Investigation of transition ion ratio variation for liquid chromatography-tandem mass spectrometry: A case study...

A transition ion ratio (TIR) is the ratio of one fragment over another from the same precursor and is frequently monitored in liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays for analyte identification. The Clinical and Laboratory Standards Institute (CLSI) C50-A guidelines give a static percent allowable TIR deviation based on the TIR level. Anecdotally, we observed failures of these rules for some of our LC-MS/MS assays. We determined what parameters may affect TIRs in a clinical setting and whether TIR variations may be analyte, matrix, instrument service, and/or concentration dependent.

Data was collected from the validation and selected periods after implementation for urine benzodiazepines (7 analytes) and plasma azole antifungals (6 analytes). TIRs for the calibrators and quality control materials on a Thermo TSQ™ Quantum Ultra from July 2016 to February 2017 for benzodiazepines in urine and Thermo TSQ™ Vantage from May 2016 to Oct 2016 for azoles in serum were monitored.

The statistically significant day-to-day TIR shift ranged from 5.7 to 27.0% of the days studies for benzodiazepines and from 5.6 to 27.8% of the days studied for azoles excluding shifts caused by instrument services. Instrument service had significant impact on all benzodiazepines except oxazepam with p-values ranging from 1.79 × 10^-6 to 1.53 × 10^-39 and 4 of the 6 azoles (fluconazole, isavuconazole, voriconazole, and itraconazole) with (p from 7.89 × 10^-3 to 1.98 × 10^-12). Lorazepam, α-hydroxyalprazolam, and hydroxyitraconazole showed significant concentration dependent TIR variations.

TIR variations may be affected by instrument services, and can be concentration and analyte dependent. Instead of using a static percent deviation rule, establishment of TIR variation criteria for each analyte during test development and validation may provide a more useful tool for analyte identification.