Standard addition and stable isotope dilution methods were used in quantifying plant hormones [2022]. Stable isotope-labelled plant hormones were fortified with samples as well as absolute standard solutions at concentrations ranging from 1.0 ng/ml to 250.0 ng/ml. Neat standard solutions were post-spiked for standard additions with actual sample in order to construct matrix calibration curves in the range of 1.0 ng/ml to 250.0 ng/ml to equalize matrix effects among samples (S1 Fig). Accordingly, actual samples were diluted (dilution factor 1.05). Calibration curves for each phytohormone were constructed for each analyte using the same matrix. Matrix effects (ME = A—B/A*100) were then calculated using the peak areas of A and B, with A identified as a peak area of an analyte in a standard solution and B as a peak area of an analyte in a matrix [9]. A peak area of an analyte derived from the sample was subtracted from B by analyzing the sample beforehand. Recovery rate was calculated by comparing the peak area of each phytohormone present in the sample spiked before SPE and the sample spiked after SPE. Limits of detection (LOD) and limit of quantification (LOQ) were defined as a signal-to-noise ratio of 3:1 and 10:1, respectively.

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