This article introduces an example of high-speed analysis of itraconazole and its active metabolite hydroxy itraconazole in plasma using the LCMS-8050 high-sensitivity triple quadrupole mass spectrometer. Although a simple method of sample pretreatment was used that involves only deproteinization, the quantitative results obtained were excellent in terms of accuracy and precision.

The pretreatment workflow is shown in Fig. 2. Known concentrations of itraconazole and hydroxy itraconazole were solvated in a 1:1 water: methanol mixture, and used to prepare standard solutions for addition to plasma. These standard solutions for addition and an acetonitrile solution of an internal standard solution were added to human plasma, the deproteinized supernatant was diluted in an aqueous solution of 1 % acetic acid, and this mixture was used to prepare standard (STD) samples for a calibration curve and QC samples to validate the analysis results. The stable isotope-labeled compound of itraconazole (itraconazole-d3) was used as the internal standard solution.
The pretreatment method used is simple and requires no labor-intensive steps such as solid phase extraction, which allows pretreatment to be performed at low cost and in a short period of time.

 
 
 
 
Structural Formula of Itraconazole and Hydroxy Itraconazole

Fig. 1 Structural Formula of Itraconazole and Hydroxy Itraconazole

Pretreatment Workflow

Fig. 2 Pretreatment Workflow

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