The work described herein incorporates a fast and simple pass-through sample preparation, robust and reliable UPLC-RP separation with a sub-2-μm C18 column designed specifically for retention and separation of polar compounds coupled to a high sensitivity tandem quadrupole MS. This method achieves high sensitivity and accuracy, with a lower limit of quantification (LLOQ) of 0.5 ng/mL from extracted human plasma.
The method combined selective and sensitive UPLC-MS/MS analysis with a simple and straightforward sample extraction using the Ostro 96-well Sample Preparation Plate. Ostro sample cleanup provided high recovery and low matrix effects, while effectively removing phospholipids and proteins from the sample. Additionally, the sub-2-μm ACQUITY UPLC HSS T3 Column provided necessary retention and resolution itaconic acid from its isobaric structural isomer, aconitic acid. The analytical sensitivity (0.5 ng/mL) and excellent reproducibility of the method described, reliably measures endogenous and elevated plasma levels of itaconic acid.