Orbitrap Illustration

Immunosuppressant drugs quantitation

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

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Workflow Overview for Immunosuppressant Drugs Quantitation

Quantitative analysis of immunosuppressant drugs such as Tacrolimus, Everolimus, Sirolimus and Cyclosporin in whole blood needs high levels of accuracy and speed. Selected reaction monitoring (SRM)-based analysis is the standard technique for quantitation. The high-resolution full-scan MS or SIM offers a simpler alternative without any drawbacks.
The workflow described here uses simple full-scan MS at 100,000 FWHM resolution. The data is acquired in the Full Scan mode, and the quantitation is performed by using extracted ion chromatograms with 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 four above-mentioned drugs with a LLOQ of 1 ng/mL for all drugs except Cyclosporin. The Cyclosporin method has a LLOQ of 10 ng/mL. The dynamic range is up to at least 50 ng/mL for the first three drugs and up to 2,000 ng/mL for Cyclosporin.



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

Rochat B.
ASMS 2012 Presentation


Comparison between a high-resolution single-stage Orbitrap and a triple quadrupole mass spectrometer for quantitative analyses of drugs

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


Quantitative Analysis of 4 Immunosuppressant Drugs in Blood using Exactive System

Kozak M.
Slide Presentation


Quantitation of Immunosuppressant Drugs in Blood Utilizing Second Generation Exactive High Resolution Accurate Mass Spectrometer and TraceFinder Software

Van Natta K, Kozak M, Nimkar S, Clarke W.
ASMS 2012 Poster