Navigation Links
Identification of Phase I and Phase II Metabolites of Buspirone on the Q TRAP LC/MS/MS System

Purpose

This application note describes the use of the new Q TRAP LC/MS/MS system with Metabolite ID software for Phase I and Phase II metabolite identification and discusses its unique capabilities that increase your productivity and confidence in your results.

Overview

Current trends in drug development are placing more emphasis on characterization of potential drug candidates earlier in the discovery process. The goal is to take more qualified drug candidates to preclinical testing and clinical trials. This shift in focus will enable drug researchers to save time and money. Nearly 60% of the costs to develop a new drug are incurred in the clinical trials phase, so there is much to be gained by selecting only the most promising drug candidates for testing at this level. Early discovery ADME (adsorption, distribution, metabolism, and excretion) studies are critical to predict a drug candidates success in further downstream testing.

Early assessment of drug metabolism of in vitro and in vivo samples provides initial information about a potential drugs biotransformation and metabolite characterization. To obtain maximum characterization information, including chemical structure and quantity, LC/MS (liquid chromatography/mass spectrometry) is often employed because it has sufficient sensitivity and specificity to measure nanogram levels of metabolites in complex matrices. Ion trap mass spectrometers are widely used to identify metabolites, and triple quadrupole mass spectrometers are generally used to measure the amount, or concentration of metabolites detected.

Key Features of the Q TRAP System

Exceptional sensitivity for metabolite identification and characterization

Powerful advanced scan modes, including neutral loss and precursor ion scans, can be used in flexible combinations to achieve unprecedented selectivity

Broad linear dynamic range provides true triple quadrupole quantitation performance and enhances identification of ions in complex matrices

Information-rich MS/MS and MS3 spectra with important low mass structural ions provide confidence in results

Metabolite ID application software makes it easy to go straight to the answers

Experimental Conditions

Rat liver microsomal incubate samples were injected on a Hypersil- Betasil C18 column (1 x 100 mm). Samples were eluted using Shimadzu 10 AD pumps with a gradient of water/methanol, from 10 to 90% methanol over 10 minutes at a flow rate of 70 μL/min. Direct injection (5 μL) on column was performed using a PE Series 200 autosampler. A TurboIonSpray source was operated at 350 C on the Q TRAP system.

Results and Discussion

The new Q TRAP LC/MS/MS system combines superior ion trap capabilities with triple quadrupole performance to provide a new level of performance and versatility in LC/MS for metabolite identification. The patented hybrid linear design enhances sensitivity, resolution, and duty cycle for MS and MS/MS full scan modes, and combines these features with the linear dynamic range and scan functions of a triple quadrupole instrument.

Add Metabolite ID application software and get automated data acquisition and processing, with a range of options to ensure maximum productivity. Metabolite ID application software leverages the Q TRAP system technology and IDA to provide rapid, high-confidence results in the fewest number of experiments.

Metabolite ID:

Identifies predefined and userdefined Phase I and Phase II transformations

Detects unexpected metabolites by recognition of isotope ratio patterns

Triggers automatic acquisition of MS/MS structural information

Confirms metabolites by correlation comparison to parent drug

Performs fully automated data acquisition and processing in batch mode

Includes a powerful library search function

Contains an easy-to-use browser for viewing results

The result is a unique LC/MS/MS platform that can identify more metabolites in less time, and quantitate them too. In this study, the Q TRAP system was used to identify Phase I and Phase II metabolites of buspirone.

The Q TRAP system has the advantage of using highly selective scan functions such as neutral loss and precursor ion scans to facilitate detection of important biotransformation products from complex matrices. The structure and mass spectrum of buspirone (Figure 1) lead to the prediction that fragmentation that can occur on the A or B side of the structure can be identified using precursor ion scans at mass 122 and 168, respectively. Further information can be gained by using these two masses in combination with masses of expected transformations, i.e. precursor ion of 138 for hydroxylation occurring on the A side.

Figure 2 compares a normal full scan TIC trace from a liquid chromatography gradient run to a precursor ion scan of mass 122 and demonstrates the gain in selectivity. Up to two precursor ion scans can be combined in the Information Dependent Acquisition (IDA) survey step, minimizing the number of injections required to gain maximum information.

Figure 3 shows XIC traces of two metabolites that were identified with a precursor ion scan of mass 122 plus buspirone. An enhanced product ion (EPI) scan of a peak at 6.86 minutes from the XIC 402 trace confirms hydroxy-buspirone as a major metabolite and provides important information on the site of modification. The IDA method also contained an enhanced resolution scan that improves the monoisotopic mass assignment, as well as provides isotope ratio information for further confirmation (Figure 4).

