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LC/MS/MS Research Method for 14 Antidepressants Utilizing Dried Blood Spots
Overview
Purpose:
To develop an LC-MS/MS research method for measuring the
concentration of antidepressants from dried blood spots (DBS).
Methods:
An LC-MS/MS method was developed to extract 14 antidepressants
from dried blood spots for quantitation.
Results:
The analytes of interest were extracted successful from dried blood
spots showing good linearity, accuracy, and precision.
Introduction
Important factors in the analysis of drugs in whole blood are accurate
measurements, storage capabilities, small sample volume, and easy extraction.
Dried blood spots are becoming an adopted clinical research technique for the
analysis of drugs in biological matrices. Due to the complexity of the solution
resulting from dissolving the blood spot, the sample must undergo further
cleanup by chromatographic separation before introduction into the mass
spectrometer. A research application is demonstrated using the Thermo
Scientific™ Prelude SPLC™ system and the Thermo Scientific™
TSQ Endura
™ triple
quadrupole mass spectrometer (Figure 1) to quantitatively
analyze 14 antidepressant drugs collected from dried blood spots.
Instrumentation
A Prelude SPLC system was used
Scientific™
TurboFlow
™
Fluoro X
2.1 x 50 mm, 2.6 µm particle size
column. The detector for the syst
spectrometer with a heated electr
mode. Thermo Scientific™
TraceF
quantitation. The liquid chromatog
Method Parameters
Mobile phases were (A) 10 mM a
(B) 10 mM ammonium formate, 0.
45/45/10 acetonitrile/isopropanol/a
The mass spectrometer quantifier
(SRM) transitions are shown in Ta
was 10
–
750 ng/mL.
FIGURE 1. Prelude SPLC system with
TSQ Endura MS
Methods
Sample Preparation
The analytes of interest were spiked into human whole blood at various
concentrations to make calibrators and controls. The samples were spotted at a
fixed volume onto Whatman
®
paper. Then, the analyte was extracted by solvent
containing isotopically labeled internal standards and transferred into clean vials
for LC-MS/MS analysis. The entire workflow is depicted in Figure 2.
SpotDBS card with
standardsandQCs
Extract solutio
wellsandadd
Punchcardwith
3mm holepunch
FIGURE 2. Workflow of preparin
FIGURE 3. Online sample cleanu