Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS
Camptothecin (CPT) has strong antitumor activity and is used as an anticancer therapeutic agent. To better understand and decipher the pathway of CPT biosynthesis in <i>Camptotheca acuminata</i>, the main purpose here was focused on creating an effective extraction strategy for a rich in...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2019-02-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/24/4/815 |
| _version_ | 1818999785427828736 |
|---|---|
| author | Zhaoxia Jin Ruyi Wan Ruxue Yan Yingying Su Honglan Huang Lihan Zi Fang Yu |
| author_facet | Zhaoxia Jin Ruyi Wan Ruxue Yan Yingying Su Honglan Huang Lihan Zi Fang Yu |
| author_sort | Zhaoxia Jin |
| collection | DOAJ |
| description | Camptothecin (CPT) has strong antitumor activity and is used as an anticancer therapeutic agent. To better understand and decipher the pathway of CPT biosynthesis in <i>Camptotheca acuminata</i>, the main purpose here was focused on creating an effective extraction strategy for a rich intermediate metabolite profile. In the present study, a 70% aqueous acetonitrile was verified as an optimal extraction solvent for microwave-assisted extraction (MAE) of metabolites by spiking experiments. Based on multi-objective optimization, the best extraction conditions of a solid-liquid ratio of 1:20, microwave power of 230 W, and a time of 4 min were achieved using a full factorial 3<sup>4</sup> experimental design. Crude extracts obtained from the shoot apex of <i>C. acuminata</i> using MAE have been qualitatively profiled by high-performance liquid chromatography coupled with linear ion trap quadrupole-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap-MS/MS) and a HPLC triple quadrupole-MS (HPLC-TSQ-MS) analysis was conducted for their metabolite content in different tissues. CPT, and ten related metabolites and their isomers, including tryptamine, loganic acid, secologanic acid, strictosidinic acid, strictosamide, strictosamide epoxide, strictosamide diol, strictosamide ketolactam, pumiloside, and deoxypumiloside, were detected and tentatively identified. Scanning electron microscopy (SEM) imaging of the shoot apex demonstrated that severe cell disruption was evident after intensified extraction processes. The study showed the difference of metabolite profiles and the enhancement of metabolite content after microwave-pretreated techniques, and the established MAE procedure is an effective methodology to preserve valuable metabolite compounds for analysis. |
| first_indexed | 2024-12-20T22:22:56Z |
| format | Article |
| id | doaj.art-d3494db6f1ee4b469063a724e280b8f6 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-20T22:22:56Z |
| publishDate | 2019-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-d3494db6f1ee4b469063a724e280b8f62022-12-21T19:24:54ZengMDPI AGMolecules1420-30492019-02-0124481510.3390/molecules24040815molecules24040815Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MSZhaoxia Jin0Ruyi Wan1Ruxue Yan2Yingying Su3Honglan Huang4Lihan Zi5Fang Yu6School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, ChinaSchool of Biological Engineering, Dalian Polytechnic University, Dalian 116034, ChinaSchool of Biological Engineering, Dalian Polytechnic University, Dalian 116034, ChinaSchool of Biological Engineering, Dalian Polytechnic University, Dalian 116034, ChinaCollege of Basic Medical Sciences, Jilin University, Changchun 130021, ChinaSchool of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, ChinaSchool of Biological Engineering, Dalian Polytechnic University, Dalian 116034, ChinaCamptothecin (CPT) has strong antitumor activity and is used as an anticancer therapeutic agent. To better understand and decipher the pathway of CPT biosynthesis in <i>Camptotheca acuminata</i>, the main purpose here was focused on creating an effective extraction strategy for a rich intermediate metabolite profile. In the present study, a 70% aqueous acetonitrile was verified as an optimal extraction solvent for microwave-assisted extraction (MAE) of metabolites by spiking experiments. Based on multi-objective optimization, the best extraction conditions of a solid-liquid ratio of 1:20, microwave power of 230 W, and a time of 4 min were achieved using a full factorial 3<sup>4</sup> experimental design. Crude extracts obtained from the shoot apex of <i>C. acuminata</i> using MAE have been qualitatively profiled by high-performance liquid chromatography coupled with linear ion trap quadrupole-orbitrap mass spectrometry (HPLC-LTQ-Orbitrap-MS/MS) and a HPLC triple quadrupole-MS (HPLC-TSQ-MS) analysis was conducted for their metabolite content in different tissues. CPT, and ten related metabolites and their isomers, including tryptamine, loganic acid, secologanic acid, strictosidinic acid, strictosamide, strictosamide epoxide, strictosamide diol, strictosamide ketolactam, pumiloside, and deoxypumiloside, were detected and tentatively identified. Scanning electron microscopy (SEM) imaging of the shoot apex demonstrated that severe cell disruption was evident after intensified extraction processes. The study showed the difference of metabolite profiles and the enhancement of metabolite content after microwave-pretreated techniques, and the established MAE procedure is an effective methodology to preserve valuable metabolite compounds for analysis.https://www.mdpi.com/1420-3049/24/4/815microwave assisted extractionTaguchi experimental designmetabolitescamptothecinHPLC-LTQ-MS/MS |
| spellingShingle | Zhaoxia Jin Ruyi Wan Ruxue Yan Yingying Su Honglan Huang Lihan Zi Fang Yu Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS Molecules microwave assisted extraction Taguchi experimental design metabolites camptothecin HPLC-LTQ-MS/MS |
| title | Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS |
| title_full | Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS |
| title_fullStr | Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS |
| title_full_unstemmed | Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS |
| title_short | Microwave-Assisted Extraction of Multiple Trace Levels of Intermediate Metabolites for Camptothecin Biosynthesis in <em>Camptotheca acuminata</em> and Their Simultaneous Determination by HPLC-LTQ-Orbitrap-MS/MS and HPLC-TSQ-MS |
| title_sort | microwave assisted extraction of multiple trace levels of intermediate metabolites for camptothecin biosynthesis in em camptotheca acuminata em and their simultaneous determination by hplc ltq orbitrap ms ms and hplc tsq ms |
| topic | microwave assisted extraction Taguchi experimental design metabolites camptothecin HPLC-LTQ-MS/MS |
| url | https://www.mdpi.com/1420-3049/24/4/815 |
| work_keys_str_mv | AT zhaoxiajin microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT ruyiwan microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT ruxueyan microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT yingyingsu microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT honglanhuang microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT lihanzi microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms AT fangyu microwaveassistedextractionofmultipletracelevelsofintermediatemetabolitesforcamptothecinbiosynthesisinemcamptothecaacuminataemandtheirsimultaneousdeterminationbyhplcltqorbitrapmsmsandhplctsqms |