The number of metabolites identified with the precursor ion scan method was compared to the number of metabolites identified with an enhanced MS method. Table 1 illustrates that the precursor ion scan method clearly identifies and confirms more metabolites in all cases.

For identification of Phase II metabolites, an IDA method with a neutral loss scan of 176 as the survey scan was used to selectively identify three glucuronide species of buspirone (Figure 5). Prior MS/MS experiments with buspirone glucuronides indicated the aglycon species as the major fragment ion, and subsequently, the IDA method was set to include an MS3 experiment following EPI, in order to gain more structural information. The MS3 experiment run on the aglycon metabolite (m/z 402) is much richer in structural information due to the triple quadrupole fragmentation pattern of the Q TRAP system. Figure 5 shows the resulting MS3 spectra of m/z 402 from the EPI scan, which, combined with supporting chemical structural information, clearly indicates the site of hydroxylation. This powerful automated approach to identifying and characterizing metabolites all in a single LC/MS IDA analysis can be used on expected and unexpected metabolites.

Conclusions

The Q TRAP LC/MS/MS system with IDA and Metabolite ID automates every aspect of metabolite identification and provides the highest quality results. Advanced, automated data processing and reporting puts highconfidence results at your fingertips in less time. The Q TRAP LC/MS/MS system, a hybrid linear ion trap, provides superior ion trap performance and high-performance triple quadrupole functionality. Scan functions unique to ion trap MS, such as precursor ion and neutral loss scans, simplify the task of finding metabolites in complex matrices, and increase the amount of information gained per experiment. Phase I and Phase II metabolites can be identified, characterized, and quantitated on a single LC/MS/MS system. The high sensitivity full scan MS and MS/MS spectra allows the identification of more metabolites, and the comprehensive Metabolite ID application software automates the process. Identify more metabolites with more confidence with the Q TRAP LC/MS/MS system.


