Automated online liquid chromatographic/mass spectrometric metabolic study for prodrug stability

Lai, Fran; Khojasteh-Bakht, Siamak C.

doi:10.1016/j.jchromb.2004.10.040

Cited by 6 publications

(3 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In 2005, a method using similar instrumentation was published. Lai et al [9] used an automated online extraction system to study pro-drug stability. The autosampler was modified and included a heated agitator for sample incubation.…”

Section: Discussionmentioning

confidence: 99%

“…Similar approaches were presented by Xu et al [7] and by Jenkins et al [8], which also used a liquid handling robot to carry out the HLM experiments. Lai et al [9] described a system where the autosampler was modified to include a heated agitator and the samples were subsequently injected into the online extraction system. The system was used to screen the stability of a prodrug in development.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Generation of metabolites by an automated online metabolism method using human liver microsomes with subsequent identification by LC-MS(n), and metabolism of 11 cathinones

Müller

Rentsch

2012

Anal Bioanal Chem

View full text Add to dashboard Cite

Human liver microsomes (HLMs) are used to simulate human xenobiotic metabolism in vitro. In forensic and clinical toxicology, HLMs are popularly used to study the metabolism of new designer drugs for example. In this work, we present an automated online extraction system we developed for HLM experiments, which was compared to a classical offline approach. Furthermore, we present studies on the metabolism of 11 cathinones; for eight of these, the metabolism has not previously been reported. Metabolites were identified based on MS(2) and MS(3) scans. Fifty-three substances encompassing various classes of drugs were employed to compare the established offline and the new online methods. The metabolism of each of the following 11 cathinones was studied using the new method: 3,4-methylenedioxy-N-benzylcathinone, benzedrone, butylone, dimethylcathinone, ethylone, flephedrone, methedrone, methylone, methylethylcathinone, naphyrone, and pentylone. The agreement between the offline and the online methods was good; a total of 158 metabolites were identified. Using only the offline method, 156 (98.7%) metabolites were identified, while 151 (95.6%) were identified using only the online method. The metabolic pathways identified for the 11 cathinones included the reduction of the keto group, desalkylation, hydroxylation, and desmethylenation in cathinones containing a methylenedioxy moiety. Our method provides a straightforward approach to identifying metabolites which can then be added to the library utilized by our clinical toxicological screening method. The performance of our method compares well with that of an established offline HLM procedure, but is as automated as possible.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Generation of metabolites by an automated online metabolism method using human liver microsomes with subsequent identification by LC-MS(n), and metabolism of 11 cathinones

Müller

Rentsch

2012

Anal Bioanal Chem

View full text Add to dashboard Cite

show abstract

“…However, many labs do have automated HPLC and LC-MS systems, such as those used for drug development studies, combinatorial synthesis monitoring, and bioanalysis. [22][23][24][25][26][27][28][29] So far, there are no reports of using an HPLC autosampler for online sample addition, buffer addition, incubation, and injection or to quantitate the stability of drug candidates.…”

Section: Introductionmentioning

confidence: 99%

Development and Application of an Automated Solution Stability Assay for Drug Discovery

Di¹,

Kerns²,

Hong³

et al. 2006

SLAS Discovery

View full text Add to dashboard Cite

Screening of solution stability provides an early alert on potential liabilities of drug candidates so that strategies can be developed to overcome the challenges. A fully automated solution stability assay has been developed to accelerate traditional manual operation. The assay uses the advanced capabilities of a high-performance liquid chromatography instrument that is present in many pharmaceutical research laboratories. The samples are prepared automatically by a temperature-controlled autosampler. The samples are delivered to the stability matrices, mixed, incubated, and injected at selected time points during the reaction time course. This automated process occurs without operator intervention, thus allowing 96 experiments to be run with 0.5 h of a scientist's time compared to 8 h for the same study when performed manually. Automation not only eliminates the manual operation but also improves accuracy and throughput. The assay protocol has been optimized to achieve homogenous mixing and eliminate carryover. The assay is robust, flexible, and high throughput. It can be used to study stability for a large number of samples under multiple incubation conditions and has a wide range of applications in drug discovery and development, such as screening compound stability in biological assay media, obtaining a stability-pH profile, surveying compound stability in physiological fluids, and performing development forced degradation and excipient compatibility. (Journal of Biomolecular Screening 2006:40-47)

show abstract

In VivoMetabolism in Preclinical Drug Development

Rollas

2010

Pharmaceutical Sciences Encyclopedia

View full text Add to dashboard Cite

In preclinical drug discovery, investigation of the in vitro, in vivo metabolism and toxicity screening of a drug candidate together play a very important role in the drug's success before entering into clinical use. Several in vitro models, such as isolated perfused livers, liver tissue slices, freshly isolated hepatocytes in suspension, primary hepatocyte cultures, cell lines, microsomes, mitochondria, and expression systems have been developed to profile human metabolism. This article focuses on the preclinical in vivo metabolism of drug candidates and prodrugs.

show abstract

Automated online liquid chromatographic/mass spectrometric metabolic study for prodrug stability

Cited by 6 publications

References 27 publications

Generation of metabolites by an automated online metabolism method using human liver microsomes with subsequent identification by LC-MS(n), and metabolism of 11 cathinones

Generation of metabolites by an automated online metabolism method using human liver microsomes with subsequent identification by LC-MS(n), and metabolism of 11 cathinones

Development and Application of an Automated Solution Stability Assay for Drug Discovery

In VivoMetabolism in Preclinical Drug Development

Contact Info

Product

Resources

About