This application note describes a rapid method for the quantification of total testosterone in serum using the Agilent 1200 Series Rapid Resolution LC (RRLC) system with the Agilent 6430 Triple Quadrupole LC/MS in atmospheric pressure chemical ionization (APCI) mode. A two dimensional LC approach was used to minimize matrix interferences and ion suppression, while reducing the time required for analysis. Excellent linearity of quantification (R2 > 0.99) was obtained in the range of 5 to 2000 ng/dL, with analysis times of only 4.5 minutes, and day-to-day precision was below 10%. The limit of detection (LOD) and limit of quantification (LOQ) were 2 ng/dL and 5 ng/dL respectively, in 5% bovine serum albumin.
This document presents a clinical research method for the quantification of total testosterone in serum. The method utilizes the Agilent 1200 Series Rapid Resolution LC (RRLC) system coupled with an Agilent 6430 Triple Quadrupole LC/MS in atmospheric pressure chemical ionization (APCI) mode. A two-dimensional LC approach with heart-cutting is employed to minimize matrix interferences and ion suppression while reducing analysis time to 4.5 minutes.
The application details the experimental setup, including reagents, sample preparation via precipitation, and specific instrument conditions for both LC and MS. Results demonstrate excellent linearity (R² > 0.99) across a range of 5 to 2000 ng/dL, with a limit of detection (LOD) of 2 ng/dL and a limit of quantification (LOQ) of 5 ng/dL. The method was verified against a reference lab method using 50 serum samples, showing a strong correlation.
| Analysis time | 4.5 min |
|---|---|
| Linearity range | 5 to 2000 ng/dL |
| Column temperature | 50°C |
| Injection volume | 10 pL |
| Drying gas temperature | 300°C |
| Capillary voltage | 4500 V |
No, this is a clinical research application note intended for research use only and not for use in diagnostic procedures.
The limit of detection for this method is 2 ng/dL.
The total analysis time required is 4.5 minutes.