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To establish a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for determine the concentration of busulfan in plasma of hematopoietic stem cell transplantation (HSCT) patients and apply it to clinical sample detection.
A Symmetry C18 column 150.0 mm ( 4.6 mm, 3.5 μm) was used. The mobile phase consisted of phase A (10 mmol·L-1 ammonium formate solution, containing 0.1% formic acid, v/v) and phase B (acetonitrile), and was subjected to gradient elution at a flow rate of 0.8 mL·min-1, and the injection volume was 10 μL. The internal standard was carbamazepine. The quantitative analysis was ionized with atmospheric pressure chemical ionization source (APCI), positive ion mode, and multi reaction monitoring (MRM). Detecting ion pairs were busulfan 264.4→151.2, carbamazepine 237.1→194.0. The specificity, standard curve, precision, recovery rate, matrix effect and stability of the method were validated.
The retention times of busulfan and internal standard were 5.1 min and 5.3 min, respectively. The standard curve equation for busulfan in plasma was y=0.06x-0.04 (r=0.998 7), with a linear range of 0.1-10.0 μg·mL-1. The relative standard deviation (RSD) of intra-day and inter-day precision were less than 10%; The recovery rate was 95.64%-104.49%, and the matrix effect was 100.11%-118.87%. The RSD of the stability of quality control samples after low-temperature storage, three freeze-thaw cycles, or room temperature storage were less than 10%. The average drug exposure of patients undergoing HSCT was (4.99±2.76) μg·mL-1·h (range 0.66-13.77 μg·mL-1·h).
The established LC-MS/MS method has short analysis time, good specificity, and high precision, and is suitable for the therapeutic drug monitoring of busulfan in HSCT 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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