitagliptin degrades more rapidly when subjected to 0.1 NaoH. Human plasma spiking studies reported 3.02 ng/mL at 3.02+/-60 min of C and T max, respectively. Keywords: Sitagliptin, Method development, HPLC, Validation, Stress degradation studies, Human plasma spiking studies

Bioanalytical Method Development and Validation of UV Spectrophotometric Method for Estimation of Metformin Hydrochloride in Urine Sample

Purpose: To develop a simple, sensitive, accurate, precise and rapid UV spectrophotometric method for the determination of metformin in human urine. Methods: Extraction of metformin from urine samples was done by Dilute and shoot method in which urine was diluted up to 1:10 ratio to avoid interference of matrix. For estimation of Metformin hydrochloride, the solvent system employed was water. The metformin hydrochloride was scanned in range of 200nm to 400 nm and wavelength of detection (λdet) was found to be 234 nm. Result: The calibration curve was linear in the range of 1-17 μg/ml. the recovery and assay studies of metformin hydrochloride were within 93-94.85% indicating that the proposed method can be estimation of metformin hydrochloride

Bioanalytical method development and validation for the quantification of raloxifene hydrochloride from mice plasma by RP-HPLC

Raloxifene hydrochloride (RLX) belongs to the Selective estrogen receptor modulator (SERM) class of drugs, exhibiting diverse affinity to estrogen receptors. The drug is known for its anti-cancer and anti-osteoporosis activity. The current study employs a Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method that is sensitive and time-efficient for quantitative evaluation of RLX from mice plasma, using tamoxifen citrate as an internal standard (IS). The protein precipitation technique was used to extract the RLX and IS. RLX was separated using a C8 column with an isocratic elution of a mobile phase consisting of acetonitrile and Milli-Q water (pH 3) at a 1 mL/min flow rate. The ultraviolet (UV) detection of the analyte and IS were carried out at 287 nm. The linearity plot was constructed in the range of 100–3200 ng/mL with a correlation coefficient of r2 ≥ 0.9996. The inter-day and intra-day accuracy and precision were found to be within acceptable limits. Stability studies revealed that the analyte remained stable. The validated RP-HPLC method was successfully applied to quantify RLX after oral administration in mice, and the pharmacokinetic (PK) parameters were established. The present RP-HPLC method could be a beneficial tool for the fast and reliable estimation of future PK interactions, PK analysis of formulations, and therapeutic drug monitoring.

Bioanalytical method development and validation for quantification of amivantamab in rat plasma by LC-MS/MS

BackgroundAmivantamab is a monoclonal bispecific anti-EGFR-MET antibody used to treat non-small cell lung cancer. There were no published methods using a liquid chromatographic—tandem mass spectrometric approach to develop and validate a feasible, novel, and thoroughly validated method for quantifying amivantamab in rat plasma.ResultsThe liquid–liquid extraction method was used to extract the analyte from rat plasma. The analyte was separated using acetonitrile–ammonium formate buffer (40:60) as a mobile phase on waters, alliance e-2695 model high-pressure liquid chromatographic system having Agilent eclipse C18, 150 mm × 4.6 mm, 3.5 µm column. The overall runtime was 6 min at a 1.0 ml/min flow rate. The method showed significant sensitivity and acceptable linearity over the 5.00–100.00 ng/ml concentration range. Accuracy was proved by mean percent recovery ranging from 98.03 to 99.99%. The intraday precision coefficient of variation (%) ranged between 0.31 and 5.43. Also, the findings such as Cmax, tmax, AUC0− t, AUC0− ∞, and half-life values of amivantamab showed that the technique was helpful for pharmacokinetic studies.ConclusionsAll the validated parameters were found to be within the acceptable range. The validated method was found to be simple, accurate, precise, and reproducible and hence can be used for the routine analysis of amivantamab, such as in-process quality control by liquid chromatographic—tandem mass spectrometry.

Review on bioanalytical and analytical method development of two potassium-sparing diuretics, namely eplerenone and spironolactone

Review on bioanalytical and analytical method development of two potassium-sparing diuretics, namely eplerenone and spironolactone

Molnupiravir Bioanalytical Method Development and Validation in Rat Plasma by LC-MS/MS Detection and Application to a Pharmacokinetic

Molnupiravir Bioanalytical Method Development and Validation in Rat Plasma by LC-MS/MS Detection and Application to a Pharmacokinetic

Bioanalytical Method Development and Validation of UV Spectrophotometric Method for Estimation of Sitagliptin in Urine sample

Bioanalytical Method Development and Validation of UV Spectrophotometric Method for Estimation of Sitagliptin in Urine sample

A RP-HPLC approach to Bioanalytical Method Development and Validation: A Review

A RP-HPLC approach to Bioanalytical Method Development and Validation: A Review