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. 2025 Aug 2;14(15):2395.
doi: 10.3390/plants14152395.

HPLC-ESI-HRMS/MS-Based Metabolite Profiling and Bioactivity Assessment of Catharanthus roseus

Affiliations

HPLC-ESI-HRMS/MS-Based Metabolite Profiling and Bioactivity Assessment of Catharanthus roseus

Soniya Joshi et al. Plants (Basel). .

Abstract

A comprehensive metabolic profiling of Catharanthus roseus (L.) G. Don was performed using tandem mass spectrometry, along with an evaluation of the biological activities of its various solvent extracts. Among these, the methanolic leaf extract exhibited mild radical scavenging activity, low to moderate antimicrobial activity, and limited cytotoxicity in both the brine shrimp lethality assay and MTT assay against HeLa and A549 cell lines. High-performance liquid chromatography-electrospray ionization-high-resolution tandem mass spectrometry (HPLC-ESI-HRMS/MS) analysis led to the annotation of 34 metabolites, primarily alkaloids. These included 23 indole alkaloids, two fatty acids, two pentacyclic triterpenoids, one amino acid, four porphyrin derivatives, one glyceride, and one chlorin derivative. Notably, two metabolites-2,3-dihydroxypropyl 9,12,15-octadecatrienoate and (10S)-hydroxypheophorbide A-were identified for the first time in C. roseus. Furthermore, Global Natural Products Social Molecular Networking (GNPS) analysis revealed 18 additional metabolites, including epoxypheophorbide A, 11,12-dehydroursolic acid lactone, and 20-isocatharanthine. These findings highlight the diverse secondary metabolite profile of C. roseus and support its potential as a source of bioactive compounds for therapeutic development.

Keywords: Catharanthus roseus; GNPS; antimicrobial assay; radical scavenging activity; secondary metabolites.

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Conflict of interest statement

Author Salyan Bhattarai was employed by the company Paraza Pharma, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
(a) MIC values and (b) MBC values of different C. roseus extracts against Staphylococcus aureus and Shigella sonnei. A01 and A02 represent the methanolic and ethyl acetate extracts of the leaves, respectively, while A05 and A06 represent the methanolic and ethyl acetate extracts of the stem, respectively. Neomycin was used as the positive control. All data are presented as mean ± standard deviation (n = 3).
Figure 2
Figure 2
Cytotoxic activity of methanolic extracts of C. roseus and doxorubicin against (a) HeLa and (b) A549 cell lines. All data are presented as mean ± standard deviation (n = 3).
Figure 3
Figure 3
Total ion chromatogram (TIC) of the methanolic leaf extract (A01) of C. roseus obtained via HPLC-ESI-HRMS/MS in positive ion mode, showing annotated metabolites. (* Unidentified peaks).
Figure 4
Figure 4
Chemical structures of annotated compounds from the methanolic leaf extract (A01) of C. roseus, identified by HPLC-ESI-MS/MS analysis.
Figure 4
Figure 4
Chemical structures of annotated compounds from the methanolic leaf extract (A01) of C. roseus, identified by HPLC-ESI-MS/MS analysis.
Figure 5
Figure 5
Chemical structures of annotated specialized metabolites from the methanolic leaf extract (A01) of C. roseus identified via GNPS analysis.
Figure 6
Figure 6
GNPS analysis and annotated specialized metabolites from the methanolic leaf extract (A01) of C. roseus: (A) Overview of the GNPS and classification of annotated compounds. (B) Enlarged view of the cluster containing pheophorbide A. (C) Enlarged view of the triterpenoid cluster. (D) Enlarged view of the tryptophan alkaloid cluster. (E) Enlarged view of the catharanthine cluster.

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