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To establish a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the determination of blood drug concentrations of nirmatrelvir and ritonavir in human plasma and apply it to therapeutic drug monitoring (TDM).
Plasma samples were precipitated, and the mobile phase consisted of 0.1% formic acid aqueous solution (phase A) and acetonitrile (phase B). Isocratic elution was used with a flow rate of 0.3 mL·min-1, a column temperature of 40 ℃, and an EC-C18 chromatographic column. Multiple reaction monitoring was conducted in positive ion mode.
The ion channels used for quantitative analysis were m/z 500 → 110 (nirmatrelvir), m/z 721.2 →296.3 (ritonavir), and m/z 614.4 → 421 (internal standard remdesivir). In human plasma, nirmatrelvir showed good linearity in the range of 0.01 to 10 µg·mL-1, while ritonavir exhibited good linearity in the range of 0.01 to 2.0 µg·mL-1. The extraction recoveries were 79.56% to 89.91% for nirmatrelvir and 82.03% to 89.56% for ritonavir. The RSD values for intra-day and inter-day precision were all less than 15%, and the stability was satisfactory. The developed method was applied to measure the blood drug concentrations of 34 patients after oral administration of Paxlovid, which ranged from 1.04 to 18.0 µg·mL-1.
The HPLC-MS/MS method established in this study is rapid, highly sensitive, and accurate, and it can be applied to monitor the therapeutic drug concentration monitoring of COVID-19 patients after taking Paxlovid.
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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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