Development of a sensitive bioanalytical method for determination of PNU-83757 in rat, monkey and human plasma: from LC-UV to LC-MS/MS

The aim of the present study is to develop and demonstrate the correlation between in vitro and in vivo Plasma Protein Binding (PPB) using the ultracentrifugation method for its validation by using marketed compounds like atenolol, theophylline and phenytoin. In this study, in vitro PPB is carried out using ultracentrifugation, by spiking the selected marketed compounds at concentrations of 5 and 15 μM in plasma. In an in vivo study, rats (n = 3) were given a single oral dose (10 mg kg(-1)) and post-dose samples were subjected to ultracentrifugation to obtain the protein-free fraction. A rapid and highly sensitive method was developed and validated for determining the free fraction of marketed compounds in rat plasma using protein precipitation and analysis using an ultra performance liquid chromatography electrospray ionization (ESI) tandem mass spectrometer system (UPLC-MS/MS). The in vitro free fraction (fup) values were 0.93 ± 0.07 for atenolol, 0.31 ± 0.03 for theophylline and 0.09 ± 0.02 for phenytoin which correlated well with the corresponding in vivo values of 0.91 ± 0.03 for atenolol, 0.25 ± 0.02 for theophylline and 0.09 ± 0.01 for phenytoin with a coefficient of variation less than 11.06%, 11.45% and 13.67%, respectively. Therefore the validated high-throughput in vitro PPB study is expected to have a powerful impact on reducing the cost as well as time in the drug discovery process.

show abstract

“…17 The precision was determined by one-way ANOVA as within and between %RSD. 18 The accuracy was expressed as %bias:…”

Section: Optimization and Validation Of Lc-ms/ms Parameters For Marke...mentioning

confidence: 99%

Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

Srikanth

Chaira

Sampathi

et al. 2013

Analyst

View full text Add to dashboard Cite

show abstract

“…The criteria for acceptability of the data included accuracy (inter‐day and intra‐day) within ±15% standard deviation (SD) from nominal values and precision (inter‐day and intra‐day) of within ±15% relative standard deviation (RSD), except for LLOQ, where it should not exceed ±20% of accuracy and as well as precision (FDA., 2001). The precision was determined by one‐way ANOVA as within and between %RSD (Williams et al ., 2002). The accuracy was expressed as % bias: …”

Section: Methodsmentioning

confidence: 99%

Bioanalytical method development and validation of novel antithrombotic agent S002‐333 by LC‐MS/MS and its application to pharmacokinetic studies

Bhatta¹,

Kumar²,

Chhonker³

et al. 2010

Biomedical Chromatography

View full text Add to dashboard Cite

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method using an electrospray ionization (ECI) source for the quantification of novel anti-thrombotic agent S002-333 [2-(4-methoxy-benzenesulfonyl)-2,3,4,9-tetrahydro-1H-β-carboxylic acid amide] in rabbit plasma was developed and validated. The extraction from plasma was carried out by simple protein precipitation extraction method. The chromatographic separation was performed on an Ultramex Cyno, (150 x 4.6 mm, 5 µm) with a guard column, using acetonitrile-water (75:25,v/v) with flow rate of 0.6 mL/min as the mobile phase. The tandem mass spectrometer was tuned in the multiple reaction monitoring mode to monitor the m/z transitions 386.4/215.4 for S002-333 and m/z 393.4/171 for the internal standard dexamethasone, using positive ion mode. The MS/MS response was linear over the concentration range from 1.56 to 200 ng/mL, with a lower limit of detection of 0.78 ng/mL. The accuracy and precision of the method were within the acceptable limit of ±20% at the lower limit of quantitation and ±15% at other concentrations and showed no significant matrix effect. The validated method can be used in most or all stages of the screening and optimizing process for future method validation of pharmacokinetic studies.

show abstract

“…Often, when the molecule advances to early drug development, a labeled compound of the given NCE (+2 to +4 differential mass units) is typically synthesized for an IS. This approach has been used for PNU-83757 (Williams et al, 2002), whereas the authors have switched from an LC-UV method to a more sensitive and reliable LC-MS/MS method. In order to accommodate this approach, a deuteriated internal standard, [2H(4)] PNU-83757 has been used.…”

Section: Introductionmentioning

confidence: 99%

Simple method for the determination of rosiglitazone in human plasma using a commercially available internal standard

Mamidi

Benjamin

Mullangi

et al. 2003

Biomedical Chromatography

View full text Add to dashboard Cite

To the best of our knowledge, bioanalytical methods to determine rosiglitazone in human plasma reported in literature use internal standards that are not commercially available. Our purpose was to develop a simple method for the determination of rosiglitazone in plasma employing a commercially available internal standard (IS). After the addition of celecoxib (IS), plasma (0.25 mL) samples were extracted into ethyl acetate. The residue after evaporation of the organic layer was dissolved in 750 microL of mobile phase and 50 microL was injected on to HPLC. The separation was achieved using a Hichrom KR 100, 250 x 4.6 mm C(18) with a mobile phase composition potassium dihydrogen phosphate buffer (0.01 m, pH 6.5):acetonitrile:methanol (40:50:10, v/v/v). The flow-rate of the mobile phase was set at 1 mL/min. The column eluate was monitored by fluorescence detector set at an excitation wavelength of 247 nm and emission wavelength of 367 nm. Linear relationships (r(2) > 0.99) were observed between the peak area ratio rosiglitazone to IS vs rosiglitazone concentrations across the concentration range 5-1000 ng/mL. The intra-run precision (%RSD) and accuracy (%Dev) in the measurement of rosiglitazone were <+/-10.69 and <-12.35%, respectively across the QC levels (50-1000 ng/mL). The extraction efficiency was >80% for both rosiglitazone and IS from human plasma. The lower limit of quantitation of the assay was 5 ng/mL. In summary, the methodology for rosiglitazone measurement in plasma was simple, sensitive and employed a commercially available IS.

show abstract

Development of a sensitive bioanalytical method for determination of PNU-83757 in rat, monkey and human plasma: from LC-UV to LC-MS/MS

Cited by 6 publications

References 13 publications

Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

Correlation of in vitro and in vivo plasma protein binding using ultracentrifugation and UPLC-tandem mass spectrometry

Bioanalytical method development and validation of novel antithrombotic agent S002‐333 by LC‐MS/MS and its application to pharmacokinetic studies

Simple method for the determination of rosiglitazone in human plasma using a commercially available internal standard

Contact Info

Product

Resources

About