Analysis of 17-Hydroxygeranyllinalool Diterpene Glycosides in Nicotiana tabacum by Using Heart-Cutting 2D-LC Coupled with Tandem MS Technique

Nicotiana tabacum was found to have the potent anti-herbivore defense capacity by secondary metabolites of 17-hydroxygeranyllinalool diterpene glycosides (HGL-DTGs). A heart-cutting two-dimensional LC coupled with tandem MS method was developed to analysis the HGL-DTGs. And 10 types of HGL-DTGs were determined qualitatively and quantitatively. In the novel method, reversed-phase liquid chromatography (RPLC) had been employed in the first dimension for separation of HGL-DTGs with other interference compounds. Then target elution, including HGL-DTGs, was cut to the trap column by online dilution from the additional pump. The analytes of HGL-DTGs were reserved and enriched on the trap column. After switch of the 6-port valve, the analytes were eluted to the second dimensional separation. The HGL-DTGs were then determined by the tandem MS under the multiple reaction monitoring mode (MRM). The intra-day and inter-day RSD of the HGL-DTGs were between 0.24 and 7.21%. Consequently, the new method was successfully utilized to detect the HGL-DTGs levels in a wide range of the tobacco samples, which showed fairly good accuracy and reproducibility. The new strategy offered a simple and fast way for further investigation of the HGL-DTGs in Nicotiana tabacum .