Development of UPLC-MS Combined with HPLC Technique to Explore the Effects of Exogenous Substances on the Saponin Contents and Growth of Panax notoginseng Leaves

Purpose We aimed to explore the effect of exogenous substances, including gibberellin (GA₃), 6-benzyladenine (6-BA), abscisic acid (ABA), methyl jasmonate (MeJA), salicylic acid (SA), naphthylacetate (NAA), ethylene (ETH), melatonin (MT), spermidine (Spd), sodium nitroprusside (SNP) and hydrogen peroxide (H₂O₂) on saponin contents of Panax notoginseng leaves.

Method We developed a new method of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-MS) together with high performance liquid chromatography (HPLC), and determined 7 saponin monomer contents and growth indexes.

Results (1) The established HPLC method could accurately measure 7 saponin monomer contents, including ginsenoside Rb₁, Rc, Rb₂, Rb₃, notoginsenoside Fa, Fe and gypenoside XVII in P. notoginsen leaves within 52 minutes within a single run; (2) MeJA treatment significantly increased the contents of 7 monomer saponins, including ginsenoside Rb₁, Rc, Rb₂, Rb₃, notoginsenoside Fa, Fe and gypenoside XVII in leaves, and had no effect on P. notoginseng growth. Other exogenous substances, including NAA, 6-BA, H₂O₂, SNP and Spd significantly increased the content of notoginsenoside Fa, while ABA significantly improved the content of gypenoside XVII, but all of them significantly decreased the total saponin contents, and had effect on P. notoginseng growth.

Conclusion A rapid and simple, stable and reliable detection method for saponin monomer of P. notoginseng leaves was established. Based on this method, it is clear that exogenous substance MeJA could significantly induce saponin monomer contents of P. notoginseng leaves.