Experimental design for the optimization and robustness testing of a liquid chromatography tandem mass spectrometry method for the trace analysis of the potentially genotoxic 1,3-diisopropylurea

Gyorgy Szekely*, Bruno Henriques, Marco Gil, Carlos Alvarez

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

This paper discusses a design of experiments (DoE) assisted optimization and robustness testing of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method development for the trace analysis of the potentially genotoxic 1,3-diisopropylurea (IPU) impurity in mometasone furoate glucocorticosteroid. Compared to the conventional trial-and-error method development, DoE is a cost-effective and systematic approach to system optimization by which the effects of multiple parameters and parameter interactions on a given response are considered. The LC and MS factors were studied simultaneously: flow (F), gradient (G), injection volume (Vinj), cone voltage (Econ), and collision energy (Ecol). The optimization was carried out with respect to four responses: separation of peaks (Sep), peak area (Ap), length of the analysis (T), and the signal-to-noise ratio (S/N). An optimization central composite face (CCF) DoE was conducted leading to the early discovery of carry-over effect which was further investigated in order to establish the maximum injectable sample load. A second DoE was conducted in order to obtain the optimal LC-MS/MS method. As part of the validation of the obtained method, its robustness was determined by conducting a fractional factorial of resolution III DoE, wherein column temperature and quadrupole resolution were considered as additional factors. The method utilizes a common Phenomenex Gemini NX C-18 HPLC analytical column with electrospray ionization and a triple quadrupole mass detector in multiple reaction monitoring (MRM) mode, resulting in short analyses with a 10-min runtime. The high sensitivity and low limit of quantification (LOQ) was achieved by (1) MRM mode (instead of single ion monitoring) and (2) avoiding the drawbacks of derivatization (incomplete reaction and time-consuming sample preparation). Quantitatively, the DoE method development strategy resulted in the robust trace analysis of IPU at 1.25ng/mL absolute concentration corresponding to 0.25ppm LOQ in 5g/l mometasone furoate glucocorticosteroid. Validation was carried out in a linear range of 0.25-10ppm and presented a relative standard deviation (RSD) of 1.08% for system precision. Regarding IPU recovery in mometasone furoate, spiked samples produced recoveries between 96 and 109 % in the range of 0.25 to 2ppm.

Original languageEnglish (US)
Pages (from-to)898-908
Number of pages11
JournalDrug Testing and Analysis
Volume6
Issue number9
DOIs
StatePublished - Sep 1 2014

Keywords

  • 1,3-diisopropylurea
  • Carry-over effect
  • LC-MS/MS; design of experiments
  • Mometasone furoate
  • Potentially genotoxic impurities

ASJC Scopus subject areas

  • Analytical Chemistry
  • Environmental Chemistry
  • Pharmaceutical Science
  • Spectroscopy

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