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To establish liquid chromatography-mass spectrometry (LC-MS/MS) quantitative method for determining the blood concentration of Upatinib in human plasma.
Upatinib 15N-d2 was used as the internal standard, and acetonitrile was used for protein precipitation pretreatment. Separate using a Waters BEH C18 (2.4 mm×50.0 mm, 1.7 μm) chromatographic column; acetonitrile (containing 0.1% formic acid) and 0.1% formic acid aqueous solution were used as mobile phases, with gradient elution at a flow rate of 0.3 mL·min-1 and column temperature of 40 ℃. The ion pair used for quantitative analysis was m/z 388.90→255.90 (Upatinib) and m/z 384.10→256.20 (Upatinib-15N-d2, internal standard) respectively by using an electrospray ion source and positive ion multi reaction monitoring mode scanning. The specificity, standard curve, quantitative cutoff, precision, recovery rate, dilution reliability, matrix effect and stability of the method were examined.
There is no mutual interference between endogenous components, internal standards and Upatinib in plasma. The linear relationship between the mass concentration of 0.75-60.00 ng·mL-1 Upatinib and peak area is good (r=0.998 6), the standard curve equation was y=8.24×10-2x+0.61×10-2, with a lower limit of quantification of 0.75 ng·mL-1. The intra-day and inter-day batch precision coefficients of variation of the quality control samples are both ≤5.3%. The accuracy of the sample concentration after 5-fold dilution is 1.1%, and the coefficient of variation is 4.6%. The matrix effect factors of low and high-quality plasma quality control samples are 1.1 and 1.0, respectively. The stability of whole blood samples and plasma samples meets the requirements of biological sample analysis.
The method used is simple, rapid, highly accurate, and sensitive, and is suitable for determining the concentration of Upatinib in human plasma. It can be used for pharmacokinetic and bioequivalence studies of Upatinib in healthy individuals.
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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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