The determination of trace plant growth regulator (PGR) has received more and more attentions in the field of phytophysiology and food safety. But the simple and sensitive method for simultaneously analysing multiple classes of PGR remains poorly investigated. In this study, a new pre-column fluorescence labelling method using 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methylbenzenesulfonate (BCETS) as the labelling reagent has been developed for simultaneous determination of seven PGRs (i.e., indole-3-acetic acid, 3-indolybutyric acid, 3-indolepropionic acid, jasmonic acid, gibberellin A3, 1-naphthylacetic acid and 2-naphthaleneacetic acid) by HPLC with fluorescent detection (FLD). The proposed method offered the LOD of 0.34-0.73 ng/mL for seven PGRs, which were significantly lower than the reported methods. The crude extract without complex pre-treatments and purification was directly labelled by BCETS and analysed by HPLC-FLD, which facilitates the high-throughput sample screening. This method was proven to be inexpensive, simple, selective, sensitive, accurate and reliable for trace PGR determination.