Introduction: Strigolactones (SLs) are the most representative germination stimu-
lants for seeds of root parasitic plants, and they show activity even at concentrations
below 10−10 M. The low amounts of stimulants produced by the host and their rapid
degradability make it crucial to develop analytical methods with very low limits of quantification.
Objective: To develop a sensitive and validated analytical method for the
simultaneous quantification of seven SLs [7‐oxoorobanchyl acetate (1), solanacol (2),
orobanchol (4), strigol (5), fabacyl acetate (6), orobanchyl acetate (7), and 5‐
deoxystrigol (8)].
Methods: SLs were analysed using ultra‐high performance liquid chromatography
coupled to tandem mass spectrometry (UHPLC‐MS/MS), with (±)‐GR24 (3) employed
as internal standard (IS). Validation was based on selectivity, linearity, precision of the
peak areas (repeatability and intermediate precision), detection and quantification
limits, and stability.
Results: A simple, rapid and reliable UHPLC‐MS/MS method has been validated for
the routine analysis of seven SLs and has been successfully applied to quantify them
in exudates and extracts from tomato roots (Solanum lycopersicum). The limits of quan-
tifications range from 0.05 μg/L for 5‐deoxystrigol to 0.96 μg/L for solanacol.
Conclusion: The method provides a useful tool for research in all the fields related
to SLs, both for studies related to their function as hormones, and signalling molecules
in the rhizosphere, without sample preparation required for extracts and root exudates in less than 11 minutes.