A HIGHLY SENSITIVE POLYCLONAL ANTIBODY-BASED ELISA FOR THERAPEUTIC MONITORING AND PHARMACOKINETIC STUDIES OF LENALIDOMIDE

Aim: To support pharmacokinetic studies and therapeutic monitoring of darunavir (DRV), a highly sensitive ELISA was developed for the determination of DRV in plasma samples at picogram levels. Results: The assay LOD and LOQ were 15 and 30 pg ml-1, respectively. The working range of the assay was 20–2000 pg ml-1. Analytical recoveries of DRV from spiked plasma were in the ranges of 98.4–113.0 and 86.0–99.1% for intra-assay and inter-assay runs, respectively. The precision of the assay was satisfactory. Conclusion: The ELISA is characterized by high throughput and it is expected to significantly contribute to routine analysis of DRV in its pharmacokinetic studies and therapeutic monitoring.

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A Novel Simultaneous Estimation of Sofosbuvir and Velpatasvir in Human Plasma by Liquid Chromatography Tandem-Mass Spectrometry after Protein Precipitation Method

Current Pharmaceutical Analysis ◽

10.2174/1573412914666180910102353 ◽

2019 ◽

Vol 15 (7) ◽

pp. 710-715

Author(s):

S.T. Narenderan ◽

Basuvan Babu ◽

T. Gokul ◽

Subramania Nainar Meyyanathan

Keyword(s):

Mass Spectrometry ◽

Tandem Mass Spectrometry ◽

Human Plasma ◽

Multiple Reaction Monitoring ◽

Simultaneous Estimation ◽

Pharmacokinetic Studies ◽

Tandem Mass ◽

Mass Spectrometric Method ◽

Highly Sensitive ◽

Tandem Mass Spectrometric

Objective: The aim of the present work is to achieve a novel highly sensitive chromatographic method for the simultaneous determination of hepatitis C agents, sofosbuvir and velpatasvir from human plasma using ritonavir as an internal standard. Methods: Chromatographic separation was achieved using Hypersil C18 column (50mm x 4.6mm, 3μm) with an isocratic elution mode using the mobile phase composition 10 mM ammonium formate buffer (pH 5.0): acetonitrile (20:80 v/v) pumped at a flow rate of 0.5 ml/min. The detection was carried out by tandem mass spectrometry using Multiple Reaction Monitoring (MRM) positive Electrospray Ionization (ESI) with proton adducts at m/z 530.10 > 243.10, 883.40 > 114.0 and 721.25 > 197.0. Results: The method validated as per USFDA guidelines with respect to linearity, accuracy, and precision was found to be acceptable over the concentration range of 0.2–2000 ng/ml and 5-2000 ng/ml for sofosbuvir and velpatasvir respectively and the method was found to be highly sensitive and selective. Conclusion: The developed tandem mass spectrometric method is robust and can be applied for the monitoring of plasma levels of the analyzed drug in preclinical and clinical pharmacokinetic studies.

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Highly Sensitive LC–MS-MS Method for the Determination of Tacrine in Rat Plasma: Application to Pharmacokinetic Studies in Rats

Journal of Chromatographic Science ◽

10.1093/chromsci/bmv155 ◽

2015 ◽

pp. bmv155

Author(s):

Suresh Ponnayyan Sulochana ◽

Vishnuvardh Ravichandiran ◽

Ramesh Mullangi ◽

Sathesh Kumar Sukumaran

Keyword(s):

Rat Plasma ◽

Pharmacokinetic Studies ◽

Highly Sensitive

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Bio analytical method development and validation for Ezetimibe and Pitavastain and its applications to pharmacokinetic studies in Rabbit plasma by using LCMS/MS

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v11i4.4670 ◽

2020 ◽

Vol 11 (4) ◽

pp. 7854-7862

Author(s):

Potturi Ramadevi ◽

Kantipudi Rambabu

Keyword(s):

