Simultaneous Determination of Six Immunosuppressants in Human Whole Blood by HPLC-MS/MS Using a Modified QuEChERS Method

Abstract

A high-performance liquid chromatography-tandem mass spectrometry method was established for the simultaneous determination of mycophenolic acid, mycophenolate mofetil, tacrolimus, rapamycin, everolimus and pimecrolimus in human whole blood by optimizing the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) preparation method. Whole blood was extracted into ethyl acetate, salted out with anhydrous magnesium sulfate, and purified with ethylenediamine-N-propyl silane adsorbent. The supernatant was evaporated under nitrogen until dry and finally reconstituted in methanol. Chromatographic separation was performed on an Agilent Poroshell 120 EC-C18 column in methanol (mobile phase A)-water (optimized for 0.1% acetic acid and 10 mM ammonium acetate, mobile phase B) at a 0.3 mL·min−1 flow rate. Electrospray ionization and positive ion multiple reaction monitoring were used for detection. The time for of analysis was 13 min. The calibration curves range of tacrolimus, rapamycin, everolimus and pimecrolimus were in the range of 1–100 ng·mL−1, mycophenolate mofetil in the range of 0.1–10 ng·mL−1 and mycophenolic acid at 10–1000 ng·mL−1. All correlation coefficients were >0.993. The coefficients of variation (CV, %) for inter-day and intra-day precision were less than 10%, while the spiked recoveries were in the range of 92.1% to 116%. Our method was rapid, sensitive, specific, and reproducible for the simultaneous determination of six immunosuppressants in human whole blood. Importantly, our approach can be used to monitor drug concentrations in the blood to facilitate disease treatment.

QuEChERS-Based Approach to the Extraction of Five Calcium Channel Blockers from Plasma Determined by UPLC-MS/MS

“…In previous work conducted by our group, we applied the QuEChERS pre-treatment technique to assess the concentrations of multiple drug classes. Analyses of multiple tyrosine kinase inhibitors (TKI) concentrations in human plasma [ 22 ] and multiple immunosuppressant concentrations in whole human blood [ 23 ] have performed well in terms of LOQ and recovery as compared to other assays for similar drugs. In addition, other members ofour group have attempted to use the QuEChERS technique in combination with LC-MS to detect 16illicitly added drugs in capsule dietary supplements containing lipid-lowering drugs, diuretics, and appetite suppressants [ 24 ].…”

Development and Validation of HPTLC Method for Estimation of Pimecrolimus in Formulations

“…A suitable and validated method should be vacant for the drug delivery system for the analysis of bulk drug and in pharmaceutical cream formulation. Various methods are reported for the analysis of pimecrolimus alone or in combination with other drugs have been reported, which included, HPLC-MS/MS [3], RP-HPLC [2] and LC-TMS [4] but pimecrolimus has not yet been subject to the HPTLC method of analysis, as far as we are aware.…”