A rapid and sensitive assay to quantify valproyl 1-O-acyl glucuronide in supernatants of sandwich-cultured rat hepatocytes using ultra-high performance liquid chromatography–tandem mass spectrometry

Surendradoss, Jayakumar; Szeitz, András; Teng, Xiao Wei; Chang, Thomas K. H.; Abbott, Frank S.

doi:10.1016/j.jchromb.2013.06.006

Cited by 4 publications

(7 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In support of our finding that in situ generated VPA-G is nontoxic to rat hepatocytes, previous studies have shown that VPA-G is one of the least reactive and the most stable acyl glucuronides (Stachulski et al, 2006). Furthermore, as demonstrated under the cell culture conditions employed in this study, acyl migration of VPA-G was not significant (Surendradoss et al, 2013), suggesting that VPA-G did not form more reactive positional isomers. Overall, VPA glucuronidation appears to be a typical detoxification pathway, as determined in the current study in sandwich-cultured rat hepatocytes.…”

Section: Discussionsupporting

confidence: 90%

“…Each sample was transferred into a microfuge tube and stored at 280°C until analysis. VPA-G concentrations in culture supernatant and cell lysate were quantified using a validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) assay with [ 2 H 6 ]VPA-G as the internal standard (Surendradoss et al, 2013). The UHPLC-MS/MS system consisted of an Agilent 1290 Infinity Binary Pump, a 1290 Infinity Autosampler, and a 1290 Infinity Thermostatted Column Compartment (Agilent Technologies, Mississauga, Ontario, Canada), which was connected to an AB Sciex QTRAP 5500 hybrid linear ion-trap triplequadrupole mass spectrometer equipped with a Turbo Spray source (AB Sciex, Concord, Ontario, Canada).…”

Section: Methodsmentioning

confidence: 99%

“…The calibration curve of DFN-G ranged from 2.1 to 2120 nM. The mobile phases, chromatographic gradient, and mass-spectrometric conditions were the same as those described previously for VPA-G assay (Surendradoss et al, 2013). The declustering potential and collision energy settings were 260 V and 212 V for DFN-G, and 240 V and 220 V for [ 2 H 6 ]VPA-G, respectively.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Evaluation of In Situ Generated Valproyl 1-O-β-Acyl Glucuronide in Valproic Acid Toxicity in Sandwich-Cultured Rat Hepatocytes

2014

Self Cite

View full text Add to dashboard Cite

Acyl glucuronides are reactive electrophilic metabolites implicated in the toxicity of carboxylic acid drugs. Valproyl 1-O-b-acyl glucuronide (VPA-G), which is a major metabolite of valproic acid (VPA), has been linked to the development of oxidative stress in VPA-treated rats. However, relatively little is known about the toxicity of in situ generated VPA-G and its contribution to VPA hepatotoxicity. Therefore, we investigated the effects of modulating the in situ formation of VPA-G on lactate dehydrogenase (LDH) release (a marker of necrosis), BODIPY 558/568 C 12 accumulation (a marker of steatosis), and cellular glutathione (GSH) content in VPA-treated sandwich-cultured rat hepatocytes. VPA increased LDH release and BODIPY 558/568 C 12 accumulation, whereas it had little or no effect on total GSH content. Among the various uridine 59-diphospho-glucuronosyltransferase inducers evaluated, b-naphthoflavone produced the greatest increase in VPA-G formation. This was accompanied by an attenuation of the increase in BODIPY 558/568 C 12 accumulation, but did not affect the change in LDH release or total GSH content in VPA-treated hepatocytes. Inhibition of in situ formation of VPA-G by borneol was not accompanied by substantive changes in the effects of VPA on any of the toxicity markers. In a comparative study, in situ generated diclofenac glucuronide was not toxic to rat hepatocytes, as assessed using the same chemical modulators, thereby demonstrating the utility of the sandwich-cultured rat hepatocyte model. Overall, in situ generated VPA-G was not toxic to sandwich-cultured rat hepatocytes, suggesting that VPA glucuronidation per se is not expected to be a contributing mechanism for VPA hepatotoxicity.

show abstract

Section: Discussionsupporting

confidence: 90%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of In Situ Generated Valproyl 1-O-β-Acyl Glucuronide in Valproic Acid Toxicity in Sandwich-Cultured Rat Hepatocytes

