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To develop a liquid chromatography-tandem mass spectrometry method for the detection of lenvatinib in quantitative dried blood spots.
Lenvatinom-[d4] was used as the internal standard. The dried blood spot samples were extracted with acetonitrile/water (70/30) and separated on a ProChrom C18 reversed-phase column (2.1 mm×100.0 mm, 3.0 μm). The mobile phase consisted of 10 mmol·L-1 ammonium acetate solution and methanol with gradient elution. The flow rate was 0.3 mL·min-1. The ionization was carried out under the electrospray ionization positive mode. Multiple reaction monitoring was used to detect the transitions of lenvatinib at m/z 427.1→370.0 and 338.0, as well as lenvatinib-[d4] at m/z 431.1→370.0. The specificity, standard curve, carryover, precision and accuracy, extraction recovery, matrix effect and stability of the method were investigated.
Endogenous components in the dried blood spot matrix of blank human whole blood did not affect the determination. The optimized pretreatment employs a ostep extraction with acetonitrile/water (70/30). Lenvatinib in dried blood spots demonstrated good linearity in the range of 10-250 ng·mL-1. The standard curve equations from three analytical batches were y=3.81×10-2x+2.76×10-2(r=0.999 7), y=2.0×10-2x-4.87×10-3(r=0.999 9), y=2.01×10-2x-3.68×10-3(r=0.999 7), respectively. The lower limit of quantification (LLOQ) was 10 ng·mL-1. The intra- and inter-batch relative errors were within ±15%, and relative standard deviation (RSD) were below 15%. There was no significant matrix effect, and lenvatinib dried blood spots sample remained stable for at least 7 days at a high temperature of 45 ℃.
The method is convenient, efficient and accurate and it is suitable for the requirements of The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use M10 guideline "Bioanalytical Method Validation and Study Sample Analysis". It can be used for the determination of lenvatinib concentrations in dried blood spot 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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