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To establish a method for determination of zonisamide in plasma of epileptic children by micro-detection technology combined with high performance liquid chromatography-mass spectrometry (HPLC-MS/MS).
10 μL of plasma samples were pre-treated by precipitation protein method with organic solvent. Reversed-phase chromatographic columns were adopted. Flow phase: 0.075% formic acid-2 mmol·L-1 ammonium acetate-water (A) and methanol (B), gradient elution; flow speed: 0.45 mL·min-1; injection volume: 1 μL. The quantitative method adopted the internal standard method, and the isotope internal standard was zonisamide -15N-D4. Ion source was electric spray ion (ESI) source, multireaction monitoring (MRM) mode for quantitative analysis, using positive ion monitoring mode. The specificity, standard curve, lower limit of quantitation (LLOQ), residual effect, precision, matrix effects, extraction recovery and stability were investigated.
The retention times of zonisamide and its internal standard were both 1.25 min, and there was no interference from endogenous substances within the retention time. The linear range of zonisamide was 1-62.5 μg·mL-1 (r=0.999 7), and LLOQ was 1 μg·mL-1. The intra-day and inter-day precision RSD was equal or less than 4.84%, and the accuracy ranges from 97.75%-107.95%, with good stability, and this method was not affected by common matrix.
This method involves micro-blood collection, which is simple to operate, highly efficient and rapid, and can be used for the monitoring of daily therapeutic drugs in children with epilepsy in clinical practice.
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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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