2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…The use of various stabilizers such as citric acid, sodium phosphate buffer, sodium hydroxide and phosphoric acid to control the pH of blood as well as plasma samples has been evaluated and reported (Smith, Hasegawa, Langendijk, & Benet, ; Zhang et al, ). Examples of different stabilizers used for analytical assays are illustrated in Table (Brandhorst, Streit, Goetze, Oellerich, & Armstrong, ; Ebner, Heinzel, Prox, Beschke, & Wachsmuth, ; Hermening, Grafe, Baktir, Mutschler, & Spahn‐Langguth, ; Mano et al, ; Otieno et al, ; Tan et al, ; Yoshiaki, ; Zhou, Feng, Kestell, Baguley, & Paxton, ). Control of temperature or keeping the samples at a lower temperature during sample collection, processing (centrifugation) and storage is another variable which can help minimize the degradation of glucuronide.…”

Section: Sample Processingmentioning

confidence: 99%

Bioanalytical challenges and strategies for accurately measuring acyl glucuronide metabolites in biological fluids

Patel

2019

Biomedical Chromatography

View full text Add to dashboard Cite

Bioanalysis of unstable compounds such as acyl glucuronide metabolites represents a great analytical challenge owing to poor analyte stability in biological matrices. The primary goal for bioanalytical assay development is to minimize the breakdown of acyl glucuronide metabolite into its parent aglycone during sample collection, transportation, storage and analysis. Samples need to be stabilized ex vivo immediately after sample collection to minimize potential breakdown and thus to ensure accurate concentration measurement of both acyl glucuronide metabolite and its parent aglycone. In this review paper, formation of acyl glucuronide metabolites, the importance of establishing acyl glucuronide exposure measurement and safety coverage, optimization of sample pretreatment to stabilize the acyl glucuronide metabolites, current analytical strategy of assaying them as well as considerations for regulatory filings are discussed. It is important to identify acyl glucuronide metabolites that are capable of undergoing hydrolysis and pH‐dependent intra‐molecular migration as well as covalently binding to plasma and tissue proteins which can cause toxicity in vivo in the early stages of drug development. Carefully planning analytical experiments, identifying structures of acyl glucuronides and monitoring their concentrations in early drug development can help assess the risks associated with their exposures and potentially predict their concentrations in human circulation.

show abstract

“…Surendradoss et al used HPLC and MS to quantify valproyl 1-O-acyl-glucuronide (VPA-G), the phase II metabolized drug of valproic acid [142, 143]. Additionally, diclofenac conjugated to GSH has been measured via HPLC to assay for GST activity [140].…”

Section: Assays Detecting Susceptibility For Idiosyncratic Reactionsmentioning

confidence: 99%

Idiosyncratic Drug-Induced Liver Injury (IDILI): Potential Mechanisms and Predictive Assays

Roth

Lee

2017

BioMed Research International

View full text Add to dashboard Cite

Idiosyncratic drug-induced liver injury (IDILI) is a significant source of drug recall and acute liver failure (ALF) in the United States. While current drug development processes emphasize general toxicity and drug metabolizing enzyme- (DME-) mediated toxicity, it has been challenging to develop comprehensive models for assessing complete idiosyncratic potential. In this review, we describe the enzymes and proteins that contain polymorphisms believed to contribute to IDILI, including ones that affect phase I and phase II metabolism, antioxidant enzymes, drug transporters, inflammation, and human leukocyte antigen (HLA). We then describe the various assays that have been developed to detect individual reactions focusing on each of the mechanisms described in the background. Finally, we examine current trends in developing comprehensive models for examining these mechanisms. There is an urgent need to develop a panel of multiparametric assays for diagnosing individual toxicity potential.

show abstract

A rapid and sensitive assay to quantify valproyl 1-O-acyl glucuronide in supernatants of sandwich-cultured rat hepatocytes using ultra-high performance liquid chromatography–tandem mass spectrometry

Cited by 4 publications

References 15 publications

Evaluation of In Situ Generated Valproyl 1-O-β-Acyl Glucuronide in Valproic Acid Toxicity in Sandwich-Cultured Rat Hepatocytes

Evaluation of In Situ Generated Valproyl 1-O-β-Acyl Glucuronide in Valproic Acid Toxicity in Sandwich-Cultured Rat Hepatocytes

Bioanalytical challenges and strategies for accurately measuring acyl glucuronide metabolites in biological fluids

Idiosyncratic Drug-Induced Liver Injury (IDILI): Potential Mechanisms and Predictive Assays

Contact Info

Product

Resources

About