This application brief describes a highly repeatable method for the quantification of estradiol in human plasma using the Agilent 7000 Triple Quadrupole GC/MS. The method provides an LOD of 1 pg/mL (2 fg on-column) and RSDs for quantification of <15% in the 1 to 600 pg/mL concentration range.
This document details a method developed on the Agilent 7890GC/7000 Triple Quadrupole GC/MS system to detect and quantify estradiol in human plasma. The method achieves a limit of detection (LOD) of 1 pg/mL (2 fg on-column) and maintains a relative standard deviation (RSD) for quantification of less than 15% across a concentration range of 1 to 600 pg/mL. The analysis utilizes the MS/MS-NCI (negative chemical ionization) mode with a specific column and injection sequence.
| Limit of Detection (LOD) | 1 pg/mL (2 fg on-column) |
|---|---|
| Quantification Range | 1 to 600 pg/mL |
| Relative Standard Deviation (RSD) | <15% |
| Analytical Column | 15m x 0.25 mm x 0.25 pm Agilent J&W DB-5MS UI |
| Carrier Gas | Helium, constant flow, 1.2 mL/min |
| MS Mode | NCI mode; 40% ammonia |
| Source Temperature | 150 °C |
| Quadrupoles Temperature | 150 °C |
| Collision Gas Flows | Nitrogen at 1.5 mL/min, Helium at 2.35 mL/min |
| Total Run Time | 2.85 minutes |
The limit of detection is 1 pg/mL, which corresponds to 2 fg injected on-column.
The method provides quantification for estradiol in the range of 1 to 600 pg/mL.
The RSD for quantification never exceeds 15%.
The instrument is operated in the MS/MS-NCI (negative chemical ionization) mode.