Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in Camptotheca acuminata and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS
作者： Zhaoxia Jin , Ruyi Wan , Ruxue Yan , Yingying Su , Honglan Huang
DOI: 10.3390/MOLECULES24040815
关键词:
摘要: Camptothecin (CPT) has strong antitumor activity and is used as an anticancer therapeutic agent. To better understand decipher the pathway of CPT biosynthesis in Camptotheca acuminata, main purpose here was focused on creating effective extraction strategy for a rich intermediate metabolite profile. In present study, 70% aqueous acetonitrile verified optimal solvent microwave-assisted (MAE) metabolites by spiking experiments. Based multi-objective optimization, best conditions solid-liquid ratio 1:20, microwave power 230 W, time 4 min were achieved using full factorial 34 experimental design. Crude extracts obtained from shoot apex C. acuminata MAE have been qualitatively profiled high-performance liquid chromatography coupled with linear ion trap quadrupole-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap-MS/MS) HPLC triple quadrupole-MS (HPLC-TSQ-MS) analysis conducted their content different tissues. CPT, ten related isomers, including tryptamine, loganic acid, secologanic strictosidinic strictosamide, strictosamide epoxide, diol, ketolactam, pumiloside, deoxypumiloside, detected tentatively identified. Scanning electron microscopy (SEM) imaging demonstrated that severe cell disruption evident after intensified processes. The study showed difference profiles enhancement microwave-pretreated techniques, established procedure methodology to preserve valuable compounds analysis.
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