This application note describes a sensitive, precise, and accurate quantitative liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the measurement of beta-estradiol acetate in human plasma. The sample preparation method employed solid phase extraction on Agilent SampliQ cartridges. The processed samples were chromatographed on an Agilent ZORBAX Eclipse Plus C18 analytical column and analyzed by triple-quadrupole tandem mass spectrometry in the multiple reaction monitoring (MRM) mode using negative electrospray ionization. The principal advantage of the LC/MS/MS method described in this application note is the simultaneous achievement of high absolute recovery (~ 80%), high sensi- tivity (LLOQ of 100 fg injected on-column or 28.6 pg/mL plasma), high precision (≤ 5.9% CV) and accuracy (81.5-118.1%), as well as excellent linearity over the concentration range 0.1 to 10000 pg/μL (R2 ≥ 0.998) with a short run time of only 1.2 min.
This document details a sensitive and accurate LC/MS/MS method for measuring B-estradiol 17-acetate in human plasma. The method utilizes solid phase extraction with Agilent SampliQ cartridges and analysis via triple-quadrupole tandem mass spectrometry in negative electrospray ionization mode.
The study achieved high absolute recovery, sensitivity with a lower limit of quantification of 28.6 pg/mL, and excellent linearity over a concentration range of 0.1 to 10000 pg/pL with a short run time of 1.2 minutes.
| Lower Limit of Quantification (LLOQ) | 100 fg injected on-column or 28.6 pg/mL plasma |
|---|---|
| Precision | < 5.9% CV |
| Accuracy | 81.5-118.1% |
| Linearity Range | 0.1 to 10000 pg/pL |
| Run Time | 1.2 min |
| Column | ZORBAX Eclipse Plus C18 analytical column |
| Extraction Cartridge | Agilent SampliQ C18 end capped (100 mg) |
| Flow Rate | 0.5 mL/min |
| Capillary Voltage | -3500 V |
| Drying Gas Temperature | 300°C |
The method achieves a lower limit of quantification (LLOQ) of 100 fg injected on-column or 28.6 pg/mL plasma.
The analysis has a short run time of only 1.2 minutes.