Method Development ◽

Extraction Process ◽

Pharmacokinetic Studies ◽

Rabbit Plasma ◽

Highly Sensitive ◽

The Us ◽

Positive Mode ◽

Method Development And Validation ◽

Development And Validation ◽

Preparation Techniques

For the gradation of Ezetimibe and Pitavastain in rabbit plasma, a highly sensitive and simple LC-MS/MS assay was developed and witnessed. The chromatographic conditions are isocratic with a waters symmetry C18 (150 x 4.6 mm, 3.5) column in isocratic mode. The detection was carried out using a mobile phase of 0.1 percent formic acid and 60:40 acetonitrile, and the detection was carried out using MS in a positive mode of electrospray ionisation. The valid approach was checked with a linear range of 10-200 ng/ml Ezetimibe and 2-40 ng/ml Pitavastain. The intraday and interday precision values were found to be within reasonable limits. The liquid extraction process is used to remove these drugs from rabbit plasma. And these drugs have been shown to be stable in freeze-thaw, autosampler, and benchtop tests in the future. The fluid chromatography coupled mass spectrometry strategy was approved by the US Food and Drug Administration for quantification of Ezetimibe and Pitavastain in rabbit plasma using D4–ezetimibe and D4–pitavastain as within norms using LC-MS consolidated with quadrupole spectrometer by electro shower ionisation process. The aim of this study is to evaluate the applicability of this approach to ezetimibe and pitavastain at different evaluation levels while taking into account various factors such as instrument stability, precision, and accuracy, sample preparation techniques, instrument calibration, recovery, and matrix effect by using Ezetimibe and Pitavastain, as well as their internal guidelines.

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Therapeutic monitoring of cyclosporine by using pharmacokinetic studies

Clinical Chemistry ◽

10.1093/clinchem/37.11.1891 ◽

1991 ◽

Vol 37 (11) ◽

pp. 1891-1892 ◽

Cited By ~ 1

Author(s):

W M Awni ◽

K L Heim-Duthoy ◽

B L Kasiske

Keyword(s):

Therapeutic Monitoring ◽

Pharmacokinetic Studies

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DEVELOPMENT AND VALIDATION OF LC-MS/MS METHOD FOR ESTIMATION OF UROCARB IN HUMAN PLASMA

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2019v11i5.33873 ◽

2019 ◽

pp. 125-130

Author(s):

IRYNA DRAPAK ◽

BORYS ZIMENKOVSKY ◽

LINA PEREKHODA ◽

SERGIY KOVALENKO ◽

LILIYA LOGOYDA

Keyword(s):

Formic Acid ◽

Human Plasma ◽

Internal Standard ◽

Sample Volume ◽

Pharmacokinetic Studies ◽

Coefficients Of Variation ◽

Highly Sensitive ◽

Regulatory Guidelines ◽

Initial Content ◽

Development And Validation

Objective: The present study was aimed to develop a rapid, specific and sensitive method based on LC-MS/MS method was developed for the determination of urocarb using etomidate as an internal standard. Methods: Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile-water–formic acid, 5: 95: 0.1 v/v), eluent B (acetonitrile–formic acid, 100: 0.1 v/v)). The initial content of the eluent B of 8%, which increases linearly to 1.0 min to 100%, is maintained up to 1.5 min and returned to the original 8% to 1.51 min. The mobile phase was delivered at a flow rate of 0.400 ml/min into the mass spectrometer ESI chamber. The sample volume was 4 μl. Results: The total chromatographic run time was 2.0 min and the elution of urocarb and IS (etomidate) occurred at ~1.53 and 1.67 min, respectively. A linear response function was established at 1-100 ng/ml for urocarb and etomidate in human plasma. The % mean recovery for urocarb in LQC, MQC and HQC was 104.1 %, 100.0 % and 97.4 %. The lowest concentration with the RSD<20% was taken as LLOQ and was found to be 1.03 ng/ml for urocarb. The within-run coefficients of variation ranged between 0.271 % and 0.478 % for urocarb. The within-run percentages of nominal concentrations ranged between 99.12 % and 100.21 % for urocarb. The between-run coefficients of variation ranged between 0.388 % and 0.601 % for urocarb. The between-run percentages of nominal concentrations ranged between 98.78 % and 101.11 % for urocarb. Conclusion: A highly sensitive, specific, reproducible, rapid and high-throughput LC-MS/MS assay was developed and validated to quantify urocarb in human plasma as per the regulatory guidelines. Due to the sensitivity of the developed method, it can be applied to the monitoring of plasma levels in the analysis of drug in preclinical and clinical pharmacokinetic studies. All the parameters and results were found within the acceptance limit as given in the validation protocol.

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