Liquid chromatography-tandem mass spectrometric quantitative determination of the HIV protease inhibitor atazanavir (BMS-232632) in human peripheral blood mononuclear cells (PBMC): practical approaches to PBMC preparation and PBMC assay design for high-throughput analysis

Jemal, Mohammed; Rao, Satish; Gatz, Michael; Whigan, Daisy B.

doi:10.1016/s1570-0232(03)00589-0

Cited by 39 publications

(41 citation statements)

References 11 publications

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“…The polyclonal anti-ATV antibodies raised in rabbits are specific for ATV since no interference due to other anti-HIV drugs (crossreactivities, Ͻ0.01%) or endogenous plasma or cell compounds was recorded. The present assay was also highly sensitive, with an LLOQ of 150 pg per ml, which compares favorably with those close to 50 ng/ml (5,7,18,25) and above 1 ng/ml (8,11,21) previously reported for the LC-UV and LC-MS-MS methods, respectively. Attempts to obtain direct measurements of the concentrations in plasma were not successful probably due to the binding of the drug to plasma proteins, in particular, ␣-1-acid glycoprotein and albumin (levels of binding, 89% and 86%, respectively) (Reyataz, Summary of product characteristic, Bristol-Myers Squibb, 2004).…”

Section: Discussionsupporting

confidence: 82%

“…Several high-performance liquid chromatographic (HPLC) assays combined with UV detection (6, 7, 18, 25) or liquid chromatography with tandem mass spectrometry (LC-MS-MS) (5,8,11,21) have been described for the quantitative determination of ATV in plasma. Only a few of these assays have been validated for use for the measurement of intracellular concentrations (5,11), and all of them involve the use of LC-MS-MS; but LC-MS-MS systems are not available in all routine laboratories that perform therapeutic drug monitoring and require expensive equipment.…”

Section: Atv Concentrations Were Measured In Peripheral Blood Mononucmentioning

confidence: 99%

“…Only a few of these assays have been validated for use for the measurement of intracellular concentrations (5,11), and all of them involve the use of LC-MS-MS; but LC-MS-MS systems are not available in all routine laboratories that perform therapeutic drug monitoring and require expensive equipment. However, no immunoassay with a sensitivity, rapidity, and cost-effectiveness superior to those of LC-MS-MS has been published to date.…”

Section: Atv Concentrations Were Measured In Peripheral Blood Mononucmentioning

confidence: 99%

See 2 more Smart Citations

Quantitative Immunoassay To Measure Plasma and Intracellular Atazanavir Levels: Analysis of Drug Accumulation in Cultured T Cells

Roucairol

Azoulay

Nevers

et al. 2007

Antimicrob Agents Chemother

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We have developed an enzyme immunoassay to measure atazanavir (ATV) levels in plasma and cells. Anti-ATV polyclonal antibodies were raised in rabbits by using a synthetic ATV derivative coupled to bovine serum albumin as the immunogen, and the enzyme tracer was prepared by chemically coupling the ATV derivative with acetylcholinesterase. These reagents were used to develop a sensitive competitive enzyme immunoassay performed in microtitration plates, and the lowest limit of quantification was 150 pg/ml, which is about 10 times more sensitive than previously published techniques. The plasma assay was performed, after a simple methanol extraction, with a minimum of 30 l of plasma. This assay showed good precision and efficiency, since the rates of recovery from human plasma and cell extracts spiked with ATV ranged form 93 to 113%, with coefficients of variation of less than 10%. ATV concentrations were measured in peripheral blood mononuclear cells incubated with various ATV concentrations and in CEM cells in the absence or presence of antiretroviral drugs and drug transporter inhibitors. The results indicated a dose-dependent uptake (intracellular concentration/extracellular concentration ratio range, 0.04 to 19). A significant increase in the accumulation of ATV was noticed in the presence of P-glycoprotein and MRP1 inhibitors (dipyridamole, inter alia).Interestingly, efavirenz significantly increased the baseline accumulation of ATV, whereas nevirapine induced a marked reduction. This new enzyme immunoassay for measuring plasma and intracellular ATV levels was fully validated and provides an inexpensive and useful tool for routine therapeutic drug monitoring. Moreover, in vitro results suggested the implication of drug transporters and interactions with other antiviral drugs that should be further explored in human immunodeficiency virus-infected patients.

