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To establish a high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) method for the simultaneous determination of fluconazole, posaconazole, and linezolid in human cerebrospinal fluid.
Cerebrospinal fluid samples were precipitated with acetonitrile containing internal standard (voriconazole) and separated by Agilent C18 column (2.7 μm, 4.6×150 mm). The mobile phase was 0.1% formic acid aqueous solution acetonitrile, gradient elution, flow rate 0.4 mL·min-1, column temperature 35 ℃, and injection volume 1μL; The spray ion source, positive ion mode and multi reaction monitoring were used for the determination. The specificity, standard curve, lowerer limit of quantification(LLOQ), precision, accuracy, recovery rate and recovery rate. This method was applied to determine the concentrations of fluconazole, posaconazole, and linezolid in clinical cerebrospinal fluid.
The methodological results showed that fluconazole (0.4-32 μg·mL-1, r=0.999 6) in cerebrospinal fluid samples, posaconazole (0.04-3.2 μg·mL-1, r=0.9992) and linezolid (0.2-16 μg·mL-1, r=0.9989) had a good linear relationship and specificity within the linear range. The LLOQ of, posaconazole, and linezolid was 0.4、0.04 and 0.2 μg·mL-1, respectively. The relative standard deviation (RSD) values of intra-day and inter-day precision were both less than 15%, with good stability and extraction recovery rates above 80%, unaffected by matrix effects. Additionally, the method exhibited good detection sensitivity for fluconazole, posaconazole, and linezolid in the cerebrospinal fluid of rats and clinical patients. The concentration of the 7 tested sample was within the linear range.
This method is simple, fast, and highly sensitive, and can be used for the determination of cerebrospinal fluid concentrations of fluconazole, posaconazole, and linezolid in clinical patients.
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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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