This application note provides a detailed procedure for developing and transferring a bupropion LC/MS/MS method on an Agilent 1290 Infinity LC system and an Agilent 6460 Triple Quadrupole LC/MS system. The developed method is comparable to the original method in terms of sensitivity and precision.
This document presents a case study demonstrating the transfer of a bioanalytical LC/MS/MS assay between laboratories using different vendor instruments. Specifically, a method originally developed on a Shimadzu LC 20A coupled to an ABI/SCIEX API 4000 was transferred to an Agilent 1290 Infinity LC System coupled to an Agilent 6460 Triple Quadrupole LC/MS System.
The study evaluated the linearity, precision, and accuracy of drug-spiked plasma samples on the Agilent system. The mobile phase, column, and multiple reaction monitoring (MRM) transitions were kept identical to the original system. The Agilent source parameters were optimized, resulting in comparable sensitivity and successful method transfer with minimal downtime.
| Column temperature | 40 ± 5 °C |
|---|---|
| Mobile phase composition | Ammonium formate (10 mM): acetonitrile (20/80, v/v) |
| Flow rate | 0.8 mL/min |
| Run time | 2 min |
| Bupropion MRM transition | 239.8 — 183 |
| Risperidone MRM transition | 411.1 — 190 |
| Drying gas flow | 13.0 L/min |
| Nebulizer pressure | 40 psig |
| Dry gas temperature | 350°C |
| Capillary voltage | 3000 V |
| Sheath gas flow | 12.0 L/min |
| Sheath gas temperature | 350°C |
| Bupropion collision energy | 7 eV |
| Bupropion fragmentor voltage | 135 V |
| Risperidone collision energy | 25 eV |
| Risperidone fragmentor voltage | 90 V |
| Calibration range | 1 to 500 ng/mL |
| Minimum R² value | > 0.998 |
The method was transferred to an Agilent 6460 Triple Quadrupole LC/MS System coupled to an Agilent 1290 Infinity LC System.
The MRM transition for bupropion was 239.8 — 183, and for the internal standard risperidone, it was 411.1 — 190.
The calibration curve was a 10-point curve plotted from 1 to 500 ng/mL using plasma extracted samples.
No, the mobile phase, column, and MRM settings were kept identical on both systems, only the source parameters were optimized.