Berube M, DiBussolo J, Lafontaine C, Shi Y Application Note 567 Blood levels of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 are commonly tested by clinical researchers to assess vitamin D sufficiency. In the last decade, liquid chromatography coupled with triple quadrupole mass spectrometry (LC-MS/MS) has become a popular technique for such measurements. Due to their higher resolving power relative to triple-stage quadrupole mass spectrometers, Orbitrap™-based mass spectrometers are better able to resolve analytes from sample matrices. In addition, the ease of initial method set up and daily use provides an advantage over triple-stage quadrupole mass spectrometers for clinical research.
A method has been created that allows precipitated serum to be injected into an HPLC system with minimal sample preparation and analyzed by an ExactiveTM Plus benchtop Orbitrap mass spectrometer. Total method time is 7.75 minutes on a Thermo Scientific Transcend TLX-1 system utilizing TurboFlow technology. Throughput can be increased to a sample every 3.7 minutes by using a Transcend™ TLX-2 multiplexed UHPLC system or 1.9 minutes with a Transcend TLX-4 system.