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Orbitrap_SciLib
Reputable Mentor II
Reputable Mentor II

Quantitative analysis of drugs in various body fluids, such as plasma, serum, and cerebrospinal fluid (CSF), is conducted in many research laboratories. SRM analysis using a triple quadrupole LC-MS/MS system has been routinely employed for quantitative analysis. Although, in most cases, SRM offers the required sensitivity and specificity, it suffers from interferences and a lack of robustness and needs expert MS users. High-resolution, accurate mass (HR/AM) mass spectrometry based on Orbitrap™ technology offers an attractive alternative with unique benefits of specificity, sensitivity and, more importantly, ease of use.
 
Here we describe the quantitative workflow solutions using the Thermo Scientific Exactive Plus system for the following classes of drugs:
 

  • Immunosuppressant drugs
  • Anticancer drugs
  • Antifungal drugs

 

For additional resources, search the Orbitrap Science Library 1GoArrow.png

Workflow Overview for Antifungal Drugs Quantitation

 
Antifungal drugs in plasma are analyzed for various research purposes. While selected reaction monitoring (SRM)-based analysis is routinely used for quantitation, high-resolution full-scan MS or selected ion monitoring (SIM)-based analysis offers a simpler alternative without any drawbacks.
 
The workflow described here quantitates Anidulafungin, Caspofungin, Fluconazole, HO-Itraconazole, Itraconazole, Posaconazole, and Voriconazole using simple full-scan MS at 70,000 FWHM resolution. The data is acquired in Full Scan mode and the quantitation is performed by using extracted ion chromatograms using a 3 ppm window around the m/z of interest. This application demonstrates the capability of the Thermo Scientific Exactive Plus MS coupled to a uHPLC system for quantitative analysis of the above mentioned drugs with a LLOQ in the low pg/mL range and ULOQ in the mid ng/mL range.



 
Workflows_Clin_DrugMon_Antifungal(2).jpg



resources

 

Quan / Qual Analyses in Clinical Research using a Second Generation Exactive High Resolution Mass Sp...

Rochat B.
ASMS 2012 Presentation

 

Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer...

Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B.
Rapid Commun Mass Spectrom. 2012 Mar 15;26(5):499-509.

Sample Preparation

Sample Preparation Workflow for Antifungal Drugs Quantitation

 
Human plasma (100 µL) was extracted with 300 µL of MeCN/formic acid solution containing the internal standard. After vortexing and centrifugation, the supernatant was diluted twice prior to injection onto the HPLC system.
 
MWorkflows_Antifungal_SP_.jpg

resources

 

Quan / Qual Analyses in Clinical Research using a Second Generation Exactive High Resolution Mass Sp...

Rochat B.
ASMS 2012 Presentation

 

Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer...

Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B.
Rapid Commun Mass Spectrom. 2012 Mar 15;26(5):499-509.

Mass Spectrometry Workflow for Antifungal Drugs Quantitation

 
The Thermo Scientific Exactive Plus or Exactive mass spectrometer with a HESI source was coupled to a Thermo Scientific Accela 1250 uHPLC or Transcend system. The MS was operated in SIM mode at a resolution of 70,000 FWHM during the entire LC run. The column eluent was diverted to waste for a specified period of time after the sample injection before being sent to the MS. The Exactive system was calibrated at the beginning of the day and used for the entire analysis. No lock mass or internal calibration was used.
 
 Scheme-ExactivePlus-Part-RGB.png

resources

 

Quan / Qual Analyses in Clinical Research using a Second Generation Exactive High Resolution Mass Sp...

Rochat B.
ASMS 2012 Presentation

 

Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer...

Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B.
Rapid Commun Mass Spectrom. 2012 Mar 15;26(5):499-509.

Data Analysis Workflow for Antifungal Drugs Quantitation

 
Thermo Scientific TraceFinder software was used for automated data processing and reporting. The quantitative processing method included the exact m/z of fragments monitored and their retention times. The extracted ion chromatograms for fragments were obtained for calibrators, QCs and samples. The area under the peak for the quantifier ion was used for creating calibration curves and calculating values of QCs and samples; the area for the other fragment was used as a qualifier. The ion ratios were measured to rule out interferences. Automatic reports were created as specified in the method.
 
MWorkflows_Endocrine1_DA_.jpg

resources

 

Quan / Qual Analyses in Clinical Research using a Second Generation Exactive High Resolution Mass Sp...

Rochat B.
ASMS 2012 Presentation

 

Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer...

Henry H, Sobhi HR, Scheibner O, Bromirski M, Nimkar SB, Rochat B.
Rapid Commun Mass Spectrom. 2012 Mar 15;26(5):499-509.

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Last update:
‎10-15-2021 11:48 AM
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