Rapamycin MRM Method


Measured Analytes

  1. Rapamycin
  2. Ascomycin (Internal Standard)

Chromatography

Liquid chromatography tandem mass spectrometry (LC-MS) was carried out with a Shimadzu Prominence HPLC system and SCIEX 6500 QTRAP mass spectrometer. An aliquot of 5 µl of each sample was injected onto an Phenomenex WidePore C18, 3 µm 100 x 2.1 mm column at 40oC for gradient separation. Mobile phases were: A, ddH2O with 0.1% Formic Acid; and B, Acetonitrile with 0.1% Formic Acid. Gradient schedule starts at 35% B and is linearly increased to 95% B at 1.2 minutes, then held at 95% B until 2.2 minutes, and then returned to 35% B at 2.3 at minutes and re-equilibration until 4.5 minutes. Time per sample injection was 4.5 minutes with a flow rate is 500µL/min.

rapamycin

Extraction Protocols:

For tissue, 100mg were processed with NextAdvance Bead Homogenization, and extracted using Phree Cartridge for lipid and protein removal. Flow through samples were dried under N2 gas and re-suspended in 50% Methanol/ddH2O for mass spectrometry.

For serum, 100uL of matrix was applied to Phree Cartridges for lipid and protein removal. Flow through samples were dried under N2 gas and re-suspended in 50% Methanol/ddH2O for mass spectrometry.

References

Development and Validation of a Bioanalytical LC-MS/MS Method for Simultaneous Determination of Sirolimus in Porcine Whole Blood and Lung Tissue and Pharmacokinetic Application with Coronary Stents. Thi-Thao-Linh Nguyen, Van-An Duong, Dang-Khoa Vo, Jeongae Jo, Han-Joo Maeng
PMID: 33467464 PMCID: PMC7829871 DOI: 10.3390/molecules26020425

Resources

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