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Section: Discussionsupporting

confidence: 82%

Section: Atv Concentrations Were Measured In Peripheral Blood Mononucmentioning

confidence: 99%

Section: Atv Concentrations Were Measured In Peripheral Blood Mononucmentioning

confidence: 99%

See 1 more Smart Citation

Quantitative Immunoassay To Measure Plasma and Intracellular Atazanavir Levels: Analysis of Drug Accumulation in Cultured T Cells

Roucairol

Azoulay

Nevers

et al. 2007

Antimicrob Agents Chemother

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“…In a majority of cases, early stage colon cancer can be treated effectively with surgery or radiotherapy. However, approximately half of patients eventually develop metastatic colon cancer, which, without effective treatment, can be fatal [2]. An alteration in susceptibility to programmed cell death is a hallmark of cancer cells and contributes to tumor development and enhances resistance to conventional anti-cancer therapies, such as radiation and cytotoxic agents.…”

Section: Introductionmentioning

confidence: 99%

P38 MAP kinase functions as a switch in MS-275-induced reactive oxygen species-dependent autophagy and apoptosis in Human colon Cancer cells

Yao

Gong

Chen

et al. 2012

Free Radical Biology and Medicine

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“…Huang et al used LC-UV to detect the down-field matrix peaks that caused ion suppression for the analyte of interest and modified the chromatographic condition [113]. Another approach for assessing consistent matrix effect among the individual matrix lots is to measure the consistency for the results obtained from sample from individual matrix lot (typically n > 6) [114][115][116][117][118][119][120][121][122][123]. One could argue that as long as the results (both back-calculated concentration and MS response) are consistent among the tested lots, matrix effects would not likely contribute negatively to the quantitation.…”

Section: Matrix Effects and Recoverymentioning

confidence: 99%

Critical Review of Development, Validation, and Transfer for High Throughput Bioanalytical LC-MS/MS Methods

Zhou¹,

Song²,

Tang³

et al. 2005

CPA

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Swift growth in the use of LC-MS/MS for the analysis of drugs in biological matrices has been compelled by the need for timely and high-quality data at many stages in drug discovery and development process: from high throughput screening of drug candidates and rapid data generation for pre-clinical studies to almost 'real-time' analysis of clinical samples. Prompt and rational method development, validation, and transfer play a pivotal role in achieving the goals of "faster, better, and cheaper" for pharmacokinetic studies since this could easily account for more than 50% of the time and labor resources for a moderate-sized project. Strategy for rational method development, validation and transfer has been largely kept as institutional knowledge but rarely appeared in literature. In this review article, strategies for developing and validating robust high throughput LC-MS/MS methods will be critically reviewed and discussed. Automated sample preparation, fast chromatography, minimization of matrix effects, and strategy of narrowing the gap between validation and incurred sample analysis are just a few topics covered in this review. Other interesting approaches for improving method efficiency and ruggedness such as direct injection SPE and liquid/liquid extracts as well as multiplexing of LC columns will also be discussed. Potential pitfalls during method development and validation are pointed out. At the end, the question "how fast is fast enough and how fast is too fast?" will be answered after considering all aspects of the method development and validation.

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Liquid chromatography-tandem mass spectrometric quantitative determination of the HIV protease inhibitor atazanavir (BMS-232632) in human peripheral blood mononuclear cells (PBMC): practical approaches to PBMC preparation and PBMC assay design for high-throughput analysis

Cited by 39 publications

References 11 publications

Quantitative Immunoassay To Measure Plasma and Intracellular Atazanavir Levels: Analysis of Drug Accumulation in Cultured T Cells

Quantitative Immunoassay To Measure Plasma and Intracellular Atazanavir Levels: Analysis of Drug Accumulation in Cultured T Cells

P38 MAP kinase functions as a switch in MS-275-induced reactive oxygen species-dependent autophagy and apoptosis in Human colon Cancer cells

Critical Review of Development, Validation, and Transfer for High Throughput Bioanalytical LC-MS/MS Methods

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