'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Identification of Differentially Expressed Gene Products with the CastAway System*
2. An Optimal DNA Microarray Substrate for the Identification of Fetal and Somatic Liver Stem Cells of the Rat
3. Identification of Low-Level Oxidation Products in Calmodulin Using NanoSpray Ion Trap Mass Spectrometry
4. Detection and Identification of Phosphorylation Sites in Proteins Using LC/MS/MS with Neutral Fragment Loss Mapping
5. Protect RNA and Improve Target Cell Identification
6. Identification of Disease Causing Mutations in Phenylketonuria by Denaturing Gradient Gel Electrophoresis Using the DCode System
7. Identification of Nonspecific Products Using Melt-Curve Analysis on the iCycler iQ Detection System, Rev A
8. Identification of cells in S phase using the Cell Proliferation Fluorescence Kit and IN Cell Analyzer 1000
9. Identification of Insecticides and Herbicides by LC/MS/MS Using the API 2000 LC/MS/MS System
10. ESR1 Gene Expression in Oncology and Metabolic Diseases Using the ASCENTATM System Target Validation and Identification of Novel Disease Indications
11. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
Post Your Comments:
(Date:11/22/2014)... Frederick, MD (PRWEB) November 21, 2014 ... sample management solutions provider, has released Limfinity® version ... more installation projects of Limfinity® than ever in ... majority configuration and minority framework enhancement. Limfinity® version ... goes far above and beyond bug fixes! , ...
(Date:11/22/2014)... Quebec , November 21, 2014 ... wechselt als Chief Commercial Officer zu KLOX ... SVP der Unternehmensentwicklung und Chief Financial Officer bestellt    ... Europa mit der Vermarktung seines jüngst zugelassenen Behandlungssystems zur ... Inc. ("KLOX" bzw. das "Unternehmen") ist sehr erfreut, die ...
(Date:11/21/2014)... Los Angeles, CA (PRWEB) November 20, 2014 ... “A Short Walk to the Other Side” ... was an “Award-Winning Finalist in the “Fiction: Short Story” ... . , Jeffrey Keen, President and CEO of USA ... entries from mainstream and independent publishers, including Simon & ...
(Date:11/21/2014)... Angeles, CA (PRWEB) November 21, 2014 ... it that makes Albert Einstein so instantly recognizable? Why ... African-born physicist, mathematician, astronomer and author Hilton Ratcliffe seeks ... they have nothing at all to do with science. ... Muse Harbor Publishing , November 21, 2014), Ratcliffe puts ...
Breaking Biology Technology:RURO Announces Limfinity® version 5.1.4 2KLOX Technologies meldet Neubesetzung von Führungspositionen 2KLOX Technologies meldet Neubesetzung von Führungspositionen 3KLOX Technologies meldet Neubesetzung von Führungspositionen 4KLOX Technologies meldet Neubesetzung von Führungspositionen 5Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 2Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 3Mystic Ink 's Short Story Collection “A Short Walk to the Other Side” Honoring Ray Bradbury Praised As Award-Winning Finalist In USA Best Book Awards 4“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 2“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 3“Stephen Hawking Smoked My Socks” Sparks Controversy in Science World 4
... 3, 2008 Lentigen Corporation today,announced an ... responsible for licensing technologies from the University ... the area of lentiviral vector,technology. The technology ... Lever at the University of Cambridge., ...
... April 3, 2008 U.S.,Preventive Medicine(R), ( http://www.USPreventiveMedicine.com ... global leader in gene discovery, today,announced they have ... testing to,expand both companies, personalized medicine services in ... letter of intent with deCODE is significant because,we ...
... Broad Array of Healthcare Products to ... Immune Deficiency and Bleeding Disorders, OTTAWA, April ... that Canadian Blood Services, and Hema-Quebec, have each ... Factor,[Recombinant]), Humate(R) P (Antihemophilic factor / von Willebrand ...
Cached Biology Technology:Lentigen Licenses Fundamental Lentiviral Vector Technology Patent From the University of Cambridge 2Lentigen Licenses Fundamental Lentiviral Vector Technology Patent From the University of Cambridge 3U.S. Preventive Medicine and deCODE Genetics Sign Letter of Intent to Expand Personalized Medicine Platform 2U.S. Preventive Medicine and deCODE Genetics Sign Letter of Intent to Expand Personalized Medicine Platform 3CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 2CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 3CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 4CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 5CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 6CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 7CSL Behring Signs Agreements With Canadian Blood Services and Hema-Quebec to Supply Life-Saving Therapies 8
(Date:11/21/2014)... 20, 2014 C-Labs LLC, a leading provider ... Things (IoT), today announced the appointment of John ... Previously a strategic advisor to the firm, Mr. Traynor ... Traynor is based out of the C-Labs office in ... Chris Muench , Chief Executive Officer. ...
(Date:11/18/2014)... , Nov. 18, 2014   EyeLock ... authentication solutions, and MorphoTrust USA (Safran), ... services, today announced a strategic partnership to offer ... motor vehicle administration (MVA), airport screening and financial ... identity-related solutions, MorphoTrust serves consumers through a nationwide ...
(Date:11/7/2014)... Auguste, associate professor, biomedical engineering, in the Grove ... New York, have identified a molecule that could ... most aggressive forms of breast cancer. , Triple ... rate owing to aggressive proliferation and metastasis and ... Auguste,s team, discovered the overexpression of intercellular adhesion ...
Breaking Biology News(10 mins):C-Labs Names Former Microsoft and Bsquare Executive as Chief Operating Officer 2EyeLock, MorphoTrust Form Strategic Partnership to Pursue Commercial Enterprise Deployments 2EyeLock, MorphoTrust Form Strategic Partnership to Pursue Commercial Enterprise Deployments 3
... /Press Release/ Toxic waste sites with elevated levels ... years of life lost" in individuals living near 373 sites ... by a Mount Sinai researcher published online today in ... MD, Pediatric Environmental Health Fellow at the Icahn School of ...
... University of London will bring researchers one-step closer to ... its function in two recently published studies. ... from the Complex Networks group at Queen Mary,s School ... can have an association despite a lack of direct ...
... important substances across their biological membranes to resist toxic ... water loss and store sugar can have profound implications ... our rapidly growing global population. , That,s the ... world whose laboratories recently discovered important properties of plant ...
Cached Biology News:Toxic waste sites cause healthy years of life lost 2Mathematicians help to unlock brain function 2New plant protein discoveries could ease global food and fuel demands 2New plant protein discoveries could ease global food and fuel demands 3New plant protein discoveries could ease global food and fuel demands 4
... These versatile vacuum cups provide flexible ... curved, or irregular surfaces. ,PFG: Flat ... Bellows cup ,PCG: Multiple Bellows cup ... from 25 to 200 mm , ...
XA7 B6...
... linear motor and ballscrew technologies, ... Cartesian solutions that include all ... and motors., , XY ... through high-flex cabling for power, ...
Total Proteins from over 50 different human Alzheimer, Diabetic, Liver Cirrhosis, and Lupus diseased tissues are available....
Biology Products: