A strategy for identification of drug metabolites from dried blood spots using triple‐quadrupole/linear ion trap hybrid mass spectrometry

Mauriala, Timo; Chauret, Nathalie; Oballa, Renata M.; Nicoll‐Griffith, Deborah A.; Bateman, Kevin P.

doi:10.1002/rcm.2013

Cited by 44 publications

(43 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Such MRM transitions were used as survey scans, and following peak detection (when a certain threshold was reached), an EPI spectrum was generated to provide structural information for metabolite assignments. Similar approaches for metabolite detection using a hybrid triple-quadrupole linear ion trap instrument have been described (43,44). A comparison between MRM chromatograms of dimethoxycurcumin incubated in the presence and absence of microsomes for 1 h is shown in Fig.…”

Section: Methodsmentioning

confidence: 94%

Metabolism and Anticancer Activity of the Curcumin Analogue, Dimethoxycurcumin

Tamvakopoulos

Dimas

Sofianos

et al. 2007

Clinical Cancer Research

180

146

View full text Add to dashboard Cite

Purpose: The plant-derived compound curcumin has shown promising abilities as a cancer chemoprevention and chemotherapy agent in vitro and in vivo but exhibits poor bioavailability. Therefore, there is a need to investigate modified curcumin congeners for improved anticancer activity and pharmacokinetic properties. Experimental Design: The synthetic curcumin analogue dimethoxycurcumin was compared with curcumin for ability to inhibit proliferation and apoptosis of human HCT116 colon cancer cells in vitro by estimating the GI 50 and LC 50 values and detecting the extent of apoptosis by flow cytometry analysis of the cell cycle. Metabolic stability and/or identification of metabolites were evaluated by recently developed mass spectrometric approaches after incubation with mouse and human liver microsomes and cancer cells in vitro. Additionally, circulating levels of dimethoxycurcumin and curcumin were determined in mice following i.p. administration. Results: Dimethoxycurcumin is significantly more potent than curcumin in inhibiting proliferation and inducing apoptosis in HCT116 cells treated for 48 h. Nearly 100% of curcumin but <30% of dimethoxycurcumin was degraded in cells treated for 48 h, and incubation with liver microsomes confirmed the limited metabolism of dimethoxycurcumin. Both compounds were rapidly degraded in vivo but dimethoxycurcumin was more stable. Conclusions: Compared with curcumin, dimethoxycurcumin is (a) more stable in cultured cells, (b) more potent in the ability to kill cancer cells by apoptosis, (c) less extensively metabolized in microsomal systems, and (d) more stable in vivo. It is likely that the differential extent of apoptosis induced by curcumin and dimethoxycurcumin in vitro is associated with the metabolite profiling and/or the extent of stability.Curcumin (diferuloylmethane) is the active yellow pigment in turmeric, a popular plant-derived coloring spice and ingredient of many cosmetics and pharmaceuticals (see refs. 1 -5 for reviews on structural and biological aspects of curcumin). In general, curcumin has been associated with a large number of biological and cellular activities/events, including antioxidative, anti-inflammatory, anticarcinogenic, and hypocholesterolemic properties (1 -5), and a large number of studies have focused on the pathways by which curcumin acts as a chemoprotective agent (1, 2, 6, 7). Further, curcumin congeners have been reported to induce apoptosis, sensitize and overcome resistance to various agents that induce apoptosis in diverse human cancer cells (8 -16), and decrease the occurrence of cardiomyocytic apoptosis after global cardiac ischemia/reperfusion (17). In general, curcumin mediates its effects by modulating several important molecular targets, including transcription factors, enzymes, cell cycle proteins, cytokines, receptors, and cell surface adhesion molecules (reviewed in refs. 2 -4).With regard to its anticancer potential, it has been shown that curcumin congeners can inhibit proliferation and/or induce apoptosis of cancer c...

show abstract

Section: Methodsmentioning

confidence: 94%

Metabolism and Anticancer Activity of the Curcumin Analogue, Dimethoxycurcumin

Tamvakopoulos

Dimas

Sofianos

et al. 2007

Clinical Cancer Research

180

146

View full text Add to dashboard Cite

show abstract

“…However, there is an increased need for the use of a single of LC/MS platform for both bioanalysis and metabolite identification, especially in small discovery organizations. Qtrap has scanning functions of MRM-, NL-, and PI-EPI as well as polarity switching that allow it to be effectively used for profiling and identification of oxidative and conjugated metabolites [41][42][43], as well as reactive metabolite screening as demonstrated in the literatures [12,13,26] and current study. Therefore, Qtrap is well suited for a small discovery or academic drug metabolism and pharmacokinetics research lab where bioanalysis or in vitro quantitative metabolism screening is the major responsibility, while metabolite screening and identification are conducted occasionally.…”

Section: Potential Applications Of the Current Workflowmentioning

confidence: 95%

Simultaneous Screening of Glutathione and Cyanide Adducts Using Precursor Ion and Neutral Loss Scans-Dependent Product Ion Spectral Acquisition and Data Mining Tools

