An ultrasensitive
analysis method for quantification of endogenous
brassinosteroids in fresh minute plants was developed based on dispersive
matrix solid-phase extraction coupled with high performance liquid
chromatography-tandem mass spectrometry. During the dispersive matrix
solid-phase extraction, plant samples were first ground with solid
sorbent (dispersant) in one microcentrifuge tube and then centrifuged
after adding extraction solvent and cleanup materials (another type
of sorbent). Three protocols based on dispersive matrix solid-phase
extraction were compared and discussed for plant samples with different
matrix complexity. The choice of any protocol was a compromise of
increasing purification efficiency and decreasing sample loss. Under
optimized conditions, the limits of detection were 1.38−6.75
pg mL–1 for five brassinosteroids in the oilseed
rape samples. The intraday and interday precisions were in the range
of 0.8%−9.8% and 4.6%−17.3%, respectively. The proposed
method was successfully applied to detection of endogenous brassinosteroids
in milligram oilseed rape (2.0 mg) and submilligram Arabidopsis
thaliana seedlings (0.5 mg). Finally, the geographical distribution
of five endogenous brassinosteroids of Brassica napus L. oilseed rape in different provinces of origin in the Yangtze River
basin was described.
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