One-pot preparation of a mixed-mode organic-silica hybrid monolithic capillary column and its application in determination of endogenous gibberellins in plant tissues

• An improved one-pot method to prepare C8/PO(OH)2-silica hybrid monolithic capillary column was proposed.
• A PT-SPE method was developed for purifying GAs.
• Chemical derivatization was employed for enhancing the MS response of GAs.
• A method of nano-LC–MS coupled to PT-SPE was developed for determination of GAs.

A newly improved one-pot method, based on “thiol-ene” click chemistry and sol–gel approach in microemulsion system, was developed for the preparation of C8/PO(OH)2-silica hybrid monolithic capillary column. The prepared monolith possesses large specific surface area, narrow mesopore size distribution and high column efficiency. The monolithic column was demonstrated to have cation exchange/reversed-phase (CX/RP) mixed-mode retention for analytes on nano-liquid chromatography (nano-LC). On the basis of the developed nano-LC system with MS detector coupled to pipette tip solid phase extraction (PT-SPE) and derivatization process, we then realized simultaneous determination of 10 gibberellins (GAs) with low limits of detection (LODs, 0.003–0.025 ng/mL). Furthermore, 6 endogenous GAs in only 5 mg rice leaves (fresh weight) were successfully detected and quantified. The developed PT-SPE-nano-LC–MS strategy may offer promising applications in the determination of low abundant bioactive molecules from complex matrix.