Jian

Liu²,

Zhao

et al. 2012

J. Am. Soc. Mass Spectrom.

View full text Add to dashboard Cite

Drugs can be metabolically activated to soft and hard electrophiles, which are readily trapped by glutathione (GSH) and cyanide (CN), respectively. These adducts are often detected and structurally characterized using separate tandem mass spectrometry methods. We describe a new method for simultaneous screening of GSH and CN adducts using precursor ion (PI) and neutral loss (NL) scansdependent product ion spectral acquisition and data mining tools on an triple quadrupole linear ion trap mass spectrometry. GSH, potassium cyanide, and their stable isotope labeled analogues were incubated with liver microsomes and a test compound. Negative PI scan of m/z 272 for detection of GSH adducts and positive NL scans of 27 and 29 Da for detection of CN adducts were conducted as survey scans to trigger acquisition of enhanced resolution (ER) spectrum and subsequent enhanced product ion (EPI) spectrum. Post-acquisition data mining of EPI data set using NL filters of 129 and 27 Da was then performed to reveal the GSH adducts and CN adducts, respectively. Isotope patterns and EPI spectra of the detected adducts were utilized for identification of their molecular weights and structures. The effectiveness of this method was evaluated by analyzing reactive metabolites of nefazodone formed from rat liver microsomes. In addition to known GSH-and CN-trapped reactive metabolites, several new CN adducts of nefazodone were identified. The results suggested that current approach is highly effective in the analysis of both soft and hard reactive metabolites and can be used as a high-throughput method in drug discovery.

show abstract

“…25,26 Control sample background subtraction has never been reported as a strategy for in vivo metabolite identification, although dynamic background subtraction 12,13 has been reported to assist knowledge-guided metabolite screening strategies in plasma and urine. The main objective of this study is to evaluate the utility of our accurate mass-based background-subtraction algorithm for unbiased profiling of drug metabolites from in vivo samples.…”

mentioning

confidence: 98%

“…This technique, along with prediction of masses of potential drug metabolites, has become an important method for the overall drug metabolite detection process. 2,6,10 Advanced algorithms, such as component detection algorithm (CODA) for noise and background reduction 11 and dynamic background subtraction to trigger sensitive product ion scans, 12,13 have also been developed and utilized. However, despite the advances of LC/MS and data process technologies, the identification of drug metabolites in biological samples continues to rely on the prediction of metabolite masses or their mass spectrometric fragmentation patterns through either data acquisition or post acquisition data mining.…”

mentioning

confidence: 99%

An algorithm for thorough background subtraction from high‐resolution LC/MS data: application to the detection of troglitazone metabolites in rat plasma, bile, and urine

Zhang

et al. 2008

J. Mass Spectrom.

View full text Add to dashboard Cite

Interferences from biological matrices remain a major challenge to the in vivo detection of drug metabolites. For the last few decades, predicted metabolite masses and fragmentation patterns have been employed to aid in the detection of drug metabolites in liquid chromatography/mass spectrometry (LC/MS) data. Here we report the application of an accurate mass-based background-subtraction approach for comprehensive detection of metabolites formed in vivo using troglitazone as an example. A novel algorithm was applied to check all ions in the spectra of control scans within a specified time window around an analyte scan for potential background subtraction from that analyte spectrum. In this way, chromatographic fluctuations between control and analyte samples were dealt with, and background and matrix-related signals could be effectively subtracted from the data of the analyte sample. Using this algorithm with a+/-1.0 min control scan time window, a+/-10 ppm mass error tolerance, and respective predose samples as controls, troglitazone metabolites were reliably identified in rat plasma and bile samples. Identified metabolites included those reported in the literature as well as some that had not previously been reported, including a novel sulfate conjugate in bile. In combination with mass defect filtering, this algorithm also allowed for identification of troglitazone metabolites in rat urine samples. With a generic data acquisition method and a simple algorithm that requires no presumptions of metabolite masses or fragmentation patterns, this high-resolution LC/MS-based background-subtraction approach provides an efficient alternative for comprehensive metabolite identification in complex biological matrices.

show abstract

A strategy for identification of drug metabolites from dried blood spots using triple‐quadrupole/linear ion trap hybrid mass spectrometry

Cited by 44 publications

References 18 publications

Metabolism and Anticancer Activity of the Curcumin Analogue, Dimethoxycurcumin

Metabolism and Anticancer Activity of the Curcumin Analogue, Dimethoxycurcumin

Simultaneous Screening of Glutathione and Cyanide Adducts Using Precursor Ion and Neutral Loss Scans-Dependent Product Ion Spectral Acquisition and Data Mining Tools

An algorithm for thorough background subtraction from high‐resolution LC/MS data: application to the detection of troglitazone metabolites in rat plasma, bile, and urine

Contact Info

Product

Resources

About