Chemometrics-assisted validation of a high-throughput method for quantitative analysis of multiclass phytohormones across plant matrices by UHPLC-QQQ-MS/MS for targeted metabolomics

The quantification of phytohormones in plant samples is significantly challenging because of their trace concentrations and the complex plant matrix. This study establishes and optimizes a simple and fast method for simultaneous quantification of multiclass phytohormones using ultra high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-QQQ-MS) for high-throughput targeted metabolomics analysis. It is characterized by simplicity and operational feasibility, and it involves sequential steps of ultrasound-assisted extraction, centrifugation, and nitrogen flow-drying. The assessment of sample preprocessing method employs the technique for order preference by similarity to an ideal solution (TOPSIS) chemometrics-assisted approach, and methanol containing 1% formic acid was selected as the optimal extraction solvent. Quantitative analysis is performed via a multiple reaction monitoring method. The method was validated satisfactory for 31 phytohormones, the specific concentration levels used to estimate recovery and precision were 0.25-200 ng/mL, and the results showed good linearity (correlation coefficients: 0.9910-0.9999), sensitivity (LOD: 0.01-2.87 ng/g; LOQ: 0.02-8.70 ng/g), accuracy (recovery rates: 64.50-129.67%), precision (relative standard deviations