Blood samples were collected in lithium heparinate tubes and centrifuged at 2250 g for 10 min before storage at −20°C. After thawing, a protein precipitation was carried out by mixing 200 μl of the blood sample and 400 μl of acetonitrile spiked with the internal standard (cabozantinib d-4) in a 1 ml Eppendorf tube. This mix was vortexed for 15 s and then centrifuged for 10 min at 4000 g. The supernatant was then injected on an Acquity UPLC I-Class system (Waters, Milford, MA) where the separation was carried out using an Acquity BEH C18 1.7 × 2.1 × 100 mm analytical column with a vanguard pre-column. The detection was carried out by a Xevo TQ-D MS/MS (Waters, Milford, MA) system using electrospray ionization and multiple reaction monitoring detection with mass transition (m/z) of 502.2 → 391.1 and 506.3 → 391.2 for cabozantinib and its internal standard, respectively. This method was fully validated according to the EMA guidelines with a range of calibration from 25 to 2500 ng/ml, accuracy of 95.5%-105.3%, intra-day precision 2.3%-5.1% and inter-day precision 7.0%-8.8% for four QC samples (50, 187.5, 1000 and 2250 ng/ml).
Xevo tqd ms ms
Manufactured by Waters Corporation
The Xevo TQD MS/MS is a triple quadrupole mass spectrometer designed for sensitive and selective quantitative analysis. It provides high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) capabilities for a wide range of applications.
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Lab products found in correlation
3 protocols using xevo tqd ms ms
1
Quantification of Cabozantinib in Blood
2
Quantitative HPLC-MS/MS Analysis of Compounds
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Separation was achieved on an ACQUITY BEH-C18 column (2.1 × 50 mm, 1.7 m) at a temperature of 40°C using a linear gradient of mobile phase A (0.1% of formic acid in water) and mobile phase B (0.1% of formic acid in methanol) as follows: 0 min, 10% B (0.4 mL/min); 1.5 min, 45% B (0.3 mL/min); 2.5 min, 75% B (0.3 mL/min); 3 min, 95% B (0.3 mL/min); 4–5 min, 10% B (0.3 mL/min). The mass spectrometer was operated in positive ion mode on a Waters Xevo TQD MS/MS with a 1.2 kV capillary voltage. The source and desolvation gas temperature were 150 and 550°C, respectively. The data were collected with multiple reaction monitor (MRM).
3
Quantification of Piperidine Metabolites in DBS, Plasma, and Urine
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Separation was achieved on an ACQUITY BEH-C18 column (2.1 × 50 mm, 1.7 m) at a temperature of 40 °C using a linear gradient of mobile phase A (0.1% of formic acid in water) and mobile phase B (0.1% of formic acid in methanol) as follows: 0 min, 10% B (0.4 mL/min); 1.5 min, 45% B (0.3 mL/min); 2.5 min, 75% B (0.3 mL/min); 3 min, 95% B (0.3 mL/min); 4–5 min, 10% B (0.3 mL/min). The mass spectrometer was operated in positive ion mode on a Waters Xevo TQD MS/MS with a 1.2 kV capillary voltage. The source and desolvation gas temperature was 150 °C and 550 °C, respectively. The data were acquired in with multiple reaction monitoring (MRM) mode. The cone and collision energy for the detection of a-AASA, P6C, a-AAA, d3-a-AAA, PA and d9-PA were shown in Table S1 . The examples of extract ion chromatogram (EIC) of a-AASA, P6C, PA, a-AAA, d9-PA and d3-AAA in control and PDE patient in DBS, plasma and urine samples were shown in Figs 1 , S1–2 , respectively.![]()
The classic extract ion chromatogram of a-AASA, P6C, PA, a-AAA, d9-PA and d3-AAA in control and PDE patient in DBS samples.
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