MSACL 2019 EU Abstract

Integration of Mycophenolate and its Metabolite Analysis in Plasma Using LC-MS/MS with Full-Automated Sample Preparation

Toshikazu Minohata (1, 2), Stéphane Moreau (3), Fanny Dayot (1), Jean-François Hoeffler (1)
(1) Alsachim SAS, Illkirch, France (2) Shimadzu Corporation, Kyoto, Japan (3) Shimadzu Europa GmbH, Germany

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Fanny Dayot (Presenter) Alsachim >> POSTER (PDF)

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Introduction
Currently sample preparation for the detection of drugs in biological samples by liquid chromatography-mass spectrometry (LC-MS/MS) involves complex offline extraction methods such as solid phase extraction or liquid/liquid extraction, all of which require additional sample concentration and reconstitution in an appropriate solvent. These sample preparation methods are time-consuming, often taking 1 hour or more per sample, and are more vulnerable to variability due to errors in manual preparation. Our approach to offering a high sensitivity drug detection method and timely, automated analysis of multiple samples is to use the automated sample preparation system coupled to the detection capabilities of a high-sensitivity triple stage quadrupole mass spectrometer.

Method
Mycophenolic acid and its glucuronide in plasma were verified using DOSIMYCOTM (Alsachim, France). plasma sample was loaded directly into the automated sample preparation system (CLAM-2000 Shimadzu, Japan). The CLAM-2000 was programmed to perform protein precipitation using methanol followed by filtration and sample collection. The sample is then transported using an arm from the CLAM-2000 to the HPLC without human intervention for LC-MS/MS analysis.
The treated samples were trapped using a DOSIMYCOTM C8 column and then separated by DOSIMYCO® C18 column at 65°C in 2 min.

Results
We evaluated this system using calibrator and control plasma spiked with mycophenolic acid and its glucuronide in DOSIMYCOTM and carried out concurrent analysis over a range of concentrations in 0.1 to 50 mg/L for mycophenolic acid and 1 to 250 mg/L for its glucuronide. The calibration curves that were generated had linear regression values of r2 >0.99 for each curve. As for reproducibility (N=3) at three concentrations, the deviation between theoretical values and actual measurements was less than 10%, and the CV value was also less than 10%.

Conclusion
We completed mycophenolate analysis using the automated sample preparation system coupled to LC-MS/MS. The results show the capability of the system for large sample set analyses with improved accuracy and precision by eliminating human error associated with manual sample handling.