Development and evaluation of a multiple-plate fraction collector for sample processing: Application to radioprofiling in drug metabolism studies

Barros, Anthony; Ly, Van T.; Chando, Theodore J.; Ruan, Qian; Donenfeld, Scott L.; Holub, David P.; Christopher, Lisa J.

doi:10.1016/j.jpba.2010.11.029

Cited by 6 publications

(4 citation statements)

References 15 publications

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“…Liquid scintillation counting (LSC) has been routinely used to analyze the total radioactivity of samples. HPLC with online radioactivity detection or fractionation with off-line radioactivity counting have been used for metabolite radio-profiling. , With these radiometric detection techniques, administration of radioactive doses in the range of 50 to 200 μCi (1.85 to 7.4 MBq) or higher generally provides the best opportunity to achieve adequate sensitivity for the plasma assessments. Over the last two decades, accelerator mass spectrometry (AMS) has been utilized in an increasing number of ADME studies to measure 14 C for PK of total DRM, to determine mass balance, and to generate 14 C-metabolite profiles.…”

Section: Other Scenariosmentioning

confidence: 99%

The Importance of Tracking “Missing” Metabolites: How and Why?

Wang,

Ballard,

Christopher

et al. 2023

J. Med. Chem.

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Technologies currently employed to find and identify drug metabolites in complex biological matrices generally yield results that offer a comprehensive picture of the drug metabolite profile. However, drug metabolites can be missed or are captured only late in the drug development process. This could be due to a variety of factors, such as metabolism that results in partial loss of the molecule, covalent bonding to macromolecules, the drug being metabolized in specific human tissues, or poor ionization in a mass spectrometer. These scenarios often draw a great deal of attention from chemistry, safety assessment, and pharmacology. This review will summarize scenarios of missing metabolites, why they are missing, and associated uncovering strategies from deeper investigations. Uncovering previously missed metabolites can have ramifications in drug development with toxicological and pharmacological consequences, and knowledge of these can help in the design of new drugs. ■ SIGNIFICANCEMetabolism plays an important role in mediating drug exposure and potential toxicity, and ultimate approval. Nevertheless, identifying metabolic pathways and weighing their relevance can be challenging. This Perspective highlights a myriad of real-world situations where a complete accounting of drug metabolites is achieved only after deeper investigations were made. This review should serve as a new valuable resource, not only to drug metabolism and medicinal chemists but also to toxicologists, regulators, and others involved in drug discovery and early development.

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Section: Other Scenariosmentioning

confidence: 99%

The Importance of Tracking “Missing” Metabolites: How and Why?

Wang,

Ballard,

Christopher

et al. 2023

J. Med. Chem.

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“…Recently, a multi-microplate fraction collector has been evaluated which made it possible to collect fractions in up to 24 96-or 384-well plates without manual intervention. Combined with UPLC this system provided high-resolution radiochromatograms with minimum manual input [63]. Online radioflow detection (HPLC-RFD) also requires minimal manual input, but suffers from poor sensitivity due to the short residence times of the radioactive peaks in the detection cell [44].…”

Section: Review | Dubbelman Rosing Schellens and Beijnenmentioning

confidence: 99%

“…This direction maintains the conventional radioactive doses of around 100 µCi, but uses high-resolution and high-throughput techniques, such as the multi-microplate fraction collector combined with microplate scintillation counting for metabolite profiling [63], to reduce the labor intensity of a mass balance study. Regarding the detection of unknown metabolites, data mining methods that can be applied to mass spectrometry data have high potency, especially in combination with high-resolution mass data.…”

Section: Future Perspectivementioning

confidence: 99%

Bioanalytical Aspects of Clinical Mass Balance Studies in Oncology

Dubbelman¹,

Rosing

Schellens

et al. 2011

Bioanalysis

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Clinical mass balance studies aim to investigate the absorption, distribution, metabolism and excretion (ADME) of a(n) (often radiolabeled) drug, following a single administration to humans. They are perfectly suited to determine the disposition and major metabolic pathways of a drug, the exposure to the parent drug and its metabolites, and the rate and route of elimination. A mass balance study, however, poses interesting challenges to the analysis of parent drug and metabolites in different biological matrices. Using recent clinical mass balance studies in oncology as an example, this review focuses on the aspects of mass balance studies, from bioanalytical assay development, analysis of clinical samples to reporting of study results. Along the way, it discusses bioanalytical problems and practical solutions.

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“…Because of the large number of wells per plate, it is widely used for combinatorial chemistry and high throughput screening. Coupled with various chemical analysis techniques, such as ultraviolet–visible spectroscopy, fluorescence spectroscopy, liquid chromatography (LC), capillary electrophoresis (CE), and mass spectrometry (MS), it is extensively used for enzyme-linked immunosorbent assays (ELISA), enzyme activity or inhibition assays, , and drug metabolism studies. , Compared to spectroscopy techniques, MS can offer direct qualification and quantification of a wide range of analytes, providing accurate and specific characterization to avoid false positive or negative results, and thereby is highly suitable for the studies of enzymatic assays, inhibitor screening assays, and drug metabolism assays. − …”

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confidence: 99%

Electrostatic Spray Ionization from 384-Well Microtiter Plates for Mass Spectrometry Analysis-Based Enzyme Assay and Drug Metabolism Screening

et al. 2017

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We have realized the direct ionization of samples from wells of microtiter plates under atmospheric conditions for mass spectrometry analysis without any liquid delivery system or any additional interface. The microtiter plate is a commercially available 384-well plate without any modification, working as a container and an emitter for electrostatic spray ionization of analytes. The approach provides high throughput for the large batches of reactions and both the qualitative and quantitative analysis of a single compound or mixture. The limits of detection in small drug molecules, peptides, and proteins are similar in comparison with standard direct infusion electrospray ionization. The analysis time per well is only seconds. These analytical merits benefit many microtiter plate-based studies, such as combinatorial chemistry and high throughput screening in enzyme assay or drug metabolism. Herein, we illustrate the application in enzyme assay using tyrosine oxidation catalyzed by tyrosinase in the presence or absence of inhibitors. The potential application in drug development is also demonstrated with cytochrome P450-catalyzed metabolic reactions of two drugs in microtiter plates followed with direct ESTASI-MS/MS-based characterization of the metabolism products.

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Development and evaluation of a multiple-plate fraction collector for sample processing: Application to radioprofiling in drug metabolism studies

Cited by 6 publications

References 15 publications

The Importance of Tracking “Missing” Metabolites: How and Why?

The Importance of Tracking “Missing” Metabolites: How and Why?

Bioanalytical Aspects of Clinical Mass Balance Studies in Oncology

Electrostatic Spray Ionization from 384-Well Microtiter Plates for Mass Spectrometry Analysis-Based Enzyme Assay and Drug Metabolism Screening

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