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To establish a high-efficiency and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative analysis of cyclosporine A in human whole blood.
Protein precipitation with methanol was employed for sample pretreatment, and cyclosporine A-d4 was used as the internal standard. Chromatographic separation was performed on a Kinetex C18 column (3.0 mm×50.0 mm, 2.6 μm, 100 Å) with a mobile phase consisting of 0.1% formic acid in water (phase A) and 0.1% formic acid in methanol (phase B) at a flow rate of 0.55 mL·min-1. The column temperature was maintained at 40 ℃. Quantitative detection was carried out using electrospray ionization in positive ion mode and multiple reaction monitoring (MRM). The method was validated for specificity, calibration curve and lower limit of quantitation (LLOQ), accuracy and precision, matrix effect, and sample stability.
The retention time of cyclosporine A was 1.19 min, with a total run time of 2.0 min per sample. A good linear relationship was observed in the concentration range of 10-1 000 ng·mL-1 (r=0.998 5), and the LLOQ was 10.0 ng·mL-1. The recovery rates ranged from 99.53% to 102.83%, with intra-batch precision ≤3.58% and inter-batch precision ≤3.24%. The sample stability was satisfactory, and the matrix effect was negligible.
The established method features short analysis time, high sensitivity, and low sample volume requirement, making it suitable for therapeutic drug monitoring and accurate quantification of cyclosporine A in human whole blood samples.
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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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