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To establish and validate a method for determining the plasma concentration of isavuconazole and to apply this method to clinical monitoring of plasma drug concentration.
The plasma samples of isavuconazole were subjected to protein precipitation using methanol. The quantification was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS), with stable isotope voriconazole-d4 serving as the internal standard. An Ultimate AQ-C18 chromatographic column was used with a gradient elution of methanol (containing 0.1% formic acid)-water (0.1% formic acid) with a flow rate of 0.3 mL·min-1. The column temperature was set at 40 ℃ and the injection volume of pretreated sample was 5.0 μL. The ion source was positive electrospray ion source, with multiple reaction monitoring mode for positive ion scanning (MRM+). The ion pairs used for quantitative analysis were m/z 438.2→224.1 (isavuconazole) and m/z 354.2→285.1 (voriconazole-d4). After administering a loading dose to five patients, plasma samples were collected on days 4, 5, 6, 12, 13, and 14 post-initiation of treatment to measure the trough concentation of the drug.
The linear range of the isavuconazole detection quality concentration was 0.1~10 μg·mL-1 (r=0.999 6), with a quantification limit of 0.25 μg·mL-1. The within-batch precision and between-run precision were not higher than 11.9%, and the relative errors were between -4.83% and 6.20%. The stability relative errors were between -2.04% and 6.89%. The extraction recoveries rates of isavuconazole and voriconazole-d4, as well as the matrix effects and residual effects, do not influence the quantitative analysis of the analytes. All trough concentrations of the samples from the five patients were within the linear range of the method.
The established LC-MS/MS method for assaying of isavuconazole is simple and accurate. It can be used for the clinical monitoring the plasma concentration of isavuconazole in patients with fungal infections.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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